4SC-202_LCMS_13886_MedChemExpress
香烟烟雾提取物对大鼠肺成纤维细胞生长的影响
香烟烟雾是有众多化学成分的复杂混合物，超过6000种成分，会对肺脏和全身产生有害影响。
吸烟已被公认与多种肺部疾病的发生发展有关，吸烟是肺癌的主要病因，也是慢性阻塞性肺疾病（COPD）等疾病的主要危险因素。
如今，吸烟还被认为与某些间质性肺病及肺纤维化有关[1]，但尚不清楚吸烟引起间质性肺病的发病机制，对吸烟与肺纤维化关系的研究也很少。
为此本研究利用体外培养的正常大鼠及肺纤维化大鼠的肺成纤维细胞，观察不同浓度香烟烟雾提取物（cigarette smoking extract，CSE）对两种细胞的影响，探讨香烟烟雾在肺纤维化中所起的作用。
1 材料与方法1.1 实验材料1.1.1 主要试剂 博来霉素购自天津太河制药有限公司；RPMI-1640培养基、胰蛋白酶、胎牛血清购自美国GIBCO 公司；凋亡检测试剂盒购自美国BD 公司；兔抗大鼠波动蛋白、纤维粘连蛋白、α-平滑肌肌动蛋白（α-SMA）单克隆抗体及SABC 免疫组化试剂盒购自武汉博士德生物工程有限公司、第二抗体为羊抗兔IgG 购自美国Jackson 公司；化学试剂购自美国Sigma 公司。
1.1.2 仪器 二氧化碳培养箱（美国FORMA 公司【摘要】 目的 通过观察不同浓度的香烟烟雾提取物（cigarette smoking extract，CSE）对正常及肺纤维化大鼠肺成纤维细胞的影响，探讨香烟烟雾与肺纤维化的可能关系。
方法 体外培养的正常及博莱霉素造模的肺纤维化大鼠的肺成纤维细胞，分别以不同浓度（25％、50％、100％）的CSE 作用12h 和24h。
噻唑蓝还原法（MTT 法）检测吸光值；流式细胞仪法观察细胞坏死与凋亡情况及细胞周期S 期（DNA 合成期）和G 1期（DNA 合成前期）的比值。
结果 高浓度（100％）的CSE 主要引起两种大鼠肺成纤维细胞的坏死。
较低浓度（25％、50％）的CSE 对正常大鼠肺成纤维细胞无明显影响（P >0.05）。
Gelucire-14-44-SDS-MedChemExpress
Inhibitors, Agonists, Screening LibrariesSafety Data Sheet Revision Date:Nov.-23-2018Print Date:Nov.-23-20181. PRODUCT AND COMPANY IDENTIFICATION1.1 Product identifierProduct name :Gelucire 14/44Catalog No. :HY-Y1892CAS No. :121548-04-71.2 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:NoneFormula:N/AMolecular Weight:N/ACAS No. :121548-04-74. 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:Pure form-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 (Oil)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 2018 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。
Pimecrolimus_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 :PimecrolimusCatalog No. :HY-13723CAS No. :137071-32-01.2 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 mixtureGHS Classification in accordance with 29 CFR 1910 (OSHA HCS)Acute toxicity, Oral (Category 4),H302Acute aquatic toxicity (Category 1),H400Chronic aquatic toxicity (Category 1),H4102.2 GHS Label elements, including precautionary statementsPictogramSignal word WarningHazard statement(s)H302 Harmful if swallowed.H410 Very toxic to aquatic life with long lasting effects.Precautionary statement(s)P264 Wash skin thoroughly after handling.P270 Do not eat, drink or smoke when using this product.P273 Avoid release to the environment.P301 + P312 IF SWALLOWED: Call a POISON CENTER or doctor/ physician if you feel unwell.P330 Rinse mouth.P391 Collect spillage.P501 Dispose of contents/ container to an approved waste disposal plant.2.3 Other hazardsNone.3. COMPOSITION/INFORMATION ON INGREDIENTS3.1 SubstancesSynonyms:SDZ⁻ASM 981Formula:C43H68ClNO11Molecular Weight:810.45CAS No. :137071-32-04. 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.IMDGUN number: 3077Class: 9Packing group: IIIEMS-No: F-A, S-FProper shipping name: ENVIRONMENTALLY HAZARDOUS SUBSTANCE, SOLID, N.O.S.Marine pollutant: Marine pollutant.IATAUN number: 3077Class: 9Packing group: IIIProper shipping name: Environmentally hazardous substance, solid, n.o.s.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。
美国Medchemexpress化合物库(小分子库)_Medchemexpress_(MCE中国)
美国Medchemexpress化合物库（小分子库）-原装进口，现货供
应，提供组合定制服务
品牌：Medchemexpress （MCE）
保存条件：-20℃
供应商：MCE中国
数量：大量
保质期：2年
Size：
Pre-dissolved DMSO/Solid（Or dry solid）
100 uL/well (10 mM solution)
200 uL/well (10 mM solution)
MedChemExpress (MCE)专注于各种抑制剂、调节剂、API、天然产物及化合物库，总部位于美国新泽西。
MCE经过十年努力已成为全球生物活性小分子领域的一流供应商。
MedChemExpress(MCE)产品涵盖近20个热门研究领域，1000多个细分靶点，超过3000个现货抑制剂、拮抗剂和激动剂。
相关的应用成果已发表于Nature、Cell等国际知名杂志，在全球20余个国家地区设有代理机构。
