School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China; Jiangsu Key Laboratory of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing 210023, China.
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China; Jiangsu Key Laboratory of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing 210023, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Mar 15;1079:34-40. doi: 10.1016/j.jchromb.2018.02.005. Epub 2018 Feb 7.
A sensitive UHPLC-MS/MS approach was developed and validated for the quantification of genkwanin, 3'-hydroxygenkwanin, apigenin, luteolin, yuanhuacine and genkwadaphnin in biological samples after oral administration of raw and vinegar-processed Daphne genkwa. Liquiritin and glycyrrhetinic acid were employed as internal standards. Six components were extracted by using protein precipitation with acetonitrile. Chromatographic separation was achieved on a Waters BEH C column (50 mm × 2.1 mm, 1.7 μm) by using a mobile phase composed of water (containing 0.1% formic acid) and acetonitrile. Mass spectrometric detection was conducted using multiple reaction monitoring (MRM). The intra- and inter-day precisions of the six analytes were below 4.87%, and the accuracies were within ±5.0%. The extraction recoveries of the six constituents were determined between 97.5% and 105.4% and the matrix effects ranged from 97.3% to 103.7%. All the samples showed satisfactory precision and accuracy after various stability tests. The established approach was successfully applied to the comparative pharmacokinetic study. Compared to the raw group, the parameters of C and AUC of genkwanin, 3'-hydroxygenkwanin, apigenin and luteolin elevated remarkably (p < 0.05) after oral delivery of vinegar-processed Daphne genkwa while the parameters of C and AUC of yuanhuacine and genkwadaphnin decreased significantly (p < 0.05). The results revealed that vinegar-processing could enhance bioavailability of genkwanin, 3'-hydroxygenkwanin, apigenin and luteolin but reduce the bioavailability of yuanhuacine and genkwadaphnin.
建立并验证了一种灵敏的 UHPLC-MS/MS 法,用于测定生品和醋制芫花中芫花素、3'-羟基芫花素、芹菜素、木樨草素、芫花定碱和芫花瑞亭的含量。采用内标法,以甘草苷和甘草酸为内标,乙腈沉淀蛋白后提取。采用 Waters BEH C 柱(50mm×2.1mm,1.7μm),以水(含 0.1%甲酸)-乙腈为流动相进行色谱分离。采用多反应监测(MRM)进行质谱检测。六种分析物的日内和日间精密度均低于 4.87%,准确度均在±5.0%范围内。六种成分的提取回收率在 97.5%105.4%之间,基质效应在 97.3%103.7%之间。经过各种稳定性试验,所有样品均显示出满意的精密度和准确度。该方法成功应用于比较药代动力学研究。与生品组相比,醋制芫花给药后芫花素、3'-羟基芫花素、芹菜素和木樨草素的 C 和 AUC 参数显著升高(p<0.05),而芫花定碱和芫花瑞亭的 C 和 AUC 参数显著降低(p<0.05)。结果表明,醋制可提高芫花素、3'-羟基芫花素、芹菜素和木樨草素的生物利用度,但降低芫花定碱和芫花瑞亭的生物利用度。