双重保留机理下液相色谱-质谱法直接分析18种氨基酸
|
摘要
采用反相和阳离子交换双重保留机理,无需添加离子对和衍生化试剂,建立了18种游离氨基酸的液相色谱-质谱联用直接分析方法。采用高效液相色谱-三重四极杆串联质谱(HPLC-MS/MS)系统,使用OSAKA SODA CAPCELL PAK CR 1∶4 (150 mm×2.1 mm, 5μm; SCX∶C_(18)=1∶4)色谱柱,以0.1%甲酸(A)-乙腈(含0.1%甲酸)(B)-50 mmol/L甲酸铵(C)为流动相进行三元梯度洗脱,对18种氨基酸进行分离检测,可得到良好的分析结果。色谱峰面积精密度(RSD)范围为0.7%～5.9%;标准曲线线性关系良好(R~2=0.993～0.999);对SD大鼠血清和尿液样品加样回收率范围为92.2%～113.6%。本方法流动相条件简单,灵敏度高,可用于血清和尿液中游离氨基酸分析,为生物样品中氨基酸检测或其它复杂基质中氨基酸的检测提供了新的方法和思路。
Amino acids are usually analyzed by derivatization or adding ion-pair reagents. In this work, a new method was established base on a mixed-mode retention mechanism(reversed-phase and cation-exchange) to determinate 18 kinds of amino acids, using simple mobile phase without the ion-pair and derivatization. In this method, 18 kinds of amino acids could be retained on a OSAKA SODA CAPCELL PAK CR 1∶4(150 mm×2.1 mm, 5 μm; SCX∶C_(18)=1∶4) column with a mobile phase of 0.1% formic acid(A)-acetonitrile(0.1% formic acid)(B)-50 mmol/L ammonium formate(C) using a gradient elution program. Mass spectrometry(MS) detection was performed with multiple reaction monitoring(MRM) using positive electrospray ionization. The relative standard deviations(RSDs) of peak area ranged from 0.7% to 5.9%. The correlation coefficients(R~2) of 18 kinds of compounds were greater than 0.993. And the recoveries of serum and urine samples from SD rats ranged from 92.2% to 113.6%. This method was proved to be simple with good reproducibility. It provides a new method and idea for determination of amino acids in biological samples or other complex matrices.
Amino acids are usually analyzed by derivatization or adding ion-pair reagents. In this work, a new method was established base on a mixed-mode retention mechanism(reversed-phase and cation-exchange) to determinate 18 kinds of amino acids, using simple mobile phase without the ion-pair and derivatization. In this method, 18 kinds of amino acids could be retained on a OSAKA SODA CAPCELL PAK CR 1∶4(150 mm×2.1 mm, 5 μm; SCX∶C_(18)=1∶4) column with a mobile phase of 0.1% formic acid(A)-acetonitrile(0.1% formic acid)(B)-50 mmol/L ammonium formate(C) using a gradient elution program. Mass spectrometry(MS) detection was performed with multiple reaction monitoring(MRM) using positive electrospray ionization. The relative standard deviations(RSDs) of peak area ranged from 0.7% to 5.9%. The correlation coefficients(R~2) of 18 kinds of compounds were greater than 0.993. And the recoveries of serum and urine samples from SD rats ranged from 92.2% to 113.6%. This method was proved to be simple with good reproducibility. It provides a new method and idea for determination of amino acids in biological samples or other complex matrices.
引文
1 Piraud M, Ruet S, Boyer S, Acquaviva C, Clerc-Renaud P, Cheillan D, Vianey-Saban C. Methods Mol. Biol., 2011, 708: 25-53
2 Kiriyama Y, Nochi H. Scientifica (Cairo), 2016, ID6494621
3 Adriana I, Maxime L, Diana L V, Dana C, Marie-Anne L, Bruno O V, Maria A M. Drug Discov. Today, 2017, 22(2): 366-376
4 Zimmerman E S, Heibeck T H, Gill A, Li X F, Murray C J, Madlansacay M R, Cuong Tran, Nathan T U, Yin G, Rivers P J, Yam A Y, Wang W D, Steiner A R, Bajad S U, Penta K, Yang W, Hallam T J. Bioconjugate Chem., 2014, 25: 351-361
