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Simultaneous determination of aconitum alkaloids in rat body fluids by high-performance liquid chromatography

机译:高效液相色谱法同时测定大鼠体液中乌头生物碱

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A sensitive, reliable and accurate high-performance liquid chromatography (HPLC) method coupled with photodiode array detector (DAD) were developed for the simultaneous quantitative determination of aconitine, mesaconitine and hypaconitine in rat plasma and urine by optimizing the extraction, separation and analytical conditions. The analyses were chromatographed on a ZORBAX Eclipse XDB-C18?column (150 mm × 4.6 mm?i.d.; 5 μm particle size) with gradient elution using solvents of acetonitrile and ammonium acetate buffer (pH 10.0).?The detection wavelength was 240 nm.?Intra-assay and inter-assay precision of the analyses were less than?10% and the average recovery rates obtained were in the range of?85.63 -?90.94% for all analysis of the three aconitum alkaloids with relative standard deviations (RSD) below?14%. Positive linear relationships were observed in correlation coefficients that exceeded 0.95. The limits of detection (signal-to-noise ratio of 3) were 2.64, 1.58 and 2.75 ng for aconitine, mesaconitine and hypaconitine, respectively.?The method can provide a scientific and technical platform to determine the concentration of aconitum alkaloids in plasma during a pilot pharmacokinetic study in rats.
机译:建立了灵敏,可靠,准确的高效液相色谱(HPLC)方法与光电二极管阵列检测器(DAD)结合,通过优化提取,分离和分析条件,同时定量测定了大鼠血浆和尿液中乌头碱,美康尼汀和猪笼草碱的含量。使用ZORBAX Eclipse XDB-C18?色谱柱(150 mm×4.6 mm?id; 5μm粒径)进行色谱分离,并使用乙腈和乙酸铵缓冲液(pH 10.0)进行梯度洗脱。检测波长为240 nm。分析的三个批间乌头生物碱的相对标准偏差(RSD)的分析内和分析间精密度均小于?10%,获得的平均回收率在?85.63-?90.94%范围内。 )低于14%。在超过0.95的相关系数中观察到正线性关系。乌头碱,中康尼汀和羟乙炔的检出限(信噪比为3)分别为2.64、1.58和2.75 ng。该方法可提供一个科学和技术平台,可测定乌头碱在血浆中乌头碱的浓度。在大鼠中进行的初步药代动力学研究。

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