在線固相萃取—高效液相色譜法測定人體尿液中的

劉丹華等

摘 要 建立了在線固相萃取高效液相色譜法測定人體尿液中的N乙酰S（N甲基甲氨酰）半胱氨酸（AMCC）的分析方法。左泵以AminoPac PA柱作為在線固相萃取柱、

References

1 Imbriani M， Maestri L， Marraccini P， Saretto G， Alessio A， Negri S， Ghittori S. Int Arch Occup Environ Health， 2002， 75（7）： 445-452

2 He J， Wang P， Zhu J Q， Wu G， Ji J M， Xue Y. Int. Arch. Occup. Environ. Health， 2010， 83（4）： 399-406

3 Kafferlein H， Mraz J， Ferstl C， Angerer J. Int. Arch. Environ. Health， 2004， 77（6）： 427-432

4 Imbriani M， Negri S， Ghittori S， Maestri L. J. Chromatogr. B， 2002， 778： 231-236

5 Perico A， Baglioni S， Bavazzano P. Rapid Commun. Mass Spectrom.， 2004， 18（16）： 1865-1868

6 LIU DanHua， TANG HongFang， WANG Han， ZHU HaiBao， RUAN Zheng， QIAN YaLing. Chinese Journal of Health Laboratory Technology， 2011， 22（11）： 2541-2543

劉丹華， 唐紅芳， 王 晗， 朱海豹， 阮 征， 錢亞玲. 中國衛生檢驗雜志， 2011， 22（11）： 2541-2543

7 GBZ/T 210.52008， Guide for Establishing Occupational Health StandardsPart 5： Determination Methods of Chemicals in Biological Materials. National Standards of the People′s Republic of China

職業衛生標準制定指南第5部分： 生物材料中化學物質測定方法. 中華人民共和國國家標準.GBZ/T 210.52008

Determination of NAcetylS（Nmethylcarbamoyl）cysteine in

Human Urine by Online Solid Phase Extraction

High Performance Liquid Chromatography

LIU DanHua1， TANG HongFang1， LIU LüYe2， JIN Yan2， ZHU HaiBao1， RUAN Zheng1， QIAN YaLing*1

1（Zhejiang Academy of Medical Science， Hangzhou 310013， China）

2（Thermofisher Scientific Corporation， Shanghai 201203， China）

Abstract A method was developed for the determination of NacetylS（Nmethylcarbamoyl）cysteine （AMCC） in human urine by online solidphase extraction（SPE）high performance liquid chromatography （HPLC）. The separation of AMCC from the urine matrix was performed on AmoniPac PA Solid phase Extraction （SPE） column with 5 mmol/L KH2PO4 as the mobile phase by left pump. Then the time was controlled to switch the valve to make only the section of sample containing AMCC transferred into the analytic columnAcclaim PAⅡ C18. The determination was performed using gradient elution of 0.1% H3PO4 （containing 5% acetonitrile） and acetonitrile by right pump. The results showed that AMCC present good linear correlation in the range of 1.0-100 mg/L with a correlation coefficient of above 0.999， the quantitation limit of the method was 0.2 mg/L （with the sample inject volume =10 μL）， the recoveries of spiked samples were in the range of 82.9%-85.9%， and the relative standard deviation （n=6） of retention time and peak area were 0.2% and 4.0% respectively. Compared with offline SPEHPLC， the proposed method was convenient， environmentally friendly， efficient and stable， and feasible for the detection of AMCC in 7 human urine samples.

Keywords NAcetylS（Nmethylcarbamoyl）cysteine； Online solid phase extraction； High performance liquid chromatography； Human urine

（Received 8 July 2014； accepted 14 October 2014）

This work was supported by the National Science & Technology Pillar Program during the 12th Fiveyear Plan Period （No. 2014BA111B02）

摘 要 建立了在線固相萃取高效液相色譜法測定人體尿液中的N乙酰S（N甲基甲氨酰）半胱氨酸（AMCC）的分析方法。左泵以AminoPac PA柱作為在線固相萃取柱、

References

1 Imbriani M， Maestri L， Marraccini P， Saretto G， Alessio A， Negri S， Ghittori S. Int Arch Occup Environ Health， 2002， 75（7）： 445-452

2 He J， Wang P， Zhu J Q， Wu G， Ji J M， Xue Y. Int. Arch. Occup. Environ. Health， 2010， 83（4）： 399-406

3 Kafferlein H， Mraz J， Ferstl C， Angerer J. Int. Arch. Environ. Health， 2004， 77（6）： 427-432

4 Imbriani M， Negri S， Ghittori S， Maestri L. J. Chromatogr. B， 2002， 778： 231-236

5 Perico A， Baglioni S， Bavazzano P. Rapid Commun. Mass Spectrom.， 2004， 18（16）： 1865-1868

6 LIU DanHua， TANG HongFang， WANG Han， ZHU HaiBao， RUAN Zheng， QIAN YaLing. Chinese Journal of Health Laboratory Technology， 2011， 22（11）： 2541-2543

劉丹華， 唐紅芳， 王 晗， 朱海豹， 阮 征， 錢亞玲. 中國衛生檢驗雜志， 2011， 22（11）： 2541-2543

7 GBZ/T 210.52008， Guide for Establishing Occupational Health StandardsPart 5： Determination Methods of Chemicals in Biological Materials. National Standards of the People′s Republic of China

