AOAC Official Method 995.04 Multiple Tetracycline Residues in Milk

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AOAC 2007 美国农药残留检测标准方法

AOAC Official Method 2007.01Pesticide Residues in Foods by Acetonitrile Extraction and Partitioning with Magnesium SulfateGas Chromatography/Mass Spectrometry and Liquid Chromatography/Tandem Mass SpectrometryFirst Action 2007[Applicable for the following pesticides in grapes, lettuces, and oranges: atrazine, azoxystrobin, bifenthrin, carbaryl, chlorothalonil, chlorpyrifos, chlorpyrifos-methyl, l -cyhalothrin (incurred in lettuces), cyprodinil, o,p ¢-DDD, dichlorv os, endosulfan sulfate,ethion (incurred in oranges), imazalil, imidacloprid,kresoxim-methyl (incurred in grapes), linuron, methamidophos,methomyl, permethrins (incurred in lettuces) procymidone,pymetrozine, tebuconazole, thiabendazole (incurred in oranges),tolylfluanid (degraded in lettuces), and trifluralin. These were representative pesticide analytes chosen in representative matrixes,and the method is expected to be applicable to many other similar pesticides and matrixes. Limits of quantitation were demonstrated to be <10 ng/g.]See Tables 2007.01A–E for the results of the interlaboratory study supporting acceptance of the method.A. PrincipleThe QuEChERS (quick, easy, cheap, effective, rugged, and safe)method uses a single-step buffered acetonitrile (MeCN) extraction and salting out liquid–liquid partitioning from the water in the sample with MgSO 4. Dispersiv e-solid-phase extraction (dispersive-SPE) cleanup is done to remove organic acids, excess water, and other components with a combination of primary secondary amine (PSA) sorbent and MgSO 4; then the extracts are analyzed by mass spectrometry (MS) techniques after a chromatographic analytical separation. Figure 2007.01 outlines the protocol in a box format. In brief, a well-chopped food sample alongwith 1 mL of 1% acetic acid (HOAc) in MeCN and 0.5 g anhydrous MgSO 4/NaOAc (4/1, w/w) per g sample are added to a centrifuge tube or bottle, which is shaken and centrifuged. A portion of the MeCN extract (upper layer) is added to anhydrous MgSO 4/PSA sorbent (3/1, w/w; 200 mg per 1 mL extract), mixed, and centrifuged. This final extract is transferred to autosampler vials for analysis by gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS) to identify and determine a wide range of pesticide residues. To achieve <10 ng/g detection limits in modern GC/MS, large volume injection (LVI) of 8 m L is typically needed, or the final extract can be concentrated and solvent exchanged to toluene (4 g/mL), in which case 2 m L splitless injection is used.Both GC/MS and LC/MS/MS techniques are prone to matrix effects in pesticide residue analysis, albeit for different reasons [Erney, D.R., Gillespie, A.M., Gilvydis, D.M., & Poole, C.F. (1993)J. Chromatogr. 638, 57–63; Hajslova, J., & Zrostlikova, J. (2003) J.Chromatogr. A 1000, 181–197; Alder, L., Luderitz, S., Lindtner, K.,& Stan, H.J. (2004) J. Chromatogr. A 1058, 67–79]. To account for these effects, matrix-matched calibration was conducted (calibration standards in solvent solution may also be used if matrix effects are shown not to occur). Due to the situation that some laboratories had LVI capability and others did not, the necessary amounts of matrix blank(s) and final extract volume was different for some laboratories than others. Depending on the water content of the matrix, a 15 g sample typically yields 11–14 mL of initial MeCN extract after centrifugation. In dispersive-SPE, roughly half of the extract is lost to the powders, thus about 6–7 mL of final extract can be expected for a 15 g sample. Two options were provided in the protocol to account for the different situations among the laboratories.ã 2007 AOAC IN T ER N A T IONALTable 2007.01A. Interlaboratory study results for incurred pesticides (and chlorpyrifos-methyl)AnalyteMatrix Avg. concn s raRSD r b, %S R c, ng/g Rec., %RSD R d, %HorRat No. of labsOutlierlabs e Chlorpyrifos-methylGrapes 165 14 8.535 8321 1.00116-C, 4-C Lettuces 178 20 11 30 89170.811011-SGOranges174 25 14 36 87200.9812Kresoxim-methyl Grapes 9.2 1.921f3.2NA 35f1.0912Cyprodinil Grapes 112 NAgNA 18 NA 160.7313l -Cyhalothrin Lettuces 58 6.111 11 NA 200.80 911-C Permethrins Lettuces 112 9.88.741 NA 36f1.63 96-C, 1-C Imidacloprid Lettuces 12NA NA 1.6NA 140.4411Ethion Oranges 198 23 12 36 NA 180.891111-C Thiabendazole Oranges 53 3.87.27.6NA 140.5812ImazalilOranges13NANA4.7NA35f1.1587-SGa s r = Standard deviation for repeatability (within laboratory).b RSD r = Relative standard deviation for repeatability.c s R = Standard deviation for reproducibility (among laboratories).d RSD R = Relative standard deviation for reproducibility.e C = Cochran outlier; SG = single Grubbs outlier.f RSD r >15%; 120% < Rec. < 70%; RSD R >25%; HorRat >1.2; and fewer than 8 laboratories in an assessment.gNA = Not applicable.Analyte Avg. C, ng/g s r, ng/g RSD r, %s R, ng/g Rec., %RSD R, %HorRat No. of labs Outlier labs Atrazine9.30.6 6.9 2.093210.651345 3.27.1 5.790130.4913365 23 6.271 9119 1.0413Azoxystrobin9.40.6 6.6 2.094210.641392 8.79.411 92120.51128-SG182 17 9.226 91140.70128-SG Bifenthrin7.80.811 2.37830b0.89112-C, 10-C86 5.9 6.914 86170.73126-C923 71 7.7136 92150.9113Carbaryl12 1.211 2.8104 27b0.85125-SG50 6.413 11 100 220.87131003 70 7.0189 100 19 1.18125-C Chlorothalonil 6.30.914 2.163b33b0.97 810-C59 8.314 13 79230.9310140 19 13 38 7027b 1.27b10Chloropyrifos8.1 1.519b 3.08137b 1.121268 8.312 14 84200.8413396 25 6.450 79120.681211-SG Cyprodinil c123 13 10 26 101 210.9513240 20 8.363 9226b 1.32b13581 42 7.3110 9519 1.0913o,p¢-DDD8.9 1.416b 3.28936b 1.091242 3.17.37.084170.6512445 32 7.147 89100.58116-C Dichlorvos7.2 1.014 1.372180.53118-SG85 7.48.715 85180.77114-C294 25 8.562 9821 1.1012Endosulfan sulfate8.60.910 1.586170.52 7b10-C 115 14 12 21 77180.8111415 56 14 111 8327b 1.47b11Imazalil7.60.89.8 3.17641b 1.22b1150 2.5 4.915 67b30b 1.19108-C432 53 12 161 7837b 2.06b11Imidacloprid8.80.88.9 3.08834b 1.041345 3.57.78.999200.7813218 18 8.224 97110.56128-SG Linuron9.9 1.717b 2.99929b0.901199 7.47.415 99150.6712971 65 6.7191 9720 1.23b12Methamidophos10 2.929b 3.0101 30b0.95 95-SG80 8.010 14 80180.7712852 72 8.4119 85140.85118-SG Methomyl9.3 1.212 2.99332b0.981250 3.3 6.79.3100 190.7413204 10 4.926 102 130.6313Procymidone8.20.78.2 2.082240.74115-SG64 6.09.416 8524 1.0113428 16 3.870 86160.90129-C Pymetrozine 6.2 1.220b 1.662b27b0.771147 3.1 6.79.662b200.8111341 20 5.859 68b170.9211Tebuconazole9.2 1.112 1.292130.41123&4-DG63 5.58.78.884140.5813439 29 6.784 8819 1.0613Tolylfluanid7.9 1.012 3.17939b 1.191334 4.313 13 67b37b 1.41b13144 13 8.842 7229b 1.37b13Trifluralin7.80.78.5 1.878230.681210-C58 3.7 6.414 7725 1.0213379 19 5.148 76130.69106-C, 4-C, 11-SG aC = Cochran outlier; SG = single Grubbs outlier; DG = double Grubbs outliers.bRSD r >15%; 120% < Rec. < 70%; RSD R >25%; HorRat >1.2; or fewer than 8 laboratories in an assessment.cCyprodinil was incurred in the grapes and affected quantitation.ã 2007 AOAC IN T ER N A T IONALAnalyte Avg. C, ng/g s r, ng/g RSD r, %s R, ng/g Rec., %RSD R, %HorRat No. of labs Outlier labs Atrazine9.9 1.515 1.899180.561170 7.9111593210.8812930 50 5.4166 9318 1.11115-C Azoxystrobin10 0.8 7.6 1.8102 180.561247 2.4 5.2 6.593140.5512531 32 6.188106 170.9412Bifenthrin9.10.8 9.1 1.491160.481166 8.012 9.488140.5911217 27 123387150.7711Carbaryl9.4 1.112 2.094220.671292 6.1 6.7 9.092 9.80.43118-SG589 38 6.4127 9822 1.24b12Chlorothalonil 6.20.814 2.062b32b0.93 6b28 10 37b147048b 1.77b 7b684 134 20b205 68b30b 1.77b 6bChloropyrifos9.0 2.124b 2.39026b0.79 912-SG, 10&11-DG86 9.411208623 1.011111-SG179 18 103090170.821111-SG Cyprodinil9.7 1.010 1.497140.441111-SG44 2.7 6.1 8.989200.791111-SG848 61 7.2117 85140.84108-SGo,p¢-DDD8.90.6 7.0 1.989210.66 881 4.8 5.91281150.63 911-C214 19 8.72786130.6210Dichlorvos 5.2 1.020b 2.452b45b 1.29b1258 6.6111277200.8112838 50 6.0224 8427 1.63b11Endosulfan sulfate 5.6 3.359b 2.556b45b 1.28b 2b38 9.625b157539b 1.48b 7b769 330 43b312 7740b 2.44b 7bImazalil7.60.3 3.5 3.57639b 1.18 82-C72 3.7 5.22457b33b 1.39b11589 47 7.9229 59b39b 2.25b11Imidacloprid c22 1.3 6.2 1.7100 7.90.28118-SG84 6.2 7.4 8.197 9.60.4112515 21 4.253101 100.58115-C Linuron8.6 1.112 1.586170.531146 2.2 4.9 7.491160.63102-C234 14 5.82594100.5311Methamidophos8.80.8 8.5 1.388150.46 86-C66 4.5 6.91282180.7211538 37 6.86384120.67 95-C, 8-SG Methomyl9.70.8 8.6 1.096100.321099 8.0 8.1 6.499 6.50.29102-SG997 24 2.4168 100 17 1.0511Procymidone10 0.6 6.2 2.2101 220.68 82-C92 8.5 9.21592170.7311967 118 12129 97130.8311Pymetrozine 6.90.4 6.1 1.469b200.591033 1.6 4.7 4.667b140.51 911-C127 8.5 6.71763b130.6110Tebuconazole9.70.7 6.9 1.297130.40114-C89 6.8 7.71189120.5212948 42 4.4226 9524 1.48b114-C Tolylfluanid 3.7 1.130b 2.237b59b 1.59b 4b9.3 3.740b 4.1 9.3b44b 1.37b 83-SG, 8-SG142 22 158614b61b 2.84b 812-C, 3&8-DG Trifluralin10 1.413 1.7103 170.541142 4.511 9.084220.8311169 25 153084180.8411aC = Cochran outlier; SG = single Grubbs outlier; DG = double Grubbs outliers.bRSD r >15%; 120% < Rec. < 70%; RSD R >25%; HorRat >1.2; or fewer than 8 laboratories in an assessment.cImidacloprid was incurred in the lettuces unbeknownst to the SD.ã 2007 AOAC IN T ER N A T IONALAnalyte Avg. C, ng/g s r, ng/g RSD r, %s R, ng/g Rec., %RSD R, %HorRat No. of labs Outlier labs Atrazine 8.9 1.011 1.989210.65112-C 908.29.11290130.5712187 19 10 2793140.6912Azoxystrobin 8.4 1.316b 1.884210.651165 5.28.0 8.186120.5212853 35 4.18285 9.60.591111-C Bifenthrin 9.7 2.324b 2.397240.75109-SG45 2.5 5.6 6.891150.59102-C488 51 10 7698160.8712Carbaryl 8.40.67.3 2.184250.771066 5.07.51488210.8811172 8.8 5.13486200.9512Chlorothalonil 4.80.816b 2.748b56b 1.57b 3b7014 20b297042b 1.74b 6b330 137 42b131 66b40b 2.09b 7bChloropyrifos11 1.614 5.0111 45b 1.58b 92-C82 4.5 5.61282150.64109-C953 97 10 284 9530b 1.85b1211-C Cyprodinil 8.70.910 2.087230.721256 4.58.0 9.075160.6512199 12 6.23580180.8612o,p¢-DDD 9.10.67.2 1.891200.60 99-C74 5.1 6.9 9.899130.561010-C967 81 8.41919720 1.22b11Dichlorvos 9.30.88.1 1.093110.35 7b12-C43 2.2 5.2 8.085190.73 812-SG446 22 5.05489120.6810Endosulfan sulfate12 5.444b 5.4124b43b 1.40b 4b3-SG 8319 23b198323 1.0110240 35 15 618025 1.28b10Imazalil c22 1.77.7 6.29628b0.98 87-C58 4.37.41392220.91 9186 9.7 5.2418722 1.0610Imidacloprid10 1.110 2.8104 27b0.861193 6.57.01293130.5711989 64 6.5124 99130.7811Linuron 7.8 1.317b 2.77835b 1.041160 3.0 5.01386210.8611387 26 6.64279110.59 9111-DG Methamidophos 9.2 1.112 1.592160.49 89-C42 3.58.2 5.685130.52 84-C211 12 5.53185150.73 94&9-DG Methomyl 8.50.88.9 2.88533b0.99 97-C68 4.87.0 8.791130.5412492 19 3.96098120.6912Procymidone110.98.1 3.9108 36b 1.15 812-C43 3.58.0 5.886140.531010-C170 16 9.72585150.7111Pymetrozine 7.5 1.318b 2.17528b0.821077 5.97.71077140.5710789 38 4.8117 79150.89 912-C Tebuconazole 8.70.78.0 1.287140.421141 2.2 5.4 6.282150.5812177 14 7.92888160.7612Tolylfluanid 5.8 1.220b 1.458b240.69 911-SG 467.516b1461b31b 1.21b119-C356 54 15 134 7138b 2.02b12Trifluralin 8.60.4 4.5 2.48628b0.87 99-C 928.69.41192120.5412915 60 6.5194 9221 1.31b116-CaC = Cochran outlier; SG = single Grubbs outlier; DG = double Grubbs outliers.bRSD r >15%; 120% < Rec. < 70%; RSD R >25%; HorRat >1.2; or fewer than 8 laboratories in an assessment.cImazalil was incurred in the oranges unbeknownst to the SD.ã 2007 AOAC IN T ER N A T IONALIn Option A, if the laboratory had LVI capability, then 1 or 2 mL extracts were taken for dispersive-SPE (the volume depended on the analyst preference and the type of centrifuge and tubes available in the laboratory). The final extract volume was 0.5 mL if 1 mL was taken for dispersive-SPE, and 1 mL if 2 mL underwent the cleanup step. In either case, two 15 g blank samples were used for the matrix blank (0-standard) and 6 matrix-matched calibration standards (5, 10, 50, 100, 250, and 1000 ng/g equiv alent concentrations). For dispersive-SPE of the matrix blanks, either 7 separate tubes using the same 1–2 mL extract volumes as the test samples could have been used, or 1–2 dispersive-SPE tube(s) with 7-fold greater extract volume(s).In Option B, if LVI is not available for GC/MS, then »30 mL of matrix blank extract was needed after dispersive-SPE cleanup to prepare the matrix-matched calibration standards (or $60 mL initial extract). In this case, 6 matrix blanks of 15 g each were extracted along with the test samples to provide enough blank extract volume, which were combined, and seven 8 mL aliquots were distributed to 7 dispersive-SPE tubes containing 0.4 g PSA + 1.2 g anhydrous MgSO4.B. Apparatus and ConditionsNote: Tables 4 and 5 of the collaborative study [J. AOAC Int. 90, 485(2007)] list the analytical instrumentation and sources of sample preparation materials used by each laboratory in the study. Further information appears in the full report. Since the time of the collaborativ e study, at least 3 v endors, United Chemical Technologies (Bristol, PA, USA), Restek (Bellefonte, PA, USA) and Supelco (Bellefonte, PA, USA) hav e introduced commercial dispersive-SPE products for QuEChERS and other applications. See Table 4 [J. AOAC Int. 90, 485(2007)] for sources of analytical instruments.(a) Gas chromatograph/mass spectrometer.—An ion trap, quadrupole, time-of-flight (TOF), or other GC/MS instrument may be used with electron impact (EI) ionization, an autosampler (AS), and computerized instrument control/data collection. Either LVI of 8 m L for a 1 g/mL MeCN extract (e.g., 75°C ramped to 275°C at 200°C/min) or 2 m L splitless injection of 4 g/mL extracts in toluene a t250°C m a y b e u s e d.A3–5m,0.25m m i d, phenylmethyl-deactivated guard column must be used as a retention gap in either case. The analytical column is a 30 m, 0.25 mm id, 0.25 m m film thickness (5%phenyl)-methylpolysiloxane (low bleed) analytical column (DB-5ms or equiv alent). Set He head pressure on the column to be 10 psi or constant flow to be 1.0 mL/min with systems capable of electronic pressure/flow control. After an appropriate time for solv ent delay, use an appropriate ov en temperature program, for example, starting at 75°C for MeCN extracts or 100°C for toluene ramped to 150°C at 25°C/min, then to 280°C at 10°C/min, and hold for 10 min. All collaborators had much experience in pesticide residue analysis and were free to use their own analytical conditions provided that peak shapes were Gaussian, peak widths at half heights were <5 s, and signal-to-noise ratio (S/N) of the quantitative ion for the pesticides at 10 ng/g equivalent concentrations in the sample were >10. For qualitative purposes (which were not the focus of this study), at least 3 ions yielding relativ e abundances that reasonably match a contemporaneously analyzed reference standard are typically needed to make an analyte identification.(b) Liquid chromatograph/tandem mass spectrometer.— A triple quadrupole, ion trap, or other LC/MS/MS instrument may be used provided it is capable of electrospray ionization (ESI) in the positive mode with computerized instrument control/data collection and has an AS. An injection volume (5–100 m L) will be determined for each instrument to achieve S/N > 10 for the quantitation ion for a 10 ng/g equivalent sample concentration. As in GC/MS, the collaborators had much experience in the analysis of pesticides and were free to use their own conditions. Suggested LC conditions, howev er, include a 15 cm long, 3.0 mm id, 3 m m particle size C18 column, flow rate of 0.3 mL/min, and gradient elution with an initial condition of 25% MeOH in 5 mM formic acid solution taken linearly in 15 min to 90% MeOH in 5 mM formic acid solution and held for 15 min. A short C18 guard column must be used to protect the analytical column, and a bypass valve must be used before the MS instrument to av oid introduction of the early and late eluting nonanalyte components into the detector. The MS/MS conditions were optimized in each laboratory using direct infusion into the ESI source to prov ide highest S/N for the quantitation ion of each LC-type analyte from a single MS/MS transition. A second transition with reasonably matching relative abundance ratios vs a contemporaneously analyzed reference standard is typically needed for qualitative purposes.(c) Centrifuge(s).—Capable of holding the 50 mL centrifuge tubes or bottles used for extraction and 10–15 mL graduated centrifuge tubes or 2 mL mini-tubes used in dispersiv e-SPE. Determine the rpm settings that yield a given relative centrifugal force (RCF), and ensure that maximum ratings of the centrifuge, tube/bottles, and rotors for the instrument are not exceeded.(d) Balance(s).—Capable of accurately measuring weights from0.05 to 100 g within ±0.01 g.(e) Freezer.—Capable of continuous operation <–20°C.(f) Furnace/oven.—Capable of 500°C operation.ã 2007 AOAC IN T ER N A T IONALTable 2007.01E. Averaged interlaboratory study results for the fortified and incurred pesticides aMatrix Recovery, %RSD r, %RSD R, %HorRat No. of labs (n) Grapes86 ± 1110 ± 422 ± 80.90 ± 0.2912 ± 1 Lettuces87 ± 1210 ± 720 ± 90.83 ± 0.4510 ± 1 Oranges87 ± 1510 ± 620 ± 80.84 ± 0.3710 ± 2 Overall87 ± 1110 ± 621 ± 80.86 ± 0.3711 ± 2Incurred NA b12 ± 422 ± 80.92 ± 0.3011 ± 2aData from fewer than 7 laboratories in an assessment were excluded.bNA = Not applicable.(g ) Food chopper and/or bl ender.—Preferably an s-blade vertical cutter (e.g. Stephan, Robotcoupe) and and probe blender (e.g. Ultra-Turrax, Propsep).(h ) Sol vent evaporator (optional ).—For the ev aporation of MeCN extracts, if LVI is not used in GC/MS.C. Reagents[See Table 5 [J. AOAC Int. 90, 485(2007)] for sources of chemicals.](a ) Anhydrous magnesium sul fate (MgSO 4).—Powder form;purity >98%; heated in bulk to 500°C for >5 h to remove phthalates and residual water.(b ) Acetonitril e (MeCN).—Quality of sufficient purity that is free of interfering compounds.(c ) Acetic acid (HOAc).—Glacial; quality of sufficient purity that is free of interfering compounds.(d ) 1% HOAc in MeCN.—Prepared on a v/v basis (e.g., 10 mL glacial HOAc in a 1 L MeCN solution).(e ) Anhydrous sodium acetate (NaOAc).—Powder form (NaOAc·3H 2O may be substituted, but 0.17 g per g sample must be used rather than 0.1 g anhydrous NaOAc per g sample).(f ) Primary secondary amine (PSA) sorbent.—40 m m particle size (Varian Part No. 12213024 or equiv alent). (Note : Premade dispersive-SPE tubes are now available from at least 3 vendors.)(g ) C 18 sorbent (optional ).—40 m m particle size, if samples contain >1% fat.(h ) G r a p h i t i z e d c a r b o n b l a c k (G C B ) s o r b e n t (optional ).—120/400 mesh size, if no structurally planar pesticides are included among the analytes.(i ) Helium.—Purity that has been demonstrated to be free of interfering compounds in GC/MS.(j ) Toluene (optional ).—Quality of sufficient purity that is free of interfering compounds; only needed if LVI is not used in GC/MS.(k ) Methanol (MeOH).—Quality of sufficient purity that is free of interfering compounds in LC/MS/MS prepared in mobile phase solution.(l ) Water.—Quality of sufficient purity that is free of interfering compounds in LC/MS/MS.(m ) Formic acid.—Quality of sufficient purity that is free of interfering compounds in LC/MS/MS prepared in mobile phase solution.(n ) Pesticide standards .—High purity reference standards of the pesticide analytes, and quality control (QC) and internal standards (ISs) prepared at highly concentrated stock solutions (e.g.,2000 ng/m L) in MeCN with 0.1% HOAc. Stored in dark vials in the freezer. Check annually for stability.(o ) Standard sol utions.—Prepared in MeCN for all collaborators: IS solution = 40 ng/m L of both d 10-parathion and d 6-a -HCH in MeCN; triphenylphosphate (TPP) solution = 2 ng/m L TPP in 1% HOAc in MeCN solution; QC-spike solution = 40 ng/m L of the 27 pesticide analytes in 0.1% HOAc in MeCN; and individual test solutions = 10 ng/m L of each of the 30 compounds to be detected (except 40 ng/m L TPP) in 0.1% HOAc in MeCN solution.Collaborators prepared a test mix and calibration standard spike solutions from those provided as described in E .(p ) Bl ank sampl e .—Verified to be free of analytes abov e the detection limit.(q ) Other reagents.—Certain instruments may require nitrogen or other materials/devices for their operation.D. Materials(a ) Fl uorinated ethyl ene propyl ene (FEP) centrifugetubes.—50 mL; e.g., Nalgene Part No. 3114-0050 or equivalent for <16 g sample (or 250 mL FEP centrifuge bottles for 16–75 g sample size).(b ) Spatul a/spoon and funnel .—For transferring sample into centrifuge tubes.(c ) Solvent dispenser and 1–4 L solvent bottle.—For transferring 15 mL 1% HOAc in MeCN per 15 g sample in FEP centrifuge tubes or bottles.(d ) Centrifuge tubes (optional ).—10–15 mL graduated. For evaporation and/or dispersive-SPE.(e ) Mini-centrifuge tubes (optional).—2 mL. For dispersive-SPE (use tubes with o-ring-sealed caps to avoid leaks).(f ) Syringes/pipets.—Capable of accurate sample introduction of 2 or 8 m L volume into GC/MS and appropriate volumes of matrix spike, IS, and calibration standard solutions (12.5–300 m L).(g ) Repeating or vol umetric pipets.—Capable of accurately transferring 0.5–8 mL solvent.(h ) Containers .—Graduated cylinders, volumetric flasks, weigh boats, v ials, and/or other general containers in which to contain samples, extracts, solutions, standards, and reagents.E. Preparation of Reagent Materials and Comminuted Sample(1) Prepare the necessary number of sealable vials/cups containing 6.0 ± 0.3 g anhydrous MgSO 4 + 1.5 ± 0.1 g anhydrous NaOAc (or 2.5ã 2007 AOAC IN T ER N A TIONALFigure 2007.01. Outline of the QuEChERS protocol used in the collaborative study.± 0.2 g NaOAc·3H2O) per 15 g sample. Scoops of appropriate volume can be used to speed the process, but weighing should still be done to check consistency. The containers should be sealed during storage and can be refilled and re-used without cleaning in between usages.(2) Prepare the necessary number of appropriate centrifuge tubes (2 mL mini-centrifuge tubes or 10–15 mL centrifuge tubes) containing 0.05 ± 0.01 g PSA sorbent + 0.15 ± 0.03 g anhydrous MgSO4 per 1 mL extract taken for dispersive-SPE cleanup. (Note: At least United Chemical Technologies, Restek, and Supelco now provide dispersive-SPE products commercially to replace this step.) If LVI is not available for GC/MS, then evaporation of the extracts will be needed, and 8 mL extract will be transferred to 10–15 mL sealable centrifuge tubes containing 0.40 ± 0.08 g PSA sorbent + 1.20 ± 0.24 g anhydrous MgSO4. For matrixes that contain >1% fat, add an additional 0.05 ± 0.01 g C18 sorbent per mL extract to the container. If no planar pesticides are among the analytes (e.g., thiabendazole, terbufos, quintozene, and hexachlorobenzene), then 0.05 ± 0.01 g GCB sorbent per mL extract can also be added to the tube. (Note: Final extract volume may have to be reduced to 0.4 mL per 1 mL aliquot in dispersive-SPE if all 4 powders are used.) (3) Prepare 1% HOAc in MeCN in dispenser bottle by adding 10 mL HOAc to 990 mL v olume of MeCN or different desired amount in the same ratio.(4) Label all vials and tubes appropriately that will be used in the method.(5) Note: Step 5 was conducted by the Study Director (SD) when preparing the test samples. An appropriate chopper must be used to comminute large, representative sample portions. An uncommon or deuterated pesticide standard may be spiked into the sample during homogenization to determine the effectiv eness of the procedure. Blend the sample until it gives a consistent texture. Transfer .200 g to a sealable container for freezer storage after further homogenization with a probe blender. Blend this subsample with the mixer until it is homogeneous. The test portion (e.g., 15 g) is taken for extraction immediately, and the container is then sealed and stored in the freezer in case re-analysis is necessary. The advantages of this approach are that the 15 g portion is highly representative of the original sample, the sample is well-comminuted to improv e extraction by shaking, less time is spent on the ov erall homogenization process than trying to prov ide equiv alent homogenization of the large initial sample with the chopper, and a frozen subsample is available for re-analysis if needed.To provide the most homogeneous comminuted samples, frozen conditions, sufficient chopping time, and appropriate sample size to chopper volume ratio should be used. Use of frozen samples also minimizes degradative and volatilization losses of certain pesticides. In this case, cut the sample into 2–5 cm3 portions with a knife and store the sample in the freezer prior to processing. Cryogenic blending devices, liquid nitrogen, or dry ice may also be used (but make sure all dry ice has sublimed before weighing samples and ensure that water condensation is minimal, especially in a humid environment). (6) For laboratories with LVI in GC/MS, prepare a test mix of the pesticides in MeCN + 0.1% HOAc to determine the retention times (t R) and MS quantitation/diagnostic ions at the particular GC/MS conditions to be used in the analysis [see Table 2 of the collaborative study (J. AOAC Int. 90, 485(2007)].The preparation of the test mix and calibration spiking standards are described as follows:(1) Test mix in MeCN + 0.1% HOAc.—4 ng/m L in 10 mL of all 30 compounds to be analyzed. Add 1 mL each of QC-spike solution + IS solution + TPP test solution + 1% HOAc in MeCN and fill to 10 mL with MeCN. Calibration spike standards in MeCN for 27 pesticide analytes (make 10 mL each in volumetric flasks, then transfer to 15 mL dark glass vials and store in freezer).(2) Cal-standard-1000.—20 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 5 mL QC-spike solution + 1 mL IS solution + 1 mL 1% HOAc in MeCN and fill to the mark with MeCN.(3) Cal-standard-250.—5 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 1.25 mL QC-spike solution + 1 mL IS solution + 1 mL 1% HOAc in MeCN and fill to the mark with MeCN.(4) Cal-standard-100.—2 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 500 m L QC-spike solution + 1 mL IS solution + 1 mL 1% HOAc in MeCN and fill to the mark with MeCN.(5) Cal-standard-50.—1 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 250 m L QC-spike solution + 1 mL IS solution + 1 mL 1% HOAc in MeCN and fill to the mark with MeCN.(6) Cal-standard-10.—0.2 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 50 m L QC-spike solution + 1 mL IS solution + 1 mL 1% HOAc in MeCN and fill to the mark with MeCN.(7) Cal-standard-5.—0.1 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 25 m L QC-spike solution + 1 mL IS solution + 1 mL 1% HOAc in MeCN and fill to the mark with MeCN. For laboratories without LVI in GC/MS, the preparation of the test mix and the calibration spiking standards are described below: (1a) Test mix for GC in tol uene.—4 ng/m L in 10 mL of all 30 compounds to be analyzed. Add 1 mL QC-spike solution + 1 mL IS solution + 1 mL TPP test solution and fill to 10 mL with toluene. Calibration spike standards in MeCN for LC/MS/MS (in dark glass AS vials stored in freezer).(2a) Cal-standard-1000.—20 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 500 m L QC-spike solution + 100 m L IS solution + 100 m L 1% HOAc in MeCN + 320 m L MeCN.(3a) Cal-standard-250.—5 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 125 m L QC-spike solution + 100 m L IS solution + 100 m L 1% HOAc in MeCN + 695 m L MeCN.(4a) Cal-standard-100.—2 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 50 m L QC-spike solution + 100 m L IS solution + 100 m L 1% HOAc in MeCN + 770 m L MeCN.Dilute QC-spike solution.—4 ng/m L. Transfer 100 m L QC-spike solution to AS vial and add 900 m L MeCN.(5a) Cal-standard-50.—1 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 250 m L dilute QC-spike solution + 100 m L IS solution + 100 m L 1% HOAc + 570 m L MeCN.(6a) Cal-standard-10.—0.2 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 50 m L dilute QC-spike solution + 100 m L IS solution + 100 m L 1% HOAc and 770 m L MeCN.(7a) Cal-standard-5.—0.1 ng/m L of each pesticide + 4 ng/m L IS in MeCN + 0.1% HOAc. Add 25 m L dilute QC-spike solution + 100 m L IS solution + 100 m L 1% HOAc + 795 m L MeCN.Calibration spike standards in toluene.—Make 10 mL each in volumetric flasks, then transfer to 15 mL dark glass vials and store in freezer.(8a) Cal-standard-1000-tol.—20 ng/m L of each pesticide + 4 ng/m L IS in toluene. Add 5 mL QC-spike solution + 1 mL IS solution and fill to the mark with toluene.ã 2007 AOAC IN T ER N A T IONAL。

