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Simultaneous determination of dl-lactic acid and dl-3-hydroxybutyric acid enantiomers in saliva of diabetes mellitus patients by high-throughput LC–ESI-MS/MS

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Abstract

A simultaneous determination method for the enantiomers of chiral carboxylic acids by the combination of ultraperformance liquid chromatography and mass spectrometry (UPLC–MS/MS) has been developed. (S)(+)-1-(2-Pyrrolidinylmethyl)-pyrrolidine (S-PMP) was used as the derivatization reagent for the high-throughput determination of biological chiral carboxylic acids, i.e., lactic acid (LA) and 3-hydroxybutyric acid (HA). The S-PMP efficiently reacted with the carboxylic acids under mild conditions at room temperature in the presence of 2,2′-dipyridyl disulfide and triphenylphosphine. The resulting S-PMP derivatives were highly responsive in the electrospray ionization (ESI)-MS operating in the positive-ion mode and gave characteristic product ions during the MS/MS, which enabled the sensitive detection using selected reaction monitoring. The derivatization was effective for the enantiomeric separation of the chiral carboxylic acids, and the resolution values of dl-LA and dl-HA were 4.91 and 9.37, respectively. Furthermore, a rapid separation of the derivatives of dl-LA and dl-HA within 7 min was performed using the UPLC system. The limits of detection on the column were in the low femtogram range (5–12 fg). The proposed procedure was successfully applied for the determination of the d- and l-isomers of LA and HA in the saliva of diabetes mellitus (DM) patients and healthy volunteers. The d-LA in DM patients was clearly higher than that in normal subjects. The derivatization followed by UPLC–ESI-MS/MS enabled the enantiomeric separation and detection of trace amounts of LA and HA in human saliva with a simple pretreatment and small sample volume.

Concentration of HA and LA enantiomers in saliva of DM patients and healthy volunteers (n = 10). DM diabetes mellitus patients, C healthy volunteers

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References

  1. Toyo’oka T (2008) J Chromatogr Sci 46:233–247

    Google Scholar 

  2. Gagné S, Crane S, Huang Z, Li CS, Bateman KP, Lévesque JF (2007) J Lipid Res 48:252–259

    Article  Google Scholar 

  3. Tsukamoto Y, Santa T, Saimaru H, Imai K, Funatsu T (2005) Biomed Chromatogr 19:802–808

    Article  CAS  Google Scholar 

  4. Pettinella C, Lee SH, Cipollone F, Blair IA (2007) J Chromatogr B 850:168–176

    Article  CAS  Google Scholar 

  5. Kretschmer A, Giera M, Wijtmans M, de Vries L, Lingeman H, Irth H, Niessen WMA (2011) J Chromatogr B 879:1393–1401

    Article  CAS  Google Scholar 

  6. Guo K, Li L (2010) Anal Chem 82:8789–8793

    Article  CAS  Google Scholar 

  7. Bollinger JG, Thompson W, Lai Y, Oslund RC, Hallstrand TS, Sadilek M, Turecek F, Gelb MH (2010) Anal Chem 82:6790–6796

    Article  CAS  Google Scholar 

  8. Higashi T, Ichikawa T, Inagaki S, Min JZ, Fukushima T, Toyo’oka T (2010) J Pharm Biomed Anal 52:809–818

    Article  CAS  Google Scholar 

  9. Li X, Franke AA (2011) Anal Chem 83:3192–3198

    Article  CAS  Google Scholar 

  10. Nozawa Y, Sakai N, Arai K, Kawasaki Y, Harada K (2007) J Microbiol Methods 70:306–311

    Article  CAS  Google Scholar 

  11. Fujii K, Ikai Y, Mayumi T, Oka H, Suzuki M, Harada K (1997) Anal Chem 69:3346–3352

    Article  CAS  Google Scholar 

  12. Tsutsui H, Fujii S, Sakamoto T, Min JZ, Todoroki K, Toyo’oka T (2012) J Sep Sci 35:1551–1559

    Article  CAS  Google Scholar 

  13. Zimmet P, Alberti KGMM, Shaw J (2001) Nature 414:782–787

    Article  CAS  Google Scholar 

  14. Yoon KH, Lee JH, Kim JW, Cho JH, Choi YH, Ko SH (2006) Lancet 368:1681–1688

    Article  Google Scholar 

  15. Knowler WC, Barrett-Connor E, Fowler SE, Hamman RF, Lachin JM, Walker EA (2002) N Engl J Med 346:393–403

    Article  CAS  Google Scholar 

  16. Tuomilehto J, Lindstrom J, Eriksson JG, Valie TT, Hamalainen H, Ilanne-Parikka P (2001) N Engl J Med 344:1343–50

    Article  CAS  Google Scholar 

  17. Racker E (1951) J Biol Chem 190:685–696

    CAS  Google Scholar 

  18. Thornalley PJ (1990) Biochem J 269:1–11

    CAS  Google Scholar 

  19. Kondoh Y, Kawase M, Kawamaki Y, Ohmori S (1992) Res Exp Med 192:407–414

    Article  CAS  Google Scholar 

  20. Christopher MM, Broussard JD, Fallin CW, Drost NJ, Peterson ME (1995) Metab Clin Exp 44:287–290

    Article  CAS  Google Scholar 

  21. McLellan AC, Plillips SA, Thornalley PJ (1992) Anal Biochem 206:12–16

    Article  CAS  Google Scholar 

  22. Thornalley PJ, Hooper NI, Jennings PE, Florkowski CM, Jones AF, Lunec J, Barnett AH (1989) Diabetes Res Clin Prac 7:115–120

