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目的:建立大鼠血浆中5种大黄蒽醌同时测定的反相离子对HPLC定量方法。方法:以1,8-二羟基蒽醌为内标,血浆预处理采用4倍样本量的甲醇沉淀,上清液在37℃水浴中用N2柔和吹干后流动相复溶进样。分析柱为Scienhome kromasil ODS-1(C18,250mm×4.6 mm,5μm);流动相为:A:甲醇,B:水相(其中含20mM磷酸二氢钾,含6mM十六烷基三甲基溴化铵,pH=4.86),A:B=(80:20);流速:1.00 ml/min;检测波长:254nm;柱温:30℃;内标法定量。结果:本法测得血浆中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的线性范围分别为0.061~29.687μg/ml、0.050~24.450μg/ml、0.062~30.300μg/ml、0.170~33.100μg/ml、0.074~14.494μg/ml,r值均大于0.999,各成分提取回收率均大于69%,准确度93~101%之间,日内与日间精密度RSD均小于10%。结论:本测定方法各成分专属性强,分离度好,分析时长适宜,样本需求量少,灵敏度高,准确、精密、稳定,适合大黄游离蒽醌的多成分药物动力学研究。
Objective: To establish a method for the simultaneous determination of five anthraquinones in rat plasma by reversed-phase ion-pair HPLC. Methods: The 1, 8-dihydroxyanthraquinone was used as the internal standard. The plasma pretreatment was precipitated with 4 times the sample volume of methanol. The supernatant was dissolved in a mobile phase of 37 ° C in a soft water bath after drying with N2. The analytical column was Scienhome kromasil ODS-1 (C18, 250 mm × 4.6 mm, 5 μm); the mobile phase was A: methanol, B: aqueous phase containing 20 mM potassium dihydrogen phosphate containing 6 mM cetyl trimethyl bromide Ammonium chloride, pH = 4.86), A: B = (80:20); flow rate: 1.00 ml / min; detection wavelength: 254 nm; Results: The linear ranges of aloe-emodin, rhein, emodin, chrysophanol and physcion were 0.061 ~ 29.687μg / ml, 0.050 ~ 24.450μg / ml, 0.062 ~ 30.300μg / ml , 0.170 ~ 33.100μg / ml, 0.074 ~ 14.494μg / ml, r values were greater than 0.999, the recovery of each component were greater than 69%, the accuracy of 93 ~ 101%, intraday and interday precision RSD were less than 10 %. Conclusion: The method has the characteristics of strong specificity, good resolution, suitable analysis duration, small sample requirement, high sensitivity, accuracy, precision and stability. It is suitable for the multi-component pharmacokinetic study of anthraquinones in rhubarb.