AOAC 991.43

Revised: March 1996 32.1.17AOAC Official Method 991.43Total, Soluble, and Insoluble Dietary Fiber in FoodsEnzymatic-Gravimetric Method, MES–TRIS BufferFirst Action 1991Final Action 1994Codex-Adopted—AOAC Method*(Applicable to processed foods, grain and cereal products, fruits, and vegetables.)Method Performance:See Table 991.43A for method performance data.A. PrincipleDuplicate samples of dried foods, fat-extracted if containing >10%fat, undergo sequential enzymatic digestion by heat stable α-amylase,protease, and amyloglycosidase to remove starch and protein. For total dietary fiber (TDF), enzyme digestate is treated with alcohol to precipi-tate soluble dietary fiber before filtering, and TDF residue is washed with alcohol and acetone, dried, and weighed. For insoluble and soluble dietary fiber (IDF and SDF), enzyme digestate is filtered, and residue (IDF) is washed with warm water, dried and weighed. For SDF,combined filtrate and washes are precipitated with alcohol, filtered,dried, and weighed. TDF, IDF, and SDF residue values are corrected for protein, ash, and blank.B. Apparatus(a ) Beakers.—400 or 600 mL tall-form.Table 991.43A Method Performance for Total, Soluble, and Insoluble Dietary Fiber in Foods (Fresh Weight Basis),Enzymatic-Gravimetric Method, MES-TRIS BufferFood Mean, g/100 g s r s R RSD r%RSD R % Barley 12.250.360.85 2.88 6.89High-fiber cereal 33.730.700.94 2.08 2.79 Oat bran 16.92 1.06 2.06 6.2612.17 Soy bran 67.14 1.01 1.06 1.50 1.58 Apricots 1.120.010.010.890.89 Prunes 9.290.130.40 1.404.31 Raisins 3.130.090.15 2.88 4.79 Carrots 3.930.130.13 3.31 3.31 Green beans 2.890.070.07 2.42 2.42 Parsley 2.660.070.14 2.635.26Soluble dietary fiber (SDF) Barley 5.020.400.628.0112.29 High-fiber cereal 2.780.440.5615.83 20.14 Oat bran 7.170.72 1.1410.04 15.90 Soy bran6.900.300.60 4.358.70 Apricots 0.530.020.02 3.77 3.77 Prunes 5.070.110.31 2.17 6.11 Raisins 0.730.050.16 6.8521.92 Carrots 1.100.070.18 6.3616.36 Green beans 1.020.080.117.8410.78 Parsley 0.640.030.10 4.6915.63 Insoluble dietary fiber (IDF) Barley 7.050.610.618.628.62 High-fiber cereal 30.520.440.71 1.44 2.33 Oat bran9.730.85 1.178.7412.02 Soy bran 60.530.700.70 1.16 1.16 Apricots 0.590.020.02 3.39 3.39 Prunes 4.170.070.09 1.68 2.16 Raisins 2.370.040.07 1.69 2.95 Carrots 2.810.090.16 3.20 5.69 Green beans 2.010.080.08 3.98 3.98 Parsley 2.370.120.24 5.0610.13 Total dietary fiber (SDF + IDF) Barley 12.140.390.70 3.21 5.77 High-fiber cereal 33.300.630.90 1.89 2.70 Oat bran 16.900.99 1.49 5.868.82 Soy bran 67.560.560.940.83 1.39 Apricots 1.120.020.02 1.79 1.79 Prunes 9.370.120.30 1.28 3.20 Raisins 3.100.050.18 1.61 5.81 Carrots 3.920.110.13 2.81 3.32 Green beans 3.030.090.12 2.97 3.96 Parsley 3.010.120.23 3.997.64(b ) Filtering crucible.—With fritted disk, coarse, ASTM 40–60µm pore size, Pyrex 60 mL (Corning No. 36060 Bchner, Corning,Inc., Science Products, Corning, NY 14831, USA, or equivalent).Prepare as follows. Ash overnight at 525° in muffle furnace. Let furnace temperature fall below 130° before removing crucibles.Soak crucibles 1 h in 2% cleaning solution at room temperature.Rinse crucibles with H 2O and then deionized H 2O; for final rinse,use 15 mL acetone and then air-dry. Add ca 1.0 g Celite to dry crucibles, and dry at 130° to constant weight. Cool crucible ca 1h in desiccator, and record weight, to nearest 0.1 mg, of crucible plus Celite.(c ) V acuum system.—V acuum pump or aspirator with regulating device. Heavy walled filtering flask, 1 L, with side arm. Rubber ring adaptors, for use with filtering flasks.(d ) Shaking water baths.—(1) Capable of maintaining 98 2°,with automatic on-and-off timer. (2) Constant temperature, adjust-able to 60°.(e ) Balance.—Analytical, sensitivity 0.1 mg.(f ) Muffle furnace.—Capable of maintaining 525 5°.(g ) Oven.—Capable of maintaining 105 and 130 3°.(h ) Desiccator .—With SiO 2 or equivalent desiccant. Biweekly,dry desiccant overnight at 130°.(i ) pH meter .—Temperature compensated, standardized with pH 4.0, 7.0, and 10.0 buffer solutions.(j ) Pipetters .—With disposable tips, 100–300 µL and 5 mL capacity.(k ) Dispensers.—Capable of dispensing 15 0.5 mL for 78%ethanol, 95% ethanol, and acetone; 40 0.5 mL for buffer.(l ) Magnetic stirrers and stir bars.C. ReagentsUse deionized water throughout.(a ) Ethanol solutions.—(1) 85%. Place 895 mL 95% ethanol into 1 L volumetric flask, dilute to volume with H 2O. (2) 78%. Place 821 mL 95% ethanol into 1 L volumetric flask, dilute to volume with H 2O.(b ) Heat-stable α-amylase solution.—Catalog Number A 3306,Sigma Chemical Co., St. Louis, MO 63178, USA, or Termamyl 300L, Catalog Number 361-6282, Novo-Nordisk, Bagsvaerd, Den-mark, or equivalent.(c ) Protease.—Catalog Number P 3910, Sigma Chemical Co, or equivalent. Prepare 50 mg/mL enzyme solution in MES/TRIS buff-er fresh daily.(d ) Amyloglucosidase solution .—Catalog Number AMG A9913,Sigma Chemical Co, or equivalent. Store at 0–5°.(e ) Diatomaceous earth.—Acid washed (Celite 545 AW, No.C8656, Sigma Chemical Co. or equivalent).(f ) Cleaning solution .—Liquid surfactant-type laboratory cleaner, designed for critical cleaning (Micro ®, International Prod-ucts Corp., Burlington, NJ 08601, USA, or equivalent). Prepare 2%solution in H 2O.(g ) MES .—2-(N -Morpholino)ethanesulfonic acid (No. M-8250,Sigma Chemical Co., or equivalent.)(h ) TRIS.—Tris(hydroxymethyl)aminomethane (No. T-1503,Sigma Chemical Co., or equivalent).(i ) MES–TRIS buffer solution.—0.05M MES, 0.05M TRIS, pH 8.2 at 24°. Dissolve 19.52 g MES and 12.2 g TRIS in 1.7 L H 2O.Adjust pH to 8.2 with 6N NaOH, and dilute to 2 L with H 2O.(Note: It is important to adjust pH to 8.2 at 24°. However, if buffer temperature is 20°, adjust pH to 8.3; if temperature is 28°,adjust pH to 8.1. For deviations between 20 and 28°, adjust by interpolation.)(j ) Hydrochloric acid solution.—0.561N . Add 93.5 mL 6N HCl to ca 700 mL H 2O in 1 L volumetric flask. Dilute to 1 L with H 2O.D. Enzyme PurityTo ensure absence of undesirable enzymatic activities and pres-ence of desirable enzymatic activities, run standards listed in Table 991.43B each time enzyme lot changes or at maximum interval of 6months.E. Sample Preparation and DigestionPrepare samples as in 985.29E (see 45.4.07) (if fat content of sample is unknown, defat before determining dietary fiber). For high sugar samples, desugar before determining dietary fiber by extract-ing 2–3 times with 85% ethanol, 10 mL/g, decanting, and then drying overnight at 40°.Run 2 blanks/assay with samples to measure any contribution from reagents to residue.Weigh duplicate 1.000 0.005 g samples (M 1 and M 2), accurate to 0.1 mg, into 400 mL (or 600 mL) tall-form beakers. Add 40 mL MES–TRIS buffer solution, pH 8.2, to each. Stir on magnetic stirrer until sample is completely dispersed (to prevent lump formation,which would make test material inaccessible to enzymes).Add 50 µL heat-stable α-amylase solution, stirring at low speed.Cover beakers with Al foil, and incubate in 95–100° H 2O bath 15min with continuous agitation. Start timing once bath temperature reaches 95° (total of 35 min is normally sufficient).Remove all beakers from bath, and cool to 60°. Remove foil.Scrape any ring from inside of beaker and disperse any gels in bottom of beaker with spatula. Rinse beaker walls and spatula with 10 mL H 2O.Add 100 µL protease solution to each beaker. Cover with Al foil,and incubate 30 min at 60 1° with continuous agitation. Start timing when bath temperature reaches 60°.Remove foil. Dispense 5 mL 0.561N HCl into beakers while stirring. Adjust pH to 4.0–4.7 at 60°, by adding 1N NaOH solution or 1N HCl solution. (Note: It is important to check and adjust pH while solutions are 60° because pH will increase at lower tempera-tures.) (Most cereal, grain, and vegetable products do not require pH adjustment. Once verified for each laboratory, pH checking procedure can be omitted. As a precaution, check pH of blank routinely; if outside desirable range, check samples also.)Add 300 µL amyloglucosidase solution while stirring. Cover with Al foil, and incubate 30 min at 60 1° with constant agitation. StartTable 991.43B Standards for Testing Enzyme Activity Standard Activity Tested Weight of Standard, gExpected Recovery, (%)Citrus pectin Pectinase 0.1–0.295–100 Arabinogalactan Hemicellulase 0.1–0.295–100β-Glucan β-Glucanase 0.1–0.295–100Wheat starch α-Amylase + AMG 1.0 0–1 Corn starch α-Amylase + AMG 1.00–1Casein Protease0.30–1timing once bath reaches 60°.F. Determination of Total Dietary FiberTo each digested sample, add 225 mL (measured after heating) 95% ethanol at 60°. Ratio of ethanol to sample volume should be 4:1. Remove from bath, and cover beakers with large sheets of Al foil. Let precipitate form 1 h at room temperature.Wet and redistribute Celite bed in previously tared crucible B(b), using 15 mL 78% ethanol from wash bottle. Apply suction to crucible to draw Celite onto fritted glass as even mat.Filter alcohol-treated enzyme digestate through crucible. Using wash bottle with 78% ethanol and rubber spatula, quantitatively transfer all remaining particles to crucible. (Note: If some samples form a gum, trapping the liquid, break film with spatula.)Using vacuum, wash residue 2 times each with 15 mL portions of 78% ethanol, 95% ethanol, and acetone. Dry crucible containing residue overnight in 105° oven. Cool crucible in desiccator ca 1 h. Weigh crucible, containing dietary fiber residue and Celite, to near-est 0.1 mg, and calculate residue weight by subtracting weight of dry crucible with Celite, B(b).Use one duplicate from each sample to determine protein, by method 960.52 (see 12.1.07), using N× 6.25 as conversion factor. For ash analysis, incinerate second duplicate 5 h at 525°. Cool in desiccator, and weigh to nearest 0.1 mg. Subtract weight of crucible and Celite, B(b), to determine ash weight.G. Determination of Insoluble Dietary FiberWet and redistribute Celite bed in previously tared crucible, B(b), using ca 3 mL H2O. Apply suction to crucible to draw Celite into even mat. Filter enzyme digestate, from E, through crucible into filtration flask. Rinse beaker, and then wash residue 2 times with 10 mL 70° H2O. Combine filtrate and water washings, transfer to pretared 600 mL tall-form beaker, and reserve for determination of soluble dietary fiber, H. Using vacuum, wash residue 2 times each with 15 mL portions of 78% ethanol, 95% ethanol, and acetone. (Note: Delay in washing IDF residues with 78% ethanol, 95% ethanol, and acetone may cause inflated IDF values.)Use duplicates to determine protein and ash as in F.H. Determination of Soluble Dietary FiberProceed as for insoluble dietary fiber determination through in-struction to combine the filtrate and water washings in pretared 600 mL tall-form beakers. Weigh beakers with combined solution of filtrate and water washings, and estimate volumes.Add 4 volumes of 95% ethanol preheated to 60°. Use portion of 60° ethanol to rinse filtering flask from IDF determination. Alterna-tively, adjust weight of combined solution of filtrate and water washings to 80 g by addition of H2O, and add 320 mL 60° 95% ethanol. Let precipitate form at room temperature 1 h.Follow TDF determination, F, from “Wet and redistribute Celite bed . . . .”I. CalculationsBlank (B, mg) determination:B = [(BR1 + BR2)/2] – P B – A Bwhere BR1 and BR2 = residue weights (mg) for duplicate blank determinations; and P B and A B = weights (mg) of protein and ash, respectively, determined on first and second blank residues. Dietary fiber (DF, g/100 g) determination:DF = {[(R1 + R2)/2] – P – A – B}/[(M1 + M2)/2] × 100 where R1 and R2 = residue weights (mg) for duplicate samples; P and A = weights (mg) of protein and ash, respectively, determined on first and second residues; B = blank weight (mg); and M1 and M2 = weights (mg) for samples.Total dietary fiber determination: Determine either by independent analysis, as in F, or by summing IDF and SDF, as in G and H. Reference: J. AOAC Int. 75, 395(1992).*Adopted as a Codex Defining Method for gravimetry/enzymatic di-gestion of total dietary fiber in infant formula and follow-up for-mula.。

