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Simultaneous LC–UV–MS–MS Analysis of Nine Pivotal Metabolites in Human Serum: Application to Studies of Impaired Glucose Tolerance

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Abstract

A rapid, sensitive, and selective LC–UV–MS–MS method for simultaneous quantification of uric acid, creatinine, xanthine, creatine, hypoxanthine, adenosine, inosine, thymidine, and uridine in serum from patients with impaired glucose tolerance (IGT) has been developed and validated. After precipitation of protein in the serum with methanol samples were evaporated with a stream of nitrogen then reconstituted with aqueous ammonia (0.002 mol L−1). Chromatographic separation was performed on a C18 column with methanol–buffer (8 mmol L−1 ammonium acetate adjusted to pH 6.1 with glacial acetic acid) as mobile phase at a flow rate of 0.8 mL min−1. UV and MS–MS detection were used to quantify the metabolites on the basis of different concentrations and detector response. Linearity was excellent, with r 2 no <0.998. Recovery, and intra-day and inter-day relative standard deviation (RSD) were >85, <10, and <10%, respectively. This reliable bioanalytical method enables evaluation of the levels of purines and pyrimidines in serum for IGT studies and diagnosis.

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Acknowledgments

These studies were supported by grants from the National Basic Research Development Program of China (no. 2005CB523503), the National Eleventh Five-Year Technology Support Program (no. 2006BAI04A04-02), and Heilongjiang Baoquan Pharmaceutical Co. Ltd.

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Authors and Affiliations

  1. School of Pharmaceutics, East-China University of Science and Technology, Shanghai, 200237, China

    Hongfu Zhong & Jianfei Xia

  2. Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology (Ministry of Education), Department of Chemistry, Tsinghua University, Beijing, 100084, China

    Hongfu Zhong, Qionglin Liang, Yiming Wang & Guoan Luo

  3. College of Chemical and Molecular Engineering, East-China University of Science and Technology, Shanghai, 200237, China

    Ping Hu

  4. China Academy of Chinese Medical Sciences, Guang’anmen Hospital, Beijing, 100053, China

    Xiaolin Tong

Authors
  1. Hongfu Zhong
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  2. Qionglin Liang
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  3. Jianfei Xia
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  4. Ping Hu
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  5. Yiming Wang
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  6. Xiaolin Tong
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  7. Guoan Luo
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Corresponding authors

Correspondence to Ping Hu, Xiaolin Tong or Guoan Luo.

Additional information

H. Zhong and Q. Liang contributed equally to this work.

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Zhong, H., Liang, Q., Xia, J. et al. Simultaneous LC–UV–MS–MS Analysis of Nine Pivotal Metabolites in Human Serum: Application to Studies of Impaired Glucose Tolerance. Chromatographia 73, 149–155 (2011). https://doi.org/10.1007/s10337-010-1833-1

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  • DOI: https://doi.org/10.1007/s10337-010-1833-1

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