毛细管电泳间接紫外法分离分析海藻和酱油中游离氨基酸方法研究
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摘要
目的建立1种快速检测海藻和酱油中游离氨基酸的毛细管电泳间接紫外法分离分析方法。方法在分离电压为-20kV,毛细管柱温25℃的条件下,40 mmol/L三羟甲基氨基甲烷(Tris)为分离缓冲液,20mmol/L烟酸为背景添加剂(NaOH调pH 11.00),质量体积分数0.015%的溴化己二甲铵(HDB)为电渗流(EOF)反转剂,考察分离缓冲液pH、浓度、HDB浓度、电压等参数对分离的影响,在最佳优化条件下13min内13种未衍生氨基酸得到基线分离。结果方法线性良好(r≥0.997 2),日内及日间精密度分别低于4.25%和7.41%,加标回收率为94.6%～105.5%。建立的方法成功应用于海藻和酱油中游离氨基酸的定性定量分析。结论结果表明,所开发的方法可用于快速测定不同样品中游离氨基酸的组成及含量,为海藻和酱油样品中游离氨基酸的快速检测奠定基础。
Objective A rapid analytical method for the determination of amino acids in seaweed and soy sauce was developed by capillary electrophoresis(CE)with indirect ultraviolet detection.Methods The experiments were set up at a separation voltage of-20 kV and a capillary column temperature of 25℃.20 mmol·L-1 nicotinic acid and 40 mmol·L-1 tris(hydroxymethyl)aminomethane(Tris)(NaOH adjusts pH 11.00)were added as background and electrolyte buffer,respectively.Meanwhile,the mass fraction of 0.015% hexadimethrine bromide(HDB)was applied as electro-osmotic flow(EOF)modifier.The effects of separation buffer pH,ionic strength,HDB concentration,and voltage on separation were investigated.Under the optical conditions,13 underivatized amino acids achieved baseline separation in 13 minutes.Results The proposed method provided a good linearity(r≥0.997 2),inter-day and intra-day precision were lower than 4.25%and 7.41%respectively,and the recovery was in the range of 94.6%～105.5%.The method has been successfully applied to the detection and analysis of amino acids in seaweed and soy sauce.Conclusion The results indicated the developed method could quickly determine the composition and content of free amino acids in different samples.The method offered a foundation for the rapid detection of free amino acids in seaweed and soy sauce.
Objective A rapid analytical method for the determination of amino acids in seaweed and soy sauce was developed by capillary electrophoresis(CE)with indirect ultraviolet detection.Methods The experiments were set up at a separation voltage of-20 kV and a capillary column temperature of 25℃.20 mmol·L-1 nicotinic acid and 40 mmol·L-1 tris(hydroxymethyl)aminomethane(Tris)(NaOH adjusts pH 11.00)were added as background and electrolyte buffer,respectively.Meanwhile,the mass fraction of 0.015% hexadimethrine bromide(HDB)was applied as electro-osmotic flow(EOF)modifier.The effects of separation buffer pH,ionic strength,HDB concentration,and voltage on separation were investigated.Under the optical conditions,13 underivatized amino acids achieved baseline separation in 13 minutes.Results The proposed method provided a good linearity(r≥0.997 2),inter-day and intra-day precision were lower than 4.25%and 7.41%respectively,and the recovery was in the range of 94.6%～105.5%.The method has been successfully applied to the detection and analysis of amino acids in seaweed and soy sauce.Conclusion The results indicated the developed method could quickly determine the composition and content of free amino acids in different samples.The method offered a foundation for the rapid detection of free amino acids in seaweed and soy sauce.
引文
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[10]Wang H Y,Pei H U,Jiang J.Determination of 19kinds of free amino acids in human serum by HPLC-MS/MS[J].JIns Anal,2007,26(S1):1063-1066.
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[12]Lorenzo M P,Villase1or A,Ramamoorthy A,et al.Optimization and validation of a CE-LIF method for amino acids determination in human plasma application to bipolar disorder study[J].Horm Metab Res,2013,26(26):265-269.