AOAC 试验方法 Method AOAC method 985.22

(i) Standard solutions.—Prepare all stock solutions and dilutions in glass-distilled acetone. Prepare GC standard solutions so 4 µL injection causes 30–70% full scale deflection in properly functioning system. Suggested concentrations are given in Table 985.22. Check responses before beginning analysis. Store all standard solutions in tightly stoppered containers in refrigerator. Let equilibrate 1 h at room temperature before using.
ganic
layer
in
first
separatory
funnel
by
passing
through
ca
14 supported on washed glass wool in 4 in(10 cm)
funnel, collecting in 500 mL K–D concentrator fitted with volumet-
Instrument used
B(g)(2)
E
0.5
0.4
0.1
0.1
0.5
0.5
0.2
0.2
0.5
0.5
0.2
0.4
2.0
0.4

AOAC 992.30

17.3.07AOAC Of f i c ial Method 992.30Con f irmed To t al Coliform and Escherichia coliin All FoodsSub s trate Sup p orting Disc Method (ColiComplete®)First Ac t ion 1992Fi nal Ac tion1996(Ap pli ca ble to si mul ta neous enu mer a tion of con firmed to tal coliform and E. coli in all foods. Not ap p li c a b le to E. coli strain O157:H7.)See Ta b le 992.30 for the results of the interlaboratory study sup p ort i ng ac c ep t ance of the method.A.Prin ci pleS u b s t r a t e s u p p o r t i n g d i s c s(S S D),c o n t a i n i n g 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-Gal) and 4-methylumbelliferyl-β-D-glucuronide (MUG), are used to de t er m ine lev e ls of to t al coliforms and E. coli. Discs are added to lauryl sul f ate tryptose broth (LST) in o c u l ated with se l ected di l u t ions of foods. Test samples are in c u b ated at 35°C and ex a m i ned af t er 24 and 48 h for con f irmed to t al coliform count and af t er 30 h for con f irmed E. coli pres e nce.β-Galactosidase, from coliforms pres e nt in foods, cleaves X-Gal into5-bromo-4-chloro-indoxyl in ter me di ate which un der goes ox i d a t ion to yield wa t er-insoluble blue dimer, vi s u a lly de t ect a ble on disc or in sur r ound i ng me d ium as con f irmed pos i t ive re s ult of to t al coliform ac tiv ity.β-Glucuronidase, from E. coli pres e nt in foods, cleaves MUG into glucuronide and methylumbelliferone which flu o r esces un d er longwave UV light (366 nm) as con f irmed pos i t ive re s ult of E. coli pres ence.β-Glucuronidase has been found spe c ific to ge n us Esch e richia (Esch e richia and Shigella) and some Sal mo nella. Prac t i c ally, from most foods, only E. coli yields pos i t ive re s ults. This method will not de t ect E. coli O157:H7.B. Ap p a r a t us(a) Pipets.—1.0 and 10.0 mL, with 0.1 mL grad u a t ions.(b) Blender.—2-speed model with ex p lo s ion-proof mo t or, ca p a b le of high-speed op e r a t ion 16 000–18 000 rpm. One glass or metal blender jar (1 L ca p ac i ty) with cover for each an a l yt i c al unit.(c) Vor t ex mixer.(d)In c u b a t or.—Ca p a b le of main t ain i ng 35°–37°C.(e)UV light source.—366 nm, handheld, 6 watts. Wear safety glasses when light is on.C. Me d ia and Re a gentsIn g re d i e nts and re a gents used to pre p are the fol l ow i ng me d ia may be ob t ained from any man u f ac t urer. For con v e n ience, com m er c ial de h y d rated me d ia equiv a l ent to for m u l a t ion may be used. Com m er c ial me d ia should be tested with known coliforms (Enterobacter, Klebsiella spp.) and E. coli prior to use to ver i fy growth pro m o t ion prop e r t ies. Tubes used for me d ia prep a r a t ions should be ex a m i ned un d er UV light (366 nm) to con f irm lack of autofluorescence prior to use.Be f ore autoclaving, ad j ust me d ia pH, as nec e s s ary, by add i ng 1M NaOH or 1M HCl, to at t ain stated post-autoclaving pH. Use pH me t er stan d ard i zed against stan d ard buff e rs, 964.24 (see A.1.04).(a) LST.—Dis s olve 20.0 g trypticase or tryptose (pan c re a tic di g est of ca s ein), 5.0 g NaCl, 5.0 g lac t ose, 2.75 g K2HPO4, 2.75 g KH2PO4, and 0.1 g so d ium lauryl sul f ate in l L H2O, mix i ng over low heat, if nec e s s ary. Dis p ense 10 mL por t ions into 20 × 150 mm tubes. Au t o c lave 15 min at 121°C. Fi n al pH must be 6.8 ± 0.1.(b) SSD.—5mm di am e ter poly eth yl ene discs con tain ing 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside and 4-methylumbelliferyl-β-D-glucuronide (MUG). Avail a ble as ColiComplete Sub s trate Sup p orting Disc from BioControl Sys t ems, Inc. (12822 SE 32nd St, Bellevue, WA 98005, USA; Tel: +1-425-603-1123).(c)Butterfield’s buf fered phos phate dil u ent.—(1) Stock so lu tion.—Dis s olve 34.0 g KH2PO4 in 500 mL H2O, ad j ust to pH 7.2 with ca 175 mL 1M NaOH, and di l ute to 1 L. Store in re f rig e r a t or.(2) Dil u ent.—Di l ute 1.25 mL stock so l u t ion to 1 L with H2O. Pre p are di l u t ion blanks with this so l u t ion, dis p ens i ng enough to al l ow for losses dur i ng autoclaving. Au t o c lave 15 min at 121°C.D. Prep a r a t ion of Test Suspensions(Pre p are all dec i m al di l u t ions with 90 mL ster i le dil u e nt plus 10 mL pre v i o us di l u t ion un l ess oth e r w ise spec i f ied. Shake all di l u t ions 25 times in 30 cm arc. Pipets must ac c u r ately de l iver re q uired vol u me; do not use to de l iver <10% of their to t al vol u me.)(a) Frozen and/or pre p ared foods and raw in g re d i e nts.—Use bal ance with ca pac ity of≥2 kg and readability of 0.1 g to aseptically weigh 50 g unthawed (if frozen) test sam p le into ster i le blender jar. Add 450 mL dil u e nt, (c)(2), and blend 2 min at high speed. (If nec e s s ary to tem p er frozen test sam p le to re m ove 50 g test por t ion, hold ≤18 h at 2°–5°C.) Not more than 15 min should elapse from time test portion is blended un t il all di l u t ions are made in ap p ro p ri a te me d ia.If en t ire test sam p le con s ists of <50 g, weigh por t ion equiv a lent to p le and add vol u me of ster i le dil u e nt re q uired to make l u t ion. To t al vol u me in blender jar must com p letely cover blades.(b)Tree nut meat halves and larger pieces.—Asep tically weigh50 g test portion into ster i le jar. Add 50 mL dil u e nt, (c)(2), and shake vig o r o usly (50 times through 30 cm arc) to ob t ain 100 di l u t ion. Let stand 3–5 min and shake just be f ore mak i ng se r ial di lu tions and in oc u la tions.(c) Nut meal.—Asep t ically weigh 10 g test portion into ster i le jar. Add 90 mL dil u e nt, C(c)(2), and shake vig o r o usly (50 times through 30 cm arc) to ob t ain 101 di l u t ion. Let stand 3–5 min and shake to re s us p end just be f ore mak i ng se r ial di l u t ions and in o c u l a t ions.E.Anal y sisIn o c u l ate LST tubes with ap p ro p ri a te test portion di l u t ion se r ies se l ected to de t er m ine lev e ls or pres e nce/ab s ence of to t al coliforms and E. coli in test sam p le. Asep t ically add 1 SSD/tube. In c u b ate tubes at 35°C.(1)For to t al coliforms.—Af ter≥24 h in c u b a t ion, ex a m i ne tubes for vi s u a lly de t ect a ble blue color on disc or in sur r ound i ng me d ium. Pres ence of de tect able blue color in di cates con firmed pos i tive to tal coliform re s ult. (Note: A wide range of blue color in t en s ity may be ex p ected, de p end i ng on test sam p le com p o s i t ion and microflora. All blue re ac tions are pos i tive re gard less of in ten sity of color.) Reincubate at 35°C for ad d i t ional 24 h; re e x a m i ne. Con t inued ab s ence of blue in d i c ates neg a t ive re s ult; pres e nce of blue color in di cates con firmed pos i tive re sult for to tal coliforms.(2) For E. coli.—Ex a m i ne tubes from step 1 un d er UV light (366 nm) af t er 30 ± 2 h from start of ini t ial in c u b a t ion. (Cau t ion: Do not look di r ectly into light source.) Flu o r es c ent tubes in d i c ate con f irmed pos i t ive re s ult for E. coli.(3)Con t rols.—Known pos i t ive E. coli and neg a t ive con t rol tubes must be in c luded to fa c il i t ate in t er p re t a t ion of MUG flu o r es c ent© 2005 AOAC IN T ER N A T IONALTa b le 992.30. Interlaboratory study re s ults for con f irmed to t al coliform and E. coli in all foods, Colicomplete®sub s trate sup p ort i ng disc methodProd uct Level Mean log/g s r s R RSD r, %RSD R, %To tal coliformsDry egg Low 1.6810.6010.506 35.8 30.1Me dium 2.8010.3960.567 14.1 20.2High 2.9760.5590.626 18.8 21.0 Frozen veg e ta bles a Low 1.0070.3640.491 36.1 48.8Me dium 1.6800.3980.568 23.7 33.8High 2.1780.4580.592 21.0 27.2Nut meats Low 1.1890.4360.562 36.7 47.3Me dium 1.8140.3560.650 19.6 35.8High 1.7200.4430.621 25.8 36.1Raw liq u id milk Lot 1 2.8980.3080.952 10.6 32.9Lot 2 2.6050.3340.941 12.8 36.1Lot 3 2.8340.3990.833 14.1 29.4 Ground beef Lot 1 3.2700.2870.596 8.78 18.2Lot 2 3.3510.2500.596 7.46 17.8Lot 3 2.6930.3430.757 12.7 28.1Pork sau s age (run 1)Lot 1 2.5710.3410.522 13.3 20.3Lot 2 2.3250.3310.500 14.2 21.5Lot 3 3.0100.1470.147 4.88 4.88Pork sau s age (run 2)Lot 10.9310.1940.347 20.8 37 3Lot 2 2.3630.3270.457 13.8 19.3Lot 3 3.2420.3190.667 9.84 20.6E. coliDry egg Low 1.0740.7670.725 71.4 67.5Me dium 1.9460.8290.679 42.6 43.9High 2.2880.6630.666 29.0 29.1 Frozen veg e ta bles a Low0.7510.3910.433 52.1 57.7Me dium 1.3610.2210.631 16.2 46.4High 2.0510.5700.577 27.8 28.1Nut meats Low0.0840.2910.423346 504Me dium0.6710.1260.302 18.8 45.0High0.6610.2270.248 34.3 37.5Raw liq u id milk Lot 1 1.6190.3420.367 21.1 22.7Lot 2 2.9010.3150.498 10.9 17.2Lot 3 3.7250.2530.398 6.79 10.7 Ground beef Lot 1 1.3020.1730.280 13.3 21.5Lot 2 1.5870.6710.696 42.3 43.9Lot 3 1.1140.3280.418 29.4 37.5Pork sau s age (run 1)Lot 1 2.2650.3550.551 15.7 24.3Lot 2 2.3620.3250.558 13.8 23.6Lot 3 1.9880.4030.436 20.3 21.9Pork sau s age (run 2)Lot 10.8170.2270.354 27.8 43.3Lot 2 1.3280.3250.309 24.5 23.3Lot 3 1.9390.2260.332 11.7 17.1aFrozen broc c oli.© 2005 AOAC IN T ER N A T IONALre a c t ion. Neg a t ive con t rols must in c lude both non-E. coli/coliformtube (Enterobacter or Klebsiella spp.) and uninoculated me d ia tube.Ref er ence:J. AOAC Int.76, 988(1993).Re v ised: June 2000© 2005 AOAC IN T ER N A T IONAL。

