水解法与超声法检测保健品中支链氨基酸粉的支链氨基酸比对
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摘要
目的比较水解法与超声法测定保健品中支链氨基酸粉的支链氨基酸(亮氨酸、异亮氨酸及缬氨酸)含量的分析方法。方法样品分别经水解法和超声法进行前处理后,经Agilent Eclipse XDB-phenyl柱分离,以甲醇和0.1%甲酸溶液为流动相,梯度洗脱,流速为0.4mL/min,柱温40℃,采用液相色谱-质谱法检测。结果亮氨酸、异亮氨酸及缬氨酸在0.01～0.05 mmol/L的浓度范围内,线性关系良好,相关系数r均大于0.99。样品经2种方法处理后,都能够准确检测支链氨基酸的含量,2种方法的实验结果比对的相对误差均<5%,水解法处理的精密度相对标准偏差为4.0%～5.3%,而超声方法的精密度相对标准偏差均小于5.0%。结论与水解法相比,超声法前处理方法操作简便,分离效果明显,更适用于提取保健食品中支链氨基酸粉。
Objective To compare the determination of branched chain amino acids in branched chain amino acid powders in health products by hydrolysis method and ultrasound method. Methods The samples were pretreated by hydrolysis and ultrasound, respectively, and then separated with Agilent Eclipse xdb-phenyl column at the flow rate of 0.4 mL/min by gradient elution using methanol and 0.1% formic acid as the mobile phase, and the temperature of the column oven was maintained at 40 ℃. The sample was determined by liquid chromatography-mass spectrometry. Results Leucine, isoleucine and valine had a good linear relationship within the concentration range of 0.01-0.05 mmol/L, and the correlation coefficient was all greater than 0.99. After being treated by 2 methods, the branched chain amino acids content of all samples could be accurately detected. The relative error of the comparison of the experimental results of the two methods was <5%, the relative standard deviations of precision treated by the hydrolysis method were 4.0%-5.3%, and the relative standard deviations of precision treated by the ultrasonic method were less than 5.0%. Conclusion Compared with hydrolysis method, the ultrasonic pre-process is simple, effective and more suitable to extract branched chain amino acid powder from health food.
Objective To compare the determination of branched chain amino acids in branched chain amino acid powders in health products by hydrolysis method and ultrasound method. Methods The samples were pretreated by hydrolysis and ultrasound, respectively, and then separated with Agilent Eclipse xdb-phenyl column at the flow rate of 0.4 mL/min by gradient elution using methanol and 0.1% formic acid as the mobile phase, and the temperature of the column oven was maintained at 40 ℃. The sample was determined by liquid chromatography-mass spectrometry. Results Leucine, isoleucine and valine had a good linear relationship within the concentration range of 0.01-0.05 mmol/L, and the correlation coefficient was all greater than 0.99. After being treated by 2 methods, the branched chain amino acids content of all samples could be accurately detected. The relative error of the comparison of the experimental results of the two methods was <5%, the relative standard deviations of precision treated by the hydrolysis method were 4.0%-5.3%, and the relative standard deviations of precision treated by the ultrasonic method were less than 5.0%. Conclusion Compared with hydrolysis method, the ultrasonic pre-process is simple, effective and more suitable to extract branched chain amino acid powder from health food.
引文
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[15]李涛,王飞久,孙修涛,等.黄官1号海带在不同海域的营养成分分析与评价[J].食品科技,2012,37(11):70-75.Li T,Wang FJ,Sun XT,et al.Nutrition assessment on Saccharina Huangguan NO.1 cultivated in different sea area[J].Food Sci Technol,2012,37(11):70-75.
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[17]任晓宇,裴晓静,张少云,等.微波水解衍生高效液相色谱法测定饲料中的氨基酸[J].分析测试学报,2017,36(10):1254-1259.Ren XJ,Pei XJ,Zhang SY,et al.Determination of amino acids in feed by high performance liquid chromatography with microwave hydrolysis derivatization[J].J Instrum Anal,2017,36(10):1254-1259.
[18]冯娅,陈红,张林,等.饲料中氨基酸检测的常规水解法与微波水解法比对[J].中国饲料,2006,(10):34-35.feng Y,Chen H,Zhang L,et al.Feed of amino acid detection in conventional hydrolysis method and microwave hydrolysis method[J].China Feed,2006,(10):34-35.
[19]张凡,周远征,赵远,等.HPLC异硫氰酸苯酯柱前衍生化测定黄柏及其不同炮制品中17种氨基酸的含量[J].中华中医药学刊,2017,35(9):2259-2262.Zhang F,Zhou YZ,Zhao Y,et al.Content determination of 17 kinds of amino acids in different processed products of phellodendron by HPLCwith pre-column PITC derivation[J].Chin Arch Tradit Chin Med,2017,35(9):2259-2262.
[20]GB 5009.124-2016食品安全国家标准食品中氨基酸的测定[S].GB 5009.124-2016 National food safety standard-Determination of amino acids in food[S].
[2]胡晴晴.支链氨基酸的功效[J].上海中医药报,2012,9(3):1.Hu QQ.The effect of branched chain amino acids[J].Shanghai J Trait Chin Med,2012,9(3):1.
[3]夏志,刘艳,夏贵霞,等.支链氨基酸与运动的关系[J].首都体育学院学报,2006,18(2):125-128.Xia Z,Liu Y,Xia GX,et al.The relationship beteen branched-chain amino acids and exercise[J].J Capit Inst Phys Educ,2006,18(2):125-128.
[4]李玉珍,聂舟,吴韶敏,等.亲水作用色谱-蒸发光散射检测保健食品中三种支链氨基酸的含量[J].分析科学学报,2014,30(4):537-539.Li YZ,Nie Z,Wu SM,et al.Detection of three branched chain amino acid content in the health food by hydrophilic effect chromatography with evaporative light scattering[J].J Anal Sci,2014,30(4):537-539.
