响应面优化-HPLC-ELSD法测定明胶型凝胶糖果中的磷脂酰丝氨酸

HPLC-ELSD determination of phosphatidylserine in glutin gelatinized candy with response surface methodology

建立了测定明胶型凝胶糖果中磷脂酰丝氨酸的高效液相色谱分析方法。样品经酸法水解,石油醚萃取,浓缩吹干后,用氯仿-甲醇溶液(9:1,V:V)定容。在单因素试验基础上,以水解温度、水解时间和盐酸浓度为影响因素,通过Box-Benhnken响应面优化法确定了最佳酸法水解条件:水解温度60℃,水解时间17 min,盐酸浓度1.8 mol/L。色谱柱选用硅胶柱(4.6 mm×250 mm,5μm),以正己烷/异丙醇/水/乙酸/三乙胺(5:81:14:1.5:0.08,体积比)为流动相,流速1.0 mL/min,柱温40℃。蒸发光散射检测器参数:漂移管温度100℃,载气(N2)流速2.0 L/min。磷脂酰丝氨酸在25~300 mg/L范围内相关系数为0.9999,回收率为93.5%~96.0%,检出限为0.05 mg/g,定量限为0.17 mg/g。该方法适用于明胶型凝胶糖果中磷脂酰丝氨酸的含量测定。

A new method for the determination of phosphatidylserine(PS)in glutin gelatinized candy by high performance liquid chromatography(HPLC)coupled with evaporative light scattering detection(ELSD)was developed.The sample was hydrolyzed by hydrochloric acid,extracted by petroleum ether.The extract was evaporated to dryness,and then volumed by chloroform/methanol(9:1,V:V).Based on one-factor analysis,Box-Benhnken response surface methodology(RSM)was employed to optimize acid hydrolysis conditions,including hydrolysis temperature,hydrolysis time and hydrochloric acid concentration.The experiment results indicated that the optimal conditions were hydrolysis temperature of 60℃,hydrolysis time of 17 minutes and hydrochloric acid concentration of 1.8 mol/L.HPLC separation was conducted on a silica gel column(4.6 mm×250 mm,5μm)with a mobile phase of hexane/isopropanol/water/acetic acid/triethylamine(5:81:14:1.5:0.08,volume ratio)at a flow rate of 1.0 mL/min.The column temperature was 40℃.For ELSD parameters:drift tube temperature and nebulizing gas(N2)flow rate were 100°C and 2.0 L/min,respectively.PS showed a good linearity in the concentration range of 25-300 mg/L,with the correlation coefficient of 0.9999.The accuracy of the method was desirable,with recoveries of PS between 93.5%and 96.0%.The method was also sensitive,with the limit detection of 0.05 mg/g and the limit quantitation of 0.17 mg/g.This method was suitable for the determination of PS in glutin gelatinized candy.