AOAC

45.3.05AOAC Official Method982.30Protein Efficiency RatioCalculation MethodFirst Action1982A.PrincipleProtein efficiency ratio is calculated from the essential amino acid composition of sample protein(DC-PER)or from both essential amino acid composition and enzymatic digestibility of sample pro-tein(C-PER).Used together,C-PER and DC-PER models are capa-ble of providing reliable estimates of protein quality for majority of foods and food ingredients currently in use.Rat bioassay,936.14 (see45.3.02),remains official method for determining protein qual-ity;C-PER and DC-PER assays are alternative methods for routine quality control screening of foods and food e of both assays is recommended when estimating protein quality to provide internal check.Experience indicates that,in rare cases,the2models will report quite different estimates of protein quality.When this oc-curs,it should be regarded as a warning that the sample under analy-sis is probably:(1)Single-cell protein or protein surrounded by heavy cell walls(e.g.,yeast or wheat bran),where DC-PER will overestimate protein quality,or(2)Partially or completely predigested proteins(e.g.,liquid protein supplements),where C-PER will underestimate protein quality or (3)Protein sources known to possess significant quantities of proteolytic inhibitors(e.g.,improperly heat-treated soy protein), where DC-PER will overestimate protein quality.For major discrepancies in PER predictions of the2models,use rat assay as assay of choice to estimate protein quality. Computational procedures for obtaining C-PER and DC-PER esti-mates are too lengthy for repetitive hand calculation.For routine use of assay,it is recommended that algorithm be placed on computer.B.Apparatus(a)Amino acid analyzer.—Able to accurately measure individ-ual amino acids at concentrates as low as20nmolar.Must be stan-dardized using known amino acid standards at least once every24h.(b)Hydrolysis tubes.—Any standard Kimax/Pyrex test tube or ampule≥15mL capacity.(c)Water-jacketed chamber.—To fit on stir plate and connected to37°C circulating H2O bath.(d)Water bath.—55°C.(e)pH meter.—Having combination pH electrode and capable of reading to0.01pH unit.C.Reagents(a)ANRC reference casein.—Available from New Zealand Milk Products,1269N McDowell,PO Box80816,Petaluma,CA 94975-8016,USA;or Teklad,a Harlan Sprague Dawley,Inc.Co., PO Box4220,Madison,WI53711,USA.(b)Amino acid standards.—ASP,THR,SER,GLU,PRO,GLY, ALA,VAL,MET,ILE,LEU,TRY,PHE,LYS,HIS,AMM,ARG, CYS,and TRP.Available from any amino acid analyzer supply house(e.g.,Beckman Instruments,Inc.,No.338088;Pierce Chemi-cal Co.,Amino Acid Standard Kit22,No20065).(c)Performic acid.—Add1mL30%H2O2to9mL formic acid (88%).Let stand1h and cool to0°C.(d)Buffer solution.—Use buffer recommended for sample dilu-tion for amino acid analyzer.(e)Enzyme solutions.—Use the following enzymes(Sigma Chemical Co.)or their equivalent:porcine pancreatic trypsin (Type IX),porcine intestinal peptidase(Grade I),bovine pancreatic α-chymotrypsin(Type II),bacterial protease(Pronase P or E).Solu-tion A.—Dissolve227040BAEE units of trypsin+1860BAEE units ofα-chymo-trypsin+0.520L-leucineβ-naphthylamide units of peptidase in10mL H2O.Solution B.—Dissolve65casein units of bacterial protease in10mL H2O.Store both solutions on ice. (f)Control protein.—Suspend10g ANRC sodium caseinate, (a),in200mL H2O and adjust to pH8with NaOH.Maintain at pH8≥1h.Freeze-dry and determine N content by Kjeldahl method.D.Nitrogen DeterminationDetermine N by955.04C(see2.4.03),920.39A(see4.5.01), 976.05A(see4.2.05),or other appropriate Kjeldahl method.E.Test Portion Hydrolysis(a)Acid hydrolysis.—Place ca0.1g(weigh to0.1mg accuracy) test portion in hydrolysis tube,add10mL6M HCl,and mix. Freeze in dry ice-alcohol bath.Draw and hold vacuum of≤50mm for1min;seal tube under vacuum.Hydrolyze24h at110±1°C. Cool,open tube,and filter hydrolysate through Whatman No.1pa-per;rinse tube3times with H2O and filter each rinse.Dry filtrate at 65°C under vacuum.Dissolve dry hydrolysate in volume of buffer appropriate for amino acid analyzer.Store hydrolysate not>1week before e this hydrolysate to determine all amino acids except methionine,cystine and/or cysteine,and tryptophan. (b)Performic acid oxidation followed by acid hydroly-sis.—Place ca0.1g(0.1mg accuracy)test portion in hydrolysis tube,add2mL cold performic acid,and let sit overnight at0–5°C. Add3mL cold HBr+0.04mL1-octanol(antifoam);immediately mix contents30s in ice-H2O bath and evaporate to dryness at40°C under vacuum.Add10mL6M HCl to tube and perform acid hydro-lysis as described above.This treatment will quantitatively convert methionine to methionine sulfone and cystine and/or cysteine to cysteic e this hydrolysate to determine methionine(MET) and cystine/cysteine(CYS).(c)Alkaline hydrolysis.—Place ca0.1g(0.1mg accuracy)test portion into glass hydrolysis tube having Nalgene polypropylene centrifuge tube as internal liner.Add25mg hydrolyzed potato starch (omit if product is high in starch).Add0.6mL fresh4.2M NaOH+ 0.04mL1-octanol.Mix contents2min under partial vacuum.Freeze tube contents in dry ice–alcohol bath.Draw and hold vacuum ≤50mm1min;seal tube while under vacuum.Hydrolyze22h at 110±1°C.Cool,open tube,and transfer contents to5mL volumet-ric flask containing sufficient cold6M HCl to neutralize hydrolysate;dilute to volume using buffer appropriate for amino acid analyzer.Centrifuge or filter hydrolysate and store e this hydrolysate to determine tryptophan(TRP).F.Amino Acid AnalysisAnalyze each of the3hydrolysates using parameters optimal for amino acid analyzer being e standard amino acid solutions to calibrate analyzer at least every24h.Each amino acid peak should have≥85%resolution.When alkaline hydrolysate is analyzed, tryptophan must be separated from pute for each of the following amino acids,the uncorrected g/16g N:ASP,THR, SER,GLU,PRO,GLY,ALA,VAL,MET,ILE,LEU,TRY,PHE, LYS,HIS,AMM,ARG,CYS,and TRP according to:g Amino acid(uncorrected)/16g test portion N=[ηmoles aa×initial test solution volume(mL)×MW aa]/[volume test solution injected(mL)×test portion weight(g)×%N for test portion×6.25×105] Compute percentage recovery by determining N content for each amino acid:g N contributed by each aa/16g test portion N=(14×number of N atoms in aa/mW aa)×(uncorrected g aa/16g test portion N)%Recovery=Σ(g aa N for each aa/16g test portion N)×100 Note:If percent recovery is<86or>105,error was made in hydroly-sis procedure(weighing errors,dilution,instrument calibration)or in computational process of percent recovery.Hydrolysis,analysis, and/or computation of percent recovery must be repeated until per-cent recovery falls within86–105tolerance before proceeding fur-ther.Adjust amino acid profile to normalize to95%hydrolysis:Correction factor=95% percent recoveryNote:For each amino acid,compute the corrected g/100g protein by:g Amino acid/16g N(corrected)=correction factor×g aa/16g NG.In Vitro Protein Digestion—For C-PERUse test portion or control weight containing10mg N.Place appropriate quantity of control protein,ANRC sodium caseinate(f)or test portion,in labeled vial containing magnetic stir-ring bar.Add10mL H2O and let ing pH meter,37°C bath,and stirrer,equilibrate and control to pH8±0.03at37°C by ad-ditions of dilute HCl and NaOH.At this time also equilibrate en-zyme solutions to pH8±0.03at37°C.Replace enzymes on ice;hold test portion and control at37°C.To equilibrated control vial,add1mL enzyme solution A while stirring.Exactly10min after addition of solution A,add1mL en-zyme solution B,and then transfer vial to55°C H2O bath.Exactly 19min after adding solution A,transfer vial back to37°C bath,insert pH electrode,and read pH at20min.pH of casein control should read6.42±0.05at20min.After proper pH reading is obtained for control,carry each test portion through identical procedure and read 20min pH(X)for each.Calculate%protein digestibility asDigestibility,%=234.84−22.56(X)puting the C-PERCompute C-PER using%digestibility and g amino acid/16g N of: LYS,MET+CYS,THR,ILE,LEU,VAL,PHE+TRY,and TRP. When combining ME+CYS and PHE+TYR,the CYS and TYR can be no>50%of MET+CYS and PHE+TYR totals,respectively. For example,if g amino acid/16g N were:MET=2and CYS=3,use 4for MET+CYS total,because maximum CYS can only be50%of MET+CYS total.Step1:Express each essential amino acid as percentage of FAO/WHO standard:FAO,%=(g aa/16g N)FAO/WHO standard×%digestibilitywhere FAO/WHO standard is assumed to be:LYS=5.44, MET+CYS=3.52,TH=4.00,ILE=4.00,LEU=7.04,VAL=4.96, PHE+TYR=6.08,TRP=0.96.Step2:Examine each percentage of FAO/WHO standard and ad-just as follows:(a)If all percentages are>90%(before rounding to nearest integer)of FAO/WHO standard,and LEU is<135%(before rounding to nearest integer)proceed to Step3;otherwise,(b)if any percentage is>100,reduce to100and proceed to Step3.Step3:Compute the following for test sample protein and refer-ence casein:X=Σ[(1/%FAO/WHO for each aa)(weight)]Y=Σweights usedWeights to be used in Step3computations are shown in Ta-ble982.30.Step4:Divide the sum of weights(Y)by sum of reciprocols(X) for both test sample protein and reference casein.Results will be termed essential amino acid scores for test sample and casein. Step5:Divide score of test sample by score of reference casein. Result expresses test sample as the ratio of reference casein,and is termed RATIO.If RATIO is>0.99and<1.01,then PER of test sam-ple is2.5and program should terminate at this point,i.e.,the test sample is casein or its equivalent.Step6:Compute the following:Z=RATIO×2.5.Step7:Compute4discriminant values to determine group into which sample is to be classified.Discriminant equations are: Group1=–671.8418–6.57689(LYS)+3.56696(MET+CYS) +13.10145(THR)+2.54503(ILE)+16.9981(LEU)–0.43395(VAL)–11.5244(PHE+TYR)+31.55321(TRP)+14.59278(Digestibility)Group2=–666.4492–2.78584(LYS)+5.17441(MET+CYS) +13.08564(THR)+4.61808(ILE)+16.22603(LEU)– 1.63223(VAL)–10.13673(PHE+TYR)+32.60196(TRP)+14.11668(Digestibility)Group3=–619.0813–3.13909(LYS)+4.26918(MET+CYS) +10.00988(THR)–1.42144(ILE)+15.7547(LEU)+ 5.6604(VAL)–11.28705(PHE+TYR)+30.49168(TRP)+13.79953(Digestibility)Group4=−744.7122−0.37674(LYS)+6.03697(MET+CYS) +11.51527(THR)+1.63251(ILE)+17.29687(LEU)+ 3.0294(VAL)−11.5033(PHE+TYR)+37.88725(TRP)+14.68169(Digestibility)Step8:Compute C-PER by examining the4group values com-puted in Step7.Choose group number that has largest value and use that number to pick correct C-PER equation.For PER predictions, use following group equations when digestibility was estimated by the4enzyme procedure:Group1:C-PER=1.12683−1.61426(Z)+0.99306(Z2)Group2:C-PER=−7.25391+8.14063(Z)−1.79517(Z2)Group3:C-PER=4.30469−1.99609(Z)+0.45996(Z2)Group4:C-PER=12.75−8.21484(Z)+1.66016(Z2)puting the DC-PERDC-PER is computed using steps just described in computing C-PER,with one additional step—percent protein digestibility is computed from amino acid profile instead of being determined via in vitro procedure.Coefficients for discriminant equations(Step7)and PER predictive equations(Step8)are also changed.Compute digestibility from amino acid profile as follows:Step1:Compute the3group discriminant values for test sample and reference casein.Group1=–203.7537–2.59402(LYS)+9.27153(LEU)+19.36964(ASP)+4.19676(PRO)+12.46035(CYS)+34.3075(AMM)Group2=–150.3707–0.78115(LYS)+7.6239(LEU)+15.46558(ASP)+3.8947(PRO)+12.79949(CYS)+29.74493(AMM)Group3=–155.9532+4.61135(LYS)+7.85429(LEU)+13.25949(ASP)+4.68431(PRO)+13.2907(CYS)+19.89403(AMM)Examine resulting discriminant values for test sample protein and reference casein,and choose group number associated with highest discriminant e group number to determine which digest-ibility equation to use.If for test sample,group equation No.3has highest value,then use digestibility equation No.3below to com-pute sample digestibility.If reference casein had highest value from group No.2equation,then use digestibility equation No.2below to compute digestibility for casein.Group1:Digestibility=67.8263+0.60144(LYS)−1.73309(LEU)+2.48377(ASP)+2.03523(PRO)−0.97312(CYS)−6.44299(AMM)Group2:Digestibility=160.5607+5.7998(LYS)−2.20744(LEU)−7.35627(ASP)−0.85275(PRO)+6.11058(CYS)−14.54944(AMM)Group3:Digestibility=116.5451+0.99537(LYS)−4.37473(LEU)−0.10243(ASP)−0.06304(PRO)−0.14005(CYS)+3.48679(AMM)Previous Step1(C-PER procedure)now becomes1-A.Steps2–6 remain as before.Step7:Substitute following discriminant group equations:Group1=−350.9675+2.34642(LYS)−8.60862(MET+CYS)−13.80721(THR)+11.71013(ILE)+11.7984(LEU)−12.10787(VAL)+9.68089(PHE+TYR)+46.88927(TRP)+7.291(Digestibility)Group2=−454.6516+7.83575(LYS)−14.3054(MET+CYS)−15.64592(THR)+13.32306(ILE)+14.1817(LEU)−17.40405(VAL)+12.36894(PHE+TYR)+64.39914(TRP+8.00712(Digestibility)Group3=−405.9275+5.01252(LYS)−8.46439(MET+CYS)−15.014(THR)+10.1986(ILE)+11.91023(LEU)−9.50181(VAL)+9.46879(PHE+TYR)+49.43095(TRP)+7.78124(Digestibility)Group4=−488.5569+9.3207(LYS)−11.36379(MET+CYS)−15.24675(THR)+10.60119(ILE)+13.93578(LEU)−12.14625(VAL)+10.15707(PHE+TYR)+63.1489(TRP)+8.22588(Digestibility)Step8:Substitute the following predictive equations: Group1:DC-PER=1.254−2.04932(Z)+1.30629(Z2)Group2:DC-PER=−4.08594+5.125(Z)−1.08398(Z2)Group3:DC-PER=4.66406−2.29297(Z)+0.50586(Z2)Group4:DC-PER=10.44141−5.93359(Z)+1.13281(Z2) Reference:JAOAC65,798(1982).Table982.30Weights to be used in step3computations %FAO/WHO a Weight≥100191–99281–90 2.8371–80461–70 5.6651–60841–5011.3131–401621–3022.6311–20320–1045.25a Round to nearest integer.。

3GPP 5G基站(BS)R16版本一致性测试英文原版(3GPP TS 38.141-1)

4.2.2
BS type 1-H.................................................................................................................................................. 26
4.3
Base station classes............................................................................................................................................27
1 Scope.......................................................................................................................................................13
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AOAC 30.1.23A AOAC Official Method 995.13-国外标准规范

30.1.23AAOAC Official Method995.13Carbohydrates in Soluble(Instant)CoffeeAnion-Exchange Chromatographic Methodwith Pulsed Amperometric DetectionFirst Action1995(Applicable for determination of free and total carbohydrates[ex-cept total fructose,which is degraded]in soluble[instant]coffee.) See Tables995.13A–J for the results of the interlaboratory study supporting the acceptance of the method.A.PrincipleFree carbohydrates.—Coffee is dissolved in H2O.Solution is fil-tered through C18disposable cartridge,and then through0.2µm membrane filter.Filtered solution is injected onto LC system.Car-bohydrates are separated on pellicular anion-exchange column and measured by pulsed amperometric detector.Total carbohydrates.—Coffee is hydrolyzed with1M HCl.Solu-tion is filtered and then passed through cation-exchange disposable cartridge in the Ag form to neutralize solution and to eliminate the Cl anion prior to injection onto LC system.B.Apparatus(a)Balance.—Analytical,weighing to0.1mg.(b)Flasks.—250mL round-bottom.(c)Volumetric flasks.—100and1000mL.(d)Pipets.—Delivering200–1000µL and5µL;with disposable tips.(e)Cylinders.—50and1000mL,tall-form,graduated.(f)Funnels.—Analytical,60°C.(g)Vacuum filtering system.—Aspirator with regulating device. System should include:heavy-walled filtering flask with ground cone neck,1L;funnel with ground glass joint,300mL;aluminum assembly clip;connection with vacuum outlet;filter holder,47mm id;and low-water extractable membrane filters,0.2µm porosity, 47mm diameter.(h)Filter papers.—Qualitative,folded,medium fast.(i)C18cartridges.—Octadecylated silica(ca10%C);6mL car-tridge volume;capable of holding500mg test portion;disposable.Con-dition and use cartridges according to manufacturer’s instructions. (j)Cation-exchange cartridges.—Styrene-based resin,Ag form, 1.8–2.0milliequivalent capacity/cartridge;disposable.Condition and use cartridges according to manufacturer’s instructions. (k)Membrane filters.—0.2µm porosity,25mm diameter;dis-posable;polypropylene.(l)Water bath.—Maintaining98±2°C.(m)Liquid chromatograph(LC).—Metal-free,compatible with 300mM NaOH.Operating conditions:mobile phase,isocratic(see Table995.13for mobile phase conditions);column temperature, ambient;flow rate,1.0mL/min;post-column solvent,300mM NaOH at0.6mL/min;detector settings,optimum parameters as pro-vided by e with computing integrator.(n)LC column.—250×4mm id;packed with polystyrene divinylbenzene substrate(10µm diameter)agglomerated with microbead quaternary amine functionalized latex(350nm diame-ter);5%cross-linked.(o)Pulsed amperometric detector.—With gold electrode.Fill reference cell with300mM NaOH.Select the detector range to avoid saturation of the major peak in chromatogram.(p)Guard column.—50×4mm id,packed with the same mate-rial as analytical column,(n).(q)Post-column solvent delivery system.—Compatible with 300mM NaOH.C.ReagentsUse18MΩ⋅cm demineralized H2O throughout.(a)Sodium hydroxide.—50%(w/w)aqueous solution,contain-ing minimum amount of Na2CO3and Hg.Do not shake or stir solu-tion before use.(b)Hydrochloric acid solution.—1.00M standard volumetric so-lution(83.3mL HCl/L).(c)Eluent A.—18MΩ⋅cm demineralized water.Filter through0.2µm membrane filter.Degas by sparging with He20–30min.Pre-pare fresh eluent A daily.(d)Eluent B.—300mM NaOH.Pipet15.6mL50%NaOH into 985mL eluent A.Degas by sparging with He20–30min.Eluent B is stable2days if stored at room temperature under He.(Note:It is crit-ical to remove dissolved CO2from eluents.Carbonate acts as strong “pusher”on LC column,resulting in drastic reduction in resolution.)(e)Carbohydrates standard solutions.—(1)Standard stock solu-tions.—1mg/mL aqueous stock solutions of arabinose,fructose, fucose,galactose,glucose,mannose,rhamnose monohydrate, ribose,xylose,sucrose,and mannitol.Weigh100mg each carbohy-drate to the nearest0.1mg into separate100mL volumetric flasks, dissolve in H2O,and dilute to volume with H2O.(2)Mixed carbohy-drates standard solution.—Further dilute and mix carbohydrates stock solutions to reach carbohydrate concentrations similar to those found in nonhydrolyzed or hydrolyzed soluble coffee test solutions. Pass diluted carbohydrates standard solution through0.2µm mem-brane filter prior to injection onto LC column.(Note:If resolution of rhamnose from arabinose is difficult to achieve,do not add rhamnose to mixed standard solution.)D.Isolation of CarbohydratesUse soluble coffee as is without grinding or homogenization.(a)Free carbohydrates.—Weigh300mg coffee to the nearest0.1mg into100mL volumetric flask.Add70mL H2O and shake until dissolution is complete.Dilute solution to volume with H2O.Filter 5–10mL solution through C18cartridge.Discard the first1mL.Pass fil-trate through0.2µm membrane filter prior to LC injection.(b)Total carbohydrates.—Weigh300mg coffee to the nearest0.1mg into100mL volumetric flask.Add50mL1.00M HCl and swirl.Place flask in boiling water bath for2.5h.(Note:Always keep the level of solution in flask below that of H2O in bath.)Swirl flaskTable995.13Conditions of mobile phase for determinationof free and total carbohydrates in solublecoffee by anion-exchange chromatographicmethod with pulsed amperometric detection Time,min Eluent A,%Eluent B,%01000(start acquisition) 50.01000(stop acquisition) 50.10100(start cleanup) 65.00100(stop cleanup) 65.11000(start re-equilibrium) 80.01000(stop re-equilibrium)Test sample N a Mean,%RSD r RSD R r R 17(2)0.024 1.6590.0010.041 1′11(0)0.1797.0500.0350.021 211(1)0.0600.91580.0020.10 2′11(0)0.1517.6460.0320.20 311(0) 1.5 2.89.90.120.44 3′11(1) 1.85 2.2180.120.94 411(0)0.619 3.6240.0630.42 4′11(0)0.782 4.6210.100.45 59(1)0.1928.2340.0450.18 5′11(0)0.30011370.0910.32 68(1)0.0597.1490.0120.083 6′10(0)0.1799.8500.050.25 a N=No.of laboratories after removal of outliers(in parentheses).Table995.13B Interlaboratory study results of the determination of free arabinose in soluble coffeeTest sample N a Mean,%RSD r RSD R r R 111(0)0.891 3.7140.0920.035 1′11(0) 3.54 6.6210.66 2.1 29(2) 1.320 1.6 5.10.0600.19 2′11(0) 4.83 3.3170.45 2.4 311(0)0.464 3.8110.0490.14 3′11(0) 4.76 3.1130.42 1.7 411(0)0.7477.3120.150.25 4′11(0) 4.54 4.618.40.59 2.4 510(1)0.505 4.47.70.0630.11 5′9(2) 4.08 3.0 4.90.340.56 611(0)0.629 4.18.90.0730.16 6′11(0) 3.79 5.7200.61 2.2a N=No.of laboratories after removal of outliers(in parentheses).Table995.13C Interlaboratory study results of the determination of free galactose in soluble coffeeTest sample N Mean,%RSD r RSD R r R 111(0)0.562 3.0 5.30.0470.084 1′10(1)17.88.18.9 4.1 4.8 211(0)0.339 4.18.00.0390.077 2′10(1)18.5 2.312 1.2 6.2 311(0)0.1919.8130.0530.070 3′11(0)8.08 2.78.00.6208.0 411(0)0.438 5.98.30.0740.10 4′11(0)13.3 3.913 1.8 5.9 59(2)0.475 3.1 4.10.0410.055 5′9(2)18.40 1.77.50.87 3.9 611(0)0.362 5.0120.0510.13 6′10(1)17.7 4.38.5 2.2 4.3a N=No.of laboratories after removal of outliers(in parentheses).Test sample N a Mean,%RSD r RSD R r R 111(0)0.1059.9210.0290.062 1′11(0)0.6848.7170.170.32 29(1)0.0421020.00.0120.024 2′10(1)0.8267.4220.170.50 310(1) 2.04 2.5 6.20.140.360 3′11(0)16.6 5.924.0 2.811.0 410(1) 1.66 4.1 6.10.190.29 4′11(0) 4.38 3.8240.47 2.9 510(1)0.18610210.0530.11 5′11(0) 1.95 5.7130.310.69 611(0)0.18610240.0520.12 6′11(0) 1.027.9140.230.40 a N=No.of laboratories after removal of outliers(in parentheses).Table995.13E Interlaboratory study results of the determination of free mannose in soluble coffeeTest sample N a Mean,%RSD r RSD R r R 111(0)0.583 4.9240.0800.40 1′10(1)17.9 5.811 2.9 5.7 210(0)0.1558.2360.0130.16 2′11(0)14.4 2.615 1.1 6.2 311(0)0.470 4.2180.0560.23 3′11(0) 2.60 2.0140.15 1.0 411(0)0.3297.0190.06517 4′11(0) 5.60 3.0150.48 2.4 510(1)0.277 4.2400.0330.31 5′11(0)7.65 2.8110.60 2.3 611(0)0.991 3.8170.110.48 6′10(0)19.1 2.222 1.212.00 a N=No.of laboratories after removal of outliers(in parentheses).Table995.13F Interlaboratory study results of the determination of free fructose in soluble coffeeTest sample N a Mean,%RSD r RSD R r R 19(0)0.17117310.0820.15 1′6(0)0.18924680.130.37 28(1)0.05421340.0320.052 39(2) 3.62 2.9180.30 1.9 3′9(0) 2.01 5.7720.32 4.1 410(1) 3.12 2.9180.26 1.6 4′7(1) 1.37 5.5700.21 2.7 510(0)0.2829.0450.0720.36 5′7(0)0.24420590.140.41 68(2)0.460 5.6160.0670.2 6′9(1)0.3637.3680.0750.70 a N=No.of laboratories after removal of outliers(in parentheses).Test sample N a Mean,%RSD r RSD R r R 17(2)0.07315740.0310.15 27(l)0.04528600.0350.077 58(0)0.10215980.0430.28 55(1)0.08016180.0360.041 67(l)0.1209.1810.0310.28 a N=No.of laboratories after removal of outliers(in parentheses).Table995.13H Interlaboratory study results of the determination of free xylose in soluble coffeeTest sample N a Mean,%RSD r RSD R r R 18(2)0.09723380.0630.10 29(0)0.1469.8200.0400.084 311(0) 1.86 4.6230.240.42 411(0)0.736 3.7280.0770.56 57(0)0.02925230.0200.023 511(0) 1.837.4220.38 1.2 69(0)0.13314240.0530.090 a N=No.of laboratories after removal of outliers(in parentheses).Table995.13I Interlaboratory study results of the determination of free sucrose in soluble coffeeTest sample N a Mean,%RSD r RSD R r R 210(0)0.149 4.8380.0200.16 310(1) 1.32 1.8100.0660.370 410(1)0.746 6.8120.140.25 511(0)0.18115420.0770.21 69(1)0.158 3.4330.0150.15 a N=No.of laboratories after removal of outliers(in parentheses).Table995.13J Interlaboratory study results of the determination of free fucose in soluble coffeeTest sample N a Mean,%RSD r RSD R r R 27(1)0.01620.0460.0090.021 38(0)0.0547.0670.0110.10 47(1)0.03623770.0230.074 57(1)0.011 5.4450.0020.014 66(1)0.02510360.0070.025 a N=No.of laboratories after removal of outliers(in parentheses).by hand every30min.Cool to room temperature under tap water. Dilute solution to100mL with H2O and filter through folded filter paper.Pass3mL filtrate through cation-exchange cartridge.Discard the first1mL.Filter neutralized solution through0.2µm membrane filter prior to LC injection.E.LC DeterminationInject equal volumes(10–20µL)of standard,C(e),and test solu-tions from D(a)or(b)onto LC column.[Note:Retention times and resolution tend to vary from column to column.Start clean-up and re-equilibration sequence only when the last monosaccharide (ribose)has been eluted.It may be necessary to perform2–3injec-tions of carbohydrates standard solution or to increase the re-equilibrium time in order to achieve a good separation of glucose, sucrose,and xylose.]Under normal conditions,approximate retention times of carbo-hydrates are:mannitol,4min;fucose,7min;rhamnose,15min;arabinose,16min;galactose,22min;glucose,25min;sucrose, 27min;xylose,30min;mannose,32min;fructose,40min;and ribose,43min.See Figure995.13for chromatogram of mixed carbo-hydrates standard solution.F.CalculationsCalculate concentration of carbohydrate in sample solution as follows:Carbohydrate,%=(R1/R2)×(W0/V0)×(V/W)×100 where R1=peak response of carbohydrate in test solution;R2=peak response of carbohydrate in carbohydrate standard solution;W0= mass of carbohydrate in the standard solution,mg;V0=total volume of the standard solution,mL;V=volume of standard solution,mL; and W=weight of test portion,mg.Express results as percent free or percent total carbohydrates (as is).References:J.AOAC Int.78,768(1995);79,1400(1996). Revised:June2000Figure995.13—HPAE-PAD chromatogram of mixed carbohydrates standard solution:mannitol,15m g;fucose,15m g/mL;rhamnose,35m g/mL;arabinose,40m g/mL;galactose,50m g/mL;glucose,55m g/mL;sucrose,45m g/mL; xylose,55m g/mL;mannose,45m g/mL;fructose,90m g/mL;and ribose,90m g/mL.。

