BAY_41-2272_LCMS_22843_MedChemExpress
BAY_41-2272_SDS_MedChemExpress
Inhibitors, Agonists, Screening LibrariesSafety Data Sheet Revision Date:May-24-2017Print Date:May-24-20171. PRODUCT AND COMPANY IDENTIFICATION1.1 Product identifierProduct name :BAY 41-2272Catalog No. :HY-12376CAS No. :256376-24-61.2 Relevant identified uses of the substance or mixture and uses advised againstIdentified uses :Laboratory chemicals, manufacture of substances.1.3 Details of the supplier of the safety data sheetCompany:MedChemExpress USATel:609-228-6898Fax:609-228-5909E-mail:sales@1.4 Emergency telephone numberEmergency Phone #:609-228-68982. HAZARDS IDENTIFICATION2.1 Classification of the substance or mixtureNot a hazardous substance or mixture.2.2 GHS Label elements, including precautionary statementsNot a hazardous substance or mixture.2.3 Other hazardsNone.3. COMPOSITION/INFORMATION ON INGREDIENTS3.1 SubstancesSynonyms:BAY41⁻2272; BAY⁻41⁻2272Formula:C20H17FN6Molecular Weight:360.39CAS No. :256376-24-64. FIRST AID MEASURES4.1 Description of first aid measuresEye contactRemove any contact lenses, locate eye-wash station, and flush eyes immediately with large amounts of water. Separate eyelids with fingers to ensure adequate flushing. Promptly call a physician.Skin contactRinse skin thoroughly with large amounts of water. Remove contaminated clothing and shoes and call a physician.InhalationImmediately relocate self or casualty to fresh air. If breathing is difficult, give cardiopulmonary resuscitation (CPR). Avoid mouth-to-mouth resuscitation.IngestionWash out mouth with water; Do NOT induce vomiting; call a physician.4.2 Most important symptoms and effects, both acute and delayedThe most important known symptoms and effects are described in the labelling (see section 2.2).4.3 Indication of any immediate medical attention and special treatment neededTreat symptomatically.5. FIRE FIGHTING MEASURES5.1 Extinguishing mediaSuitable extinguishing mediaUse water spray, dry chemical, foam, and carbon dioxide fire extinguisher.5.2 Special hazards arising from the substance or mixtureDuring combustion, may emit irritant fumes.5.3 Advice for firefightersWear self-contained breathing apparatus and protective clothing.6. ACCIDENTAL RELEASE MEASURES6.1 Personal precautions, protective equipment and emergency proceduresUse full personal protective equipment. Avoid breathing vapors, mist, dust or gas. Ensure adequate ventilation. Evacuate personnel to safe areas.Refer to protective measures listed in sections 8.6.2 Environmental precautionsTry to prevent further leakage or spillage. Keep the product away from drains or water courses.6.3 Methods and materials for containment and cleaning upAbsorb solutions with finely-powdered liquid-binding material (diatomite, universal binders); Decontaminate surfaces and equipment by scrubbing with alcohol; Dispose of contaminated material according to Section 13.7. HANDLING AND STORAGE7.1 Precautions for safe handlingAvoid inhalation, contact with eyes and skin. Avoid dust and aerosol formation. Use only in areas with appropriate exhaust ventilation.7.2 Conditions for safe storage, including any incompatibilitiesKeep container tightly sealed in cool, well-ventilated area. Keep away from direct sunlight and sources of ignition.Recommended storage temperature:Powder-20°C 3 years4°C 2 yearsIn solvent-80°C 6 months-20°C 1 monthShipping at room temperature if less than 2 weeks.7.3 Specific end use(s)No data available.8. EXPOSURE CONTROLS/PERSONAL PROTECTION8.1 Control parametersComponents with workplace control parametersThis product contains no substances with occupational exposure limit values.8.2 Exposure controlsEngineering controlsEnsure adequate ventilation. Provide accessible safety shower and eye wash station.Personal protective equipmentEye protection Safety goggles with side-shields.Hand protection Protective gloves.Skin and body protection Impervious clothing.Respiratory protection Suitable respirator.Environmental exposure controls Keep the product away from drains, water courses or the soil. Cleanspillages in a safe way as soon as possible.9. PHYSICAL AND CHEMICAL PROPERTIES9.1 Information on basic physical and chemical propertiesAppearance White to off-white (Solid)Odor No data availableOdor threshold No data availablepH No data availableMelting/freezing point No data availableBoiling point/range No data availableFlash point No data availableEvaporation rate No data availableFlammability (solid, gas)No data availableUpper/lower flammability or explosive limits No data availableVapor pressure No data availableVapor density No data availableRelative density No data availableWater Solubility No data availablePartition coefficient No data availableAuto-ignition temperature No data availableDecomposition temperature No data availableViscosity No data availableExplosive properties No data availableOxidizing properties No data available9.2 Other safety informationNo data available.10. STABILITY AND REACTIVITY10.1 ReactivityNo data available.10.2 Chemical stabilityStable under recommended storage conditions.10.3 Possibility of hazardous reactionsNo data available.10.4 Conditions to avoidNo data available.10.5 Incompatible materialsStrong acids/alkalis, strong oxidising/reducing agents.10.6 Hazardous decomposition productsUnder fire conditions, may decompose and emit toxic fumes.Other decomposition products - no data available.11.TOXICOLOGICAL INFORMATION11.1 Information on toxicological effectsAcute toxicityClassified based on available data. For more details, see section 2Skin corrosion/irritationClassified based on available data. For more details, see section 2Serious eye damage/irritationClassified based on available data. For more details, see section 2Respiratory or skin sensitizationClassified based on available data. For more details, see section 2Germ cell mutagenicityClassified based on available data. For more details, see section 2CarcinogenicityIARC: No component of this product present at a level equal to or greater than 0.1% is identified as probable, possible or confirmed human carcinogen by IARC.ACGIH: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by ACGIH.NTP: No component of this product present at a level equal to or greater than 0.1% is identified as a anticipated or confirmed carcinogen