经典名方黄连汤UPLC指纹图谱及6种成分含量测定研究

Establishment of the UPLC fingerprint of classical herbal prescription Huanglian Decoction and determination of the contents of 6 components

目的:建立15批黄连汤物质基准UPLC指纹图谱同时测定盐酸小檗碱、盐酸巴马汀、桂皮醛、6-姜辣素、甘草苷、甘草酸的含量,为黄连汤的质量标准研究提供参考。方法:利用ACQUITY UPLC?CSHTM C18(100 mm×2.1 mm,1.7μm)色谱柱,乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱,建立了15批黄连汤物质基准指纹图谱,利用"中药色谱指纹图谱相似度评价系统"2012年版对15批黄连汤指纹图谱进行相似度评价。结果:采集了15批黄连汤物质基准指纹图谱,其相似度均>0.9,标定了26个共有峰,指认了12个专属峰。盐酸小檗碱、盐酸巴马汀、甘草酸铵、甘草苷、6-姜辣素和桂皮醛的平均加样回收率分别为100.56%,101.51%,102.41%,103.76%,97.66%和96.74%,RSD均<3%,各成分在相应浓度范围内线性关系良好(R2>0.9999),15批黄连汤物质基准含量测定范围:盐酸小檗碱、盐酸巴马汀、甘草酸、甘草苷、6-姜辣素和桂皮醛质量分数分别为13.58~23.29,3.46~4.69,2.14~6.74,1.61~4.52,0.83~1.46和1.61~6.67 mg·g^(-1)。结论:建立的黄连汤指纹图谱方法简便快捷、含量测定准确,为经典名方黄连汤物质基准的质量评价及深入研究奠定实验基础。

Objective:To establish the UPLC fingerprints of 15 batches of Huanglian Decoction standard preparation and determine the contents of berberine hydrochloride,palmatine hydrochloride,cinnamic aldehyde,6-gingerol,glycyrrhizin and glycyrrhizic acid simultaneously by UPLC,providing reference for the quality standard research of Huanglian Decoction.Methods:ACQUITY UPLC?CSHTM C18(100 mm×2.1 mm,1.7μm)column used with acetonitrile(A)-0.1%phosphoric acid aqueous solution(B)as the gradient elution mobile phase to establish the fingerprints of 15 batches of Huanglian Decoction standard preparation,and the similarity evaluation of the fingerprints of 15 batches of Huanglian Decoction was carried out using the"Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System"(2012).Results:The fingerprints of 15 batches of Huanglian Decoction standard preparation were collected,and their similarities were all>0.9;26 common peaks were calibrated,and 12 exclusive peaks were identified.The average recovery rates of berberine hydrochloride,palmatine hydrochloride,ammonium glycyrrhizinate,glycyrrhizin,6-gingerol and cinnamaldehyde were 100.56%,101.51%,102.41%,103.76%,97.66%and 96.74%,respectively.The RSDs were all<3%.The linear relationship of the calibration curve of each component was good within the corresponding concentration range(R2>0.9999).The mass fractions of berberine hydrochloride,palmatine hydrochloride,glycyrrhizic acid,glycyrrhizin,6-gingerol and cinnamaldehyde in 15 batches of Huanglian Decoction standard preparation were determined as 13.58 to 23.29,3.46 to 4.69,2.14 to 6.74,1.61 to 4.52,0.83 to 1.46 and 1.61 to 6.67 mg·g^(-1),respectively.Conclusion:The established fingerprint method of Huanglian Decoction is simple,rapid and accurate,which lays an experimental foundation for the quality evaluation and in-depth study of the material reference of the classic formula Huanglian Decoction.