AOAC饲料中药物官方分析法

AOAC饲料中药物官方分析法（2000）第5章40页2000AOAC国际版5．3．02杆菌肽预混剂AOAC官方分析方法杯碟法（本法应用于含≥10g杆菌肽/454g的预混剂）A原理：杆菌肽从已酸化有机溶剂系统中的饲料提取。

提取物离心，上清液用磷酸盐缓冲液稀释，然后用杯碟法用藤黄微球菌检定菌分析。

B试剂与装置(a)微生物—藤黄微球菌ATCC号10240 培养基（见）(b)提取溶剂—混合溶剂，27%乙腈（H3CN），27%甲醇，3%pH6.0的磷酸盐缓冲剂（见）,41%水和2%磷酸(85%)，加入0.5gEDTA/L（提取溶剂用EDTA饱和的）(c)磷酸盐缓冲剂—5%，pH6.5，见957.23B(d)C（见）(d)稀释剂—甲醇—5%pH6.5磷酸盐缓冲剂（12+88）(e)稀盐酸—小心加89ml盐酸至适量水中，并稀释到1L（1M）。

进一步稀释溶液1﹕100（0.01M）。

(f)钢管见957.23 C (a) (见)(g)钢管分布器：随机的见957.23(d) (见)C标准液见957.24*(a)和(b)(见)。

也即制备0.3和0.16u/ml溶液以平皿作成供试样品，用于监测定量用。

D杯碟准备采用1个约15ml的琼脂抗生素培养基1，957.23A(a)(见)。

通过实验皿最适的浓度（通常为0.02—0.05%）的藤黄微球菌ATCC No 1024，加到琼脂培养基上，使获得抑菌圈为15-17mm，用于0.2u/ml杆菌肽。

注入4个钢管内，标准曲线上的每点（即16个杯点），另外，4个杯为供试液。

标准曲线与杯碟两次（即32个钢圈），将作为检查样品0.3、0.16u/ml。

因此，总的将需要48个杯、两个曲线，对于每个预混剂样品加4个附加杯。

使琼脂培养基铺平，保持水平和一定硬度。

使用前转移到冰箱中，冷却≥1小时，所用碟当天配制。

E 提取准确称取含有约4600单位的杆菌肽饲料量装到300ml 的容量瓶中，或相当的容量瓶，用100ml 移液管加100ml 的提取溶剂，用振荡器振摇，提取饲料≥5分钟，转移上清液到塑料离心管中，2000转/分离心10分钟，通过玻璃棉过滤上清液至刻度量器中，稀释到制取的终稀释度为0.2±0.05u/ml 。

AOAC 991.43

JBI_Critical_Appraisal-Checklist_for_Systematic_Reviews

The Joanna Briggs InstituteIntroductionThe Joanna Briggs Institute (JBI) is an international, membership based research and development organization within the Faculty of Health Sciences at the University of Adelaide. The Institute specializes in promoting and supporting evidence-based healthcare by providing access to resources for professionals in nursing, midwifery, medicine, and allied health. With over 80 collaborating centres and entities, servicing over 90 countries, the Institute is a recognized global leader in evidence-based healthcare.JBI Systematic ReviewsThe core of evidence synthesis is the systematic review of literature of a particular intervention, condition or issue. The systematic review is essentially an analysis of the available literature (that is, evidence) and a judgment of the effectiveness or otherwise of a practice, involving a series of complex steps. The JBI takes a particular view on what counts as evidence and the methods utilized to synthesize those different types of evidence. In line with this broader view of evidence, the Institute has developed theories, methodologies and rigorous processes for the critical appraisal and synthesis of these diverse forms of evidence in order to aid in clinical decision-making in health care. There now exists JBI guidance for conducting reviews of effectiveness research, qualitative research, prevalence/incidence, etiology/risk, economic evaluations, text/opinion, diagnostic test accuracy, mixed-methods, umbrella reviews and scoping reviews. Further information regarding JBI systematic reviews can be found in the JBI Reviewer’s Manual on our website.JBI Critical Appraisal ToolsAll systematic reviews incorporate a process of critique or appraisal of the research evidence. The purpose of this appraisal is to assess the methodological quality of a study and to determine the extent to which a study has addressed the possibility of bias in its design, conduct and analysis. All papers selected for inclusion in the systematic review (that is –those that meet the inclusion criteria described in the protocol) need to be subjected to rigorous appraisal by two critical appraisers. The results of this appraisal can then be used to inform synthesis and interpretation of the results of the study. JBI Critical appraisal tools have been developed by the JBI and collaborators and approved by the JBI Scientific Committee following extensive peer review. Although designed for use in systematic reviews, JBI critical appraisal tools can also be used when creating Critically Appraised Topics (CAT), in journal clubs and as an educational tool.JBI Critical Appraisal Checklist for Systematic Reviewsand Research SynthesesReviewer DateAuthor Year Record NumberYes No UnclearNot applicable1.Is the review question clearly and explicitly stated? □□□□2.Were the inclusion criteria appropriate for the reviewquestion? □□□□3.Was the search strategy appropriate? □□□□4.Were the sources and resources used to search forstudies adequate? □□□□5.Were the criteria for appraising studies appropriate? □□□□6.Was critical appraisal conducted by two or morereviewers independently? □□□□7.Were there methods to minimize errors in dataextraction? □□□□8.Were the methods used to combine studies appropriate? □□□□9.Was the likelihood of publication bias assessed? □□□□10.Were recommendations for policy and/or practicesupported by the reported data? □□□□11.Were the specific directives for new researchappropriate? □□□□ Overall appraisal: Include □Exclude □Seek further info □Comments (Including reason for exclusion)JBI Critical Appraisal Checklist for Systematic Reviewsand Research SynthesesHow to cite:Aromataris, E., Fernandez, R., Godfrey, C., Holly, C., Kahlil, H. & Tungpunkom, P. Summarizing systematic reviews: methodological development, conduct and reporting of an Umbrella review approach Int J Evid Based Healthc. 2015,13(3):132-40.When conducting an umbrella review using the JBI method, the critical appraisal instrument for Systematic Reviews should be used.The primary and secondary reviewer should discuss each item in the appraisal instrument for each study included in their review. In particular, discussions should focus on what is considered acceptable to the aims of the review in terms of the specific study characteristics. When appraising systematic reviews this discussion may include issues such as what represents an adequate search strategy or appropriate methods of synthesis. The reviewers should be clear on what constitutes acceptable levels of information to allocate a positive appraisal compared with a negative, or response of “unclear”. This discussion should ideally take place before the reviewers independently conduct the appraisal.Within umbrella reviews, quantitative or qualitative systematic reviews may be incorporated, as well as meta-analyses of existing research. There are 11 questions to guide the appraisal of systematic reviews or meta-analyses. Each question should be answered as “yes”, “no”, or “unclear”. Not applicable “NA” is also provided as an option and may be appropriate in rare instances.1. Is the review question clearly and explicitly stated?The review question is an essential step in the systematic review process. A well-articulated question defines the scope of the review and aids in the development of the search strategy to locate the relevant evidence. An explicitly stated question, formulated around its PICO (Population, Intervention, Comparator, Outcome) elements aids both the review team in the conduct of the review and the reader in determining if they review has achieved its objectives.Ideally the review question should be articulated in a published protocol; however this will not always be the case with many reviews that are located.2.Were the inclusion criteria appropriate for the review question?The inclusion criteria should be identifiable from, and match the review question. The necessary elements of the PICO should be explicit and clearly defined. The inclusion criteria should be detailed and the included reviews should clearly be eligible when matched against the stated inclusion criteria. Appraisers of meta-analyses will find that inclusion criteria may include criteria around the ability to conduct statistical analyses which would not be the norm for a systematic review. The types of included studies should be relevant to the review question, for example, an umbrella review aiming to summarize a range of effective non-pharmacological interventions for aggressive behaviors amongst elderly patients with dementia will limit itself to including systematic reviews and meta-analyses that synthesize quantitative studies assessing the various interventions; qualitative or economic reviews would not be included.3.Was the search strategy appropriate?A systematic review should provide evidence of the search strategy that has been used tolocate the evidence. This may be found in the methods section of the review report in some cases, or as an appendix that may be provided as supplementary information to the review publication. A systematic review should present a clear search strategy that addresses each of the identifiable PICO components of the review question. Some reviews may also provide a description of the approach to searching and how the terms that were ultimately used were derived, though due to limits on word counts in journals this may be more the norm in online only publications. There should be evidence of logical and relevant keywords and terms and also evidence that Subject Headings and Indexing terms have been used in the conduct of the search. Limits on the search should also be considered and their potential impact; for example, if a date limit was used, was this appropriate and/or justified? If only English language studies were included, will such a language bias have an impact on the review? The response to these considerations will depend, in part, on the review question.4.Were the sources and resources used to search for studies adequate?A syst ematic review should attempt to identify “all” the available evidence and as such thereshould be evidence of a comprehensive search strategy. Multiple electronic databases should be searched including major bibliographic citation databases such as MEDLINE and CINAHL.Ideally, other databases that are relevant to the review question should also be searched, for example, a systematic review with a question about a physical therapy intervention should also look to search the PEDro database, whilst a review focusing on an educational intervention should also search the ERIC. Reviews of effectiveness should aim to search trial registries. A comprehensive search is the ideal way to minimize publication bias, as a result, a well conducted systematic review should also attempt to search for grey literature, or “unpublished” studies; this may involve searching websites relevant to the review question, or thesis repositories.5.Were the criteria for appraising studies appropriate?The systematic review should present a clear statement that critical appraisal was conducted and provide the details of the items that were used to assess the included studies. This may be presented in the methods of the review, as an appendix of supplementary information, or as a reference to a source that can be located. The tools or instruments used should be appropriate for the review question asked and the type of research conducted. For example, a systematic review of effectiveness should present a tool or instrument that addresses aspects of validity for experimental studies and randomized controlled trials such as randomization and blinding – if the review includes observational research to answer the same question a different tool would be more appropriate. Similarly, a review assessing diagnostic test accuracy may refer to the recognized QUADAS1 tool.6.Was critical appraisal conducted by two or more reviewers independently?Critical appraisal or some similar assessment of the quality of the literature included in a systematic review is essential. A key characteristic to minimize bias or systematic error in the conduct of a systematic review is to have the critical appraisal of the included studies completed independently and in duplicate by members of the review team. The systematic review should present a clear statement that critical appraisal was conducted by at least two reviewers working independently from each other and conferring where necessary to reach decision regarding study quality and eligibility on the basis of quality.7.Were there methods to minimize errors in data extraction?Efforts made by review authors during data extraction can also minimize bias or systematic errors in the conduct of a systematic review. Strategies to minimize bias may include conducting all data extraction in duplicate and independently, using specific tools or instruments to guide data extraction and some evidence of piloting or training around their use.8.Were the methods used to combine studies appropriate?A synthesis of the evidence is a key feature of a systematic review. The synthesis that ispresented should be appropriate for the review question and the stated type of systematic review and evidence it refers to. If a meta-analysis has been conducted this needs to be reviewed carefully. Was it appropriate to combine the studies? Have the reviewers assessed heterogeneity statistically and provided some explanation for heterogeneity that may be present? Often, where heterogeneous studies are included in the systematic review, narrative synthesis will be an appropriate method for presenting the results of multiple studies. If a qualitative review, are the methods that have been used to synthesize findings congruent with the stated methodology of the review? Is there adequate descriptive and explanatory information to support the final synthesized findings that have been constructed from the findings sourced from the original research?9.Was the likelihood of publication bias assessed?As mentioned, a comprehensive search strategy is the best means by which a review author may alleviate the impact of publication bias on the results of the review. Reviews may also present statistical tests such as Egger’s test or funnel plots to also assess the potential presence of publication bias and its potential impact on the results of the review. This question will not be applicable to systematic reviews of qualitative evidence.10.Were recommendations for policy and/or practice supported by the reported data?Whilst the first nine (9) questions specifically look to identify potential bias in the conduct of a systematic review, the final questions are more indictors of review quality rather than validity.Ideally a review should present recommendations for policy and practice. Where these recommendations are made there should be a clear link to the results of the review. Is there evidence that the strength of the findings and the quality of the research been considered in the formulation of review recommendations?11.Were the specific directives for new research appropriate?The systematic review process is recognized for its ability to identify where gaps in the research, or knowledge base, around a particular topic exist. Most systematic review authors will provide some indication, often in the discussion section of the report, of where future research direction should lie. Where evidence is scarce or sample sizes that support overall estimates of effect are small and effect estimates are imprecise, repeating similar research to those identified by the review may be necessary and appropriate. In other instances, the case for new research questions to investigate the topic may be warranted.References1.P Whiting, AWS Rutjes, JB Reitsma, PMM Bossuyt, J Kleijnen. The development of QUADAS:a tool for the quality assessment of studies of diagnostic accuracy included in systematicreviews BMC Medical Research Methodology 2003, 3:25 doi:10.1186/1471-2288-3-25。