by NTP.OSHA: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by OSHA.Reproductive toxicityClassified based on available data. For more details, see section 2Specific target organ toxicity - single exposureClassified based on available data. For more details, see section 2Specific target organ toxicity - repeated exposureClassified based on available data. For more details, see section 2Aspiration hazardClassified based on available data. For more details, see section 212. ECOLOGICAL INFORMATION12.1 ToxicityNo data available.12.2 Persistence and degradabilityNo data available.12.3 Bioaccumlative potentialNo data available.12.4 Mobility in soilNo data available.12.5 Results of PBT and vPvB assessmentPBT/vPvB assessment unavailable as chemical safety assessment not required or not conducted.12.6 Other adverse effectsNo data available.13. DISPOSAL CONSIDERATIONS13.1 Waste treatment methodsProductDispose substance in accordance with prevailing country, federal, state and local regulations.Contaminated packagingConduct recycling or disposal in accordance with prevailing country, federal, state and local regulations.14. TRANSPORT INFORMATIONDOT (US)This substance is considered to be non-hazardous for transport.IMDGThis substance is considered to be non-hazardous for transport.IATAThis substance is considered to be non-hazardous for transport.15. REGULATORY INFORMATIONSARA 302 Components:No chemicals in this material are subject to the reporting requirements of SARA Title III, Section 302.SARA 313 Components:This material does not contain any chemical components with known CAS numbers that exceed the threshold (De Minimis) reporting levels established by SARA Title III, Section 313.SARA 311/312 Hazards:No SARA Hazards.Massachusetts Right To Know Components:No components are subject to the Massachusetts Right to Know Act.Pennsylvania Right To Know Components:No components are subject to the Pennsylvania Right to Know Act.New Jersey Right To Know Components:No components are subject to the New Jersey Right to Know Act.California Prop. 65 Components:This product does not contain any chemicals known to State of California to cause cancer, birth defects, or anyother reproductive harm.16. OTHER INFORMATIONCopyright 2017 MedChemExpress. The above information is correct to the best of our present knowledge but does not purport to be all inclusive and should be used only as a guide. The product is for research use only and for experienced personnel. It must only be handled by suitably qualified experienced scientists in appropriately equipped and authorized facilities. The burden of safe use of this material rests entirely with the user. MedChemExpress disclaims all liability for any damage resulting from handling or from contact with this product.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。
3_种常用碳青霉烯类抗生素血药浓度UPLC-MS
3种常用碳青霉烯类抗生素血药浓度UPLC-MS/MS检测方法的建立Δ秦怡1＊，张瑞霞2，吕雅瑶2，翁莉莉1，张弋2 #（1.天津医科大学一中心临床学院，天津 300192；2.天津市第一中心医院药学部，天津 300192）中图分类号 R917；R978.1文献标志码 A 文章编号 1001-0408（2024）03-0343-05DOI 10.6039/j.issn.1001-0408.2024.03.14摘要目的建立3种临床常用碳青霉烯类抗生素——厄他培南（ETP）、亚胺培南（IPM）、美罗培南（MEM）血药浓度检测的超高效液相色谱-质谱联用（UPLC-MS/MS）法。
方法血浆样品经甲醇沉淀蛋白后，以3种抗生素的稳定性同位素（ETP-D4、IPM-D4、MEM-D6）为内标，采用ACQUITY UPLC BEH C18（2.1 mm×50 mm，1.7μm）色谱柱分离；流动相为98%乙腈+2%水+0.1%甲酸和98%水+2%乙腈+0.1%甲酸，梯度洗脱；流速为0.3 mL/min；柱温为40 ℃；采用正离子、多反应监测模式进行扫描分析。
结果该方法专属性良好，在ETP、IPM、MEM 0.2～200、0.1～100、0.1～100μg/mL范围内线性良好（r2≥0.993），批内、批间精密度和准确度良好（RE均≤5.14%，RSD均≤11.15%），基质效应、提取回收率较一致（RSD≤12.99%）。
结论本实验建立了一种可以同时定量ETP、IPM、MEM血药浓度的UPLC-MS/MS法，该方法样品前处理简单、检测时间短、所需样品量少，可满足临床需求。
关键词碳青霉烯类抗生素；超高效液相色谱-质谱联用；血药浓度；厄他培南；亚胺培南；美罗培南Establishment of UPLC-MS/MS method for the determination of plasma concentration of three common carbapenem antibioticsQIN Yi1，ZHANG Ruixia2，LYU Yayao2，WENG Lili1，ZHANG Yi2（1. First Central Clinical College of Tianjin Medical University，Tianjin 300192，China；2. Dept. of Pharmacy，Tianjin First Central Clinical Hospital，Tianjin 300192， China）ABSTRACT OBJECTIVE To establish a UPLC-MS/MS method for the determination of plasma concentration of three carbapenem antibiotics，i.e. ertapenem （ETP），imipenem （IPM）and meropenem （MEM）.METHODS After protein precipitation with methanol，the plasma samples were separated by ACQUITY UPLC BEH C18column （2.1mm×50mm，1.7μm）using stable isotopes of three antibiotics （ETP-D4，IPM-D4，MEM-D6）as the internal standard. The mobile phases were 98%acetonitrile +2% water +0.1%formic acid and 98%water +2%acetonitrile +0.1%formic acid，by gradient elution. The flow rate was 0.3mL/min and the column temperature was 40 ℃. Scanning analysis was performed in the positive ion and multiple reaction monitoring mode. RESULTS The method had good specificity，good linearity （r2≥0.993）in the range of 0.2-200，0.1-100and 0.1-100μg/mL of ETP，IPM and MEM，and good intra-batch and inter-batch precision and accuracy （all RE≤5.14%，all RSD≤11.15%），the matrix effect and extraction recovery were consistent （RSD≤12.99%）. CONCLUSIONS This study establishes the UPLC-MS/MS method to simultaneously quantify the plasma concentration of ETP，IPM and MEM. The method has the advantages of simple pretreatment， short detection time and small sample quantity to meet clinical requirement.KEYWORDS carbapenem antibiotics； UPLC-MS/MS； plasma concentration； ertapenem； imipenem； meropenem碳青霉烯类抗生素具有抗菌谱广、抗菌活性强、耐药率低的特点，已成为治疗重症感染的主要选择。
marked manuscript
Quality evaluation of Flos Lonicerae through a simultaneous determination of seven saponins by HPLC with ELSDXing-Yun Chai1, Song-Lin Li2, Ping Li1*1Key Laboratory of Modern Chinese Medicines and Department of Pharmacognosy, China Pharmaceutical University, Nanjing, 210009, People’s Republic of China2Institute of Nanjing Military Command for Drug Control, Nanjing, 210002, People’s Republic of China*Corresponding author: Ping LiKey Laboratory of Modern Chinese Medicines and Department of Pharmacognosy, China Pharmaceutical University, Nanjing 210009, People’s Republic of China.E-mail address: lipingli@Tel.: +86-25-8324-2299; 8539-1244; 135********Fax: +86-25-8532-2747AbstractA new HPLC coupled with evaporative light scattering detection (ELSD) method has been developed for the simultaneous quantitative determination of seven major saponins, namely macranthoidinB (1), macranthoidin A (2), dipsacoside B (3), hederagenin-28-O-β-D-glucopyranosyl(6→1)-O-β-D- glucopyranosyl