高效液相色谱法测定永川豆豉中的6种有机酸

Determination of Six Organic Acids in Yongchuan Douchi, a Chinese Traditional Fermented Soybean Product, by HPLC

建立高效液相色谱法同时测定永川豆豉中酒石酸、苹果酸、乳酸、醋酸、柠檬酸和琥珀酸6种有机酸含量的方法。通过色谱条件的优化,确定永川豆豉中6种有机酸含量测定的最佳色谱条件:色谱柱为Agilent ZORBAX SB-Aq(4.6 mm×250 mm,5μm),流动相为0.01 mol/L磷酸氢二铵(pH 2.5)和甲醇溶液,流速0.6 mL/min、柱温30℃、检测波长220 nm。在此条件下,上述6种有机酸可得到很好的分离和测定。该方法各种酸的线性范围为0.001~1.000 mg/mL,标准曲线相关系数均在0.999 9以上,精密度检测相对标准偏差为0.07%~0.36%(n=5),重复性检测相对标准偏差为0.97%~3.04%(n=5),回收率在94.89%~104.28%之间。该方法简单、快捷、准确、重复性好,可应用于永川豆豉中6种有机酸的同时快速测定。

A high performance liquid chromatographic(HPLC) method was established for the simultaneous determination of organic acids in Yongchuan douchi, a Chinese traditional fermented soybean product. The optimal chromatographic conditions were determined. An Agilent ZORBAX SB-Aq(4.6 mm × 250 mm, 5 μm) column was used to separate the organic acids. The mobile phase was a mixture of 0.01 mol/L(NH4)2HPO4(pH 2.5) and methanol solution at a flow rate of 0.6 mL/min. The column temperature and UV detection wavelength were set as 30 ℃ and 220 nm, respectively. The sixorganic acids were successfully separated and determined under the chosen experimental conditions. The calibration curves for all these organic acids displayed a good linear relationship with correlation coefficients above 0.999 9 and precision relative standard deviations(RSDs) of 0.07%–0.36%(n = 5) and repeatability RSD of 0.97%–3.04%(n = 5). The recovery rates of the six organic acids were in the range of 94.89%–104.28%. The method is convenient, rapid, accurate, reproducibleand applicable to determine organic acids in Yongchuan douchi.