HPLC法和生物测定法测定鲑鱼降钙素肠溶胶囊含量的比较

目的:采用高效液相色谱(HPLC)法测定鲑鱼降钙素肠溶胶囊中降钙素的含量,并与生物测定法进行比较。方法:HPLC法色谱柱为依利特ODS2 C18,流动相为0.4%四丁基氢氧化铵水溶液(pH 3.8)-乙腈(334∶166,V/V),流速为1 ml/min,检测波长为220 nm;生物测定法参照《中国药典》相关方法进行。结果:HPLC法降钙素检测质量浓度线性范围为5²00μg/ml(r=0.999 3),辅料对主药测定无干扰,精密度试验RSD<5%,平均回收率为96.5%,含量测定结果为99.96%;生物测定法可信限率为28.84%,虽符合要求(<45%),但含量测定结果为111.62%,估计与制剂中含吸收促进剂有关。结论:与生物测定法相比,HPLC法测定鲑鱼降钙素肠溶胶囊含量,操作简便、成本低、重复性好,结果更为可靠。

Comparison of HPLC and Bioassay Method for Content Determination of Salmon Calcitonin Enteric-coated Capsules

OBJECTIVE： To adopt HPLC method to determine the content of calcitonin in Salmon calcitonin enteric-coated cap- sule, and to compare with the results of biological assay. METHODS： Hypersil ODS2 C18 column was used with mobile phase con- sisted of 0.40 tetramethylammonium hydroxide （pH 3.8）-acetonitrile （334：166, V/V） at the flow rate of 1 ml/min. The detection wavelength was set at 220 nm. The bioassay referred to related method stated in Chinese Pharmacopoeia. RESULTS： In HPLC as- say, the linear range of calcitonin were 5-200 μg/ml （r=0.999 3） without inference from excipients. RSD of precision test was low- er than 5%, and average recovery was 96.5%. Result of content determination was 99.96%. FL of biological assay was 28.84%, which was in line with the standard （〈45%）; but content determination result was 111.62%, which was associated with accelerant in the preparation. CONCLUSIONS： Compared with bioassay, HPLC assay for determining the content of calcitonin in Salmon calcitonin enteric-coated capsule is simple, low-cost, reproducible and reliable in results.