ester (4), macranthoside B (5), macranthoside A (6), and hederagenin-3-O-α-L-arabinopyranosyl(2→1)-O-α-L-rhamnopyranoside (7)in Flos Lonicerae, a commonly used traditional Chinese medicine (TCM) herb.Simultaneous separation of these seven saponins was achieved on a C18 analytical column with a mixed mobile phase consisting of acetonitrile(A)-water(B)(29:71 v/v) acidified with 0.5% acetic acid. The elution was operated from keeping 29%A for 10min, then gradually to 54%B from 10 to 25 min on linear gradient, and then keep isocratic elution with 54%B from 25 to 30min.The drift tube temperature of ELSD was set at 106℃, and with the nitrogen flow-rate of 2.6 l/min. All calibration curves showed good linear regression (r2 0.9922) within test ranges. This method showed good reproducibility for the quantification of these seven saponins in Flos Lonicerae with intra- and inter-day variations of less than 3.0% and 6.0% respectively. The validated method was successfully applied to quantify seven saponins in five sources of Flos Lonicerae, which provides a new basis of overall assessment on quality of Flos Lonicerae.Keywords: HPLC-ELSD; Flos Lonicerae; Saponins; Quantification1. IntroductionFlos Lonicerae (Jinyinhua in Chinese), the dried buds of several species of the genus Lonicera (Caprifoliaceae), is a commonly used traditional Chinese medicine (TCM) herb. It has been used for centuries in TCM practice for the treatment of sores, carbuncles, furuncles, swelling and affections caused by exopathogenic wind-heat or epidemic febrile diseases at the early stage [1]. Though four species of Lonicera are documented as the sources of Flos Lonicerae in China Pharmacopeia (2000 edition), i.e. L. japonica, L. hypoglauca,L. daystyla and L. confusa, other species such as L. similes and L. macranthoides have also been used on the same purpose in some local areas in China [2]. So it is an important issue to comprehensively evaluate the different sources of Flos Lonicerae, so as to ensure the clinical efficacy of this Chinese herbal drug.Chemical and pharmacological investigations on Flos Lonicerae resulted in discovering several kinds of bioactive components, i.e. chlorogenic acid and its analogues, flavonoids, iridoid glucosides and triterpenoid saponins [3]. Previously, chlorogenic acid has been used as the chemical marker for the quality evaluation of Flos Lonicerae,owing to its antipyretic and antibiotic property as well as its high content in the herb. But this compound is not a characteristic component of Flos Lonicerae, as it has also been used as the chemical marker for other Chinese herbal drugs such as Flos Chrysanthemi and so on[4-5]. Moreover, chlorogenic acid alone could not be responsible for the overall pharmacological activities of Flos Lonicerae[6].On the other hand, many studies revealed that triterpenoidal saponins of Flos Lonicerae possess protection effects on hepatic injury caused by Acetaminophen, Cd, and CCl4, and conspicuous depressant effects on swelling of ear croton oil [7-11]. Therefore, saponins should also be considered as one of the markers for quality control of Flos Lonicerae. Consequently, determinations of all types of components such as chlorogenic acid, flavonoids, iridoid glucosides and triterpenoidal saponins in Flos Lonicerae could be a better strategy for the comprehensive quality evaluation of Flos Lonicerae.Recently an HPLC-ELSD method has been established in our laboratory for qualitative and quantitative determination of iridoid glucosides in Flos Lonicerae [12]. But no method was reported for the determination of triterpenoidal saponins in Flos Lonicera. As a series studies on the comprehensive evaluation of Flos Lonicera, we report here, for the first time, the development of an HPLC-ELSD method for simultaneous determination of seven triterpenoidal saponins in the Chinese herbal drug Flos Lonicerae, i.e.macranthoidin B (1), macranthoidin A (2), dipsacoside B (3), hederagenin-28-O-β-D-glucopyranosyl(6→1)-O-β-D- glucopyranosyl ester (4), macranthoside B (5), macranthoside A (6), and hederagenin-3-O-α-L-arabinopyranosyl(2→1)-O-α-L-rhamnopyranoside (7) (Fig. 1).2. Experimental2.1. Samples, chemicals and reagentsFive samples of Lonicera species,L. japonica from Mi county, HeNan province (LJ1999-07), L. hypoglauca from Jiujang county, JiangXi province (LH2001-06), L. similes from Fei county, ShanDong province (LS2001-07), L. confuse from Xupu county, HuNan province (LC2001-07), and L. macranthoides from Longhu county, HuNan province (LM2000-06) respectively, were collected in China. All samples were authenticated by Dr. Ping Li, professor of department of Pharmacognosy, China Pharmaceutical University, Nanjing, China. The voucher specimens were deposited in the department of Pharmacognosy, China Pharmaceutical University, Nanjing, China. Seven saponin reference compounds: macranthoidin B (1), macranthoidin A (2), dipsacoside B (3), hederagenin-28-O-β-D-glucopyranosyl(6→1)-O-β-D- glucopyranosyl ester (4), macranthoside B (5), macranthoside A (6), and hederagenin-3-O-α-L-arabinopyranosyl(2→1)-O-α-L-rhamnopyranoside (7) were isolated previously from the dried buds of L. confusa by repeated silica gel, sephadex LH-20 and Rp-18 silica gel column chromatography, their structures were elucidated by comparison of their spectral data (UV, IR, MS, 1H- NMR and 13C-NMR) with references [13-15]. The purity of these saponins were determined to be more than 98% by normalization of the peak areas detected by HPLC with ELSD, and showed very stable in methanol solution.HPLC-grade acetonitrile from Merck (Darmstadt, Germany), the deionized water from Robust (Guangzhou, China), were purchased. The other solvents, purchased from Nanjing Chemical Factory (Nanjing, China) were of analytical grade.2.2. Apparatus and chromatographic conditionsAglient1100 series HPLC apparatus was used. Chromatography was carried out on an Aglient Zorbax SB-C18 column（250 4.6mm, 5.0µm）at a column temperature of 25℃.A Rheodyne 7125i sampling valve (Cotati, USA) equipped with a sample loop of 20µl was used for sample injection. The analog signal from Alltech ELSD 2000 (Alltech, Deerfield, IL, USA)was transmitted to a HP Chemstation for processing through an Agilent 35900E (Agilent Technologies, USA).The optimum resolution was obtained by