柱前衍生_反相高效液相色谱法测定不同方法煮制的猪肉及其汤汁中的游离氨基酸.docx

技术与应用DOI: 10． 3724 / SP． J． 1123． 2011． 01041摘要: 建立了反相高效液相色谱( RP-HPLC) 分离检测用不同方法煮制的猪肉及其汤汁中 17 种游离氨基酸的方法。样品经 6-氨基 喹 啉 基-N-羟基琥珀酰亚氨基甲酸酯 ( AQC ) 柱 前 衍 生 后，采 用 Nova-PakTM C18 色 谱 柱 分 离，以 AccQ·Tag Eluent A稀释液、乙腈和超纯水为流动相，梯度洗脱，检测波长为 248 nm，在 47 min 内实现了 17 种氨基 酸衍生物的基线分离。各氨基酸在 1 ～ 100 μmol / L( 胱氨酸在 0. 5 ～ 50 μmol / L) 范围内呈现出良好的线性关系，相 应的线性相关系数( r2 ) 均大于 0. 99; 17 种氨基酸的检出限( 以信噪比为 3 计) 为 0. 29 ～ 0. 96 μmol / L; 在汤汁中的 加标回收率为 86. 5% ～ 101. 0%。该方法前处理过程简单，分离效果好，是检测肉及其汤汁中氨基酸的有效手段，可 应用于肉制品的质量评定和工艺优化。关键词: 反相高效液相色谱; 柱前衍生; 游离氨基酸; 猪肉; 汤汁中图分类号: O658文献标识码: A文章编号: 1000-8713( 2011) 10-1041-05Abstract: A reversed-phase high performance liquid chromatographic ( RP-HPLC ) method was devel-oped for the determination of 17 amino acids in the pork and its broth cooked by different methods． 6- Aminoquinolyl-N-hydroxyl-succinimidyl-carbamate ( AQC ) was used as pre-column derivatization rea- gent． In the RP-HPLC method，a Nova-PakTM C18 column was used with the dilution of AccQ·Tag Elu- ent A，acetonitrile and water as the mobile phases in a gradient elution mode． The 17 amino acids were baseline separated within 47 min with ultraviolet ( UV) detection at 248 nm． Each amino acid showed good linearity in the range of 1 － 100 μmol / L except cystine ( Cys2 ) in the range of 0. 5 － 50 μmol / L with the correlation coefficient ( r2 ) more than 0. 99． The detection limits ( S / N = 3 ) of 17 amino acids were ranged from 0. 29 to 0. 96 μmol / L，and the spiked recoveries in a cooked broth sample were from86. 5% to 101. 0%． The results showed that the proposed method can be applied to determine amino acids for meat quality assessment and process optimization with simple pretreatment and good separation． Key words: reversed-phase high performance liquid chromatography ( RP-HPLC) ; pre-column derivat- ization; free amino acids; pork; broth猪肉是我国居民主要的肉食品来源，其富含蛋白质、脂肪和适宜的水分。烹饪过程使其更有利于 人体消化和吸收。中式烹饪中，有水煮、高压蒸煮和炖煮 3 种基本煮制方法，而各种煮制方法都有其不