银黄胶囊HPLC特征图谱和6个成分含量测定

HPLC Characteristic Chromatogram and Content Determination of Six Components for Yinhuang Capsules

关键词:  银黄胶囊  新绿原酸  绿原酸  隐绿原酸  3-5-O-二咖啡酰基奎宁酸  黄芩苷  黄芩素  高效液相色谱法  特征图谱

Key words:  Yinhuang capsules  neochlorogenic acid  chlorogenic acid  cryptochlorogenic acid  3-5-O-dicaffeoylquinic  baicalin  baicalein  HPLC  characteristic chromatogram

摘要:

目的 采用HPLC建立银黄胶囊的特征图谱，并同时测定6个成分的含量。方法 采用Agilent ZORBAX SB-C₁₈（250 mm×4.6 mm，5 μm），柱温30℃，以乙腈-0.4%乙酸溶液为流动相，梯度洗脱，流速为1.0 mL·min^-1，检测波长为327 nm，进样量10 μL，对20批银黄胶囊进行特征图谱研究，采用“中药色谱指纹图谱相似度评价系统（2012.130723版本）”对结果进行分析，并对色谱峰进行指认。结果 建立了银黄胶囊的HPLC特征图谱，指认了其中的6个特征峰，并对20批银黄胶囊中6个成分进行含量测定。新绿原酸、绿原酸、隐绿原酸、3-5-O-二咖啡酰基奎宁酸、黄芩苷和黄芩素质量浓度分别在0.638 7~6.387 μg·mL^-1（r=0.999 3）、2.893~28.93 μg·mL^-1（r=0.999 8）、0.664 1~6.641 μg·mL^-1（r=0.999 7）、0.743 1~7.431 μg·mL^-1（r=0.999 6）、16.19~161.9 μg·mL^-1（r=0.999 7）和2.143~21.43 μg·mL^-1（r=0.999 7）内线性关系良好，平均加样回收率（n=6）分别为100.19%（RSD=1.76%）、101.14%（RSD=1.29%）、98.86%（RSD=1.20%）、98.05%（RSD=0.99%）、99.38%（RSD=1.92%）、99.70%（RSD=1.92%），20批银黄胶囊中上述6个成分含量分别为0.637 9~4.100，3.069~15.28，0.960 4~4.906，0.625 9~4.440，82.69~118.2，2.118~14.76 mg·g^-1。结论 所建立的HPLC特征图谱专属性强，结合6个主要成分含量测定能够为银黄胶囊的质量控制提供科学依据。

Abstract:

OBJECTIVE To establish HPLC characteristic chromatogram and simultaneous determination of six components for Yinhuang capsules. METHODS The HPLC analysis was carried out on a column of Agilent ZORBAX SB-C₁₈ (250 mm×4.6 mm, 5 μm) with the temperature of 30℃. The mobile phase was consisted of acetonitrile-0.4% acetic acid with gradient at the flow rate of 1.0 mL·min^-1, the detection wavelength was 327 nm and the injection volume was 10 μL. Characteristic chromatograms of 20 batches of Yinhuang capsules were studied and evaluated by "Similarity Evaluation System for Chromatographic Fingerprint of TCM Software (2012. 130723 Version)", and the characteristic peaks in the fingerprint were identified by standard substances. RESULTS The HPLC characteristic chromatograms for 20 batches of the samples were established, and six characteristic peaks were indentified. And the method of simultaneous determination for six components was established. The linear ranges of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 3-5-O-dicaffeoylquinic, baicalin and baicalein were 0.638 7-6.387 μg·mL^-1(r=0.999 3), 2.893-28.93 μg·mL^-1(r=0.999 8), 0.664 1-6.641 μg·mL^-1(r=0.999 7), 0.743 1-7.431 μg·mL^-1(r=0.999 6), 16.19-161.9 μg·mL^-1(r=0.999 7) and 2.143-21.43 μg·mL^-1(r=0.999 7), respectively. The average recoveries(n=6) of six components were 100.19% (RSD=1.76%), 101.14% (RSD=1.29%), 98.86% (RSD=1.20%), 98.05% (RSD=0.99%), 99.38% (RSD=1.92%) and 99.70% (RSD=1.92%), respectively. The contents of six components in 20 batches of Yinhuang capsules were 0.637 9-4.100, 3.069-15.28, 0.960 4-4.906, 0.625 9-4.440, 82.69-118.2 and 2.118-14.76 mg·g^-1, respectively. CONCLUSION The proposed specific HPLC characteristic chromatograms and quantitation method of six components can provide scientific basis for quality control of the Yinhuang capsules.