OBJECTIVE: To set up the method for quantitative determination of bacterial endotoxin in 4 kinds of heat-clearing and detoxifying TCM injections. METHODS: The kinetic chromogenic assay (KCA) method was used. The determination conditions included detection temperature (37±0.5) ℃, detection wavelength 405 nm, preset absorbance (A) 0.1, and reaction time 3 600 s. According to photometry method and guidance principle general rule of bacterial endotoxin test in 2015 edition of Chinese Pharmacopoeia (part Ⅳ), reliability test and interference test of standard curve were conducted. The non-interference concentration of 4 type (Qingkailing injection, Compound Banbianlian injection, Yinzhihuang injection and Shuanghuanglian injection) was determined and quantitative determination of bacterial endotoxin was also conducted. RESULTS: The correlation coefficient r of standard curve was 0.999. Qingkailing injection, Compound Banbianlian injection, Yinzhihuang injection and Shuanghuanglian injection didn’t interfere with the detection of tachypleus amebocyte lysate in 200, 125, 75 and 12.5 times and below dilution times respectively. The recoveries were 50%-200%, and the contents of bacterial endotoxin in samples were 1.66, 1.04, 0.62, 0.104 EU/mL, which were all lower than the limits of samples. CONCLUSIONS: KCA is suitable for the detection of bacterial endotoxin in 4 kinds of TCM injections. The method is simple, convenient, stable and reliable.
关键词
动态显色法清热解毒类中药注射液细菌内毒素干扰试验
Keywords
Kinetic chromogenic assayHeat-clearing and detoxifyingTCM injectionBacterial endotoxinInterference test