[关键词]
[摘要]
目的 建立清心滋肾方水煎液(Qinxin Zishen Prescription Decoction,QZPD)的HPLC指纹图谱,并测定15种成分的含量,为清心滋肾方(Qinxin Zishen Prescription,QZP)的质量控制提供依据。方法 采用Phenomenex Kinetex C18(100 mm×4.60 mm,2.6 μm)色谱柱分离,流动相为甲醇、乙腈和0.2%甲酸水溶液,梯度洗脱,检测波长为245、280 nm,柱温40℃。建立10批QZPD的HPLC指纹图谱,并进行相似度评价、对共有峰进行归属及指认,并测定指认出的15个成分的含量。结果 10批QZPD HPLC指纹图谱相似度在0.923~0.998,指认出共有峰33个,其中黄连中有7个峰(P11、P14~P16、P24、P29、P30),莲子心(P7、P19)及酸枣仁(P14、P21)中各有2个峰,丹参(P4、P5、P10、P17、P18、P28、P31~P33)及山茱萸(P1~P3、P6、P8、P9、P12、P13、P20)中各有9个峰,其余5个共有峰(P22、P23、P25~P27)来源未明确,而生地黄、钩藤及浮小麦对共有峰贡献不明显。通过与对照品比对,指认出没食子酸(P2)、5-羟甲基糠醛(P3)、丹参素(P4)、原儿茶醛(P5)、莫诺苷(P9)、咖啡酸(P10)、马鞭草苷(P12)、马钱苷(P13)、木兰花碱(P14)、黄连碱(P24)、紫草酸(P28)、小檗碱(P29)、巴马汀(P30)、丹酚酸B(P31)和丹酚酸E(P33),并对这15种成分进行含量测定,定量结果分别为158.3~248.2、233.6~321.3、45.9~166.0、24.3~38.6、800.7~1 263.6、26.6~54.9、44.5~108.2、470.4~757.3、85.6~178.6、11.1~34.2、56.2~106.4、25.9~138.9、21.0~59.2、951.6~2 244.7、38.6~92.8 μg/g。结论 所建立的QZPD HPLC指纹图谱及定量测定方法稳定性、重复性好,可为QZP质量控制和评价提供参考。
[Key word]
[Abstract]
Objective To establish an HPLC fingerprint of Qingxin Zishen Prescription Decoction (QZPD) and determine the contents of its multiple components, so as to provide a scientific basis for quality control. Methods HPLC analysis was performed on a Phenomenex Kinetex C18 column (100 mm×4.60 mm, 2.6 μm) for gradient elution with the mobile phase consisting of methanol, acetonitrile and 0.2% formic acid aqueous. The detection wavelength was set at 245 nm and 280 nm, and the column temperature was 40℃. Fingerprints of ten batches of QZPD were determined, and the similarities among fingerprints were evaluated. Attributive analysis and identification of common peaks were performed and the contents of 15 components were determined. Results The fingerprint similarities of 10 batches of QZPD were ranged from 0.923 to 0.998 compared with the reference fingerprint, and 33 common peaks were identified in the fingerprint. Among them, seven peaks (P11, P14-P16, P24, P29, P30) were identified from Coptidis Rhizoma, two peaks (P7, P19) were identified from Nelumbinis Plumula, two peaks (P14, P21) were identified from Ziziphi Spinosae Semen, nine peaks (P4, P5, P10, P17, P18, P28, P31-P33) were identified from Salvia miltiorrhiza, nine peaks (P1-P3, P6, P8, P9, P12, P13, P20) were identified from Corni Fructus, while five peaks (P22, P23, P25-P27) cannot be originated and none of the common peaks was identified from Rehmanniae Radix. Uncariae Ramulus Cum Uncis and Triticum aestivum. By comparing with the chemical reference, fifteen components, including gallic acid (P2), 5-hydroxymethylfurfural (P3), danshensu (P4), protocatechuic aldehyde (P5), morroniside (P9), caffeic acid (P10), cornin (P12), loganin (P13), magnoflorine (P14), coptisine (P24), lithospermic acid (P28), berberine (P29), palmatine (P30), salvianolic acid B (P31) and salvianolic acid E (P33), were identified and quantified. The contents of the fifteen components were 158.3-248.2, 233.6-321.3, 45.9-166.0, 24.3-38.6, 800.7-1 263.6, 26.6-54.9, 44.5-108.2, 470.4-757.3, 85.6-178.6, 11.1-34.2, 56.2-106.4, 25.9-138.9, 21.0-59.2, 951.6-2 244.7 and 38.6-92.8 μg/g, respectively. Conclusion The method established in this study is stable and highly reproducible, and can provide basis for quality control of QZPD.
[中图分类号]
R286.02
[基金项目]
国家自然科学基金资助项目(81574009);江苏省卫计委科研课题(Q201602);江苏省研究生科研与实践创新计划(KYCX19_1210);江苏省自然科学基金面上项目(BK20181504);江苏省高层次卫生人才"六个一工程"拔尖人才项目(LGY2019072)