職業衛生標準制定指南第5部分： 生物材料中化學物質測定方法. 中華人民共和國國家標準.GBZ/T 210.52008

Determination of NAcetylS（Nmethylcarbamoyl）cysteine in

Human Urine by Online Solid Phase Extraction

High Performance Liquid Chromatography

LIU DanHua1， TANG HongFang1， LIU LüYe2， JIN Yan2， ZHU HaiBao1， RUAN Zheng1， QIAN YaLing*1

1（Zhejiang Academy of Medical Science， Hangzhou 310013， China）

2（Thermofisher Scientific Corporation， Shanghai 201203， China）

Abstract A method was developed for the determination of NacetylS（Nmethylcarbamoyl）cysteine （AMCC） in human urine by online solidphase extraction（SPE）high performance liquid chromatography （HPLC）. The separation of AMCC from the urine matrix was performed on AmoniPac PA Solid phase Extraction （SPE） column with 5 mmol/L KH2PO4 as the mobile phase by left pump. Then the time was controlled to switch the valve to make only the section of sample containing AMCC transferred into the analytic columnAcclaim PAⅡ C18. The determination was performed using gradient elution of 0.1% H3PO4 （containing 5% acetonitrile） and acetonitrile by right pump. The results showed that AMCC present good linear correlation in the range of 1.0-100 mg/L with a correlation coefficient of above 0.999， the quantitation limit of the method was 0.2 mg/L （with the sample inject volume =10 μL）， the recoveries of spiked samples were in the range of 82.9%-85.9%， and the relative standard deviation （n=6） of retention time and peak area were 0.2% and 4.0% respectively. Compared with offline SPEHPLC， the proposed method was convenient， environmentally friendly， efficient and stable， and feasible for the detection of AMCC in 7 human urine samples.

Keywords NAcetylS（Nmethylcarbamoyl）cysteine； Online solid phase extraction； High performance liquid chromatography； Human urine

（Received 8 July 2014； accepted 14 October 2014）

This work was supported by the National Science & Technology Pillar Program during the 12th Fiveyear Plan Period （No. 2014BA111B02）

摘 要 建立了在線固相萃取高效液相色譜法測定人體尿液中的N乙酰S（N甲基甲氨酰）半胱氨酸（AMCC）的分析方法。左泵以AminoPac PA柱作為在線固相萃取柱、

References

1 Imbriani M， Maestri L， Marraccini P， Saretto G， Alessio A， Negri S， Ghittori S. Int Arch Occup Environ Health， 2002， 75（7）： 445-452

2 He J， Wang P， Zhu J Q， Wu G， Ji J M， Xue Y. Int. Arch. Occup. Environ. Health， 2010， 83（4）： 399-406

3 Kafferlein H， Mraz J， Ferstl C， Angerer J. Int. Arch. Environ. Health， 2004， 77（6）： 427-432

4 Imbriani M， Negri S， Ghittori S， Maestri L. J. Chromatogr. B， 2002， 778： 231-236

5 Perico A， Baglioni S， Bavazzano P. Rapid Commun. Mass Spectrom.， 2004， 18（16）： 1865-1868

6 LIU DanHua， TANG HongFang， WANG Han， ZHU HaiBao， RUAN Zheng， QIAN YaLing. Chinese Journal of Health Laboratory Technology， 2011， 22（11）： 2541-2543

劉丹華， 唐紅芳， 王 晗， 朱海豹， 阮 征， 錢亞玲. 中國衛生檢驗雜志， 2011， 22（11）： 2541-2543

7 GBZ/T 210.52008， Guide for Establishing Occupational Health StandardsPart 5： Determination Methods of Chemicals in Biological Materials. National Standards of the People′s Republic of China

職業衛生標準制定指南第5部分： 生物材料中化學物質測定方法. 中華人民共和國國家標準.GBZ/T 210.52008

Determination of NAcetylS（Nmethylcarbamoyl）cysteine in

Human Urine by Online Solid Phase Extraction

High Performance Liquid Chromatography

LIU DanHua1， TANG HongFang1， LIU LüYe2， JIN Yan2， ZHU HaiBao1， RUAN Zheng1， QIAN YaLing*1

1（Zhejiang Academy of Medical Science， Hangzhou 310013， China）

2（Thermofisher Scientific Corporation， Shanghai 201203， China）

Abstract A method was developed for the determination of NacetylS（Nmethylcarbamoyl）cysteine （AMCC） in human urine by online solidphase extraction（SPE）high performance liquid chromatography （HPLC）. The separation of AMCC from the urine matrix was performed on AmoniPac PA Solid phase Extraction （SPE） column with 5 mmol/L KH2PO4 as the mobile phase by left pump. Then the time was controlled to switch the valve to make only the section of sample containing AMCC transferred into the analytic columnAcclaim PAⅡ C18. The determination was performed using gradient elution of 0.1% H3PO4 （containing 5% acetonitrile） and acetonitrile by right pump. The results showed that AMCC present good linear correlation in the range of 1.0-100 mg/L with a correlation coefficient of above 0.999， the quantitation limit of the method was 0.2 mg/L （with the sample inject volume =10 μL）， the recoveries of spiked samples were in the range of 82.9%-85.9%， and the relative standard deviation （n=6） of retention time and peak area were 0.2% and 4.0% respectively. Compared with offline SPEHPLC， the proposed method was convenient， environmentally friendly， efficient and stable， and feasible for the detection of AMCC in 7 human urine samples.

Keywords NAcetylS（Nmethylcarbamoyl）cysteine； Online solid phase extraction； High performance liquid chromatography； Human urine

（Received 8 July 2014； accepted 14 October 2014）

This work was supported by the National Science & Technology Pillar Program during the 12th Fiveyear Plan Period （No. 2014BA111B02）