AS 2159-2009 Piling Design and Construction

AS 2159—2009Australian Standard ® Piling—Design and installationAS 2159—2009 s e d b y H A T C H A S S O C I A T E S o n 30 N o v 2009This Australian Standard® was prepared by Committee CE-018, Piling. It was approved onbehalf of the Council of Standards Australia on 19 June 2009.This Standard was published on 4 November 2009.The following are represented on Committee CE-018:• Australian Building Codes Board•Australian Geomechanics Society• AUSTROADS• Concrete Institute of Australia • Engineers Australia• Monash University• Piling and Foundation Specialists Federation • University of SydneyThis Standard was issued in draft form for comment as DR 08180.Standards Australia wishes to acknowledge the participation of the expert individuals thatcontributed to the development of this Standard through their representation on theCommittee and through the public comment period.Keeping Standards up-to-dateAustralian Standards® are living documents that reflect progress in science, technology andsystems. To maintain their currency, all Standards are periodically reviewed, and new editionsare published. Between editions, amendments may be issued.Standards may also be withdrawn. It is important that readers assure themselves they areusing a current Standard, which should include any amendments that may have beenpublished since the Standard was published.Detailed information about Australian Standards, drafts, amendments and new projects canbe found by visiting w w .auStandards Australia welcomes suggestions for improvements, and encourages readers tonotify us immediately of any apparent inaccuracies or ambiguities. Contact us via email atmail@.au , or write to Standards Australia, GPO Box 476, Sydney, NSW 2001.s e d b y H A T C H A S S O C I A T E S o n 30 N o v 2009AS 2159—2009Australian Standard ®Piling—Design and installationOriginated as AS 2159—1978.Third edition 2009. COPYRIGHT© Standards AustraliaAll rights are reserved. No part of this work may be reproduced or copied in any form or byany means, electronic or mechanical, includ ing photocopying, without the writtenpermission of the publisher.s e d b y H A T C H A S S O C I A T E S o n 30 N o v 2009AS 2159—2009 2PREFACEThis Standard was pre pare d by the Standards Australia Committe e CE-018, Piling, tosupersede AS 2159—1995.The objective of this Standard is to provide requirements for design and installation of pilesfor supporting structur e s. Th e obj e ct of this r e vision is to align with updat e dAS 1170 Standards and reflect changes in practice since the previous edition.Major changes to the previous edition are as follows:(a) Revision of the overall Standard.(b) Re vision of the se tting of stre ngth re duction factors, that is, the se le ction of the‘safety’ level appropriate to the installation being designed.(c)Revision of the negative skin friction requirements. (d) Revision of durability requirements to assist designers to achieve predicted life.(e ) Include re quire me nts for ne we r pile type s and installation me thods including ste e lscrew piles, jacking, screwing and screwed cast in place.(f)Requirement for some testing to be ‘normative’. (g) Inclusion of new types of test including rapid pile testing.The te rms ‘normative ’ and ‘informative ’ have be e n use d in this Standard to de fine theapplication of the appendix to which they apply. A ‘normative’ appendix is an integral partof a Standard, whereas an ‘informative’ appendix is only for information and guidance.Statements expressed in mandatory terms in notes to tables are deemed to be requirementsof this Standard.Notes to the text contain information and guidance and are not considered to be an integralpart of the Standard.s e d b y H A T C H A S S O C I A T E S o n 30 N o v 20093 AS 2159—2009CONTENTSPageFOREWORD (5)SECTION 1 SCOPE AND GENERAL1.1 SCOPE (6)1.2 NORMATIVE REFERENCES (6)1.3 DEFINITIONS (7)1.4 NOTATION (10)1.5 CLASSIFICATION OF PILES (13)SECTION 2 SITE INVESTIGATION2.1 GENERAL (15)2.2 INFORMATION REQUIRED (15)SECTION 3 DESIGN REQUIREMENTS AND PROCEDURES3.1 OBJECTIVE OF PILE DESIGN (16)3.2 GENERAL DESIGN REQUIREMENTS (16)3.3 ACTIONS AND COMBINATIONS FOR STRENGTH AND SERVICEABILITYDESIGN (17)SECTION 4 GEOTECHNICAL DESIGN4.1 GENERAL (20)4.2 ASSESSMENT OF GEOTECHNICAL PARAMETERS (20)4.3 GENERAL PRINCIPLES OF GEOTECHNICAL STRENGTH DESIGN (21)4.4 DESIGN REQUIREMENTS FOR STRENGTH (24)4.5 GENERAL PRINCIPLES OF GEOTECHNICAL DESIGN FORSERVICEABILITY (29)4.6 DESIGN REQUIREMENTS FOR SERVICEABILITY (29)SECTION 5 STRUCTURAL DESIGN5.1 SCOPE OF SECTION (32)5.2 GENERAL PRINCIPLES OF STRUCTURAL STRENGTH DESIGN (32)5.3 CONCRETE AND GROUT PILES (33)5.4 STEEL PILES (36)5.5 COMPOSITE STEEL AND CONCRETE PILES (36)5.6 TIMBER PILES (37)SECTION 6 DURABILITY DESIGN6.1 GENERAL (38)6.2 GENERAL PRINCIPLES OF DURABILITY DESIGN (38)6.3 ACID SULFATE SOILS (38)6.4 DESIGN FOR DURABILITY OF CONCRETE PILES (39)6.5 DESIGN FOR DURABILITY OF STEEL PILES (42)6.6 DESIGN FOR DURABILITY OF TIMBER PILES (45)SECTION 7 MATERIALS AND CONSTRUCTION REQUIREMENTS7.1 GENERAL (47)7.2 TOLERANCES AND DEFECTS (47)s e d b y H A T C H A S S O C I A T E S o n 30 N o v 2009AS 2159—2009 4Page7.3 DISPLACEMENT PILES—PREFORMED (48)7.4 DISPLACEMENT PILES—DRIVEN CAST IN PLACE (52)7.5 DISPLACEMENT PILES—SCREWED CAST IN PLACE (53)7.6 NON-DISPLACEMENT PILES (54)7.7 RECORDS OF DATA (57)SECTION 8 TESTING8.1 SCOPE (60)8.2 GENERAL REQUIREMENTS (60)8.3 PILE LOAD TESTING (62)8.4 STATIC LOAD TESTING (65)8.5 HIGH-STRAIN DYNAMIC PILE TESTING (67)8.6 BI-DIRECTIONAL LOAD TESTING (68)8.7 RAPID LOAD TESTING (69)8.8 INTEGRITY TESTING (69)APPENDICESA STATIC LOAD TEST (71)B HIGH-STRAIN DYNAMIC PILE TESTING (78)C RAPID PILE TESTING (81)D INTEGRITY TESTING (85)E LIMIT STATES—SYMBOLS AND DEFINITIONS (89)BIBLIOGRAPHY (90)s e d b y H A T C H A S S O C I A T E S o n 30 N o v 20095 AS 2159—2009FOREWORDDecisions in pile design are based on design formulae, empirical and practical experience,and the accumulated records of a large number of applications of proprietary systems (bothsuccessful and otherwise). As such, there is a great need for flexibility, experience,engineering judgement and commonsense in designing and constructing a piled footingsystem. In a real sense, these requirements are in conflict with the need to make unqualifiedmandatory statements and, as a result, many of the stipulations of this Standard are shortand simple when, in other cases, extensive arrays of multiple choices are provided. Whereapplicable, explanatory notes are added to some clauses in this Standard and additionalcommentary is provided.s e d b y H A T C H A S S O C I A T E S o n 30 N o v 2009AS 2159—2009 6STANDARDS AUSTRALIAAustralian StandardPiling—Design and installationS E C T I O N 1 S C O P E A N D G E N E R A L1.1 SCOPEThis Standard sets out minimum requirements for the design, c onstruc tion and testing ofpiled footings for c ivil engineering and building struc tures on land or immediate inshorelocations. It does not extend to offshore (deepwater) construction.NOTES:1 AS 5100 series should be considered for the design of footings for road bridges.2 Where the strength o r serviceability o f an existing structure is to be evaluated, the generalprinciples o f this Standard sho uld be applied. The actual pro perties o f the materials in thestructure should be used.3 The durability requirements are appropriate for structures with design life within ±20% of thetarget design life.1.2 NORMATIVE REFERENCESThe normative documents referenced in this Standard are the following:NOTE: Documents referenced for informative purposes are listed in the Bibliography.AS1012Methods of testing concrete (all Parts) 1163Structural steel hollow sections 1170Structural design actions 1170.4 Part 4: Earthquake actions in Australia1289 Methods of testing soils for engineering purposes 1289.6.3.1 Part 6.3.1: Soil strength and c onsolidation tests—Determination of the penetration resistance of a soil—Standard penetration test (SPT) 1289.6.5.1 Part 6.5.1: Soil strength and c onsolidation tests—Determination of the static cone penetration resistance of a soil—Field test using a mechanical and electrical cone or friction-cone penetrometer 1379 Specification and supply of concrete 1450 Steel tubes for mechanical purposes 1554 Stru c tural steel welding 1554.1 Part 1: Welding of steel structures 1579 Arc-welded steel pipes and fittings for water and waste-water 1604 Specification for preservative treatment 1604.1 Part 1: Sawn and round timber 1720 Timber stru c tures 1720.1 Part 1: Design methods s e d b y H A T C H A S S O C I A T E S o n 30 N o v 20097 AS 2159—2009AS2758 Aggregates and rock for engineering purposes2758.1 Part 1: Concrete aggregates2832Cathodic protection of metals 2832.2Part 2: Compact buried structures 2832.3 Part 3: Fixed immersed structures3600 Concrete structures3818 Timber—Heavy structural products—Visually graded3818.3 Part 3: Piles3972 Portland and blended cements4100 Steel structures5100 Bridge design5100.5 Part 5: Concrete5100.6 Part 6: Steel and composite constructionAS/NZS1170Structural design actions 1170.0Part 0: General principles 1594Hot-rolled steel flat products 3678 Structural steel—Hot-rolled plates, floorplates and slabs3679 Structural steel3679.1 Part 1: Hot-rolled bars and sections3679.2 Part 2: Welded I sections4671 Steel reinforcing materialsASTMC 566-97 Standard Test Method for Total Evaporable Moisture Content of Aggregate byDrying1.3 DEFINITIONSFor the purpose of this Standard, the definitions below apply.1.3.1 Bored cast in place pileA pile, with or without a liner, formed by excavating or boring a hole in the ground andsubsequently filling it with plain or reinforced concrete.1.3.2 Cased pile A pile formed in the ground by installing a liner and partially or wholly filling it with plain or reinforced concrete after excavation. 1.3.3 Cone penetration test (CPT) A test in accordance with AS 1289.6.5.1, to determine the penetration resistance of a soil. 1.3.4 Continuous flight auger pile (CFA) A pile formed in the ground by drilling with a hollow flight auger that is subsequently and progressively withdrawn, with the cavity below the auger tip being gradually filled with concrete or cement grout injected under pressure. 1.3.5 Design action s e d b y H A T C H A S S O C I A T E S o n 30 N o v 2009AS 2159—2009 81.3.6 Design action effect (E d )Action effect computed from the design values of the actions or design loads.1.3.7 Design geotechnical strength (R d,g )The product of the design ultimate geotechnical strength (R d,ug ) and the geotechnical strength reduction factor (φg ).1.3.8 Design lifePeriod of time during which a structure or a structural element, when designed, is assumed to perform for its intended purpose with expected maintenance but without major structural repair being necessary.1.3.9 Design serviceability load (E ds )The load on a pile corresponding to the serviceability limit state.1.3.10 Design structural strength (R d,s )The product of the design ultimate structural strength (R d,us ) and the structural strength reduction factor (φs ).1.3.11 Design ultimate geotechnical strength (R d,ug )An estimate of the ultimate geotechnical strength assessed using calculations in accordance with Section 4 of this Standard.1.3.12 Design ultimate structural strength (R d,us )The limit state at which static equilibrium is lost, or at which structural elements fail.NOTE: The design ultimate structural strength may be assessed using calculations in accordance with Section 5 of this Standard.1.3.13 Driven cast in place pileA pile formed by driving a liner, which is either permanent or temporary, and filling with plain or reinforced concrete.1.3.14 Driven preformed pileA prefabricated pile installed in the ground by driving.1.3.15 DurabilityAbility of a structure or a structural element to maintain adequate performance for a given time under expected actions and environmental influences.1.3.16 End-bearing pileA pile where the major component of the resistance of the pile is contributed by the force developed at the base of the pile. 1.3.17 Footing A part of a structure in direct contact with and transmitting load to the supporting foundation. 1.3.18 Foundation The soil, subsoil or rock, whether built-up or natural, upon which a structure is supported. NOTE: The term ‘foundation’ is commonly used to mean both the footing and the ground supporting the footing. 1.3.19 Friction pile s e d b y H A T C H A S S O C I A T E S o n 30 N o v 20091.3.20 Ground anchorA tendon anchored into the ground by bond and used to provide a reaction for test loading piles.1.3.21 Large displacement pilesPreformed or cast in place piles, generally with a solid cross-section dimension of at least 300 mm, installed by driving, screwing, pushing, vibrating or similar methods, which cause a displacement such that significant stresses are induced in the surrounding soils, which may increase the load capacity of the pile and cause displacement of the surrounding soils. 1.3.22 Limit stateCondition for which a system is designed, and beyond which it ceases to fulfil its intended function and becomes unfit for use.NOTE: There are recognized limit states, e.g., for fire, serviceability, stability and strength.1.3.23 PileA structural member that is driven, screwed, jacked, vibrated, drilled or otherwise installed in the ground so as to transmit loads to the underlying soil or rock and provide a foundation for structure. 1.3.24 Pile groupNumber of piles installed in close proximity and usually having a common pile cap. 1.3.25 Pile head Top of a pile. 1.3.26 Pile heaveDisplacement (usually vertical) of a pile caused by the driving, or by external ground movements, of piles in close proximity. 1.3.27 Raking pileA pile installed at an angle to the vertical.1.3.28 Serviceability limit state (SLS), serviceabilityA limit state beyond which specified service criteria are no longer met, such as unacceptably large displacements, vibrations, cracking, spalling and other local damage. 1.3.29 SetPermanent penetration of a driven pile or liner per blow of the hammer.1.3.30 Small displacement pilesPreformed or cast in place piles, generally with a hollow cross-section or a solid cross-section dimension less than 300 mm, installed by driving, screwing, pushing, vibrating or similar methods, which cause a small displacement such that significant stresses or displacements are not induced in the surrounding soils. 1.3.31 Standard penetration test (SPT)A test in accordance with AS 1289.6.3.1, to determine the penetration resistance of a soil. 1.3.32 Steel screw pilesPreformed small displacement piles installed by rotating a steel pipe, which has one or more spiral flights (helices) welded to it.s e d b y H A T C H A S S O C I A T E S o n 30 N o v 20091.3.33 Temporary compressionThe temporary pile-head deflection during a hammer blow, comprising elastic deflection of the pile cushion, the pile and the soil. 1.3.34 Test pilePile subjected to a loading test with the primary purpose of establishing the load deformation characteristics, and/or the ultimate structural strength of the pile, and/or the ultimate geotechnical strength of the pile/soil system. 1.3.35 Test ultimate geotechnical strength (R t,ug )An estimate of the ultimate geotechnical strength assessed from a load test carried out in accordance with Section 8 of this Standard. 1.3.36 Toe The base of the pile.1.3.37 Ultimate geotechnical strength (R ug )The resistance developed by an axially or laterally loaded pile or pile group at which static equilibrium is lost or at which the supporting ground fails. 1.4 NOTATIONThe symbols used in this Standard are listed below. Unless a contrary indication appears elsewhere, the symbols used in this Standard shall be as defined below. The notations in Clause 3.3, relating to load and combinations in AS 1170.4, have not been incorporated in this table.TABLE 1.1 NOTATIONSymbol Term Text referenceA b Plan area of pile baseClauses 4.4.1, 4.4.2 bA ′ Net area of pile base resisting uplift, i.e., the differencebetween cross-sectional areas of the pile base and the pile shaft Clause 4.4.2 A g Area of the pile cross-section Clause 5.3.3(b)ARR Average risk rating for overall designClause 4.3.2, Table 4.3.2 (C) A s Surface area of pile in intimate contact with soil Clauses 4.4.1, 4.4.2 A scCross-sectional area of compression reinforcement Clause 5.3.3(b)c Pile wave speed Paragraph C2.2, Appendix C d Pile diameterClause 5.6.3.2, Table 8.4.3.1 d b Diameter of longitudinal steel Clause 5.3.7 d t Pile base (toe) diameter Tables 8.4.3.1, 8.5.2 D d Dowel diameterClause 5.6.3.2 D Overall minimum width of pile in plane of bending Clause 5.2.2(b) EAverage Young’s modulus of pileTables 8.4.3.1, 8.5.2(continued )s e d b y H A T C H A S S O C I A T E S o n 30 N o v 2009TABLE 1.1 (continued)Symbol Term Text referenceE dDesign action effectClauses 1.3.6, 3.2.2(b),5.4.2.3, 3.2.2(d), 4.3.1, 5.2.1, 8.3.3.4, Paragraph B8,Appendix B, Tables 8.3.3.2, 8.3.3.4 and E1, Appendix E E ds Design serviceability loadClauses 1.3.9, 4.6.3(a), Paragraph B8, Appendix B, Tables 8.3.3.2, 8.3.3.3, 8.4.3.1, 8.5.2 and E1, Appendix EF eh Bending moments, shear forces and axial actions induced by heave due to unloading of ground due to excavation Clauses 3.3.1.2(d), 3.3.2(b) F em Bending moments, shear forces and axial actions induced by lateral ground movementsClauses 3.3.1.2(c), 3.3.2(b) F es Compressive and tensile actions in the pile induced by vertical ground movementsClauses 3.3.1.2(b), 3.3.2(b) F nfActions due to negative frictionClauses 3.3.1.2(a), 3.3.2(b), 4.6.3, Tables 8.3.3.3, 8.4.3.1 and E1, Appendix E f b Ultimate base pressure for compression pile Clause 4.4.1 f bt Ultimate base pressure for uplift pile Clause 4.4.2 c f ′ Characteristic concrete strengthTable 6.4.3 cmf ′ Characteristic strength of concrete at relevant age Clause 7.3.3.1(a), Table 7.3.3.1 f m,s Average skin friction for condition of full mobilization—Compression pileClause 4.4.1 f m,st Average skin friction for condition of full mobilization—Tension pileClause 4.4.2f sy Yield stress for reinforcement in concrete piles Clauses 7.3.3.1(b), 7.3.2.g Acceleration due to gravity (9.8 m/s 2) Paragraph C5.4, Appendix C h Depth to cut-offClause 7.2.1(b)IRR Individual risk rating for risk factor Clause 4.3.2, Tables 4.3.2(A), 4.3.2(B)kConcrete placement factor Clause 5.2.1, 5.3.2, 5.3.6 K Testing benefit factorClause 4.3.1 l 1 Minimum edge distance to head of pileClause 5.6.3.2 L nf Length of the test pile in contact with ground expected to undergo long-term settlement Tables 8.4.3.1, 8.5.2 L Pile lengthTables 8.4.3.1, 8.5.2,Paragraph C2.3, Appendix C M d Design bending moment Clause 5.2.2 N d Design axial loadClause 5.2.2(b) pPercentage of total piles tested that meet the specified acceptance criteriaClause 4.3.1(continued )s e d b y H A T C H A S S O C I A T E S o n 30 N o v 2009TABLE 1.1 (continued)Symbol Term Text referenceP gMaximum test load for assessment of geotechnical ultimate limit state R t,ugClauses 8.3.3.1, 8.3.3.2,8.3.3.4, 8.5.2, Paragraph A3.1, Appendix A, Paragraph B1, Appendix B, Tables 8.4.3.1, 8.5.2, A1, A2, Appendix A p o Total overburden pressure at base level Clause 4.4.1 P max Pile jacking installation forceClause 7.3.4.1P sMaximum test load for assessment of pile performance at serviceability limit state = E dsClauses 8.3.3.1, 8.3.3.2, Paragraph B1, Appendix B, Tables 8.3.3.2, 8.3.3.3, 8.4.3.1, 8.5.2, A1, A2, A3, Appendix A, B1, Appendix B P uMaximum test load for assessment of design geotechnical ultimate limit stateClauses 8.3.3.1, 8.3.3.2,Paragraph A3.1, Appendix A and Paragraph B1,Appendix B Tables 8.3.3.2, 8.3.3.3, 8.4.2, 8.4.3.1, A1, Appendix AR d,g Design geotechnical strength of pileClauses 1.3.7, 3.2.2(c), 3.2.2(d), 4.3.1, Table E1, Appendix ER d,s Design structural strength of pileClauses 1.3.10, 3.2.2(c), 3.2.2(d), 5.2.1, 5.4.2.3, Table E1, Appendix E R t,ugUltimate geotechnical strength of a pile as assessed from a load test carried out in accordance with Section 8 of this Code Clauses 1.3.35, 8.4.2.2, 8.4.3.5, Tables 8.3.3.2, 8.3.3.3, E1, Appendix E R ugUltimate geotechnical strength of pile. This is estimated either by calculation (R d,ug ) or by test (R t,ug ) Clauses 1.3.37, 7.3.4.1, Tables 8.3.3.2, 8.4.2, E1, Appendix ER us Ultimate structural strength of pileClauses 5.2.1, 5.3.1, 5.3.2, Table E1, Appendix E R d,ugDesign ultimate geotechnical strength of pile (ultimate load capacity)Clauses 1.3.7, 1.3.11, 4.3.1, 4.3.3, 4.4.2, 4.4.4, 8.2.4, Table E1, Appendix E R d,g,c Design ultimate geotechnical strength of combined pile and raft foundationClause 4.4.4, Table E1, Appendix ER d,us Design ultimate structural strength of pileClause 1.3.10, 1.3.12, R d,ug,s Design ultimate geotechnical strength of shallow or raft footing, for the net area in contact with the supporting groundClause 4.4.4, Table E1, Appendix ER d,ug,szDesign ultimate geotechnical strength of pile in stable zone, i.e., the soil strata not subject to externally imposed ground settlementsClause 4.6.3, Table E1, Appendix E S u Ultimate value of various actions appropriate for particular combinations Clause 3.3.2(b) W Weight of pileClauses 4.4.1, 4.4.2w i Weighting factor for individual risk ratings Clause 4.3.2, Tables 4.3.2(A) γCoefficient of jacked pressureClause 7.3.4.1s e d b y H A T C H A S S O C I A T E S o n 30 N o v 2009TABLE 1.1 (continued)Symbol Term Text referenceδ Pile movementsClause 3.2.3φgGeotechnical strength reduction factor for single piles or pilegroupsClauses 4.3.1, 4.4.4, 4.6.3, 8.3.3.4, Paragraph B8, Appendix B, Table 8.3.3.2 φgb Basic geotechnical strength reduction factor given in Clause 4.3.2Clauses 4.3.1, 4.3.2 φgs Geotechnical strength reduction factor for the shallow or raft footingClause 4.4.4φs Structural strength reduction factor for single piles or pile groupsClauses 1.3.10, 5.2.1, 5.3.1, 5.3.4, 5.3.5, 5.4.2.3 φtfIntrinsic test factorClause 4.3.11.5 CLASSIFICATION OF PILES 1.5.1 GeneralThe classification of pile types used in this Standard is illustrated in Figure 1.5. Pile typesare broadly classified into ‘displacement’ and ‘non-displacement’ piles and further subdivided on the basis of the method of pile installation and formation. 1.5.2 Displacement pilesDisplacement piles are defined as those that displace the ground through which they are being installed. To operate as a displacement pile, the displaced volume shall approximate the pile volume.Displacement piles may be installed by hammering, pushing, screwing, vibrating or other means to force them into the ground.Displacement piles may be one of the following: (a)Preformed Solid and hollow sections that are installed in the ground and left in position. Such piles may be extended by splicing on additional lengths of piling. Preformed piles may be fabricated from— (i)concrete, reinforced or prestressed; (ii) steel—H Section, tube and other sections; (iii) timber; or(iv) a combination of concrete, steel or timber sections. (b)Driven cast in place Pile formed in situ by driving a tubular liner to form a void, which is then wholly or partially filled with concrete or grout. The liner may be either— (i)permanent —made of concrete or steel with open or closed ends of constant or tapered section; or(ii) temporary —steel tube extracted during concreting or grouting, with or withoutan expanded base. (c)Screwed cast in place Piles formed in situ by screwing a threaded tube into the ground with concrete placement as the screw head is withdrawn.s e d b y H A T C H A S S O C I A T E S o n 30 N o v 20091.5.3 Non-displacement piles 1.5.3.1 GeneralPiles formed in situ by removing soil, using either rotary drilling, percussion, reverse circulation, grabbing, chiselling and mechanical or hand excavation methods, to form a void, which is then filled with concrete or grout. During removal of the soil, the sides of the excavated void may or may not be supported. 1.5.3.2 SupportedThe support may be either— (a) permanent —using steel, concrete or other liners; or (b)temporary —using— (i)steel, concrete or other liners or timber shoring; (ii) drilling fluids; or (iii) continuous flight augers. 1.5.3.3 UnsupportedPiles in which the ground is left exposed during excavation.1.5.4 Partial displacement, post-grouted and preloaded non-displacement piles Various techniques, such as partial displacement augers, post-grouting of the shaft or base and preloading the base of non-displacement piles, are used to improve the performance of non-displacement piles.Soil and rock displaced during installationSoil and rock removed before or duringinstallationLarge displacementSmall displacementSteelScrewOpen tubeOther sections H section PreformedCast in placePermanentlinerScrewedDrivenTemporar y linerConcreteTimberCompositeConcrete shellClosed steel tubeReinforcedPrestressedSuppor tedUnsuppor tedTemporar y Suppor tShoring or liners Drilling fluid Soil on continuousflight augerDisplacement pilesNon-displacement pilesPermanent Suppor tSteel linerConcrete linerOtherFIGURE 1.5 CLASSIFICATION OF PILE TYPESs e d b y H A T C H A S S O C I A T E S o n 30 N o v 2009S E C T I O N 2 S I T E I N V E S T I G A T I O N2.1 GENERALFor any site on which it is proposed to install piles, site investigation shall be carried out toprovide sufficient information to fulfil the requirements of Clause 2.2. When planning the site investigation, existing relevant information shall be taken into account.NOTE: The intention of this Section is to ensure that adequate information is available for design and construction.2.2 INFORMATION REQUIREDAppropriate site investigations shall provide information on geotechnical conditions according to AS 1726, as follows: (a) The geotechnical design of piles.(b) Assessment of geotechnical conditions for pile construction or installation. (c)Some additional site-specific aspects, including— (i)potential for ground heave—damage to adjacent structures or neighbouring piles;(ii) vibration effects—potential for damage to adjacent structures; (iii) expansive soil problems;(iv) potential difficulties with pile cap construction; (v)groundwater conditions; (vi) negative friction effects;(vii) near-surface conditions or lateral load design, if relevant;(viii) possible obstructions to installation, e.g., boulders or old footings or piles; (ix) potential for slope instability; (x)effects of excavation or scour; (xi) effects of contaminated sites;(xii) an assessment of the site surface for the provision of a safe work platform forpiling equipment;(xiii) potential for acid sulfate soils; and(xiv) potential for weak or compressible layers, or caverns below the pile base,including soils below lava flows. (d)Assessment of the potential effects of site conditions on pile durability.NOTE: The site investigation should obtain information on all materials that might influence the strength and serviceability performance of the structure. Due account should be taken of the range of foundation options that might apply. This should include testing of the soil and groundwater for aggressive agents, including sulphate, chloride and pH, to ensure appropriate exposure classification in regard to durability.s e d b y H A T C H A S S O C I A T E S o n 30 N o v 2009。

