目的 建立茵陈蒿汤(Yinchenhao Decoction,YCHD)含药血清指纹图谱,并对含药血清中药源性成分进行分析。方法 高效液相色谱法,采用Symmetry C18(4.6 mm×250 mm,5 μm)色谱柱,Security Guard Cartridges C18(4.0 mm×3.0 mm)保护柱,流动相为乙腈-0.1%磷酸水梯度洗脱,流速1 mL·min-1,柱温30 ℃,检测波长238和440 nm;以SD大鼠为供体,灌胃给药制备10批茵陈蒿汤含药血清,测定含药血清指纹图谱,建立指纹图谱共有模式,并进行相似度评价;与空白血清指纹图谱比对,指认含药血清中的药源性成分;与各单味药含药血清指纹图谱比对,分析各药源性成分来源;与体外指纹图谱、对照品指纹图谱色谱峰保留时间及紫外光谱图比对,对各药源性成分进行鉴别。结果 238 nm和440 nm下10批茵陈蒿汤含药血清指纹图谱与共有模式的相似度分别≥0.904和0.903;两个波长下茵陈蒿汤含药血清指纹图谱标定药源性成分共有峰20个,其中6个为体外原形成分,14个为体内代谢产物;6个原形成分中的3个分别为来源于栀子的环烯醚萜类成分京尼平苷酸、栀子苷和来源于大黄的蒽醌类成分大黄酸,另外3个中的1个为来源于大黄的蒽醌类成分,2个为来源于栀子的西红花酸类成分;14个代谢产物中的1个为环烯醚萜类成分代谢产物,4个为蒽醌类成分代谢产物,9个为西红花酸类成分代谢产物。结论 本实验建立的方法准确、可靠,可用于茵陈蒿汤含药血清中药源性成分分析。
Abstract
OBJECTIVE To establish the fingerprint of the serum containing drug of Yinchenhao Decoction (YCHD), and analyze the drug-originated constituents in serum. METHODS An HPLC method was set up on a Symmetry C18 (4.6 mm×250 mm, 5 μm) column with a Security Guard Cartridges C18 (4.0 mm×3.0 mm) guard column by gradient elution of acetonitrile-0.1% phosphoric acid in water at the flow rate of 1.0 mL·min-1. The column temperature was maintained at 30 ℃. The detection was carried out at 238 and 440 nm. SD rats were taken as serum donors. Ten batches of serum containing drugs were prepared after ig administration of YCHD. The fingerprints of these serum samples were determined, the common modes were established, and the similarities between fingerprints were evaluated. Drug-originated constituents in blood were distinguished by comparing the fingerprint of serum containing YCHD with the one of control serum. The sources of drug-originated constituents were analyzed by comparing the fingerprint of serum containing YCHD with the ones of serum containing individual herbs of YCHD. Drug-originated constituents in blood were identified by comparing the retention time and UV spectra of peaks in fingerprint of serum containing YCHD with the ones of YCHD and reference substances, respectively. RESULTS The similarities between the fingerprints of ten batches of serum containing YCHD and common models at 238 and 440 nm were greater than 0.904 and 0.903, respectively. Twenty common peaks of drug-originated constituents were identified in the fingerprints of serum containing drug of YCHD at 238 and 440 nm. Out of them, six were the original compounds, the other 14 were metabolites. Three constituents out of six original form ones were identified as geniposidic acid and geniposide (iridoids originated from GF), and rhein (anthraquinone from RRR), and the other three were identified as an anthraquinone from RRR and two crocetin derivatives from FG. Out of 14 metabolites, one was identified as the metabolite of iridoid, four were from anthraquinone, and nine were from crocetin derivatives. CONCLUSION The established method is accurate, reliable and can be used for the analysis of drug-originated constituents in serum containing drug of YCHD.
关键词
茵陈蒿汤 /
含药血清 /
高效液相色谱法 /
指纹图谱 /
血中药源性成分 /
茵陈 /
栀子 /
大黄
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Key words
Yinchenhao decoction (YCHD) /
serum containing drug /
HPLC /
fingerprint /
drug-originated constituents in blood /
Artemisiae Scopariae Herba (ASH) /
Gardeniae Fructus (GF) /
Rhei Radix et Rhizoma (RRR)
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中图分类号:
R284
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参考文献
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脚注
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基金
江苏省中医药局科技资助项目(HZ07071);江苏省高校自然科学基金资助项目(08KJB360009)
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