using a linear gradient elution. The mobile phase was composed of acetonitrile(A) and water(B) which acidified with 0.5% acetic acid. The elution was operated from keeping 29%A for 10min, then gradually to 54%B from 10 to 25 min in linear gradient, and back to the isocratic elution of 54%B from 25 to 30 min.The drift tube temperature for ELSD was set at 106℃and the nitrogen flow-rate was of 2.6 l/min. The chromatographic peaks were identified by comparing their retention time with that of each reference compound tried under the same chromatographic conditions with a series of mobile phases. In addition, spiking samples with the reference compounds further confirmed the identities of the peaks.2.3. Calibration curvesMethanol stock solutions containing seven analytes were prepared and diluted to appropriate concentration for the construction of calibration curves. Six concentrationof the seven analytes’ solution were injected in triplicate, and then the calibration curves were constructed by plotting the peak areas versus the concentration of each analyte. The results were demonstrated in Table1.2.4. Limits of detection and quantificationMethanol stock solution containing seven reference compounds were diluted to a series of appropriate concentrations with methanol, and an aliquot of the diluted solutions were injected into HPLC for analysis.The limits of detection (LOD) and quantification (LOQ) under the present chromatographic conditions were determined at a signal-to-noise ratio (S/N) of 3 and 10, respectively. LOD and LOQ for each compound were shown in Table1.2.5. Precision and accuracyIntra- and inter-day variations were chosen to determine the precision of the developed assay. Approximate 2.0g of the pulverized samples of L. macranthoides were weighted, extracted and analyzed as described in 2.6 Sample preparation section. For intra-day variability test, the samples were analyzed in triplicate for three times within one day, while for inter-day variability test, the samples were examined in triplicate for consecutive three days. Variations were expressed by the relative standard deviations. The results were given in Table 2.Recovery test was used to evaluate the accuracy of this method. Accurate amounts of seven saponins were added to approximate 1.0g of L. macranthoides,and then extracted and analyzed as described in 2.6 Sample preparation section. The average recoveries were counted by the formula: recovery (%) = (amount found –original amount)/ amount spiked ×100%, and RSD (%) = (SD/mean) ×100%. The results were given in Table 3.2.6. Sample preparationSamples of Flos Lonicerae were dried at 50℃until constant weight. Approximate 2.0g of the pulverized samples, accurately weighed, was extracted with 60% ethanol in a flask for 4h. The ethanol was evaporated to dryness with a rotary evaporator. Residue was dissolved in water, followed by defatting with 60ml of petroleum ether for 2 times, and then the water solution was evaporated, residue was dissolved with methanol into a 25ml flask. One ml of the methanol solution was drawn and transferred to a 5ml flask, diluted to the mark with methanol. The resultant solution was at last filtrated through a 0.45µm syringe filter (Type Millex-HA, Millipore, USA) and 20µl of the filtrate was injected to HPLC system. The contents of the analytes were determined from the corresponding calibration curves.3. Results and discussionsThe temperature of drift tube and the gas flow-rate are two most important adjustable parameters for ELSD, they play a prominent role to an analyte response. In ourprevious work [12], the temperature of drift tube was optimized at 90°C for the determination of iridoids. As the polarity of saponins are higher than that of iridoids, more water was used in the mobile phase for the separation of saponins, therefore the temperature for saponins determination was optimized systematically from 95°C to 110°C, the flow-rate from 2.2 to 3.0 l/min. Dipsacoside B was selected as the testing saponin for optimizing ELSD conditions, as it was contained in all samples. Eventually, the drift tube temperature of 106℃and a gas flow of 2.6 l/min were optimized to detect the analytes. And these two exact experimental parameters should be strictly controlled in the analytical procedure [16].All calibration curves showed good linear regression (r2 0.9922) within test ranges. Validation studies of this method proved that this assay has good reproducibility. As shown in Table 2, the overall intra- and inter-day variations are less than 6% for all seven analytes. As demonstrated in Table 3, the developed analytical method has good accuracy with the overall recovery of high than 96% for the analytes concerned. The limit of detection (S/N=3) and the limit of quantification (S/N=10) are less than 0.26μg and 0.88μg respectively (Table1), indicating that this HPLC-ELSD method is precise, accurate and se nsitive enough for the quantitative evaluation of major non- chromaphoric saponins in Flos Lonicerae.It has been reported that there are two major types of saponins in Flos Lonicerae, i.e. saponins with hederagenin as aglycone and saponins with oleanolic acid as the aglycone [17]. But hederagenin type saponins of the herb were reported to have distinct activities of liver protection and anti-inflammatory [7-11]. So we adoptedseven hederagenin type saponins as representative markers to establish a quality control method.The newly established HPLC-ELSD method was applied to analyze seven analytes in five plant sources of Flos Lonicerae, i.e. L. japonica,L. hypoglauca,L. confusa,L. similes and L. macranthoides(Table 4). It was found that there were remarkable differences of seven saponins contents between different plant sources of Flos Lonicerae. All seven saponins analyzed could be detected in L. confusa and L. hypoglauca, while only dipsacoside B was detected in L. japonica. Among all seven saponins interested, only dipsacoside B was found in all five plant species of Flos Lonicerae analyzed, and this compound was determined as the major saponin with content of 53.7 mg/g in L. hypoglauca. On the other hand, macranthoidin B was found to be the major saponin with the content higher than 41.0mg/g in L. macranthoides,L. confusa, and L. similis, while the contents of other analytes were