柱前衍生反相高效液相色谱法测定新疆特色食品馕中氨基酸含量

Determination of amino acids in Xinjiang special food Nang by reversed-phase high performance liquid chromatography with pre-column derivatization

目的建立柱前衍生反相高效液相色谱法(reversed-phase high performance liquid chromatography,RP-HCLP)测定新疆特色食品馕中17种氨基酸含量的检测方法。方法通过优化酸水解参数,并以6-氨基喹啉-N-羟基琥珀酰亚胺基甲酸酯为衍生试剂,用乙腈为流动相A,水为流动相B,磷酸盐缓冲液溶液为流动相C进行梯度洗脱,柱温37℃条件下用AccQ-Tag色谱柱(3.9 mm×150 mm, 4μm)进行分离,采用反相-高效液色谱法荧光检测器(激发波长250 nm、发射波长395 nm)进行测定。结果当取样量为100 mg、水解液加入体积为10 mL、水解温度为115℃、水解时间为24 h时,水解效果最好。17种氨基酸线性回归方程的相关系数在0.9985以上(n=3),检出限为0.011~0.058mg/L,回收率为81.6%~116.2%,相对标准偏差为0.41%~6.94%(n=3)。结论该方法操作简单、灵敏度高、线性关系和回收率好,适用于新疆特色食品馕中氨基酸含量的测定。

Objective To establish a method for determination of 17 kinds of amino acids in Xinjiang special food Nang by reversed-phase high performance liquid chromatography(RP-HPLC) with pre-column derivatization. Methods By optimizing the parameters of acid hydrolysis, 6-aminoquinoline-n-hydroxysuccinimide formate was used as derivatization reagent, with acetonitrile as mobile phase A, water as mobile phase B, phosphate buffer solution as mobile phase C for gradient elution. The separation was performed on AccQ tag column(3.9 mm× 150 mm, 4 μm) at 37 ℃ and fluorescence detector(excitation wavelength 250 nm, emission wavelength 395 nm) was used for detection. Results The best hydrolysis effect was obtained when the sample weight was 100 mg, the volume of hydrolysate was 10 mL, the hydrolysis temperature was 115 ℃, and the hydrolysis time was 24 h. The correlation coefficients of the linear regression equation of 17 amino acids were above 0.9985(n=3), the detection limits were 0.011-0.058 mg/L, the recoveries were 81.6%-116.2%, and the relative standard deviations were 0.41%-6.94%(n=3). Conclusion This method is simple, sensitive, and the linear and recovery are good, which is suitable for the determination of amino acid in Xinjiang special food Nang.