虎力散片中乌头类生物碱成分HPLC特征图谱及多成分含量测定

Study on Specific Chromatogram and Multi-components Determination of Alkaloids in Hulisan Tablets by HPLC

关键词:  虎力散  特征图谱  制草乌  生物碱  含量测定

Key words:  Hulisan  specific chromatogram  Radix Aconiti Kusnezoffii Preparata  alkaloid  content determination

摘要:

目的 建立虎力散片中乌头类生物碱的HPLC特征图谱，并同时测定6种生物碱含量。方法 采用Kromasil 100-5-C₁₈色谱柱（4.6 mm×250 mm，5 μm），以乙腈（A）-0.2%乙酸溶液（三乙胺调pH至6.20）（B）为流动相，梯度洗脱，流速1.0 mL·min^–1，柱温35℃，检测波长235 nm。采用中药色谱指纹图谱相似度评价系统对结果进行分析，并结合聚类分析、主成分分析等模式识别技术对样品的总体质量进行分析评价，并对主要有效成分进行定量测定。结果 11批虎力散片的乌头类生物碱特征图谱中共确认10个共有峰，与混合对照品比较确定6个生物碱：苯甲酰新乌头原碱、苯甲酰次乌头原碱、苯甲酰乌头原碱、新乌头碱、次乌头碱、乌头碱。来自3个厂家的11批虎力散片中苯甲酰新乌头原碱、苯甲酰次乌头原碱、苯甲酰乌头原碱、新乌头碱、次乌头碱、乌头碱的含量分别在231.190~1 002.977，17.022~105.744，21.962~48.406，5.436~39.678，2.692~97.854，1.707~43.342 μg·g^–1。结论 所建立的HPLC特征图谱方法和多成分含量测定方法，简便可行，结果准确可靠，重现性好，可有效评价虎力散片的质量。

Abstract:

OBJECTIVE To establish the HPLC specific chromatogram of aconitum alkaloids in Hulisan tablets and simultaneously determine the contents of 6 alkaloids. METHODS Kromasil 100-5-C₁₈ column(4.6 mm×250 mm, 5 μm) was used with gradient elution of acetonitrile(A)-0.2% acetic acid(triethylamine adjusted pH to 6.20)(B) at the flow rate of 1.0 mL·min^-1. The column temperature was 35℃ and the detection wavelength was 235 nm. The similarity evaluation system for chromatographic fingerprint of TCM was used to calculate the similar degree. Then, the further quality assessment of the drug was carried out by similarity evaluation combined with CA and PCA. And the contents of the active ingredients of sample were determined. RESULTS Ten common peaks were identified in the aconitum alkaloids characteristic chromatogram of 11 batches of Hulisan tablets. And 6 alkaloids were identified as benzoylmesaconitine, benzoylhypaconitine, benzoylaconitine, mesaconitine, hypaconitine and aconitine. The contents of benzoylmesaconitine, benzoylhypaconitine, benzoylaconitine, mesaconitine, hypaconitine and aconitine in 11 batches of Hulisan tablets from three manufacturers were 231.190-1 002.977, 17.022-105.744, 21.962-48.406, 5.436-39.678, 2.692-97.854 and 1.707-43.342 μg·g^-1. CONCLUSION The established HPLC specific chromatogram method and multi-component content determination method is simple, reliable, accurate and can effectively evaluate the quality of Hulisan tablets.