much lower.In our previous study [12], overall HPLC profiles of iridoid glucosides was used to qualitatively and quantitatively distinguish different origins of Flos Lonicerae. As shown in Fig.2, the chromatogram profiles of L. confusa, L. japonica and L. similes seem to be similar, resulting in the difficulty of clarifying the origins of Flos Lonicerae solely by HPLC profiles of saponins, in addition to the clear difference of the HPLC profiles of saponins from L. macranthoides and L. hypoglauca.Therefore, in addition to the conventional morphological and histological identification methods, the contents and the HPLC profiles of saponins and iridoids could also be used as accessory chemical evidence toclarify the botanical origin and comprehensive quality evaluation of Flos Lonicerae.4. ConclusionsThis is the first report on validation of an analytical method for qualification and quantification of saponins in Flos Lonicerae. This newly established HPLC-ELSD method can be used to simultaneously quantify seven saponins, i.e. macranthoidin B, macranthoidin A, dipsacoside B, hederagenin-28-O-β-D-glucopyranosyl(6→1)-O-β-D- glucopyranosyl ester, macranthoside B, macranthoside A, and hederagenin-3-O-α-L-arabinopyranosyl(2→1)-O-α-L-rhamnopyranoside in Flos Lonicerae. Together with the HPLC profiles of iridoids, the HPLC-ELSD profiles of saponins could also be used as an accessory chemical evidence to clarify the botanical origin and comprehensive quality evaluation of Flos Lonicerae.AcknowledgementsThis project is financially supported by Fund for Distinguished Chinese Young Scholars of the National Science Foundation of China (30325046) and the National High Tech Program（2003AA2Z2010）.[1]Ministry of Public Health of the People’s Republic of China, Pharmacopoeia ofthe People’s Republic of China, V ol.1, 2000, p. 177.[2]W. Shi, R.B. Shi, Y.R. Lu, Chin. Pharm. J., 34(1999) 724.[3]J.B. Xing, P. Li, D.L. Wen, Chin. Med. Mater., 26(2001) 457.[4]Y.Q. Zhang, L.C. Xu, L.P. Wang, J. Chin. Med. Mater., 21(1996) 204.[5] D. Zhang, Z.W. Li, Y. Jiang, J. Pharm. Anal., 16(1996) 83.[6]T.Z. Wang, Y.M. Li, Huaxiyaoxue Zazhi, 15(2000) 292.[7]J.ZH. Shi, G.T. Liu. Acta Pharm. Sin., 30(1995) 311.[8]Y. P. Liu, J. Liu, X.SH. Jia, et al. Acta Pharmacol. 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Mater., 22(1999) 366.Fig. 1 Chemical structures of seven saponins from Lonicera confusa macranthoidin B (1), macranthoidin A (2), dipsacoside B (3), hederagenin-28-O-β-D-glucopyranosyl(6→1)-O-β-D- glucopyranosyl ester (4), macranthoside B (5), macranthoside A (6), and hederagenin-3-O-α-L-arabinopyranosyl(2→1)-O-α-L-rhamnopyranoside (7)Fig. 2Representative HPLC chromatograms of mixed standards and methanol extracts of Flos Lonicerae.Column: Agilent Zorbax SB-C18 column（250 4.6mm, 5.0µm）, temperature of 25℃; Detector: ELSD, drift tube temperature 106℃, nitrogen flow-rate 2.6 l/min.A: Mixed standards, B: L. confusa, C: L. japonica, D: L. macranthoides, E: L. hypoglauca, F: L. similes.Table 1 Calibration curves for seven saponinsAnalytes Calibration curve ar2Test range(μg)LOD(μg)LOQ(μg)1 y=6711.9x－377.6 0.9940 0.56–22.01 0.26 0.882 y=7812.6x－411.9 0.9922 0.54–21.63 0.26 0.843 y=6798.5x－299.0 0.9958 0.46–18.42 0.22 0.724 y=12805x－487.9 0.9961 0.38–15.66 0.10 0.345 y=4143.8x－88.62 0.9989 0.42–16.82 0.18 0.246 y=3946.8x－94.4 0.9977 0.40–16.02 0.16 0.207 y=4287.8x－95.2 0.9982 0.42–16.46 0.12 0.22a y: Peak area; x: concentration (mg/ml)Table 2 Reproducibility of the assayAnalyteIntra-day variability Inter-day variability Content (mg/g) Mean RSD (%) Content (mg/g) Mean RSD (%)1 46.1646.2846.2246.22 0.1346.2245.3647.4226.33 2.232 5.385.385.165.31 2.405.285.345.045.22 3.043 4.374.304.184.28 2.244.284.464.024.255.204 nd1)-- -- nd -- --5 1.761.801.821.79 1.701.801.681.841.77 4.706 1.281.241.221.252.451.241.341.201.26 5.727 tr2)-- -- tr -- -- 1): not detected; 2): trace. RSD (%) = (SD/Mean) ×100%Table 3 Recovery of the seven analytesAnalyteOriginal(mg) Spiked(mg)Found(mg)Recovery(%)Mean(%)RSD(%)1 23.0823.1423.1119.7122.8628.1042.7346.1351.0199.7100.699.399.8 0.722.692.672.582.082.913.164.735.515.7698.197.6100.698.8 1.632.172.152.091.732.182.623.884.404.6598.8103.297.799.9 2.94nd1)1.011.050.980.981.101.0297.0104.8104.1102.0 4.250.880.900.910.700.871.081.561.752.0197.197.7101.898.9 2.660.640.620.610.450.610.751.081.211.3397.796.796.096.8 0.97tr2)1.021.101.081.031.111.07100.9102.799.1100.9 1.81): not detected; 2): trace.a Recovery (%) = (Amount found –Original amount)/ Amount spiked ×100%, RSD (%) = (SD/Mean) ×100%Table 4 Contents of seven saponins in Lonicera spp.Content (mg/g)1 2 3 4 5 6 7 L. confusa45.65±0.32 5.13±0.08 4.45±0.11tr1) 2.04±0.04tr 1.81±0.03 L. japonica nd2)nd 3.44±0.09nd nd nd nd L. macranthoides46.22±0.06 5.31±0.13 4.28±0.10 tr 1.79±0.03 1.25±0.03 tr L. hypoglauca11.17±0.07 nq3)53.78±1.18nd 1.72±0.02 2.23±0.06 2.52±0.04 L. similes41.22±0.25 4.57±0.07 3.79±0.09nd 1.75±0.02tr nd 1): trace; 2): not detected.. 3) not quantified owing to the suspicious purity of the peak.。
SCI_Chem 影响因子
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3.20.01489 5.62318>10.00.013647.55389.40.008828.821867 5.10.27819 4.24368.40.00872 6.542227 5.50.53637 3.497299.90.01888.1211078 4.40.37491 5.1891191 2.40.20333 4.012473 1.80.05534 3.461368.20.03818 3.07130997.70.83183 2.99465 5.90.01757 3.141169 1.40.00944 3.344569 4.20.12336 3.006377.20.00647 2.73229 1.90.00155 3.27323 3.90.00716 4.441557.70.01045 2.068458 1.40.02121 2.833576 6.70.15215 2.032167.90.00342 3.148457 3.60.07856 2.503112 4.60.01202 1.97501 6.90.05008 1.9062220.00197 2.73916>10.00.003282.792Eigenfactor®Metrics286 2.60.0208 2.0046>10.00.00142 1.13896 2.80.01517 2.301119.20.00397 3.974>10.00.00086 1.894 44940.03848 1.5233>10.00.00076 2.537437.60.01013 2.301 63220.04884 2.393 2150.003483173 4.80.28954 1.5629>10.00.00211 3.009 138 5.80.01695 1.794 1015 1.70.02966 1.597 508.20.01226 2.37 160850.18182 1.40244 5.40.01448 3.213188.30.00431 2.295 107.10.00156 2.34 674 6.40.06437 1.6 1916 4.10.1766 1.469 480 5.40.05059 1.419 284>10.00.04014 1.60356 1.20.00194 1.703 617 3.70.05475 1.603 14797.80.17236 1.533 8907.80.09182 1.35 404 4.40.04405 1.352 239.20.00403 2.705 448 4.10.05929 1.252 507 3.50.04948 1.865 480 5.30.05919 1.416 315 1.30.00585 1.6362>10.00.00057 1.543 255 2.10.01222 1.495 116 6.20.01779 2.376 953 