上海皓元生物医药科技有限公司(MCE 中国) 是MedChemExpress (MCE) 亚洲总代理。
MCE化合物库涵盖20余种不同的类型，超过2500个化合物，进口原装，
现货供应，提供详实的生物活性信息、化学结构信息、质控图谱（NMR和HPLC 等）。
还可根据您的实际研究需要，为您度身定制任意组合、规格、布板的特殊化合物库。
/screening-libraries.html
现有特色化合物库有：。
WHO International Standard 1st WHO International Standard for Human Papillomavirus (HPV) Type 16 DNA
WHO International Standard1st WHO International Standard for Human Papillomavirus (HPV)Type 16 DNA NIBSC code: 06/202 Instructions for use(Version 2.0, Dated 10/11/2010)1. INTENDED USEThe 1st International Standard for HPV Type 16 (HPV-16) DNA Nucleic Acid Amplification Techniques consists of a freeze-dried preparation of recombinant plasmid containing full-length HPV-16 DNA cloned via its unique BamH1 site (Quint et al., 2006). The standard has been formulated in a background of purified human genomic DNA, lyophilized in 0.5 ml aliquots and stored at -20 °C. The material was calibrated in an international collaborative study involving 19 laboratories (Wilkinson et al., 2008). The International Standard contains material that is proprietory to third parties and should be used for the sole purpose of calibrating in-house or working standards for the amplification and detection of HPV-16 DNA. The International Standard should not be used for any other purpose and should be discarded after use. 2. CAUTIONThis preparation is not for administration to humans .This material contains DNA derived from C33A cells. As with all materials of biological origin, this preparation should be regarded as potentially hazardous to health. It should be used and discarded according to your own laboratory's safety procedures. Such safety procedures should include the wearing of protective gloves and avoiding the generation of aerosols. Care should be exercised in opening ampoules or vials, to avoid cuts.3. UNITAGEThe 1st International Standard for HPV-16 DNA Nucleic Acid Amplification Techniques has been assigned a unitage of 5 x 106 International Units (IU) per ampoule.Traceability statement:It was proposed at a WHO meeting in January 2008 (WHO Meeting Report, 2008) that the instructions for use of the International Standard for HPV-16 DNA include the calculations and assumptions used in determining the theoretical HPV-16 qenome equivalents (GEq) of the bulk material used in formulating the International Standard, thus demonstrating that 1 IU is equivalent to 1 GEq for HPV-16 DNA . The definitive unitage of the 1st WHO International Standard for HPV-16 DNA therefore remains as IU while the traceability statement would allow users to equate IU with GEq.Assays for DNA concentration of the recombinant HPV-16 plasmid stock preparation were performed in Dr Cosette Wheeler‟s laboratory, University of New Mexico (UNM). DNA concentrations were determined by absorbance at 260 nm as well as spectrofluorometrically using the Picogreen assay (Invitrogen Corporation, USA). A correlation coefficient of 0.95 or higher was obtained between the two DNA measurements. 