    Article  CAS  Google Scholar 

  23. McLellan AC, Plillips SA, Thornalley PJ (1992) Anal Biochem 206:17–23

    Article  CAS  Google Scholar 

  24. Thornalley PJ (1994) Amino Acids 6:15–23

    Article  CAS  Google Scholar 

  25. McLellan AC, Thornalley PJ, Benn J, Sonksen PH (1994) Clin Sci 87:21–29

    CAS  Google Scholar 

  26. Thornalley PJ, McLellan AC, Lo TWC, Benn J, Sonksen PH (1996) Clin Sci 91:575–582

    CAS  Google Scholar 

  27. Haschke-Becher E, Baumgartner M, Bachmann C (2000) Clin Chim Acta 298:99–109

    Article  CAS  Google Scholar 

  28. Wellmer A, Prange J, Gerber J, Zysk G, Lange P, Michel U, Eiffert H, Nau R (2001) Scand J Infect Dis 33:909–913

    Article  CAS  Google Scholar 

  29. Marti R, Varela E, Segura RM, Alegre J, Surinach JM, Pascual C (1997) Clin Chem 43:1010–1015

    CAS  Google Scholar 

  30. Gutheil WG (1998) Anal Biochem 259:62–67

    Article  CAS  Google Scholar 

  31. McLellan AC, Plillips SA, Thornalley PJ (1992) Anal Biochem 206:12–16

    Article  CAS  Google Scholar 

  32. Avramescu A, Noguer T, Magearu V, Marty JL (2001) Anal Chim Acta 433:81–88

    Article  CAS  Google Scholar 

  33. Oi N, Kitahara H, Aoki F (1994) J Chromatogr A 666:457–462

    Article  CAS  Google Scholar 

  34. Olieman C, DeVries ES (1988) Neth Milk Dairy J 42:111–120

    CAS  Google Scholar 

  35. Norton D, Crow B, Bishop M, Kovalcik K, George J, Bralley JA (2007) J Chromatogr B 850:190–198

    Article  CAS  Google Scholar 

  36. Fukushima T, Adachi S, Ichihara H, Al-Kindy S, Imai K (1999) Biomed Chromatogr 13:418–424

    Article  CAS  Google Scholar 

  37. Fukushima T, Lee J-A, Korenaga T, Ichihara H, Kato M, Imai K (2001) Biomed Chromatogr 15:189–195

    Article  CAS  Google Scholar 

  38. Hasegawa H, Fukushima T, Lee J-A, Tsukamoto K, Moriya K, Ono Y, Imai K (2003) Anal Bioanal Chem 377:886–891

    Article  CAS  Google Scholar 

  39. Toyo’oka T, Ishibashi M, Takeda Y, Nakashima K, Imai K (1991) J Chromatogr 588:61–71

    Article  Google Scholar 

  40. Chiappin S, Antonelli G, Gatti R, De Palo EF (2007) Clin Chim Acta 383:30–40

    Article  CAS  Google Scholar 

  41. Gröschl M (2008) Clin Chem 54:1759–1769

    Article  Google Scholar 

  42. Avogaro A, Toffolo G, Miola M, Valerio A, Cobelli C, Del Prato S (1996) J Clin Invest 98:108–115

    Article  CAS  Google Scholar 

  43. Avogaro A, Crepalgi C, Miola M, Maran A, Pengo V, Tiengo A, Del Prato S (1996) J Endocrinol Invest 19:99–105

    CAS  Google Scholar 

Download references

Acknowledgments

This study was supported in part by a Grant-in-Aid for Scientific Research (C) from the Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Laboratory of Analytical and Bio-Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, 422-8526, Japan

    Haruhito Tsutsui, Toshiki Mochizuki, Jun Zhe Min, Kenichiro Todoroki, Koichi Inoue & Toshimasa Toyo’oka

  2. Laboratory of Clinical Pharmaceutics and Pharmacy Practice, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, 422-8526, Japan

    Toshio Maeda

  3. Department of Pharmaceutical, Numazu City Hospital, Harunoki Higasi-Shiizi, Numazu, 410-0302, Japan

    Ichiro Noge

  4. Department of Clinical and Molecular Endocrinology, Numazu City Hospital, Harunoki Higasi-Shiizi, Numazu, 401-0302, Japan

    Yutaka Kitagawa

Authors
  1. Haruhito Tsutsui
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  2. Toshiki Mochizuki
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  3. Toshio Maeda
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  5. Yutaka Kitagawa
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  6. Jun Zhe Min
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  7. Kenichiro Todoroki
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  8. Koichi Inoue
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  9. Toshimasa Toyo’oka
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Correspondence to Toshimasa Toyo’oka.

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Tsutsui, H., Mochizuki, T., Maeda, T. et al. Simultaneous determination of dl-lactic acid and dl-3-hydroxybutyric acid enantiomers in saliva of diabetes mellitus patients by high-throughput LC–ESI-MS/MS. Anal Bioanal Chem 404, 1925–1934 (2012). https://doi.org/10.1007/s00216-012-6320-0

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  • DOI: https://doi.org/10.1007/s00216-012-6320-0

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