HPLC检测黑皮鸡枞菌中的水溶性维生素

Determination of Water Soluble Vitamins in Black Termitomyces spp. by High Performance Liquid Chromatography

研究建立一种简单、快速、可同时检测新鲜黑皮鸡枞菌和菌粉中7种水溶性维生素(维生素C、维生素B_1、维生素B_2、维生素B_6、维生素B_(12)、烟酸和叶酸)的高效液相色谱(high performance liquid chromatographic,HPLC)方法,为黑皮鸡枞菌的深度开发和加工提供科学依据。采用Inertsil ODS-SP(5μm,4.6 mm×250 mm)色谱柱,流动相A为0.05 mol/L的KH_2PO_4溶液(pH=6.0),流动相B为甲醇,流速0.8 mL/min,柱温为室温,检测波长为268 nm,梯度洗脱,反相高效液相色谱,紫外检测器。7种维生素标准曲线的线性相关系数R2在0.999 6～1.000 0之间,相对标准偏差RSD在0.31%～2.17%之间,加标回收率在88.07%～106.01%之间,最低检出限为0.15μg/mL～1.52μg/mL,定量限为0.49μg/mL～5.08μg/mL。利用所建方法检测出新鲜黑皮鸡枞菌中含有4种水溶性维生素,菌粉中含有5种水溶性维生素且都以维生素C和烟酸为主,水溶性维生素总量分别达到22.25μg/g和242.41μg/g。

This study had developed a high-performance liquid chromatographic （HPLC） method which was simple and rapid for the detection of seven water-soluble vitamins （Vitamin C, Vitamin B1, Vitamin B2, Vitamin BG, Vitamin B12, niacin, and folic acid） simultaneously in fresh black Termitomyces spp. and powder, and it provides a scientific basis for the in-depth development and processing on black Termitomyces spp..The compounds were separated on a Inertsil ODS-SP （5 μm,4.6 mm×250 mm）column at room temperature. The mobile phase used was a mixture of phase A containing 0.05 moll/potassium dihydrogen phosphate （pH6.0） and phase B containing methanol through the gradient elution at a flow rate of 0.8 mL/min. Detection was performed using a UV detector at 268 nm. The method showed good linearity with coefficients （R2） of determination varying between 0.999 6 and 1.000 0 and precision （relative standard deviation, RSD） ranging from 0.31% to 2.17 %, and recovery rates were in the range between 88.07 % and 106.01% for 7 water soluble vitamins. The limits of detection and quantification were 0.15 μg/mL- 1.52 μg/mL and 0.49 μg/mL-5.08 μg/mL, respectively.Using the established method detected fresh black Termitomyces spp. contained 4 kinds of water soluble vitamins and powder contained 5 kinds of water soluble vitamins and vitamin C and niacin primarily. The total water soluble vitamins reached 22.25 μg/g and 242.41μg/g respectively.