aoac 2002.02

45.4.15AOAC Official Method2002.02Resistant Starch in Starch and Plant MaterialsEnzymatic DigestionFirst Action2002[Applicable to plant and starch materials containing resistant starch(RS)contents ranging from2.0to64%on an“as is”basis.] See Table2002.02for the results of the interlaboratory study sup-porting acceptance of the method.A.PrincipleNonresistant starch is solubilized and hydrolyzed to glucose by the combined action of pancreaticα-amylase and amyloglucosidase (AMG)for16h at37°C.The reaction is terminated by addition of ethanol or industrial methylated spirits(IMS)and RS is recovered as a pellet by centrifugation.RS in the pellet is dissolved in2M KOH by vigorously stirring in an ice–water bath.This solution is neutral-ized with acetate buffer and the starch is quantitatively hydrolyzed to glucose with AMG.Glucose is measured with glucose oxidase–peroxidase reagent(GOPOD),which is a measure of RS content.Nonresistant starch(solubilized starch)is determined by pooling the original supernatant and the washings and measuring the glucose content with GOPOD.B.Apparatus(a)Grinding mill.—Centrifugal,with12-tooth rotor and1.0mm sieve,or similar device.Alternatively,a cyclone mill can be used for small test samples.(b)Meat mincer.—Hand-operated or electric,fitted with4mm screen.(c)Bench centrifuge.—Holding16×100mm glass test tubes, operating at ca1500×g.(d)Shaking water bath.—Grant OLS200[Grant Instruments (Cambridge)Ltd.,Royston Hertfordshire SG86GB,UK,Tel.:+44 (0)1763260811;Fax:+44(0)1763262410;E-mail: paulp@],or equivalent.Set in linear motion at 100rpm on the dial(equivalent to a shake speed of200strokes/min), a stroke length of35mm,and37°C.(e)Water bath.—Maintaining50±0.1°C.(f)Vortex mixer(g)Magnetic stirrer(h)Magnetic stirrer bars.—5×15mm.(i)pH Meter(j)Stop-clock timer.—Digital.(k)Analytical balance.—Weighing to0.1mg.(l)Spectrophotometer.—Operating at510nm,preferably fitted with flow-through10mm path length cell.(m)Pipets.—Delivering100µL;with disposable tips.Alterna-tively,use motorized hand-held dispenser.(n)Pipetter.—Delivering2.0,3.0,and4.0mL.(o)Culture tubes.—Corning,glass screw-cap,16×125mm. (p)Glass test tubes.—16×100mm,14mL.(q)Test tube racks.—Holding16×100mm tubes.(r)Thermometer.—37±0.1and50±0.1°C.(s)Volumetric flask s.—100,200,and500mL;1and2L.C.Reagents(a)Sodium maleate buffer.—100mM,pH6.0.Dissolve23.2g maleic acid in1600mL water and adjust pH to6.0with4M(160g/L)NaOH solution.Add0.6g CaCl2⋅2H2O and0.4g sodium azide,and adjust volume to2L.Solution is stable at4°C for12months. (b)Sodium acetate buffer.—1.2M,pH3.8.Add70mL glacial acetic acid to800mL water and adjust to pH3.8with4M NaOH so-lution.Adjust volume to1L with water.Solution is stable at room temperature for12months.(c)Sodium acetate buffer.—100mM,pH4.5.Pipette5.8mL gla-cial acetic acid to900mL water and adjust to pH4.5with4M NaOH solution.Adjust volume to1L with water.Solution is stable at4°C for2months.(d)Potassium hydroxide solution.—2M.Add11.2g KOH to 150mL water and dissolve by stirring.Adjust volume to200mL with water.Stable at room temperature for at least12months. (e)Aqueous ethanol or IMS.—Approximately50%(v/v).Dilute 500mL ethanol(95or99%)or IMS(denatured ethanol;ca95%eth-anol plus5%methanol)to1L with water.Stable at room tempera-ture for at least12months.(f)Stock amyloglucosidase stock solution.—3300units(U)/mL in50%e directly without dilution.Solution is viscous; dispense from positive displacement dispenser.AMG solution is stable for up to5years when stored at4°C.(Note:One unit enzyme activity is amount of enzyme required to release1µmol glucose from soluble starch per minute at40°C and pH4.5.)AMG solution should be devoid of detectable levels of free glucose.(g)AMG solution.—300U/mL.Dilute2mL concentrated AMG solution,(f),to22mL with100mM sodium maleate buffer(pH6.0), (a).Divide into5mL aliquots and store frozen in polypropylene containers between use.Stable to repeated freeze–thaw cycles for >5years at–20°C.(h)Pancreatic a-amylase suspension.—10mg(30U/mL)plus AMG(3U/mL).Immediately before use,suspend1g pancreatic α-amylase in100mL sodium maleate buffer,(a),and stir for5min. Add1mL AMG solution(300U/mL),(g),and mix well.Centrifuge at>1500×g for10min,and carefully decant the e this solution on the day of preparation.(i)GOPOD–aminoantipyrine buffer mixture.—Mixture of glu-cose oxidase,>12000U/L;peroxidase,>650U/L;and 4-aminoantipyrine,0.4mM.Prepare buffer concentrate by dissolv-ing136g KH2PO4,42g NaOH,and30g4-hydroxybenzoic acid in 900mL water.Adjust to pH7.4with either2M HCl or2M NaOH. Dilute solution to1L,add1g sodium azide,and mix well until dis-solved.Buffer concentrate is stable for up to3years at4°C.To prepare GOPOD–aminoantipyrine buffer mixture,dilute 50mL buffer concentrate e part of diluted buffer to dis-solve entire contents of vial containing freeze-dried GOPOD–aminoantipyrine mixture.Transfer contents of vial to1L volumetric flask containing diluted buffer,and adjust to volume (GOPOD).Reagent is stable2–3months when stored at4°C and 2–3years when stored at–20°C.Check color formation and stability of GOPOD–aminoantipyrine buffer mixture by incubating(in dupli-cate)3.0mL GOPOD–aminoantipyrine buffer mixture with certi-fied glucose standard(100µg dried crystalline glucose in0.2mL 0.2%sodium benzoate solution).After15,20,30,and60min incu-bation,read absorbance,A,of solution at510nm.Maximum color should be reached within20min,and color should be stable for at least60min at50°C after maximum color is achieved.(j)Glucose standard solution.—1mg/mL.Dissolve1.00g anhy-drous,analytical reagent grade crystalline D-glucose(99.5%)in900mL of0.2%benzoic acid solution in water.Adjust volume to1L in volumetric flask and store in well-sealed glass container.Stable at room temperature>5years.Items(f)and(h)–(j)are supplied in the Resistant Starch Assay Kit available from Megazyme International Ireland Ltd.(Bray Business Park,Bray,County Wicklow,Ireland),but preparations of reagents and buffers which meet these criteria may also be used.D.Preparation of Test SamplesGrind ca50g test sample of grain or lyophilized plant material in grinding mill,B(a),to pass1.0mm sieve.Transfer all material to wide-mouthed plastic jar and mix well by shaking and inversion. Grinding is not required with industrial starch preparations supplied as a fine powder.E.Measurement of Resistant Starch(a)Hydrolysis of nonresistant starch.—Accurately weigh100±5mg test portion directly into each screw-cap tube,B(o),and gently tap the tube to ensure that material falls to the bottom.Add4.0mL pancreaticα-amylase(10mg/mL)containing AMG(3U/mL), C(h),to each tube.Tightly cap the tubes,mix on a Vortex mixer,and attach them horizontally,under water,in a shaking water bath,B(d), aligned in the direction of motion.Incubate at37°C with continuous shaking(200strokes/min for16h).(Note:For linear motion,a set-ting of100on the water bath is equivalent to200strokes/min; 100forward and100reverse.)Remove tubes from water bath and remove excess water on tubes with paper towel.Remove tube caps and add4.0mL IMS(99%,v/v) or ethanol(95–99%).Mix tube contents vigorously on Vortex mixer.Centrifuge tubes at ca1500×g for10min(noncapped).Care-fully decant supernatants and resuspend pellets in2mL50%IMS, C(e),with vigorous mixing on Vortex mixer,B(f).Add additional6 mL50%IMS,C(e),mix tubes,and centrifuge again at1500×g for 10min.Repeat this suspension and centrifugation step once more. Carefully decant supernatants and invert tubes on absorbent paper to drain excess liquid.(b)Measurement of RS.—Add magnetic stirrer bar(5×15mm) and2mL2M KOH,C(d),to each tube and resuspend the pellets. Dissolve RS by stirring for ca20min in an ice–water bath over a magnetic stirrer(do not mix on a Vortex mixer as this may cause the starch to emulsify).In this step,ensure that tube contents are being vigorously stirred when KOH solution is added to avoid formation of a lump of starch material which would be difficult to dissolve. Add8mL1.2M sodium acetate buffer(pH3.8),C(b),to each tube with stirring on the magnetic stirrer.Immediately add0.1mL AMG (3300U/mL),C(f),mix well on magnetic stirrer,and then place tubes in a water bath at50°C.Incubate tubes for30min with inter-mittent mixing on a Vortex mixer.For test samples containing>10%RS,quantitatively transfer con-tents of tube to100mL volumetric flask using water wash bottle. Use external magnet to retain stirrer bar in the tube while washing the solution from the tube with a water wash bottle.Adjust to100mL with water.Centrifuge an aliquot of the solution at1500×g for 10min.For test samples containing<10%RS,directly centrifuge tubes at1500×g for10min without dilution.For such products,the final volume in the tube is10.3±0.05mL.Transfer0.1mL aliquots(in duplicate)of either diluted or undi-luted supernatants into glass test tubes(16×100mm),B(p),add 3.0mL GOPOD reagent,C(i),mix tube contents on Vortex mixer, and incubate at50°C for20min.Prepare reagent blank solutions by mixing0.1mL0.1M sodium acetate buffer(pH4.5),C(c),and 3.0mL GOPOD reagent.Prepare glucose standards(in quadrupli-cate)by mixing0.1mL glucose(1mg/mL),C(j),and3.0mL GOPOD reagent,C(i).Incubate at50°C for20min,cool,and set spectrophotometer to0with the reagent blank.Measure the absorbance of each solution at510nm against the reagent blank.Av-erage duplicate absorbance values.The GOPOD color response with glucose is linear over the absorbance range0.0–1.5absorbance units.F.CalculationsCalculate RS(%,“as is”basis)in test samples as follows: (1)For products containing>10%RS.—RS(g/100g sample)=∆A×F×(100/0.1)×(1/1000)×(100/W)×(162/180)=∆A×F/W×90(2)For products containing<10%RS.—Table2002.02.Interlaboratory study results for measurement of resistant starch by enzymatic digestion in starch samples and se-lected plant materialsSample Mean RS a,%No.of labs b,c s r s R RSD r,%RSD R,%r d R e HORRATHylon VII(HAMS)f46.2937(0) 1.91 3.87 4.128.37 5.3410.84 3.72 Green banana43.5636(1) 1.39 3.69 3.188.47 3.8810.34 3.74 Native potato starch63.3935(2) 2.66 3.77 4.20 5.947.4510.54 2.77 CrystaLean(retrograded HAMS)39.0434(3)0.77 2.00 1.97 5.13 2.15 5.61 2.23 ActiStar(RS)48.2836(1) 1.12 2.81 2.32 5.83 3.147.87 2.61 Kidney beans(canned) 4.6635(2)0.110.21 2.42 4.580.320.60 1.44 Corn flakes 2.2034(3)0.080.24 3.4310.90.210.67 3.08a Calculated on“as is”basis(“as is”for banana,kidney beans,and corn flakes means on a lyophilized basis).b,c b=Number of collaborating laboratories(number of outlier laboratories).d r=2.8×sr .e R=2.8×sR .f High amylose maize starch.RS(g/100g sample)=∆A×F×(10.3/0.1)×(1/1000)×(100/W)×(162/180)=∆A×F/W×9.27where∆A=averaged absorbance(reaction)read against the reagent blank;F=conversion factor from absorbance to micrograms[the absorbance obtained for100µg glucose in the GOPOD reaction is determined and F=100(micrograms of glucose divided by the GOPOD absorbance for this100µg glucose];100/0.1=volume ad-justment(0.1mL taken from100mL);1/1000=conversion from micrograms to milligrams;W=“as is”weight of test portion ana-lyzed;100/W=factor to present starch as a percentage of test portion weight;162/180=factor to convert from free glucose,as deter-mined,to anhydro-glucose as occurs in starch;10.3/0.1=volume adjustment(0.1mL taken from10.3mL)for test portion containing 0–10%RS where the incubation solution is not diluted and the final volume is10.3±0.05mL.Reference:J.AOAC Int.85,1103(2002).。

AOAC_2007英文版_01内部培训资料

AOAC_2007英文版_01内部培训资料AOAC_2007英文版_01 Internal Training MaterialIntroduction:In this training material, we will discuss the AOAC (Association of Official Analytical Chemists) 2007 English version and its significance in quality assurance and food safety. This guide aims to provide a comprehensive understanding of the AOAC methods and their importance in laboratory testing and analysis.1. Background of AOAC:The Association of Official Analytical Chemists (AOAC) is a globally recognized organization that develops and validates analytical methods for various industries, including food, pharmaceuticals, agriculture, and environmental analysis. Founded in 1884, AOAC ensures the reliability and accuracy of analytical data through rigorous scientific standards.2. Purpose of AOAC Methods:The AOAC methods serve multiple purposes in the field of analytical chemistry. They provide standardized procedures for testing and analysis, ensuring consistency and comparability of results across different laboratories. By establishing a common ground for measurement, the AOAC methods enable accurate determination of various parameters, such as nutrient content, contaminants, and adulterants in food and other matrices.3. Benefits of AOAC Methods:The utilization of AOAC methods offers several advantages in laboratory testing and analysis:a) Reliability: AOAC methods are thoroughly validated and have undergone extensive peer review, ensuring their reliability in generating accurate and precise results.b) Consistency: The standardized procedures outlined in AOAC methods allow for consistent sample preparation, instrument calibration, and data interpretation, reducing variability between different laboratories and analysts.c) Traceability: AOAC methods provide a clear documentation trail, including detailed instructions, calculations, and quality control measures. This traceability enhances transparency and facilitates result comparison and verification.d) Regulatory Compliance: Many regulatory bodies and organizations, including the US Food and Drug Administration (FDA) and the European Food Safety Authority (EFSA), recommend or require the use of AOAC methods for the analysis of food and other products. Adhering to these methods ensures compliance with regulatory standards.4. AOAC Performance Tested Methods:AOAC offers a program called "Performance Tested Methods" (PTM) to verify the accuracy and reliability of analytical methods developed by manufacturers and laboratories. The PTM certification assures users that a particular method has met the stringent criteria set by AOAC regardingmethod performance and ruggedness. PTM-certified methods provide an extra level of confidence in the accuracy of results.5. AOAC Official Methods of Analysis:AOAC Official Methods of Analysis (OMA) are widely recognized standards used by laboratories worldwide. These methods undergo extensive validation and review processes, involving collaborative studies across multiple laboratories to ensure their accuracy and reproducibility. OMA methods cover a wide range of parameters, including microbiological analysis, pesticide residue determination, proximate analysis, and mycotoxin detection.Conclusion:The AOAC 2007 English version plays a crucial role in ensuring the quality and safety of food and other products through standardized and validated analytical methods. By following these methods, laboratories can generate reliable and accurate results, contributing to consumer protection and regulatory compliance. The AOAC methods provide a foundation for consistent and traceable measurement practices, enabling effective quality assurance and reliable product testing in various industries.。

