恤彤注射液中间体丹参提取液的指纹图谱分析及多指标含量测定

Fingerprint and Multi-component Quantification Analysis of Salvia Miltiorrhiza Extract Intermediate from Xutong Injection

关键词:  恤彤注射液  丹参提取液  指纹图谱  含量测定  5-羟甲基糠醛  酚酸类物质

Key words:  Xutong injection  Salvia miltiorrhiza extract  fingerprint  content determination  5-HMF  phenolic acids

摘要:

目的 建立恤彤注射液中间体丹参提取液的HPLC-UV指纹图谱，并对其中的5-羟甲基糠醛、丹参素、原儿茶醛、迷迭香酸、紫草酸、丹酚酸A及丹酚酸B共7种指标成分进行定量分析。方法 采用Waters Cortecs C₁₈色谱柱（100 mm×4.6 mm，2.7 μm），流动相0.4%甲酸水溶液（A）-0.4%甲酸乙腈（B），流速0.8 mL·min^-1，梯度线性程序洗脱，进样量3 μL，柱温25℃，检测波长280 nm。结果 在建立的色谱条件下，指纹图谱峰型对称，分离度较好，能较全面地反映丹参提取液的化学信息。共确定了24个共有峰，10批丹参提取液的指纹图谱相似度均>0.960；在一定质量范围内，7种指标成分的峰面积与质量浓度均呈良好的线性关系，加样回收率为96.49%~100.18%，RSD为1.24%~2.27%（n=9），所测定的7种指标成分总量为8 202.2~12 078.7 μg·g^-1。结论 建立的分析方法准确、简单、重复性好，HPLC-UV指纹图谱结合定量测定能更全面地反映丹参提取液的质量，可用于恤彤注射液中间体丹参提取液的质量评价。

Abstract:

OBJECTIVE To establish the HPLC-UV fingerprint of Salvia miltiorrhiza extract intermediate from Xutong injection, and simultaneously to determine the contents of 7 indicators including 5-HMF, Danshensu, protocatechualdehyde, rosmarinic acid, lithospermic acid, salvianolic acid A, salvianolic acid B. METHODS The separation was achieved on a Waters Cortecs C₁₈ column(100 mm×4.6 mm, 2.7 μm) with a flow rate of 0.8 mL·min^-1 in line gradient mode at 25℃. The 0.4% formic acid solution-0.4% formic acid acetonitrile were used as mobile phases A and B, respectively. The sample injection volume was set at 3 μL and detection wavelength was 280 nm. RESULTS The established fingerprint with good resolution and symmetrical peak shape could comprehensively reflect the chemical information of Salvia miltiorrhiza extract. The 24 common peaks were identified and the fingerprint similarity among 10 batches of extract was >0.960. Within a certain mass range, the peak area and mass concentration of 7 indicators had a good linear relationship. The average recovery of 7 kinds of compounds was 96.49%-100.18% and the RSD was 1.24%-2.27%(n=9), the total amount of the 7 indicators in the 10 batches of Salvia miltiorrhiza extract ranged was 8 202.2-12 078.7 μg·g^-1. CONCLUSION The established HPLC-UV method is accurate, simple and reproducible. HPLC-UV fingerprint combined with quantitative determination of indicators can more fully reflect the quality of Salvia miltiorrhiza extract to provide reference for quality evaluation of Salvia miltiorrhiza extract immediate from Xutong injection.