药用辅料抗坏血酸棕榈酸酯的细菌内毒素检查法研究

Detection of bacterial endotoxin in pharmaceutical excipient ascorbyl palmitate

目的:建立"三氯甲烷溶解后加水振荡萃取"抗坏血酸棕榈酸酯细菌内毒素检查方法。方法:抗坏血酸棕榈酸酯,加入三氯甲烷使溶解,加入不同体积的BET水,旋涡混匀30 s以上,5000 r·min^(-1)离心2 min,取上清液,制成供试品溶液和供试品阳性对照,抗坏血酸棕榈酸酯浓度在相当于1.0 mg·mL^(-1)以下时,对λ为0.25 EU·mL^(-1)的鲎试剂与细菌内毒素的反应均无干扰。结果:同法制备浓度为0.5 mg·mL^(-1)的供试品溶液,用上述供试品溶液和BET水各做阴性对照管,结果表明两个厂家的供试品在相当于0.5 mg·mL^(-1)或更低浓度时对2个不同厂家的鲎试剂与细菌内毒素的反应无干扰作用。结论:萃取后的水相溶液做内毒素检查能够较准确反映抗坏血酸棕榈酸酯中的内毒素含量,可为不溶于水的原料药和药用辅料进行细菌内毒素检查提供参考。

Objective:To establish a bacterial endotoxin test(BET)method of ascorbic palmitate(AP)by shaking extraction with water following AP dissolved by trichloromethane.Methods:After AP was dissolved in trichloroform,BET water was added at various ratios and vortexed for more than 30 s.After centrifugation at 5000 r·min^(-1)for 2 min,the supernatant was with drawn for the preparation of test solution and positive control.When the concentration of AP was less than 1.0 mg·mL^(-1),no interference was observed for the reaction between bacterial endotoxin andλ0.25 EU·mL^(-1)limulus lysate.Results:The test solution with concentration of 0.5 mg·mL^(-1)was prepared by the same method.The above test solution and BET water were used as corresponding negative control.The results showed that the test solutions of two manufacturers had no interference on the reaction of bacterial endotoxin with limulus lysate reagent of the two manufacturers when the concentration was 0.5 mg·mL^(-1)or lower.Conclusion:The endotoxin test of aqueous solution after extraction can accurately reflect the endotoxin content of AP,which can provide reference for bacterial endotoxin test of water-insoluble active pharmaceutical ingredients and pharmaceutical excipients.