2.30.03563 1.2844>10.00.00016 2.2160 5.10.006940.708137.90.00319 2.303 3283 3.40.34957 1.353 2798.70.02859 1.299 773 3.90.057280.943 493 5.30.04212 1.304 11447.10.090460.92115617.50.124780.98 259 5.70.03346 1.295 372 2.80.02555 1.219 1289>10.00.12482 1.03 7277.10.06585 1.03830 5.80.00343 1.39794 6.80.01439 1.685 303 6.60.019140.892 2119 6.70.21487 1.164 9987.10.059110.76263 3.40.002370.436140.000190.962 475 5.50.03101 1.1898.60.00225 1.225 194 2.50.00680.908 817 6.60.031350.985 68170.09849 1.714 1358 2.50.073 1.334 1194 2.60.032550.749 10720.00309 1.26 1021 4.40.12987 1.194 459 3.20.02416 1.192 2618.90.03446 1.396 1804 3.70.15067 1.296 1418.60.01298 1.609 1709 4.50.08330.842 327 1.30.00320.94181 4.90.00904 1.23 327 4.70.03579 1.258 17750.0131 1.044 2597.10.02119 1.078 877 4.30.068460.99799 1.40.001410.915 449 6.30.028910.786 16157.50.18356 1.081 22860.020170.96535 1.90.00176 1.224 1160 3.80.055440.914 201 6.30.017610.949 426 4.10.025740.772 2120 3.70.074070.708 1016 5.80.054290.757 321 5.50.013530.63690 3.30.006210.726 184 4.80.010070.73123>10.00.00228 1.513 610 3.80.032170.72 842 5.40.057730.829 213 5.40.01270.77439 2.40.001731002 4.80.033670.729 208 4.30.00930.537 5247.60.043570.93 116>10.00.02104 1.812 173>10.00.013330.936 589 5.30.032210.807 108 6.10.009190.931 487 4.70.03951 1.142 76450.047770.779 1666 5.20.091160.854 193 1.80.004620.921 120 2.20.004670.894 3338.70.027140.756 4327.10.027040.625 19520.003610.533 194 6.90.011710.909 2597.30.013360.716 266 6.80.026850.752 87080.070140.869 2187.70.01310.75610 5.60.00062 1.012 4189.20.026590.815 622 5.20.034970.666 1709.90.01712 1.222 178 5.80.009120.836 907.70.00320.65258>10.00.01221 1.371 4877.30.041850.896 187 5.60.009820.662 338 6.50.019280.711 443 6.30.026840.651 20850.01330.806 409 4.50.027060.735 774 3.20.034350.762 15287.90.107180.745 739 5.40.053960.715 227.60.001180.774 105 5.10.007480.647 129 4.90.00780.74475 3.50.002990.639212 3.70.013920.784 201 5.60.009290.599 500 6.30.06138 1.238 >10.00.00042 1.049 290 6.10.019640.625 251 5.60.017330.895 12498.40.075460.68 241 2.20.005560.628 1341 6.70.120620.829 651 6.90.026690.572 224 5.10.010210.672 290 5.30.013220.702 194 1.60.002660.7278>10.00.000620.792 314>10.00.019270.649 32830.001390.381 520 6.30.034550.62153 5.20.004350.904 200 1.30.002460.856 109 3.70.00563143 5.80.013670.825 439 4.20.025470.66 441>10.00.03210.853 17290.80.002020.494 1527.40.009010.741 624 2.40.016810.714 337 6.40.02850.71 173 1.90.002010.614 124 6.30.008010.593 2207.60.010810.602 4708.20.028730.596 221 5.20.00853 1.29 4957.50.036490.714 967.20.003150.52311>10.00.000450.543 184 3.50.006730.588 1064 2.80.017560.666 2437.60.014140.744 257.10.002380.868 105130.022770.574 10130.003550.834 2518.20.017620.63 16669.90.081330.508 1478 4.10.10840.548 14830.008560.8753427.80.015630.587 559.80.002470.6496 1.50.001250.668648.10.002270.561 178 6.20.010850.6 247>10.00.011750.526 202 5.20.013550.701 1484 5.40.069550.546 116 6.40.005740.60572 2.10.00140.58954>10.00.004010.856 351 6.60.011110.57 960 3.90.021660.681 1549.50.006150.597 5790.008180.646 6720.0008353 2.30.00061176 2.60.005320.633 1947.80.016490.569 223 5.60.014960.968 25280.006360.457 1626 5.50.049760.537 1977.10.008730.585 128 5.40.00580.57919>10.00.001080.734 650 3.70.017210.673 9419.10.050830.59113>10.00.00130.598 21090.007690.475 906>10.00.066080.687 350 3.90.005590.396 2247.80.01640.479 1789 5.50.074220.539 419>10.00.014550.491 579 6.80.034280.585 11717.30.084950.73 918.60.003920.471 458.60.00220.655 100 6.20.007370.666 335 5.10.012850.644 964 5.50.038290.59199 5.20.004810.478 302 6.60.012460.48640 2.10.000610.345 392>10.00.021350.719339>10.00.01560.489 5138.90.025150.615 50 4.50.001620.659 1368.50.005580.639 353 5.90.018730.713 495 4.80.011170.321 1147.60.003960.391 156 6.70.022520.706 508 6.50.027410.476 82 5.10.003380.472 410>10.00.020760.421 400>10.00.012770.531 463 4.70.008250.53 728 5.70.012880.253 2367.10.012090.669 180 3.80.004490.45 360>10.00.018270.679 2957.80.035210.653 950 3.40.026580.586 218 5.20.006840.446 2547.50.007820.487。
不同酶消化法提取猪原代肝细胞的效果比较
532024.4·试验研究0 引言猪圆环病毒（PCV ）是Circoviridae 科Circovirus 属的一种无囊膜的单链环状DNA 病毒。
在已知的4个血清型中，PCV2为猪易感的致病性病毒[1]。
PCV2感染会诱导宿主免疫抑制引起猪圆环病毒病（PCVD ），包括断奶仔猪多系统衰竭综合征、新生仔猪先天性脑震颤、皮炎与肾病综合征、猪呼吸道病综合征、母猪繁殖障碍等，给全世界养猪业带来较大的经济损失，是世界各国的兽医与养猪业者公认的造成重大影响的猪传染病[2]。
PCV2的感染在猪生长发育的不同阶段有不同的组织嗜性。
但无论是胎儿阶段还是出生后，肝细胞都是PCV2感染和复制的靶细胞。
因此，PCV2也被视为一种能够诱导猪肝炎的病毒[3]。
且PCV2诱导的肝细胞凋亡在PCV2引发的相关病变和疾病的发病机制中具有关键性作用[4]。
因此，方便、快捷地获取大量有活性的猪肝细胞对于研究PCVD 的致病机制具有重大意义。
目前获取肝细胞常用的方法主要包括机械分离细胞法、非酶分离细胞法、离体酶消化法和酶灌流法等[5]。
因此，本试验采用简便、经济、无需特殊设备、仅需部分肝组织的离体酶消化法，比较不同酶消化分离猪原代肝细胞的效果，为一般实验室提取分离大量有活性的猪肝细胞提供参考。
1 材料与方法1.1 材料1.1.1 主要试剂新鲜猪肝组织，Hank's 平衡盐溶液（HBSS ），磷酸盐缓冲液（无菌PBS ），4%多聚甲醛（PFA ），收稿日期：2024-01-27基金项目：国家自然科学基金项目：复杂器官与组织在脾脏内的功能性再生（32230056）作者简介：周徐倩（1999-），女，汉族，浙江温州人，硕士在读，研究方向：组织工程与再生医学。
*通信作者简介：董磊（1978-），男，汉族，安徽阜阳人，博士，教授，研究方向：组织工程与再生医学、生物材料。
周徐倩，董磊．不同酶消化法提取猪原代肝细胞的效果比较[J]．现代畜牧科技，2024，107（4）：53-55． doi ：10.19369/ki.2095-9737.2024.04.014． ZHOU Xuqian ，DONG Lei ．Comparison of the Effect of Different Enzyme Digestion Methods on Extraction of Porcine Primary Hepatocytes[J]．Modern Animal Husbandry Science & Technology ，2024，107（4）：53-55．不同酶消化法提取猪原代肝细胞的效果比较周徐倩，董磊*（南京大学，江苏 南京 210023）摘要：猪肝细胞是猪圆环病毒的靶细胞，简单快速地提取猪原代肝细胞对于研究猪圆环病毒病的致病机制具有重要意义。
有机化学实验中关于过柱的实验方法和技巧
关于过柱的实验方法和技巧(注意：有机溶剂对身体特有害别是心肺；肝脏等所有过柱操作都要在通风橱里进行！！常说的过柱子应该叫柱层析分离，也叫柱色谱。
我们常用的是以硅胶或氧化铝作固定相的吸附柱。
由于柱分的经验成分太多，所以下面我就几年来过柱的体会写些心得，希望能有所帮助。
1、柱子可以分为：加压，常压，减压压力可以增加淋洗剂的流动速度，减少产品收集的时间，但是会减低柱子的塔板数。
所以其他条件相同的时候，常压柱是效率最高的，但是时间也最长，比如天然化合物的分离，一个柱子几个月也是有的。
减压柱能够减少硅胶的使用量，感觉能够节省一半甚至更多，但是由于大量的空气通过硅胶会使溶剂挥发（有时在柱子外面有水汽凝结），以及有些比较易分解的东西可能得不到，而且还必须同时使用水泵抽气（很大的噪音，而且时间长）。
以前曾经大量的过减压柱，对它有比较深厚的感情，但是自从尝试了加压后，就几乎再也没动过减压的念头了。
加压柱是一种比较好的方法，与常压柱类似，只不过外加压力使淋洗剂走的快些。
压力的提供可以是压缩空气，双连球或者小气泵（给鱼缸供气的就行）。
特别是在容易分解的样品的分离中适用。
压力不可过大，不然溶剂走的太快就会减低分离效果。
个人觉得加压柱在普通的有机化合物的分离中是比较适用的。
2、关于柱子的尺寸，应该是粗长的最好柱子长了，相应的塔板数就高。
柱子粗了，上样后样品的原点就小（反映在柱子上就是样品层比较薄），这样相对的减小了分离的难度。
试想如果柱子十厘米，而样品就有二厘米，那么分离的难度可想而知，恐怕要用很低极性的溶剂慢慢冲了。
而如果样品层只有厘米，那么各组分就比较容易得到完全分离了。
当然采用粗大的柱子要牺牲比较多的硅胶和溶剂了，不过这些成本相对于产品来说也许就不算什么了（有些不环保的说，不过溶剂回收重蒸后也就减小了部分浪费）。
现在见到的柱子径高比一般在1：5～10，书中写硅胶量是样品量的30～40倍，具体的选择要具体分析。
如果所需组分和杂质分的比较开（是指在所需组分rf 在～，杂质相差以上），就可以少用硅胶，用小柱子（例如200毫克的样品，用2cm×20cm的柱子）；如果相差不到，就要加大柱子，我觉得可以增加柱子的直径，比如用3cm的，也可以减小淋洗剂的极性等等。
贯叶金丝桃二氯甲烷部位的化学成分研究
贯叶金丝桃二氯甲烷部位的化学成分研究张亦云;韩竹箴;张倩;王佳;程明赛;王振中;杨颖博;肖伟【期刊名称】《中成药》【年(卷),期】2024(46)5【摘要】目的研究贯叶金丝桃Hypericum perforatum L.二氯甲烷部位的化学成分。