AOAC标准目录

AOAC电子版标准目录一、AOAC方法1. AOAC Official Method 993.31.Phosphorus (Available) in Fertilizers. Direct Extraction Method2. AOAC Official Method 993.31.Nitrogen (Total) in Fertilizers. Combustion Method.3. AOAC Official Method 995.01.Dirthianon in Technical Products.and Fornmulations.4. AOAC Official Method 995.14.Methomy in insecticidal Formulations.Reversed-phase liquid Chromatographic Method5.AOAC Official Method 993.02.bentazon in pesticide formulations liquid Chromatographic Method6.AOAC Official Method 995.02.Cyfiuthrin in Pesticide formulations liquid Chromatographic Method7.AOAC Official Method 993.01 Phosphamidon in Technical and Formulated Products8.AOAC Official Method 996.03 Acephate in Technical and Soluble Powder Formulations9.AOAC Official Method 995.08.Atrazine in Water Magnetic Particle lmmunoassay10.AOAC Official Method 2000.05. Determination of Glyphosate and AminomethyphonicAcid(ACMPA)in Crops11.AOAC Official Method 993.15 1,2-Dibromoethane and 1,2-Dibromo-3-chloropropane in Water12.AOAC Official Method 994.19 Total nitrogen in Urine Pyrochemiuminescence Method13.AOAC Official Method 995.21 Yeast and mold counts in Foods hydrophobic Grid Membranefilter14.AOAC Official Method 996.02 Coliform Count in Dairy Products15.AOAC Official Method 2000.15 Rapid enumeration of Coliforms in Foods Dry RehydratableFilm Method16.AOAC Official Method 2000.13 Reveal for E.coli O157:H7 Test System in selected foods17.AOAC Official Method 2000.14 Reveal for E.coli O157:H7 Test in selected foods andEnvironmental Swabs18.AOAC Official Method 993.06 Staphylococal enterotoxins in selected foods19.AOAC Official Method 995.12 Staphylococcus autrus lsolated from foods latexAggluTINATION Test Method20.AOAC Official Method 2001.05 Petrifilm S.aureus count Platr Method for the RapidEnumeration of Staphylococcus aureus in Seleced Foods21.AOAC Official Method 993.10 Clistridium Perfringrns from Shellfish lron Milk Method22.AOAC Official Method 995.20Salmonella in Raw,Highly contaminated Foods and poultryFeed23.AOAC Official Method 993.08 Salmonella in foods24.AOAC Official Method 993.07 Sa;monella Cocoa and Chocolate motility Enrichment onmodified Sem-Solld25.AOAC Official Method 995.07 Salmonella in dried Milk Products Motility Enrichment onModified Sem-Solld26.AOAC Official Method 994.04 Salmonella in drt Foods Refrigerated Pre-Enrichmenr27.AOAC Official Method 2000.07 Salmonella in Fooods Rapid Cocorimetric28.AOAC Official Method 2000.07 Salmonella in Fooods with a Low Microbial Load Detecition29.AOAC Official Method 2001.07 Salmonella in Selected Foods lmmumno-ConcentrationSalmonella (ICS)30.AOAC Official Method 2001.08 Salmonella in Selected Foods lmmumno-ConcentrationSalmonella (ICS)31. AOAC Official Method 2001.09 Salmonella in Selected Foods lmmumno-ConcentrationSalmonella (ICS)32. AOAC Official Method 2001.08 Salmonella in Selected Foods lmmumno-ConcentrationSalmonella (ICS)33.AOAC Official Method 993.12 Listeria monceytogenes in milk ang dairy Products34.AOAC Official Method 993.09 Listeria in Dairy Pruducts, Seafoods, and Meats. ColorimetricDeoxyribonucleic Acid Hybridization Method ( GENE-TRAK Listeria Assay)35. AOAC Official Method 994.03. Listeria monocytogenes in Dairy Pruducts, Seafoods, andMeats. Colorimetric Deoxyribonucleic Acid Hybridization Method (Listeria-Tek)36.AOAC Official Method 994.06. Vibrio vulnificus. Gas Chromatographic dentification Methodby Microbial Fatty Acid profile37.AOAC Official Method 993.32.Multiple sulfonamide Residues in Raw Bovine MilkLiquid Chromatographic Method First Action 199338.AOAC Official Method 2001.14 Determination of Nitrogen(Total)in Cheese kieldahl Method39.AOAC Official Method 993.O5 l-Malic/Total Malic Acid Ratio in Apple Juice40.AOAC Official Method 995.06 D-Malic Acid in Apple Juice Liquid Chromatographic Method41. AOAC Official Method 994.11 Benzoic Acid in Orange Juice Liquid ChromatographicMethod42.AOAC Official Method 995.17 Beet Sugar in fruit Juices43.AOAC Official Method 993.20 Lodine Value of Fates and Oils Wijs(Cyclohexane-AceticSolvent)Method44.AOAC Official Method 994.02 Lead in Edible Oils and Fats Direct Graphite Furnace45.AOAC Official Method 994.18 Mon-and Diglycerides in Fats and Oils Gas ChromatographicMethod46.AOAC Official Method 2000.17 Determination of Trace Glucose and Fructose Determinationof Trace Glucose and Fructose in Raw Cane Sugar47.AOAC Official Method 994.09 Glucoamylase Activity in Lndustrial Enzyme Preparations48.AOAC Official Method 995.11 Phosphorus (total)in Foods Colorimetric Method49.AOAC Official Method 2001.13 Determination of Vitamin A(Retinol)in Foods LiquidChromatography50.AOAC Official Method 2000.11 Polydextrose in Foods lon Chromotography51.AOAC Official Method 2000.01 Determination of 3-Chloro-1,2-Propanediol in Foods andFood Ingredients52.AOAC Official Method 993.16 Total Aflato xins(B1,B2,and G1)in Corn Enzyme-Linkedlmmunosorbent Assay Method53.AOAC Official Method 993.17 Aflatoxins in Corn and Peanuts Thin-Layer ChromatographicMethod54. AOAC Official Method 994.08 Aflatoxins in Corn,Almonds, brazil Nuts,Peanuts,andPeanuts,and Pistachio Nuts55.AOAC Official Method 2000.16 Aflatoxin B1 in Baby Foood lmmmunoaffinty Column HplcMethod56.AOAC Official Method 2000.08 Aflatoxin M1in Liquid Milk lmmmunaffinity Column byLiquid Chromatography57.AOAC Official Method 995.15 Fumonisins B1,B2,and B3in Corn liquid ChromatographicMethod58.AOAC Official Method 2001.04 Determination of Fumonisins B1,and B2,in Corn and CornFlakes59.AOAC Official Method 2001.06 Determination of Total Fumonisins in Corn Competitive ofDirecet Enzyme-Linked Immunosorbent Assay60.AOAC Official Method 2000.09 Ochratoin A in Roaseted Coffee Immunoaffinity ColumnHPLC Method61.AOAC Official Method 2000.03 Ochratoin A in Barley Immunoaffinity by Column HPLC62.AOAC Official Method 2001.01 Determination of Ochratoxin A in Wine and Beer63.AOAC Official Method 995.10 Patulin in Appple Juice Liquid Chromatoraphic Method64.AOAC Official Method 2000.02Patulin in Clear and Cloudy Apple Juices and Apple Puree65.AOAC Official Method 994.01 Zearlenone in Corn Wheat,and Feed Enzyme LinkedImmunosorbent (Agri-screen)Method66.AOAC Official Method 993.03 Nitrate in Baby Foods spectrophotometric Method67.AOAC Official Method 999.12 Taurine in Pet Food68.AOAC Official Method 996.16 Selenium in Feeds and Premixes69.AOAC Official Method 999.13 Ethoxyquin in Feeds Liquid Chromatographic Method70.AOAC Official Method 999.16 Sulfamethazine in Animal Feeds71.AOAC Official Method 997.04 Monensin in Premix and Animal Feeds LiquidChromatographic Method72.AOAC Official Method 998.02 Neomycin in Feeds Stahl Microbiological Agar73.AOAC Official Method 997.01 Tebuconazole in Fungicide and Technical formulations74.AOAC Official Method 997.14 Thiodicarb in Technical Products and Formulations75.AOAC Official Method 999.04 Determination of Chlorothalonil and Hexachlorobenzene76.AOAC Official Method 997.07 N-octyl Bicycloheptene Dicarboximide (MGK 264),Pyrethrinsand Piperonyl Butoxide (PB)77.AOAC Official Method 996.12 Glyphosate in Water-Soluble Granular Formulations78.AOAC Official Method 997.12 Imidacloprid in Liquid and Solid Formulations Reversed-PhaseLiquid Chromatographic Method79.AOAC Official Method 999.17 Lead and Cadmium Extracted from Ceramic Foodware80.AOAC Official Method 999.10 Lead,Cadmium,Zinc, Copper,and lron in foods81.AOAC Official Method 999.11 Determination of Lead,Cadmium,Copper, Iron,and Zincin Foods82.AOAC Official Method 997.15 Lead in Sugars Graphite Furnace Atomic Absorption Method83.AOAC Official Method 2000.04 Iodine-131 in Milk Radiochemical Separtion Method84.AOAC Official Method 998.11 Screening Test for Nitrate in Forages With a Test Strip85.AOAC Official Method 997.02 Yeast and Mold Cunts in Foods Dry Rehydratable Film Method(Petrifilm TM Method)86.AOAC Official Method 996.09 Escherichia coli O157:H7 in Selected Foods VisualImmunoprecipitate Assay (VIP TM)87.AOAC Official Method 996.10 Enterohemorrhagic Escherichia coli(EHEC)O157:H7Detection in Selected Foods88.AOAC Official Method 997.11Escherichia coli O 157:H7 Counts in Foods HydrophobicGrid Membrane Filter (ISO-GRID)Method Using89.AOAC Official Method 996.08 Salmonella in Foods Enzyme-Linked ImmunofluoressentAssay90.AOAC Official Method 997.16 LOCA TE Enzyme-Linked Immmunosorbent Assay forldentification of Salmonella in Foods91.AOAC Official Method 998.09 Salmonella in Foods Coloric Polyclonal Enzyme92.AOAC Official Method 999.09 Vlp for Salmonella for the Detection of Motile and Non-MotileSalmonella in All Foods93.AOAC Official Method 995.22 Listeria in Foods Colorimetric Polyclonal Enzyme94.AOAC Official Method 996.14 Detection of Listeria Monocytogenes and Related ListeriaSpecies in Selected Foods and from Environmental surfaces95.AOAC Official Method 997.03 Detection of Listeria Monocytogenes and Related ListeriaSpecies in Selected Foods and from Environmental surfaces96.AOAC Official Method 999.06 Listeria in Foods Enzyme-Linked ImmunofluorescentAssay(ELFA)97.AOAC Official Method 995.09 Chortetracycline,Oxytetracycline,and Tetracycline in Edible Animal Tissues98.AOAC Official Method 997.09 Nitrogen in Beer,Wort,and Brewing Grains Protein (Total)by Calculation99.AOAC Official Method 996.11 Starch (Total)in Cereal Products Amylonglucosidass-α-Amylase Method100.AOAC Official Method 995.04 Multiple Tetracline Residues in Milk Metal Chelate Affinity-Liquid Chromatographic Method101.AOAC Official Method 998.04 Neutral Lactase (β-Galactosidase)Activity in Industrial Enzyme Preparations102.AOAC Official Method 999.05 Naringin and Neohesperidin in Orange Juice103.AOAC Official Method 998.03 Aflatoxins in Peanuts Aflatonxins in Peanuts Alternative BF Method104.AOAC Official Method 999.07 Aflatoxin B1 and Total Aflatoxins in Peanut Butter,Pistachio Paste,Fig Paste,and Paprika Powder105.AOAC Official Method 997.05 Taurine in Powdered Milk and Powdered Infant Formulae 106.AOAC Official Method 995.05 Vitamin D in Infant Formulas and Enteral Products Liquid Chromatographic Method107.AOAC Official Method 999.15 Vitamin K in Milk and Infant Formulas Liquid Chromatographic Method108.AOAC Official Method 986.07 Fensulfothion in Chromatographic Method。

欧洲药典7.5版

EUROPEAN PHARMACOPOEIA 7.5
INDEX
To aid users the index includes a reference to the supplement in which the latest version of a text can be found. For example : Amikacin sulfate...............................................7.5-4579 means the monograph Amikacin sulfate can be found on page 4579 of Supplement 7.5. Note that where no reference to a supplement is made, the text can be found in the principal volume.
English index ........................................................................ 4707
Latin index ................................................................................. 4739
EUROPEAN PHARMACOPபைடு நூலகம்EIA 7.5
Index
Numerics 1. General notices ................................................................... 7.5-4453 2.1.1. Droppers...................

方法确认

Method Validation 方法确认
Excellence on a Global Scale
Method Validation - Objectives 方法确认 – 目标
By the end of this session you will be able to通过学习你将能够:

Evaluate a new method with respect to what type of validation data are required获取相关确认数据，对新方法进行评估 Assess validation data to determine specific requirements that must be met for the validation to be verified and accepted 评价确认数据，以确定方法确认应满足的特定要求已经验证并可接受

Expertise and training专业技能和培训
Consideration of safety and hazardous waste disposal issues对实验安全和有害废物处理的考虑 Demonstration of the laboratory’s ability to meet the requirements of the customer实验室对消费者需求的满足能力的展示
5
Excellence on a Global Scale
ISO 17025:
Validation is the confirmation by examination and the provision of objective evidence that the particular requirements for a specific intended use are fulfilled.方法确认是通过检查和提供客观证据确证某一特定用途的特殊要求已得到满足3

GrangerCausalityAnalysis（GCA）-IndexofAnonymous

1. IntroductionGranger causality analysis (GCA) is a method for investigating whether one time series can correctly forecast another (Granger, 1969). This method is based on multiple regression analysis. At individual level, many studies performed F statistics on the residuals (Goebel et al., 2003, Londei et al., 2009). A recent study (Chen et al., 2009) used signed path coefficients to perform t-test at group level statistics. The negative path coefficients were explained as an inhibitory effect (McFarlin et al., 2009). A GCA toolkit which output signed path coefficients has been implemented on AFNI (/sscc/gangc/V AR.html). Here we implemented a MA TLAB GUI toolkit named REST-GCA which was based on REST ().2. Model and MethodsIf we have two time series X and Y , the paired model is as following:∑∑==--+++=p n p n t t p t n p t n t E CZ Y B X A Y 11)()(∑∑==--+++=p n p n t t p t n p t n t E Z C X B Y A X 11''')('')('t X and t Y represent the two time series at time t . )(p t X - and )(p t Y - represent the time series at time t-p, p representing the number of lagged time points (order).n A and 'n A are signed path coefficients. n B and 'n B are autoregression coefficients.t E and 't E are residual. We followed Chen ’s (Chen et al., 2009) extended V ector Autoregression model which took the covariables t Z at time t into account, instead of regressing them out before GCA.Considering the complicated significance of high order, the current version of REST-GCA supports only the lagged time points=1 (order=1) condition.age3.1.ROI-wise GCA:The ROI-wise GCA in the current version of REST-GCA supports only two ROIs. Users need to set two ROIs. The procedure of adding subjects and ROIs definition is the same as Functional Connectivity analysis in REST. REST-GCA will export two text files. The text file with the suffix name ‘x2y’ saves the signed path coefficient of X to Y and the autoregressive coefficient of Y The text file with the suffix name ‘y2x’saves the signed path coefficient of Y to X and the autoregressive coefficient of X.3.2.Voxel-wise GCA:V oxel-wise GCA calculates causal effects of an ROI time series on that of every voxel in the brain (x to y) and the time series of every voxel in the brain on that of the ROI (y to x). Users need to setA an ROI. Covariables could be added. The output results are two signed path coefficient maps (nA). The voxel-wise GCA in the current version of REST-GCA does not export and 'nautoregressive coefficients.4.Examples of results:4.1.ROI-wise:This ROI-wise GCA is at individual level. Two spherical ROIs were placed in the right frontal-insular cortex (x=37, y=25, z=-4, radius=5mm) and the dorsal anterior cingulate cortex (x=1, y=33, z=25, radius=5mm). The results are two text files.GCA_SubjectName_x2y.txt:2.7507440376311693e-001 7.4028188213001311e-001GCA_SubjectName_y2x.txt:-9.7560878619967109e-002 9.6053881226426607e-001The first column is the signed path coefficient. The second column is the autoregressive coefficient.4.2.Voxel-wise:We processed a dataset of resting-state fMRI from 40 healthy subjects (3T Siemense, TR=2s, Order=1, 230 volumes). A region of interest (ROI) was placed in the right frontal-insular cortex (centered at x=37, y=25, z=-4 with radius=5mm, Sridharan et al., 2008). Then a voxel-wise GCA without covariates was performed. Two signed path coefficient maps were produced. One-sample t-tests (SPM8) were performed. The right FIC showed significantly positive causal effect on the dorsal anterior cingulate cortex (dACC) (Fig. 1a). This result is consistent with that in a previous study (Sridharan et al., 2008). Interestingly, the dACC showed significantly negative effect on the right FIC (Fig. 1b).Figure1:Results of the one-sample t test (FEW corrected, p<0.05). The right FIC showed significantly positive causal effect on dACC (Figure-1a). The dACC showed significant negative causal effect on the right IFC (Figure-1b).Reference:(1)C.W.J. Granger, (1969) Investigating Causal Relations By Econometric Models AndCross-spectral Methods. Econometrica ,37 (3), 424–438(2) R.Gobel et al., (2003) Investigating directed cortical interactions in time-resolved fMRI datausing vector autoregressive modeling and Granger causality mapping. MagneticResonance Imaging, 21, 1251–1261(3)G. Chen et al., (2009) Granger Causality via V ector Auto-Regression Tuned for FMRI DataAnalysis. Proc Intl Soc Mag Reson Med, 17(4) McFarlin, D.R et al., (2009) The Effect of Controllability on Context Dependent GrangerCausality in Snake Phobia fMRI Data at 400-ms Resolution. Society for Neuroscience,Chicago, 289.15(5) Sridharan, D et al., (2008), A critical role for the right fronto-insular cortex in switchingbetween central-executive and default-mode networks, Proc Natl Acad Sci U S A,105(34):12569-74.Acknowledgment:Many thanks to Dong Zhangye, Y an Chaogan and Huang Jian for their help!。

aoac标准

aoac标准AOAC国际是一个非营利性科学组织，致力于发展分析测试方法和标准。

AOAC标准是一种被广泛认可的标准，用于验证食品、饲料、环境、药品等样品的分析测试方法的准确性和可靠性。

AOAC标准的制定和认可需要经过严格的科学评审和实验验证，以确保测试方法的准确性和可靠性，从而保障公众健康和安全。

AOAC标准的制定和认可过程需要遵循一定的程序和规定。

首先，提交测试方法的组织或个人需要向AOAC国际提交申请，申请材料需要包括测试方法的详细描述、实验数据、验证报告等。

接下来，AOAC国际会组织专家对提交的测试方法进行评审，评审的内容包括方法的准确性、可靠性、重复性等。

经过评审通过后，测试方法需要进行多中心实验验证，以确保在不同实验室和条件下的适用性和可靠性。

最终，经过实验验证的测试方法会被提交给AOAC国际的官方委员会进行最终的认可和发布。

AOAC标准的制定和认可需要严格遵循科学原则和程序，以确保测试方法的准确性和可靠性。

AOAC国际的专家团队对提交的测试方法进行评审，确保方法符合科学规范和要求。

多中心实验验证可以检验测试方法在不同实验室和条件下的适用性和可靠性，从而保证测试方法的普适性和可靠性。

AOAC标准的认可和发布，为食品安全、环境保护、药品监管等领域提供了可靠的分析测试方法，对保障公众健康和安全起到了重要作用。

总的来说，AOAC标准是一种被广泛认可的标准，用于验证食品、饲料、环境、药品等样品的分析测试方法的准确性和可靠性。

AOAC标准的制定和认可需要经过严格的科学评审和实验验证，以确保测试方法的准确性和可靠性，从而保障公众健康和安全。

AOAC标准的认可和发布，为食品安全、环境保护、药品监管等领域提供了可靠的分析测试方法，对保障公众健康和安全起到了重要作用。

AOAC标准的制定和认可过程需要遵循一定的程序和规定，确保测试方法的准确性和可靠性。

AOAC国际的专家团队对提交的测试方法进行评审，确保方法符合科学规范和要求。