5 Hannelore K, Katja D, Wolfram G, Peter J O. Anal. Bioanal. Chem., 2009, 393(2): 445-452
6 LIU Chang-Jiao, YANG Yue-Yue, WANG Ni, YU Ping. China Food Additives, 2018, (1): 189-195刘长姣, 杨越越, 王妮, 余平. 中国食品添加剂, 2018, (1): 189-195
7 SU Jian-Kun, WANG Xue, LU Jian-Qiu, QIAO Yan-Jiang, GAO Xiao-Yan. Chinese Journal of Experimental Traditional Medical Formulae, 2012, 18(15): 135-138苏建坤, 王雪, 卢建秋, 乔延江, 高晓燕. 中国实验方剂学杂志, 2012, 18(15): 135-138
8 LU Ming, SUN Dai-Ni, WANG Yang, SHAO Hong. Chinese Journal of Pharmaceutical Analysis, 2010, 30(12): 2323-2327陆明, 孙黛妮, 汪杨, 邵泓. 药物分析杂志, 2010, 30(12): 2323-2327
9 HE Xin-Rui, WU Zhong-Yong, YE Ying-Li, GAO Xu-Dong, CHEN Shi-En, MA Zhong-Ren. Journal of Instrumental Analysis, 2016, 35(7): 922-928赫欣睿, 武中庸, 叶永丽, 高旭东, 陈士恩, 马忠仁. 分析测试学报, 2016, 35(7): 922-928
10 WANG Yuan, WANG Hai-Long, JIN Gao-Wa, ZHANG Fei-Fang, YANG Bing-Cheng, LIANG Xin-Miao. Modernization of Traditional Chinese Medicine and Materia Materia-World Science and Technology, 2014, 16(6): 1347-1352汪媛, 王海龙, 金高娃, 章飞芳, 杨丙成, 梁鑫淼. 世界科学技术-中医药现代化, 2014, 16(6): 1347-1352
11 DING Ya-Yun, XIE Meng-Xia, DENG Zhi-Wei. Journal of Beijing Normal University(Natural Science), 2001, 37(4): 526-529丁雅韵, 谢孟峡, 邓志威. 北京师范大学学报(自然科学版), 2001, 37 (4): 526-529
12 WANG Wei-Hao, YANG Bin. Chinese Pharmaceutical Journal, 2017, 52(5): 372-376王维皓, 杨滨. 中国药学杂志, 2017, 52(5): 372-376
13 Tapuhi Y, Schich D E, Lindner W. Anal. Biochem., 1981, (115) : 123-129
14 Ni J J, Li X, Li W, Wu L J. Biomed. Chromatogr., 2016, 30(11): 1796-1806
15 Muramoto K, Kamiya H, Kawauchi K. Anal. Biochem., 1984, 141: 446-450
16 Zhang G H, Jin W W, Fan P, Feng X N, Bai Y, Tao T, Yu L J. Int. J. Mass Spectrom., 2015, 392: 1-6
17 ZHANG Jie,GE Yong-Hui, YANG Hui, ZHU Wen-Jing, ZHANG Xiang-Min, GENG Zhao-Liang. Tobacco Science & Technology, 2017, 50(11): 58-65张婕, 葛永辉, 杨慧, 朱文静, 章向敏, 耿召良. 烟草科技, 2017, 50(11): 58-65
18 Krumpochov P, Bruyneel B, Molenaar D, Koukou A, Wuhrer M, Niessen W M. A, Giera M. J. Pharm. Biomed. Anal., 2015, 114: 398-407
19 LI Peng-Fei, TAO Pei-Pei, ZHANG Xu-De, AN Zhuo-Ling, ZHANG Qian, LI Yue, HAO Qian-Qian, LIU Li-Hong. Chinese J. Anal. Chem., 2013, 41(9): 1347-1352李鹏飞, 陶蓓蓓, 张绪得, 安卓玲, 张茜, 李悦, 郝倩倩, 刘丽宏. 分析化学, 2013, 41(9): 1347-1352
20 Waterval W A H, Scheijen J L J M. Ortmans-Ploemen M M J C, Habets-vander Poel C D, Bierau J. Clin. Chim. Acta, 2009, 407: 36-42
21 Lea A, Ngb A, Kwanb A, Cusmano-Ozoga K, Cowana T M. J. Chromatogr. B, 2014, 944: 166-174
22 HUANG Xin, LI Shuai-Ping, ZHANG Yong, LIU Shu-Ying. Chinese J. Anal. Chem., 2016, 44(11): 1652-1658黄鑫, 李帅坪, 张勇, 刘淑莹. 分析化学, 2016, 44(11): 1652-1658
23 LI Tai-Feng, XUE Wei, LI Min, SHI Ai-Xin, LI Ke-Xin. Chinese Journal of New Drugs, 2015, 24(14): 1659-1664李泰锋, 薛薇, 李敏, 史爱欣, 李可欣. 中国新药杂志, 2015, 24(14): 1659-1664
24 Vilches A P, Norstr?m S H, Bylund D. J. Sep. Sci., 2017, 40(7): 1482-1492
25 Choi M S, Rehman S U, Kima I S, Park H J, Song M Y, Yoo H H. J. Pharm. Biomed. Anal., 2017, 145: 52-58
2 Kiriyama Y, Nochi H. Scientifica (Cairo), 2016, ID6494621
3 Adriana I, Maxime L, Diana L V, Dana C, Marie-Anne L, Bruno O V, Maria A M. Drug Discov. Today, 2017, 22(2): 366-376