方法贯叶金丝桃二氯甲烷部位采用硅胶、ODS、Sephadex LH-20、半制备HPLC等进行分离纯化,根据理化性质及波谱数据鉴定所得化合物的结构。
结果从中分离得到11个化合物,分别鉴定为贯叶金丝桃降碳聚酮A(1)、α-芒柄花醇(2)、(3 R)-thunberginol(3)、2-geranyloxy-1-(2-methylpropanoyl)-phloroglucinol(4)、4,6-dihydroxy-2-O-(3″,7″-dimethyl-2″,6″-octadienyl)-1-(2′-methylbutanoyl)benzene(5)、norhyperpalum G(6)、garsubellin A(7)、garsubellin B(8)、(2″R/S)-kellerine C(9)、kobusone(10)、圣草酚(11)。
结论化合物1为新化合物,化合物2~3为首次在藤黄科植物中分离得到,化合物4~10为首次从该植物中分离得到。
【总页数】7页(P1552-1558)【作者】张亦云;韩竹箴;张倩;王佳;程明赛;王振中;杨颖博;肖伟【作者单位】南京中医药大学康缘中药学院;中药制药过程控制与智能制造技术全国重点实验室;上海中医药大学中药研究所;江苏康缘药业股份有限公司【正文语种】中文【中图分类】R284.1【相关文献】1.新疆贯叶金丝桃不同部位中金丝桃苷、芦丁和金丝桃素的含量及提取工艺对它们的影响2.不同药用部位及采收方法对贯叶金丝桃中抗抑郁活性成分的影响3.贯叶连翘不同部位金丝桃素含量分析4.贯叶金丝桃的化学成分与药理作用研究进展5.九叶青花椒二氯甲烷部位化学成分研究因版权原因，仅展示原文概要，查看原文内容请购买。
211091464_超高效液相色谱串联质谱法测定乳制品中透明质酸
杜国辉，范维江，陈玉娟，等. 超高效液相色谱串联质谱法测定乳制品中透明质酸[J]. 食品工业科技，2023，44（8）：334−340. doi:10.13386/j.issn1002-0306.2022070057DU Guohui, FAN Weijiang, CHEN Yujuan, et al. Determination of Hyaluronic Acid in Dairy Products by Ultra-high Performance Liquid Chromatography-Tandem Mass Spectrometry[J]. Science and Technology of Food Industry, 2023, 44(8): 334−340. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022070057· 分析检测 ·超高效液相色谱串联质谱法测定乳制品中透明质酸杜国辉1，范维江1，陈玉娟2, *，陈雯雯2，乔莉苹2，王萃玲3（1.山东商业职业技术学院食品工业产业学院，山东济南 250103；2.华熙生物科技股份有限公司，山东济南 250101；3.齐鲁医药学院，山东济南 250103）摘 要：本文建立一种超高效液相色谱-串联质谱（UPLC-MS/MS ）测定乳制品中透明质酸含量的分析方法。
样品经透明质酸酶降解，乙腈稀释，PRIME HLB 固相萃取柱净化，Waters BEH Amide （2.1 mm×100 mm ，1.7 μm ）酰胺柱分离，以0.2%氨水-乙腈（含0.2%氨水）为流动相梯度洗脱。
采用电喷雾离子源负离子模式扫描，多反应监测模式进行检测，外标法定量。
结果表明：乳制品中透明质酸在0.5~200 mg/kg 添加水平下回收率为91.4%~106.2%，RSD 为2.3%~6.7%。
Expression, Purification and Crystallization of
Expression, Purification and Crystallization of the Mycobacterium Tuberculosis HSP16.3 Molecular Chaperone Background of Mycobacterium Tuberculosis HSP16.3HSP16.3, a 16.3 kDa protein from Mycobacterium Tuberculosis, was originally identified as a prominent antigen (Kingston et al., 1987). During the stationary phase, HSP16.3 is maximally expressed and becomes a main protein of the latent phase (Yuan et al., 1996). Previous studies showed that HSP16.3 can make the cell structure stable and prevent stationary Mycobacterium Tuberculosis from autolysing (Cunningham et al., 1998). In previous studies, HSP16.3 was found as one of theα-crystallin-related small heat shock proteins (sHSP) with molecular chaperone activity. Experiments in vitro revealed that HSP16.3 can suppress the thermal aggregation of citrate synthase at 39.5˚C, without consumption of A TP (Chang et al., 1996).Now the Mycobacterium Tuberculosis HSP16.3 gene was cloned to the plasmid pSTE-HSP16.3, and transformed to E.Coli. BL21(DE3) strain.Material and MethodExpressionThings to have ready before Starting.-Plate or glycerol culture-Sterile LB 25ml in a 50mL shaker flasker, 250ml in a 500mL shaker flasker, all together autoclaved, antibiotic added afterword.- antibiotic and sterile water- TipsPrepare the LB and autoclave:Fomula of the LB medium for 1 Liter:Bacto Tryptone (BT) 10 gBacto Y east Extract (BYE) 10 gNaCl 10gThe LB medium, dd H2O and the tips all together autoclaved at 121 ˚C for 20 minutes.Method:1 Innoculate 25 ml LB Medium ( containing 100 ug) and grow culture overnight(37˚C, 200rpm).2 Next morning inoculate 250 ml prewarmed LB Medium ( containing 100 ug) with the 25 ml overnight culture and grow at 37 ˚C, 200rpm, HSP16.3 was overexpressed in soluble form intracellularly without IPTG induction.3 Incubate the Culture for 10 hours before havesting the cell at 4000 g for 20 minutes.4 Resuspend the cell pellet in 30 ml Butter A and freeze the Sample in -80˚C refigerator.PurificationDE52 Ion-Exchange columnThings to have ready before Starting.-Butter A: 50 mM Imidazole pH 6.5 (1 liter)-Butter B: 50 mM Imidazole pH 6.5 , 300mM NaClall together Fitrate with 0.2 um membrane.- DE52 medium , column ,Gradient maker, UV-monitor and Fractioner- TipsMethod:1 Thaw the cell pellet and vortex .2 Add 0.4ml 100 mM PMSF and sonicate (400kw, 4s-6s 50 cycle* 5 )3 Centrifuge 15000 rpm, 30 minutes to pellet debris4 Transfer supernatant to a 50 ml conicale tube and discard the pellet.5 The supernatant dilute to 50 ml with Buffer A and then load to DE52 ion-exchange columns (20ml), which was pre-equibrated with 100ml Buffer A. And then wash the unbound proteins with 100 ml Buffer A.6 Elute the protein with a linear gradient : 200ml buffer A plus 200ml buffer B, 2ml/min, 6ml each fraction.7 Run 15% SDS-PAGE to determine the HSP16.3 peak.Desalting by dialysis1 Preparation of the dialysis tubeCut the tube in a suitable length (20-30 cm)Boil the tube in solution containing 10 mM NaHCO3 for a few minutes.Boil the tube in solution containing 10 mM EDTA for a few minutes.Rasin the tube with de-ion water2 Pool the HSP16.3 peak and dialysis the Sample against 1000ml Buffer A for more than 6hours.Q-Separose (HP) Ion-Exchange Column1 load the sample to Q-Separose (HP) Ion-Exchange column (20ml), which was pre-equibrated with 100ml Buffer A. And then wash the unbound proteins with 100 ml Buffer A.2 Elute the protein with a linear gradient : 200ml buffer A plus 200ml buffer B, 2ml/min, 6ml each fraction.3 Run 15% SDS-PAGE to determine the purity of the HSP16.3 peak.Gel filtration ColumnThe HSP peak was a final volumn 0.3ml and then run though a Superdex75 (HR, 10/30mm) gel filtration column in 150mM NaCl and 5mM Imdazole, pH6.5. Crystallization1 The purified HSP16.3 was solvent-exchanged to water and concentrated to 20mg/ml before crystallization trails (Bradford). All the crystallization trials were carried out using the hanging-drop vapor-diffusion method at 291K: drops consisted of2 microlitres of HSP16.3 protein solution plus 2 microlitres of the precipitant. The drops were equilibrated against 0.2 ml precipitant at room temperature. The crystallization conditions were investigated with a PEG4000 Kit.Result and discussionThe purity of the final HSP16.3 was over 95% by SDS-PAGE. The crystallization trials of HSP16.3 yielded Cubic crystals with a size of 0.8*0.8*0.6mm in a few days.20040060080010001200mAUBuffer Tris-HCL pH 8.5 Precipitant PEG 4000 MethodV apor Diffusion Temperature 293 K Size0.8*0.8*0.6mmReferencesChang Z., Primm, T.P., Jakana J., Lee H. I., Serysheva I., Chiu W., Gilber H. F., Quiocho F. A., (1996) J Biol Chem 271:7218-7223Cunningham A. F., Spreadbury C. L., (1998) J. Bacteriol. 184:801-808Kingston A. E., Salgame P. R., Mitchison N.A., Colston M. J. (1987) Infect. Immun 55,3149-3154Yuan Y., Crane D. D., Barry C. E. III (1996) J Bacteriol178: 4484-4492。