10 ng HPV-16 plasmid DNA/μl was supplied to NIBSC for formulating the bulk material for subsequent freeze-drying. The UNM laboratory also provided NIBSC with a statement indicating that 1.0 x 1011 GEq/ml for HPV-16 is equal to 1.17 ng/μl. 10 ng HPV-16 plasmid DNA/μl plasmid stock preparation is therefore equivalent to 8.547 x 1011 HPV-16 GEq/ml. NIBSC used this data in formulating the 1st International Standard for HPV Type 16 DNA.Formulation of bulk material for the 1st International Standard for HPV Type 16 DNA (NIBSC code 06/202):At NIBSC, the bulk HPV-16 plasmid DNA material was prepared according to the formula:HPV GEq/ml of bulk material = (HPV GEq/ml of plasmid stock x volume plasmid stock) / volume bulk material.Therefore,HPV-16 GEq/ml of bulk material = (8.547 x 1011 HPV-16 GEq/ml plasmid stock) x (0.02223 ml HPV-16 plasmid stock) / 1900 ml HPV-16 bulk material = 1.0 x 107 HPV-16 GEq/ml bulk materialThe HPV-16 DNA bulk material was subsequently freeze-dried in 0.5 ml aliquots.Certain assumptions are required for equating IU to GEq for the 1st International Standard for HPV-16 DNA: 1) 1.0 x 1011 GEq/ml for HPV-16 is equal to 1.17 ng/μl. 2) There is no loss in activity of the HPV-16 DNA upon lyophilization. 3) The recombinant HPV-16 plasmid DNA accurately mimics the activity of HPV-16 viral DNA in biological samples.Independent calculation of GEq/ml for recombinant HPV-16 plasmid DNA.NIBSC also independently calculated the genome equivalence of the HPV-16 plasmid stock preparation and bulk preparation in which the molecular weights of the full-length HPV-16 genome and pBR322 DNA were based on sequence content using BioEdit Sequence Alignment Editor v7.0.5.3 (Tom Hall, Isis Pharmaceuticals Inc., USA). The sequences used for determining the molecular weights are GenBank Accession number J01749.1 for pBR322 and the reference sequence for HPV16 (Accession K02718).BioEdit dataDNA molecule: HPV16 Accession K02718 Length = 7904 base pairsMW= 4786756.00 Daltons, double strandedDNA molecule: cloning vector pBR322 Length = 4361 base pairsMW= 2653867.00 Daltons, double strandedFormulaeGEq/ml of the HPV plasmid stock was calculated according to the formula: GEq/ml of the HPV plasmid stock = (DNA concentration of HPV plasmid stock) x (MW of HPV DNA + MW of pBR322)-1 x (Avogadro‟s Number) where Avogadro‟s Number = 6.022x1023 molecules/molGEq/ml of the bulk HPV DNA materials was calculated according to the formula:HPV GEq/ml of bulk material = (HPV GEq/ml of plasmid stock x volume plasmid stock) / volume bulk material.CalculationThe recombinant HPV-16 plasmid stock preparation was supplied to NIBSC at a concentration of 10 ng/μl. Using the MW determinations shown above, the GEq/ml of the HPV-16 plasmid stock is:= (10 x 10-9 g/μl) x (mol/(7440623 g) x (6.022x1023 molecules/mol) = 8.093 x 108 molecules/μl = 8.093 x 1011molecules/ml = 8.093 x 1011 HPV-16 GEq/ml22.23μl of the recombinant HPV-16 plasmid stock was diluted to a final volume of 1900ml, therefore,HPV-16 GEq/ml of bulk material = (8.093 x 1011 HPV-16 GEq/ml plasmid stock) x (0.02223 ml HPV-16 plasmid stock) / 1900 ml HPV-16 bulk material = 0.947 x 107 HPV-16 GEq/ml bulk material4. CONTENTSCountry of origin of biological material: United Kingdom.Each ampoule contains the lyophilized equivalent of 0.5 ml HPV-16 plasmid DNA in 10mM Tris buffer pH7.4 containing 1mM EDTA, 5 mg/ml trehalose and ~1 x 106 human GEq/ml derived from C33a cells.5. STORAGEThe ampoule should be stored at -20 °C or below on receipt.Please note: because of the inherent stability of lyophilized material, NIBSC may ship these materials at ambient temperature.6. DIRECTIONS FOR OPENINGDIN ampoules have an …easy -open‟ coloured stress point, where the narrow ampoule stem joins the wider ampoule body.Tap the ampoule gently to collect the material at the bottom (labeled) end. Ensure that the disposable ampoule safety breaker provided is pushed down on the stem of