茵陈蒿汤HPLC指纹图谱研究及共有峰归属分析

Study on HPLC Fingerprints of Yinchenhao Decoction and Attributive Analysis of Common Peaks

目的建立茵陈蒿汤HPLC指纹图谱,并对其共有峰进行归属分析和指认。方法 Symmetry C18(4.6mm×250mm,5μm)色谱柱,乙腈-0.1%磷酸梯度洗脱,流速1.0mL/min,柱温30℃,检测波长238nm和440nm。测定10批茵陈蒿汤样品指纹图谱,建立共有模式,并进行相似度评价,通过与各组方药味及对照品保留时间和紫外光谱图比对,对共有峰进行归属分析和指认。结果 238nm和440nm下10批样品指纹图谱与共有模式的相似度分别≥0.931和0.951;238nm下标定共有峰56个,其中24个来源于大黄、27个来源于栀子、4个来源于茵陈、1个为栀子和茵陈共有。来源于大黄的10个成分被指认,另有8个和2个成分分别具有蒽醌类和儿茶素类化合物的紫外吸收特征;来源于栀子中的4个成分被指认,另外10个、1个和3个成分分别具有环烯醚萜类化合物、西红花酸类化合物和绿原酸类似物的紫外吸收特征;来源于茵陈和栀子共有成分被指认为绿原酸;来源于茵陈的2个成分具有绿原酸类似物的紫外吸收特征;440nm下标定共有峰32个,其中16个与238nm下为同一成分,另外16个新成分均来源于栀子,且均具有西红花酸类化合物的紫外吸收特征。结论该法建立的指纹图谱可较全面地对茵陈蒿汤进行成分分析,为该方剂质量控制提供了依据。

OBJECTIVE To establish HPLC fingerprints of Yinchenhao decoction （YCHD） and to analyze and identify their common peaks. METHODS HPLC was performed on a Symmetry C18 （250 min× 4.6 mm, 5 μm） column and detected at 238 nm and 440 nm by gradient elution at the flow rate of 1.0 mL/min by using acetonitrile-0.1% phosphoric acid in water as the mobile phase. Column temperature was 30 ℃. Fingerprints of 10 batches of YCHD were determined, the common modes were established and the similarities between fingerprints were evaluated. Attributive analysis and identification of common peaks were performed by comparing the retention time and UV spectra among 10 batches of YCHD, the formula ingredients and ref- erence substances. RESULTS Similarities between fingerprints of 10 batches of YCHD and common modes at 238 nm and 440 nm were ≥0.931 and 0.951, respectively. 56 common peaks were determined in fingerprints at 238nm, 24, 27 and 4 of them were originated from Rhei Radix et Rhizome （RRR）, Fructus Gardeniae （FG） and Artemisiae Capillaris （ AC）, respective- ly. Other 1 was originated from FG and AC in common. 10 components from RRR were determined and identified, while an other 8 and 2 components displayed ultraviolet absorption characteristics of anthraquinones and catechins compounds, respec- tively. 4 components from FG were identified, while another 10, 1 and 3 showed ultraviolet absorption characteristics of iri- doids, crocetin and chlorogenic acid analogues, respectively. The component from FG and AC in common was identified as chlorogenic acid. 2 components from AC exhibited the similar ultraviolet absorption characteristics of chlorogenic acid. 32 com- mon peaks were detected in fingerprints at 440 nm, including 16 same ones at 238 nm. 16 new ones were all originated from FG, which showed similar ultraviolet absorption characteristics of rocetin derivatives. CONCLUSION The established finger-prints can make comprehensive analysis on the chemical composition of YCHD and provide the basis for the quality control.