柴胡疏肝汤高效液相指纹图谱的建立及含量测定

Establishment of HPLC Fingerprint of Chaihushugan Decoction and Content Determination

目的建立柴胡疏肝汤高效液相指纹图谱方法,并同时测定柴胡皂苷a、汉黄芩苷、芍药苷的含量,为该方质量控制提供参考。方法采用RP-HPLP-DAD法,利用ZORBAX SB-C8色谱柱(250 mm×4.6 mm,5μm),在检测波长210nm、柱温25℃下,以乙腈(A)-0.2%磷酸水溶液(B)为流动相进行梯度洗脱。对10批柴胡疏肝汤进行指纹图谱及柴胡皂苷a、汉黄芩苷、芍药苷含量测定研究,采用"中药色谱指纹谱图相似度评估软件"(2004A)进行分析。结果建立了柴胡疏肝汤指纹图谱共有模式,标定22个共有峰,10批柴胡疏肝汤相似度较好(均大于0.96);通过与标准品的保留时间和紫外光谱图对比,确定了7个成分;柴胡皂苷a、汉黄芩苷、芍药苷重复性、稳定性良好,含量测定结果可靠。结论指纹图谱的建立和多成分含量测定,可以为该方提供较全面的质量控制依据。

Objective To establish the fingerprint of Chaihushugan decoction and establish a method for the simuhaneous determination of Saikosaponin a, wogonoside and paeoniflorin , to provide a quality evaluation method for the decoction. Methods The fingerprint was established by HPLC with Diode Array Detector （DAD） and ZORBAX SB - Cs column （ 250 mm × 4.6 mm, 5 μm） , gradient eluted with acetonitrile -0.2% phosphoric acid as mobile phase. The detection was at 210 nm, and column temperature was at 25 ℃. Ten batches of Chaihushugan decoction were analyzed by HPLC fingerprints and the content of determination methods. Data were analyzed by using Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （Version 2004A）. Results The common pattern of chromatographic fingerprint of Chaihushugan decoction was established, 22 common peaks were identified. The similarity of the batches of Chaihushugan decoction were more than 0.96. Seven components were identified by comparison with the retention times and UV spectra of standard substances. The repeatability and stability of Saikosaponin a, wogonoside and paeoniflorin were good, and the results of content determination were reliable. Conclusion The method of established HPLC fingerprint and quantitative analysis of components were established, which could reflect the overall characteristics of decoction and improve the specificity for quality control of the herbal formula.