the ampoule and against the shoulder of the ampoule body. Hold the body of the ampoule in one hand and the disposable ampoule breaker covering the ampoule stem between the thumb and first finger of the other hand. Apply a bending force to open the ampoule at the coloured stress point, primarily using the hand holding the plastic collar.Care should be taken to avoid cuts and projectile glass fragments that might enter the eyes, for example, by the use of suitable gloves and an eye shield. Take care that no material is lost from the ampoule and no glass falls into the ampoule. Within the ampoule is dry nitrogen gas at slightly less than atmospheric pressure. A new disposable ampoule breaker is provided with each DIN ampoule.7. USE OF MATERIALNo attempt should be made to weigh out any portion of the freeze-dried material prior to reconstitution.The 1st International Standard for HPV-16 DNA contains high copy number template. There is a high risk of HPV-16 plasmid DNA contamination via aerosolization upon opening of the glass ampoule. The material must be opened and handled in a separate laboratory environment, away from other pre-amplification components such as reagents, labware and samples.The material is supplied lyophilized and, before use, should be reconstituted in 0.5 ml sterile nuclease-free water. Ensure that the inside surface of the ampoule is wetted with the added water so that any particles of freeze-dried material adhering to the glass are reconstituted. The reconstituted material has a final concentration of 1 X 107 IU/ml. The reconstituted material is suitable for calibration of in-house or working standards for the amplification and detection of HPV-16 DNA.. The material is not suitable for calibrating or assessing extraction, precipitation or centrifugation procedures. The material has NOT been calibrated for human DNA nucleic acid amplification techniques.8. STABILITYReference materials are held at NIBSC within assured, temperature-controlled storage facilities. The 1st International Standard for HPV-16 DNA should be stored at -20 °C or below on receipt.Studies on the stability of reconstituted standard are underway. Users should determine the stability of the reconstituted material according to their own method of preparation, storage and use.NIBSC follows the policy of WHO with respect to its reference materials.9. REFERENCESQuint, W. G. V., Pagliusi, S. R., Lelie, N., de Villiers, E. M., Wheeler, C. M. and the World Health Organization Human Papillomavirus DNA International Collaborative Study Group. (2006). Results of the First WorldHealth Organization International Collaborative Study of Detection of Human Papillomavirus DNA. J. Clin. Microbiol. 44: 571-579.Wilkinson, D.E., Baylis, S.A., Padley, D., Heath, A.B., Ferguson, M., Pagliusi, S.R., et al. Establishment of the 1st World Health Organization international standards for human papillomavirus type 16 DNA and type 18 DNA. Int J Cancer 2010 Jun 15;126(12):2969-83.WHO meeting report, on “Standardization of HPV assays and the role of HPV LabNet in supporting vaccine introduction” Geneva, Switzerland, 23-25 January 2008, in preparation.10. ACKNOWLEDGEMENTS11. FURTHER INFORMATIONFurther information can be obtained as follows; This material: enquiries@ WHO Biological Standards:http://www.who.int/biologicals/en/JCTLM Higher order reference materials: /en/committees/jc/jctlm/ Derivation of International