大蒜素诱导肝癌细胞SMMC-7721凋亡的形态观察
大蒜素诱导肝癌细胞SMMC-7721凋亡的形态观察目的观察大蒜素对体外培养的肝癌细胞SMMC-7721的生长抑制和诱导凋亡的作用。
方法用形态观察、MTT和TUNEL检测等方法检测大蒜素对SMMC-7721细胞的生长抑制和诱导凋亡情况。
结果大蒜素作用后SMMC-7721呈现出了明显的凋亡形态，凋亡小体集中出现后又迅速消失。
凋亡指数由525上升至3061(P＜005)。
结论大蒜素对SMMC-7721细胞有显著的抑制生长和诱导凋亡的作用，光镜下的形态特征明显、独特。
标签：大蒜素；细胞凋亡；肝癌大蒜(allium sativum)为百合科多年生草本植物大蒜的鳞茎，大蒜素(allicin)是从大蒜中提取的挥发性油状物，是二烯丙基三硫化物、二烯丙基二硫化物以及甲基烯丙基二硫化物等的混合物。
中医药学认为大蒜有行滞气，暖脾胃，解毒杀虫之功效。
近来研究表明，大蒜还具有降血脂、抗动脉粥样硬化、抗肿瘤和提高免疫功能等多方面的作用。
流行病学调查显示，大蒜产区和长期食用大蒜的人群，其癌症发生率偏低［1］,因此有关于大蒜防癌、抗癌作用的研究日益受到关注。
本实验采用形态学观察、MTT等方法，进行了大蒜素对体外培养的肝癌细胞的诱导凋亡作用的研究。
1材料与方法11细胞培育细胞株：SMMC-7721肝癌细胞购自武汉冷藏中心；培养液：RPMI 1640培养基(GIBCO，USA)；新生牛血清：购自杭州四季青生物工程公司；胰蛋白酶：025%胰酶，002%EDTA；CO2培养箱：IT-41，Japan；倒置相差显微镜：Olympus，Japan。
12药品大蒜素注射液：上海禾丰制药厂；四甲基偶氮唑盐(MTT)：SIGMA公司。
用PBS配成5 mg/ml，除菌、分装后，4 ℃避光保存。
96孔培养板：SIGMA 公司；TUNEL检测试剂盒：华美生物公司。
13MTT检测将浓度为5×104 个/ml的细胞接种于96孔细胞培育板中，每孔加100 μl，待细胞处于对数生长期时加大蒜素。
检测样本中烟酰胺含量的试剂在制备诊断静脉流出障碍性疾病的试剂盒中的应用[发明专利]
![检测样本中烟酰胺含量的试剂在制备诊断静脉流出障碍性疾病的试剂盒中的应用[发明专利]](https://img.taocdn.com/s3/m/8c6ee0355e0e7cd184254b35eefdc8d376ee14f0.png)
专利名称：检测样本中烟酰胺含量的试剂在制备诊断静脉流出障碍性疾病的试剂盒中的应用
专利类型：发明专利
发明人：吉训明,卫慧敏,周陈,姜慧敏,李明,周一帆
申请号：CN202210048747.9
申请日：20220117
公开号：CN114487372A
公开日：
20220513
专利内容由知识产权出版社提供
摘要：本发明公开了检测样本中烟酰胺含量的试剂在制备诊断静脉流出障碍性疾病的试剂盒中的应用，涉及生物医学技术领域。
本发明首次提出烟酰胺可以用于判断静脉流出障碍性疾病的生物标志物，通过对受试者静脉流出障碍疾病部位中的烟酰胺进行含量的测定，能够诊断或辅助诊断出其是否患有静脉流出障碍性疾病或具体的发病程度，为静脉流出障碍性疾病的诊断检测提供了新思路。
此外，通过补充烟酰胺，使得患者大脑中烟酰胺的含量恢复正常也可以成为一种治疗或改善静脉流出障碍性疾病的途径。
申请人：北京航空航天大学
地址：100089 北京市海淀区学院路37号
国籍：CN
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合成生物学在医药领域的应用进展
·专家论坛·合成生物学在医药领域的应用进展李瀚纯王鑫瞿旭东王舒［弈柯莱生物科技（集团）股份有限公司上海 200241］摘要近20年来，合成生物学在生物回路构建、生物元件标准化，以及各种基因组/代谢工程工具和方法的开发方面不断取得突破。
合成生物学的快速发展正在改变生物技术行业的产业布局。
目前，合成生物技术已广泛应用于天然产物合成、医学、能源、工业等多个领域。
医药的需求也推动了合成生物学的发展，包括将体外催化技术应用于手性医药化学品的绿色制造，将异源途径整合到细胞中以有效生产药物等。
合成生物学凭藉更经济、环境友好等突出特点，将颠覆一部分传统医药的制造方式。
本文概要介绍合成生物学在手性医药化学品的绿色制造和植物天然产物的生物制造方面的应用进展。
关键词合成生物学 医药化学品 天然产物 萜类化合物 芳香族化合物中图分类号：Q819; O629.71 文献标志码：A 文章编号：1006-1533(2024)07-0024-08引用本文李瀚纯, 王鑫, 瞿旭东, 等. 合成生物学在医药领域的应用进展[J]. 上海医药, 2024, 45(7): 24-31; 55.Applications of synthetic biology in the pharmaceutical fieldLI Hanchun, WANG Xin, QU Xudong, WANG Shu(Abiochem Biotechnology Co., Ltd., Shanghai 200241, China)ABSTRACT In the past two decades, synthetic biology has made breakthroughs in the construction of biocircuits, the standardization of biological elements and the development of various genomic/metabolic engineering tools and approaches.Its rapid development is changing the industrial layout of biotechnology industry. At present, synthetic biotechnology has been widely used in many fields such as natural product synthesis, medicine, energy and industry. Pharmaceutical demands have also driven its development, including the application of in vitro catalytic technology in the green manufacturing of chiral pharmaceutical chemicals and the integration of heterologous pathways into designer cells to efficiently produce medicines and so on. Synthetic biology, with its more economical and environmentally friendly features, will subvert some traditional pharmaceutical manufacturing methods. This article reviews the applications of synthetic biology in the green manufacturing of chiral pharmaceutical chemicals and the biological manufacturing of natural plant products.KEY WORDS synthetic biology; pharmaceutical chemicals; natural products; terpenoids; aromatic compounds合成生物学是采用工程科学研究理念，对生物体进行定向设计、理性改造甚至创造新型生物体的一门学科。
德谷门冬双胰岛素注射液治疗2_型糖尿病的疗效及安全性研究
DOI：10.16658/ki.1672-4062.2023.19.084德谷门冬双胰岛素注射液治疗2型糖尿病的疗效及安全性研究戴卉，张开凤，朱凤丽江苏省镇江市丹徒区人民医院内分泌科，江苏镇江212000[摘要]目的探讨德谷门冬双胰岛素注射液在2型糖尿病中的效果以及安全性。
方法选取2022年1月—2023年7月江苏省镇江市丹徒区人民医院收治的62例2型糖尿病患者为研究对象，按随机数表法分为对照组（n=31）和观察组（n=31）。
对照组患者接受门冬胰岛素30注射液治疗，观察组患者接受德谷门冬双胰岛素注射治疗。
对比两组患者临床疗效、血糖变化和不良反应发生率。
结果观察组治疗有效为96.77%，高于对照组的77.42%，差异有统计学意义（χ2=5.167，P=0.023）。
治疗前，两组患者血糖水平比较，差异无统计学意义（P>0.05）；治疗后，两组患者血糖水平均改善，且观察组血糖指标低于对照组，差异有统计学意义（P< 0.05）。
观察组不良反应发生率低与对照组，差异有统计学意义（P<0.05）。
结论德谷门冬双胰岛素的应用可以明显改善2型糖尿病患者血糖水平，疗效更为确切，且安全性更高，不会增加用药后不良反应。
[关键词] 2型糖尿病；德谷门冬双胰岛素；门冬胰岛素30注射液；安全性[中图分类号] R587 [文献标识码] A [文章编号] 1672-4062（2023）10（a）-0084-04Study on the Efficacy and Safety of Insulin Degludec and Insulin Aspart Injection in the Treatment of Type 2 Diabetes MellitusDAI Hui, ZHANG Kaifeng, ZHU FengliDepartment of Endocrinology, Zhenjiang Dantu District People's Hospital, Zhenjiang, Jiangsu Province, 212000 China [Abstract] Objective To explore the effect and safety of insulin degludec and insulin aspart injection in type 2 diabe⁃tes mellitus.Methods 62 patients of type 2 diabetes mellitus patients admitted to Zhenjiang Dantu District People's Hospital, Jiangsu Province from January 2022 to July 2023 were selected as study objects and divided into the control group (n=31) and the observation group (n=31) by taking the random number table method. The patients in the control group were treated with insulin aspart 30 injection and the patients in the observation group were treated with insulin degludec and insulin aspart injection. Compared the clinical efficacy, the changes in blood glucose and the incidence of adverse reactions between the two groups of patients.Results The treatment effectiveness of the observation group was 96.77%, which was higher than that of the control group, which was 77.42%, and the difference was statistically significant (χ2=5.167, P=0.023). There was no statistically significant difference in blood glucose levels between the two groups before treatment (P>0.05). After treatment, blood glucose levels improved in both groups, and the level of blood glucose in the observation group were lower than those in the control group, and the difference was statistically significant (P<0.05). The incidence of adverse reactions in the observation group was lower than that in the control group, and the difference was statistically significant (P<0.05).Conclusion The application of insulin degludec and in⁃sulin aspart can significantly improve the blood glucose level of patients with type 2 diabetes mellitus, the efficacy is more accurate, and the safety is higher, and it will not increase the occurrence of adverse reactions after the use of medication.[作者简介]戴卉（1985-），女，本科，主治医师，研究方向为内分泌科。