HPLC-ELSD法同时测定生物转化样品中牛磺熊去氧胆酸等5种成分的含量

目的:建立同时测定生物转化样品中牛磺熊去氧胆酸、牛磺鹅去氧胆酸、熊去氧胆酸、鹅去氧胆酸、牛磺7-酮石胆酸5种成分含量的方法。方法:采用高效液相色谱法,检测器为蒸发光散射检测器。色谱柱为Agilent XBD-C18,流动相为0.05%三氟乙酸-乙腈,梯度洗脱,流速为1.0 ml/min;漂移管温度为80℃,N2流量为3.0 ml/min,柱温为40℃。结果:牛磺熊去氧胆酸、牛磺鹅去氧胆酸、熊去氧胆酸、鹅去氧胆酸、牛磺7-酮石胆酸检测质量浓度线性范围分别为10.00²00.08、10.22200.08、10.22²04.32、10.00204.32、10.00¹99.92、9.85199.92、9.85¹97.08、10.25197.08、10.25²04.92μg/ml(r=0.999 5204.92μg/ml(r=0.999 5⁰.999 8);精密度、稳定性、重复性试验的RSD均不大于2.0%。5种成分的平均回收率分别为97.9%、98.4%、97.9%、98.1%、98.3%,RSD分别为0.49%、0.63%、0.81%、0.72%、0.78%(n=3)。结论:该方法快速、准确、重复性好,可用于生物转化样品中牛磺熊去氧胆酸等5种成分的定量测定。

Simultaneous Content Determination of 5 Components as Tauroursodeoxycholic Acid in Biotransformation Samples by HPLC-ELSD

OBJECTIVE： To develop a method for the content determination of tauroursodeoxychoiic acid （TUDCA）, tauroche- nodeoxycholic acid （TCDCA）, ursodeoxycholic acid （UDCA）, chenodeoxycholic acid （CDCA） and tauro-7-keto lithocholic acid （T-7-KLCA） in biotransformation samples. METHODS： HPLC method was adopted. ELSD was used as detector. The determination was performed on Agilent XBD-C18 column with mobile phase consisted of 0.05% TFA-acetonitrile （gradient elution） at the flow rate of 1.0 ml/min. The drift tube temperature was set at 80℃, and Nz flow was 3.0 ml/min. The column temperature was 40℃. RESULTS： The linear ranges of TUDCA, TCDCA, UDCA, CDCA and T-7-KLCA were 10.00-200.08, 10.22-204.32, 10.00-199.92, 9.85-197.08, 10.25-204.92 μg/ml （r=0.999 5-0.999 8）, respectively. RSDs of precision, stability and reproducibility tests were all lower than 2.0%. The average recoveries of 5 components were 97.9% （RSD=0.49%, n=3）, 98.4% （RSD= 0.63%, n=3）, 97.9% （RSD=0.81%, n=3）, 98.1% （RSD=0.72%, n=3） and 98.3% （RSD=0.78%, n=3）, respectively. CONCLUSIONS： The method is rapid, accurate and repeatable, which can be used for the quantitative determination of 5 components as TUDCA in biotransforrnation samples.