Units:/products/biological_reference_materials/frequently _asked_questions/how_are_international_units.aspx Ordering standards from NIBSC:/products/ordering_information/frequently_asked_q uestions.aspxNIBSC Terms & Conditions:/terms_and_conditions.aspx12. CUSTOMER FEEDBACKCustomers are encouraged to provide feedback on the suitability or use of the material provided or other aspects of our service. Please send any comments to enquiries@13. CITATIONIn all publications, including data sheets, in which this material is referenced, it is important that the preparation's title, its status, the NIBSC code number, and the name and address of NIBSC are cited and cited correctly.15. LIABILITY AND LOSSInformation provided by the Institute is given after the exercise of all reasonable care and skill in its compilation, preparation and issue, but it is provided without liability to the Recipient in its application and use. It is the responsibility of the Recipient to determine the appropriateness of the standards or reference materials supplied by the Institute to the Recip ient (“the Goods”) for the proposed application and ensure that it has the necessary technical skills to determine that they are appropriate. Results obtained from the Goods are likely to be dependant on conditions of use by the Recipient and the variability of materials beyond the control of the Institute.All warranties are excluded to the fullest extent permitted by law, including without limitation that the Goods are free from infectious agents or that the supply of Goods will not infringe any rights of any third party.The Institute shall not be liable to the Recipient for any economic loss whether direct or indirect, which arise in connection with this agreement.The total liability of the Institute in connection with this agreement, whether for negligence or breach of contract or otherwise, shall in no event exceed 120% of any price paid or payable by the Recipient for the supply of the Goods.If any of the Goods supplied by the Institute should prove not to meet their specification when stored and used correctly (and provided that the Recipient has returned the Goods to the Institute together with written notification of such alleged defect within seven days of the time when the Recipient discovers or ought to have discovered the defect), the Institute shall either replace the Goods or, at its sole option, refund the handling charge provided that performance of either one of the above options shall constitute an entire discharge of the Institute‟s liability under this Condition.。
硝基咪唑类药物测定方法——液相色谱
硝基咪唑类药物测定方法——液相色谱(6)液相色谱-质谱法(liquid chromatography mass spectrometry，LC-MS) 随着LC与MS接口瓶颈解决，采纳LC-MS技术测定硝基咪唑类药物的讨论报道不断增多，主要有热喷雾电离源(TSI)、大气压化学电离源(APCI)和电喷雾电离源(ESI)三种离子化方式。
LC-MS可分析GC-MS 所不能分析的强极性、难挥发、热不稳定性的化合物，但在测定中内源性基质引起的离子抑制效应较为常见，通过转变质谱条件、提高色谱分别效能、应用合适的内标物可克服基质效应。
1)热喷雾电离源(TSI) TSI是最早应用的LC-MS接口，但存在重复性差、定量效果差的缺点，现在已很少应用。
Cannavan等建立了禽组织和蛋中DMZ的LC-MS测定办法。
色谱柱为ODS3(250mm×4mmi.d.)，流淌相为甲醇-0.05mol/L(1+1，v/v)，质谱采纳TSI离子源接口，离子源和四极杆温度分离为250℃和100℃，SIM模式检测。
办法LOD小于1ng/g，回收率为93%～102%。
2)大气压化学电离源(APCI) APCI借助电晕放电启动气相离子化，形成单电荷的准分子离子，适合测定极性较小的化合物。
Sams等讨论了液相色谱-大气压化学电离-质谱(LC-APCI-MS)办法检测禽肉和蛋中DMZ、RNZ及其共同代谢物MNZOH。
色谱柱为Prodigy ODS3柱(250mm×3.2mmid，5μm)，流淌相为-0.05mol/L乙酸铵(13+87，v/v)，流速0.5mL/min，电晕放电电压3.22kV，高压透射电压(lensvoltage)0.0V，锥孔电压10V，锥孔电压偏移距5V，源温140℃，APCI探针温度500℃，正离子、SIM模式检测，每种物质检测2个离子。
办法LOD为0.1μg/kg(DMZ、RNZ)和0.5μg/kg(MNZOH)；平均回收率为65%(DMZ)、87%(RNZ)和75%(MNZOH)，RSD分离为22%、11%和14%。