黄体酮注射液细菌内毒素检查的方法比较研究

目的： 建立黄体酮注射液细菌内毒素检查的方法，探讨油剂的细菌内毒素检查方法。 方法： 将黄体酮注射液与水混合振荡，将内毒素从注射用油萃取到水中进行测定。考察水萃取液的干扰及水油振荡萃取步骤中内毒素的绝对回收率。 结果： 黄体酮注射液与水振荡后，水萃取液对内毒素检查无干扰。振荡萃取步骤中外加20、10、5 EU·mL^-1浓度的内毒素回收率在78.5%~117.4%之间，符合药典要求。 结论： 黄体酮注射液作为油剂，经水萃取后采用动态浊度法和凝胶法进行细菌内毒素检查均可行。

Objective: To set up the bacterial endotoxin test for progesterone injection and study what should be done to confirm the method is suitable for the determination of bacterial endotoxins in oily solutions. Methods: Since the progesterone injection is an oily solution,the oily injection was shaken with water.The endotoxins were then extracted in the aqueous phase from the oil-based solutions without interference(inhibition/enhancement)of the product and the recovery of the endotoxins added into the oily-injection was determined. Results: The validation test demonstrated that the endotoxins in oily injection passed over into the aqueous phase was not influenced(inhibited/enhanced)by extraction with water.For oily injection with concentrations of 20,10,5 EU·mL^-1 of endotoxin,78.5%-117.4% of the endotoxin was recovered after extraction,which met the requirements of the Pharmacopoeia. Conclusion: The kinetic turbidimetric limulus test and the gel-clot test were suitable for detecting bacterial endotoxins in progesterone injection.

[1] ChP 2010.Vol Ⅱ(中国药典2010年版.二部)[S].2010:Appendix(附录)Ⅺ E 99
[2] HU Yu-chi(胡宇驰),LI Na(李娜),ZHOU Jian- ping(周建平).Study on gelation clot method of bacterial endotoxin test in testorerone undecanoate injection(十一酸睾酮注射液细菌内毒素检查方法学研究)[J].Chin J Pharm Anal(药物分析杂志),2010,30(12):2425
[3] PAN Wei-song(潘卫松),XIAO Ying(肖瑛).Study on the test method of bacterial endotoxin in refined olive oil(精制橄榄油细菌内毒素检查的可行性研究)[J].China Med Pharm(中国医药科学),2011,1(8):10
[4] LI Rong(李荣),WU Ling(吴玲).Determination of bacterial endotoxin for levofloxacin lactate/sodium chloride injection by kinetic turbidimetric limulus test(动态浊度法检测乳酸左氧氟沙星氯化钠注射液中的细菌内毒素)[J].China Pharm(中国药房),2012,23(9):837
[5] LU Wen-bin(卢文斌).Detection of bacterial endotoxin in Huangqi injection by the method of kinetic turbidimetric limulus test(动态浊度法定量测定黄芪注射液中细菌内毒素的含量)[J].Chin J Mod Appl Pharm(中国现代应用药学),2012,29(3):271
[6] XIAO Gui-nan(肖贵南),GUAN Yong-yuan(关永源),WU Zhao-di(吴招娣),et al. Analysis of endotoxin-specific TAL reagent on bacterial endotoxins in mannatide sodium chloride injection(特异性鲎试剂分析甘露聚糖肽氯化钠注射液的细菌内毒素)[J].Chin Pharmacol Bull(中国药理学通报),2003,19(9):1076
[7] YAO Zhi(姚治),NI Wei-fang(倪维芳),ZHU She-min(朱社敏),et al.Detection of bacterial endotoxin in iohexol by the method of kinetic turbidimetric limulus test(动态浊度法定量测定碘海醇中细菌内毒素的含量)[J].Chin J Mod Appl Pharm(中国现代应用药学),2009,26(2):150
[8] WANG Lu(王璐),ZHANG Hong-yu(张红宇),LIU Qin(刘勤).Study on bacterial endotoxins test for breviscapine injection by gel clot techniques and photometric techniques(凝胶法和光度法测定灯盏花素注射液中细菌内毒素的含量)[J].J Kunming Med Univ(昆明医学院学报),2010,31(2):39
[9] GUO Meng(郭萌),LI Guan-min(李冠民),HUANG Qing-quan(黄清泉).Advances in research of bacterial endotoxin(细菌内毒素研究进展)[J].Acta Lab Anim Sci Sin(中国实验动物学报),2009,17(5):397
[10] ZHENG Xia(郑霞),ZHAO Yu-mei(赵毓梅).Advance in the research of determination of bacterial endotoxin(细菌内毒素检查法的应用进展)[J].China Trop Med(中国热带医学),2009,9(1):170
[11] Yamamoto A,Ochiai M,Fujiwara H,et al.Evaluation of the applicability of the bacterial endotoxin test to antibiotic products[J].Biologicals,2000,28(3):155
[12] Fingola FF,Albertino SR,Abrantes Sde M,et al.Intral aboratory validation of kinetic chromogenic Limulus amebocytelysate assay for bacterial endotoxin determination in anti-bothropic serum[J].J Pharm Biomed Anal,2013,85:93