4 Zimmerman E S, Heibeck T H, Gill A, Li X F, Murray C J, Madlansacay M R, Cuong Tran, Nathan T U, Yin G, Rivers P J, Yam A Y, Wang W D, Steiner A R, Bajad S U, Penta K, Yang W, Hallam T J. Bioconjugate Chem., 2014, 25: 351-361
5 Hannelore K, Katja D, Wolfram G, Peter J O. Anal. Bioanal. Chem., 2009, 393(2): 445-452
6 LIU Chang-Jiao, YANG Yue-Yue, WANG Ni, YU Ping. China Food Additives, 2018, (1): 189-195刘长姣, 杨越越, 王妮, 余平. 中国食品添加剂, 2018, (1): 189-195
7 SU Jian-Kun, WANG Xue, LU Jian-Qiu, QIAO Yan-Jiang, GAO Xiao-Yan. Chinese Journal of Experimental Traditional Medical Formulae, 2012, 18(15): 135-138苏建坤, 王雪, 卢建秋, 乔延江, 高晓燕. 中国实验方剂学杂志, 2012, 18(15): 135-138
8 LU Ming, SUN Dai-Ni, WANG Yang, SHAO Hong. Chinese Journal of Pharmaceutical Analysis, 2010, 30(12): 2323-2327陆明, 孙黛妮, 汪杨, 邵泓. 药物分析杂志, 2010, 30(12): 2323-2327
9 HE Xin-Rui, WU Zhong-Yong, YE Ying-Li, GAO Xu-Dong, CHEN Shi-En, MA Zhong-Ren. Journal of Instrumental Analysis, 2016, 35(7): 922-928赫欣睿, 武中庸, 叶永丽, 高旭东, 陈士恩, 马忠仁. 分析测试学报, 2016, 35(7): 922-928
10 WANG Yuan, WANG Hai-Long, JIN Gao-Wa, ZHANG Fei-Fang, YANG Bing-Cheng, LIANG Xin-Miao. Modernization of Traditional Chinese Medicine and Materia Materia-World Science and Technology, 2014, 16(6): 1347-1352汪媛, 王海龙, 金高娃, 章飞芳, 杨丙成, 梁鑫淼. 世界科学技术-中医药现代化, 2014, 16(6): 1347-1352
11 DING Ya-Yun, XIE Meng-Xia, DENG Zhi-Wei. Journal of Beijing Normal University(Natural Science), 2001, 37(4): 526-529丁雅韵, 谢孟峡, 邓志威. 北京师范大学学报(自然科学版), 2001, 37 (4): 526-529
12 WANG Wei-Hao, YANG Bin. Chinese Pharmaceutical Journal, 2017, 52(5): 372-376王维皓, 杨滨. 中国药学杂志, 2017, 52(5): 372-376
13 Tapuhi Y, Schich D E, Lindner W. Anal. Biochem., 1981, (115) : 123-129
14 Ni J J, Li X, Li W, Wu L J. Biomed. Chromatogr., 2016, 30(11): 1796-1806
15 Muramoto K, Kamiya H, Kawauchi K. Anal. Biochem., 1984, 141: 446-450
16 Zhang G H, Jin W W, Fan P, Feng X N, Bai Y, Tao T, Yu L J. Int. J. Mass Spectrom., 2015, 392: 1-6
17 ZHANG Jie,GE Yong-Hui, YANG Hui, ZHU Wen-Jing, ZHANG Xiang-Min, GENG Zhao-Liang. Tobacco Science & Technology, 2017, 50(11): 58-65张婕, 葛永辉, 杨慧, 朱文静, 章向敏, 耿召良. 烟草科技, 2017, 50(11): 58-65
18 Krumpochov P, Bruyneel B, Molenaar D, Koukou A, Wuhrer M, Niessen W M. A, Giera M. J. Pharm. Biomed. Anal., 2015, 114: 398-407
19 LI Peng-Fei, TAO Pei-Pei, ZHANG Xu-De, AN Zhuo-Ling, ZHANG Qian, LI Yue, HAO Qian-Qian, LIU Li-Hong. Chinese J. Anal. Chem., 2013, 41(9): 1347-1352李鹏飞, 陶蓓蓓, 张绪得, 安卓玲, 张茜, 李悦, 郝倩倩, 刘丽宏. 分析化学, 2013, 41(9): 1347-1352
20 Waterval W A H, Scheijen J L J M. Ortmans-Ploemen M M J C, Habets-vander Poel C D, Bierau J. Clin. Chim. Acta, 2009, 407: 36-42
21 Lea A, Ngb A, Kwanb A, Cusmano-Ozoga K, Cowana T M. J. Chromatogr. B, 2014, 944: 166-174
22 HUANG Xin, LI Shuai-Ping, ZHANG Yong, LIU Shu-Ying. Chinese J. Anal. Chem., 2016, 44(11): 1652-1658黄鑫, 李帅坪, 张勇, 刘淑莹. 分析化学, 2016, 44(11): 1652-1658
23 LI Tai-Feng, XUE Wei, LI Min, SHI Ai-Xin, LI Ke-Xin. Chinese Journal of New Drugs, 2015, 24(14): 1659-1664李泰锋, 薛薇, 李敏, 史爱欣, 李可欣. 中国新药杂志, 2015, 24(14): 1659-1664
24 Vilches A P, Norstr?m S H, Bylund D. J. Sep. Sci., 2017, 40(7): 1482-1492
25 Choi M S, Rehman S U, Kima I S, Park H J, Song M Y, Yoo H H. J. Pharm. Biomed. Anal., 2017, 145: 52-58