[13]Li D,Wang Z,Wang L,et al.Separation and determination of amino acids by CE using 1-butyl-3-methylimidazolium-based ionic liquid as background electrolyte[J].Chromatogr,2009,70(5/6):825-830.
[14]Lee Y H,Lin T I.Capillary electrophoretic determination of amino acids with indirect absorbance detection[J].JChromatogr A,1994,680(1):287-297.
[15]付国妮,何友昭,王晓奎,等.采用隔离池的毛细管电泳-间接紫外吸收法测定茶叶中的氨基酸[J].色谱,2007,25(2):193-196.
[16]Fu G N,He Y Z,Wang L,et al.Determination of amino acids in tobacco samples by capillary electrophoresis/indirect absorbance detection with isolation of the electrolysis compartment and p-aminobenzoic acid as a background electrolyte[J].Japan Soc Anal Chem,2006,22(6):883-887.
[2]张晓梅,郭芮,苏红,等.羊栖菜营养成分分析与安全性评价[J].食品工业科技,2018,4:296-300.
[3]冯志强,周芳梅,黄永连,等.全自动氨基酸分析仪鉴别不同种类酱油中氨基酸的分析研究[J].中国食品添加剂,2013,5:198-205.
[4]Callejón R M,Tesfaye W,Torija M J,et al.HPLC determination of amino acids with AQC derivatization in vinegars along submerged and surface acetifications and its relation to the microbiota[J].Eur Food Res Technol,2008,227(1):93-102.
[5]李梅,杨朝霞,解彬,等.AQC柱前衍生高效液相色谱法测定啤酒中的21种游离氨基酸[J].色谱,2007,25(6):939-941.
[6]Zeng X Y,Wang M F,Zhu B K,et al.Simultaneous analysis of AY and amino acids in corn oligopeptides by HPLC-fluorescence detector with OPA/FMOC-Cl pre-column derivatization[J].Int J Food Agri Envir,2013,11(1):86-90.
[7]Moodie I M,Hough B J,Labadarios D.Determination of amino acids in urine by gas chromatography[J].J Sep Sci,2015,12(7):437-441.
[8]Mudiam M K,Ch R,Jain R,et al.Rapid and simultaneous determination of twenty amino acids in complex biological and food samples by solid-phase microextraction and gas chromatography-mass spectrometry with the aid of experimental design after ethyl chloroformate derivatization[J].JChromatogr B,2012,907(20):56-64.
[9]Yamazaki T,Eyama S,Takatsu A.Concentration measurement of amino acid in aqueous solution by quantitative 1HNMR spectroscopy with internal standard method[J].Japan Soc Anal Chem,2017,33(3):369-373.
[10]Wang H Y,Pei H U,Jiang J.Determination of 19kinds of free amino acids in human serum by HPLC-MS/MS[J].JIns Anal,2007,26(S1):1063-1066.
[11]Casella I G,Contursi M.Isocratic ion chromatographic determination of underivatized amino acids by electrochemical detection[J].Anal Chim Acta,2003,478(2):179-189.
[12]Lorenzo M P,Villase1or A,Ramamoorthy A,et al.Optimization and validation of a CE-LIF method for amino acids determination in human plasma application to bipolar disorder study[J].Horm Metab Res,2013,26(26):265-269.
[13]Li D,Wang Z,Wang L,et al.Separation and determination of amino acids by CE using 1-butyl-3-methylimidazolium-based ionic liquid as background electrolyte[J].Chromatogr,2009,70(5/6):825-830.
[14]Lee Y H,Lin T I.Capillary electrophoretic determination of amino acids with indirect absorbance detection[J].JChromatogr A,1994,680(1):287-297.
[15]付国妮,何友昭,王晓奎,等.采用隔离池的毛细管电泳-间接紫外吸收法测定茶叶中的氨基酸[J].色谱,2007,25(2):193-196.
[16]Fu G N,He Y Z,Wang L,et al.Determination of amino acids in tobacco samples by capillary electrophoresis/indirect absorbance detection with isolation of the electrolysis compartment and p-aminobenzoic acid as a background electrolyte[J].Japan Soc Anal Chem,2006,22(6):883-887.
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