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目的建立一种测定未衍生化氨基酸的方法,测定醋龟甲中羟脯氨酸。方法以高效液相色谱-蒸发光散射检测器(HPLC-ELSD)法,采用Prevail C18(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.7%三氟醋酸溶液(含5.0 mmol/L七氟丁酸)为流动相进行洗脱,洗脱时间为25 min。结果羟脯氨酸峰面积的自然对数与相应浓度的自然对数呈良好的线性关系,其方程为Y=1.212 4 X+7.803 4,r=0.999 6,羟脯氨酸的线性范围为0.1～1.0 mg/mL,平均加样回收率为101.7%,RSD为2.7%。结论该法快速、简便、准确,可作为醋龟甲药材的质量控制方法。
Objective To establish a method for the determination of underivatized amino acids and determine hydroxyproline in Methods High performance liquid chromatography with evaporative light scattering detector (HPLC-ELSD) was performed on a Prevail C18 (250 mm × 4.6 mm, 5 μm) column with acetonitrile-0.7% trifluoroacetic acid Fluorobutanoic acid) as eluent, the elution time was 25 min. Results The natural logarithm of hydroxyproline peak area showed a good linear relationship with the natural logarithm of corresponding concentration. The equation was Y = 1.212 4 X + 7.803 4, r = 0.999 6, and the linear range of hydroxyproline was 0.1 ~ 1.0 mg / mL, the average recovery was 101.7% with RSD 2.7%. Conclusion The method is rapid, simple and accurate, and can be used as a quality control method for vinegar tortoiseshell.