[5]刘颖慧,张经华,王明林,等.二维阀切换离子色谱法测定海带中游离氨基酸[J].分析化学计量,2016,25(4):23-26.Liu yh,Zhang JH,Wang ML,et al.Determination of free amino acids of kelp by 2D valve-switching ion chromatography[J].Chem Anal Meter,2016,25(4):23-26.
[6]李辉,岳先领,李皓,等.离子色谱法测定烟叶中游离氨基酸[J].湖北农业科学,2015,54(23):6023-6025,6029.Li H,Yue XL,Li H,et al.Determination of free amino acids in tobacco by ion chromatography method[J].Hubei Agric Sci,2015,54(23):6023-6025,6029.
[7]于宏晓,徐海涛,马强,等.离子色谱-积分脉冲安培法测定烟草中18种游离氨基酸[J].中国烟草科学,2012,33(4):51-54.yu HX,Xu HT,Ma Q,et al.Determination of 18 free amino acids in tobacco with high performance anion-exchange chromatography with integrated pulsed amperometric method[J].Chin Tob Sci,2012,33(4):51-54.
[8]肖小华,吴文威,李攻科.微波辅助萃取-GC/MS联用分析竹子中氨基酸[J].分析试验室,2009,28(9):1-4.Xiao XH,Wu WW,Li GK.Determination of amino acids in bamboo by microwave-assisted extraction coupled with GC/MS[J].Chin J Anal Lab,2009,28(9):1-4.
[9]刘百战,孙哲建,徐玉田.毛细管气相色谱法测定卷烟中的游离氨基酸[J].中国烟草学报,1999,5(2):1-6.Liu BZ,Sun ZJ,Xu YT.Determination of free amino acids in tobacco by ethyl chloroformate derivatization and capillary gas chromatography[J].Acta Tabac Sin,1999,5(2):1-6.
[10]周建华.毛细管气相色谱法分析氨基酸[J].山东轻工业学院学报(自然科学版),1998,(4):48-50.Zhou JH.Capillary gas chromatography analysis of amino acids[J].JShandong Instit Light Ind(Nat Sci Ed),1998,(4):48-50.
[11]Abdel-Hady D,Albishri HM.Simultaneous determination of acrylamide,asparagine and glucose in food using short chain methy imidazoloum ionic liquid based ultrasonic assisted extraction coupled with analyte focusing by ionic liquid micelle collapse capillary electrophoresis[J].Food Chem,2015,188:551,558.
[12]郑重,孙琦,石永伟,等.高效液相色谱-质谱法直接定量分析植物酵素中多种氨基酸成分[J].色谱,2015,33(3):309-313.Zheng Z,Sun Q,Shi YW,et al.Direct quantitative analysis of amino acids in fermented beverage of plant extract using high performance liquid chromatography-tandem mass spectrometry[J].Chin J Chromatogr,2015,33(3):309-313.
[13]Herbert PB,Ratola N,Alves A.HPLC determination of amino acids in musts and port wine using OPA/FMOC derivatives[J].J Food Sci,2000,65(7):1130-1133.
[14]沈锐,杨荣振,李磊.氨基酸类保健品中17种氨基酸的在线衍生-HPLC-DAD测定[J].现代食品,2017,10(19):124-128.Shen R,Yang RZ,Li L,et al.Determination of 17 amino acids in amino acid by health products by online derivatization-HPLC-DAD[J].Mod Food,2017,10(19):124-128.
[15]李涛,王飞久,孙修涛,等.黄官1号海带在不同海域的营养成分分析与评价[J].食品科技,2012,37(11):70-75.Li T,Wang FJ,Sun XT,et al.Nutrition assessment on Saccharina Huangguan NO.1 cultivated in different sea area[J].Food Sci Technol,2012,37(11):70-75.
[16]苗雨田,杨悠悠,王浩,等.全自动氨基酸分析仪法测定不同年份黄酒中游离氨基酸的含量[J].食品安全质量检测学报,2015,6(4):1154-1161.Miao YT,Yang YY,Wang H,et al.Determination of amino acids content in yellow rice wine of different years with automatic amino acid analyzer[J].J Food Saf Qual,2015,6(4):1154-1161.
[17]任晓宇,裴晓静,张少云,等.微波水解衍生高效液相色谱法测定饲料中的氨基酸[J].分析测试学报,2017,36(10):1254-1259.Ren XJ,Pei XJ,Zhang SY,et al.Determination of amino acids in feed by high performance liquid chromatography with microwave hydrolysis derivatization[J].J Instrum Anal,2017,36(10):1254-1259.
[18]冯娅,陈红,张林,等.饲料中氨基酸检测的常规水解法与微波水解法比对[J].中国饲料,2006,(10):34-35.feng Y,Chen H,Zhang L,et al.Feed of amino acid detection in conventional hydrolysis method and microwave hydrolysis method[J].China Feed,2006,(10):34-35.
[19]张凡,周远征,赵远,等.HPLC异硫氰酸苯酯柱前衍生化测定黄柏及其不同炮制品中17种氨基酸的含量[J].中华中医药学刊,2017,35(9):2259-2262.Zhang F,Zhou YZ,Zhao Y,et al.Content determination of 17 kinds of amino acids in different processed products of phellodendron by HPLCwith pre-column PITC derivation[J].Chin Arch Tradit Chin Med,2017,35(9):2259-2262.
[20]GB 5009.124-2016食品安全国家标准食品中氨基酸的测定[S].GB 5009.124-2016 National food safety standard-Determination of amino acids in food[S].