柱前衍生-高效液相色谱法测定化妆品中16种氨基酸的含量  

Determination of 16 Amino Acids in Cosmetics by High Performance Liquid Chromatography with Pre-Column Derivatization

作　　者：王丁林[1] 张娜娜 周耀斌[1] WANG Dinglin;ZHANG Nana;ZHOU Yaobin(National Center of Quality Inspection and Testing on Cosmetics&Cleaning Products,Shanghai Institute of Quality Inspection and Technical Research,Shanghai 200233,China)

机构地区：[1]上海市质量监督检验技术研究院,国家保洁产品质量检验检测中心,上海200233

出　　处：《理化检验（化学分册）》2024年第7期674-681,共8页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

摘　　要：提出了2,4-二硝基氟苯(DNFB)柱前衍生-高效液相色谱法测定化妆品中甘氨酸、亮氨酸、组氨酸、缬氨酸、甲硫氨酸、脯氨酸、半胱氨酸、苏氨酸、谷氨酸、赖氨酸、酪氨酸、精氨酸、异亮氨酸、色氨酸、苯丙氨酸、丙氨酸等16种氨基酸含量的方法。取化妆品样品5 g置于50 mL离心管中,加入5 mL水,超声10 min,加热煮沸15 min,冷却后加入15 mL 35 g·L^(−1)磺基水杨酸溶液,以转速10500 r·min^(−1)离心20 min。取全部上清液,用35 g·L^(−1)磺基水杨酸溶液定容至25 mL。取1 mL样品溶液,加入0.5 mol·L^(−1)碳酸氢钠溶液(pH 9.0)1 mL和1%(体积分数)DNFB衍生溶液1 mL,混匀,于60℃加热10 min,冷却后用0.01 mol·L^(−1)磷酸二氢钾溶液(pH 7.0)定容至5 mL,所得溶液中16种氨基酸经X Bridge C_(18)色谱柱分离,以不同体积比的含1%(体积分数)N,N-二甲基甲酰胺的0.05 mol·L^(−1)乙酸钠缓冲液(pH 6.5)-体积比1∶1的乙腈-水混合溶液为流动相进行梯度洗脱,在360 nm处检测。结果表明:16种氨基酸标准曲线的线性范围均为0.004~40.0 mg·L^(−1),检出限(3S/N)为0.1~0.2 mg·kg^(−1);对空白样品进行3个浓度水平的加标回收试验,回收率为89.0%~105%,测定值的相对标准偏差(n=7)均不大于10%。方法用于11种市售化妆品分析,检出的氨基酸成分多为组氨酸、甘氨酸、甲硫氨酸、谷氨酸。A method for the determination of 16 amino acids including glycine,leucine,histidine,valine,methionine,proline,cysteine,threonine,glutamic acid,lysine,tyrosine,arginine,isoleucine,tryptophan,phenylalanine,and alanine in cosmetics by high performance liquid chromatography with 2,4-dinitrofluorobenzene(DNFB)pre-column derivatization was proposed.5 g of cosmetic sample was taken into a 50 mL-centrifuge tube,and 5 mL of water was added.The mixture was sonicated for 10 min and boiled for 15 min.After cooling,15 mL of 35 g·L^(−1)sulfosalicylic acid solution was added and the mixture was centrifuged at a rotational speed of 10500 r·min^(−1)for 20 min.All of the supernatant was taken and its volume was made up to 25 mL with 35 g·L^(−1) sulfosalicylic acid solution.1 mL of sample solution was taken,and 1 mL of 0.5 mol·L^(−1)sodium bicarbonate solution(pH 9.0)and 1 mL of 1%(volume fraction)DNFB derivative solution were added.The mixture was heated at 60℃for 10 min and cooled.Its volume was made up to 5 mL with 0.01 mol·L^(−1)potassium dihydrogen phosphate solution(pH 7.0).16 amino acids in the obtained solution were separated on a X Bridge C_(18) chromatographic column,and gradient elution was performed using a mobile phase consisting of 0.05 mol·L^(−1)sodium acetate buffer solution(pH 6.5)containing 1%(volume fraction)DMF and a mixture of acetonitrile-water at a volume ratio of 1∶1 at different volume ratios.The detection was performed at 360 nm.As shown by the results,the linear ranges of the standard curves of 16 amino acids were 0.004-40.0 mg·L^(−1),with detection limits(3S/N)in the range of 0.1-0.2 mg·kg^(−1).Test for recovery was made on blank sample by standard addition method at 3 concentration levels,giving results in the range of 89.0%-105%,with RSDs(n=7)of the determined values not more than 10%.This method was applied to the analysis of 11 commercially available cosmetics,and it was shown that the detected amino acid components were mostly histidine,glycine,methionine,and glutamic ac

关 键 词：柱前衍生 高效液相色谱法 化妆品 氨基酸 

分 类 号：O657.7[理学—分析化学]