黄芩汤指纹图谱建立及不同相态抗皮肤癣菌活性谱效关系研究
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篇名: 黄芩汤指纹图谱建立及不同相态抗皮肤癣菌活性谱效关系研究
TITLE: Fingerprint establishment of Huangqin decoction and study on spectrum-effect relationship of its antidermatophytic activity in different phase states
摘要: 目的 建立黄芩汤(HQD)的指纹图谱,拆分相态并筛选抗皮肤癣菌活性相态,进行谱效关系研究。方法采用高效液相色谱法,以黄芩苷为参照峰,采用《中药色谱指纹图谱相似度评价系统(2012版)》绘制10批HQD的指纹图谱并进行相似度评价,确定共有峰;采用高速离心与透析相结合的方法对HQD进行相态拆分并对其进行表征,同时测定HQD及其不同相态抗皮肤癣菌的最小抑菌浓度(MIC);以37个共有峰的峰面积为自变量,MIC为因变量,采用SPSS21.0软件进行Pearson相关性分析。结果10批HQD共有37个共有峰,相似度均大于0.99;共指认出10个成分,分别为芍药内酯苷、芍药苷、芹糖甘草苷、黄芩苷、千层纸素A苷、汉黄芩苷、黄芩素、甘草酸、汉黄芩素、千层纸素A。HQD共拆分成沉淀相态(HQD-P)、真溶液相态(HQD-S)和纳米相态(HQD-N)3个相态;HQD-P冻干粉呈不规则颗粒状,平均粒径为4.670~91.522μm;HQD-S冻干粉呈均匀片状,未检测到粒径;HQD-N冻干粉呈球形,粒径为(129.0±12.9)nm。在同等给药剂量下,各相态抗皮肤癣菌MIC由低到高依次为HQD-N(4.64mg/mL)<HQD(5.85mg/mL)<HQD-P(7.37mg/mL)<HQD-S(12.89mg/mL)。Pearson相关分析结果显示,37个共有峰中,有25个共有峰(含已指认的10个成分)峰面积与MIC呈显著负相关(相关系数绝对值>0.95且P<0.05)。结论10批HQD的化学成分组成一致性较好;HQD-N为该方的活性相态,其中芍药苷、芹糖甘草苷、黄芩苷等10个成分可能是HQD抗皮肤癣菌的有效成分,且其抗皮肤癣菌作用强弱与这些成分含量及物理相态相关。
ABSTRACT: OBJECTIVE To establish the fingerprint of Huangqin decoction (HQD), to separate the phase states and screen the active phase states of antidermatophytic activity so as to study the spectrum-effect relationship. METHODS HPLC method was adopted using baicalin as reference, the fingerprints of 10 batches of HQD were drawn and the similarity evaluation was carried out using the Similarity Evaluation System of Chromatographic Fingerprint of TCM (2012 edition) to determine the common peak; the phase states of HQD were separated and characterized by high-speed centrifugation and membrane dialysis. The minimum inhibitory concentrations (MIC) of HQD and its different phase states against Trichophyton mentagrophytes were determined simultaneously. Using the peak area of 37 common peaks as independent variable, MIC as dependent variable, Pearson correlation analysis was performed by using SPSS 21.0 software. RESULTS A total of 37 common peaks were obtained in HPLC fingerprints of 10 batches of HQD, with the similarity higher than 0.99. Ten components were identified, such as albiflorin, paeoniflorin, liquiritin apioside, baicalin, melaleuca glycoside A, wogonoside, baicalein, glycyrrhizic acid, wogonin and oroxylin A. HQD was split into 3 phase states, such as precipitation phase (HQD-P), solution phase (HQD-S) and nano phase (HQD-N). The morphology of HQD-P was irregular granular, and the average particle size was 4.670-91.522 μm. The morphology of HQD-S was uniform flakes, and no particle size was detected. HQD-N was spherical in shape and the particle size was (129.0±12.9) nm. MIC values of each phase state of HQD against T. mentagrophytes in different phase states were HQD-N (4.64 mg/mL) <HQD (5.85 mg/mL) <HQD-P (7.37 mg/mL) <HQD-S (12.89 mg/mL) at the same dosage. Pearson correlation analysis showed that the peak area of 25 of the 37 common peaks (including identified components) was significantly negatively correlated with MIC (absolute values of correlation coefficient>0.95 and P<0.05). CONCLUSIONS The chemical composition of 10 batches of HQD is consistent; HQD-N is the active phase state of HQD. Ten components such as paeoniflorin, liquiritin apioside and baicalin may be the main active components of HQD. The antidermatophytic effect of HQD is closely related to its component content and physical phase state.
期刊: 2023年第34卷第06期
作者: 沈成英;吴文明;侯雄军;邓冯沂;何洁;胡建新
AUTHORS: SHEN Chengying,WU Wenming,HOU Xiongjun,DENG Fengyi,HE Jie,HU Jianxin
关键字: 黄芩汤;指纹图谱;相态;抗皮肤癣菌作用;谱效关系
KEYWORDS: Huangqin decoction; fingerprint; phase state; antidermatophytic activity; spectrum-effect relationship
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