, 70:1095 | Cite as

Alternative LC–MS–MS Method for Simultaneous Determination of Proguanil, Its Active Metabolite in Human Plasma and Application to a Bioequivalence Study

  • Satish G. Pingale
  • Ketan K. Nerurkar
  • Ajit M. Padgaonkar
  • Uttam D. Pawar
  • Kiran V. Mangaonkar


An alternative rapid and sensitive liquid chromatography–tandem mass spectrometry method has been developed and validated for simultaneous analysis of proguanil (PRO) and cycloguanil (CYC) in human plasma. The analytes were extracted from human plasma by solid phase extraction. Riluzole (RIL) was used as an internal standard for proguanil and cycloguanil. A HyPURITY Advance C18 column provided chromatographic separation of analytes followed by detection with mass spectrometry. The method involves simple isocratic chromatography conditions and mass spectrometric detection in the positive ionization mode using an API-4000 system. The proposed method has been validated with linear range of 1.5–150.0 ng mL−1 for PRO and 0.5–50.0 ng mL−1 for CYC. The inter-run and intra-run precision values are within 2.54, 9.19% for PRO and 1.99, 10.69% for CYC at LOQ levels. The overall recoveries for PRO and CYC were 102.52 and 106.72%, respectively. Total elution time was as low as 2.50 min. This validated method was used successfully for analysis of plasma samples from a bioequivalence study.


Column liquid chromatography Tandem mass spectrometry Proguanil in human plasma Cycloguanil and riluzole 



The authors are indebted to Prof. R. T. Sane, Dr. Vikas Vaidya, Mr. Nelson Varghese and Mr. Sudhir Pawar for their continuous support and for providing the laboratory facilities required for this assay.


  1. 1.
    GlaxoSmithKline-Malarone (atovaquone and proguanil hydrochloride) tablets prescribing information (2007) 1–4Google Scholar
  2. 2.
    Wangboonskul J, White NJ, Nosten F, ter Kuile F, Moody RR, Taylor RB (1993) Eur J Clin Pharmacol 44:247–251. doi: 10.1007/BF00271366 CrossRefGoogle Scholar
  3. 3.
    De Aguiar PF, Heyden YV, Oost YV, Coomber TJ, Massart DL (1997) J Pharm Biomed Anal 15:1781–1787. doi: 10.1016/S0731-7085(96)01967-X CrossRefGoogle Scholar
  4. 4.
    Kelly JA, Fletcher KA (1986) J Chromatogr A 381:464–471Google Scholar
  5. 5.
    Edstein MD (1986) J Chromatogr A 380:184–189Google Scholar
  6. 6.
    Taylor RB, Moody RR, Oobekpe NA (1987) J Chromatogr A 416:394–399. doi: 10.1016/0378-4347(87)80526-1 CrossRefGoogle Scholar
  7. 7.
    Bergqvist Y, Funding L, Krysen B, Leek T, Yvell K (1998) Ther Drug Monit 20:325–330. doi: 10.1097/00007691-199806000-00014 CrossRefGoogle Scholar
  8. 8.
    Taylor RB, Reid RG, Low AS (2001) J Chromatogr A 916:201–206. doi: 10.1016/S0021-9673(00)01035-9 CrossRefGoogle Scholar
  9. 9.
    Taylor RB, Reid RG (1995) J Pharm Biomed Anal 13:21–26. doi: 10.1016/0731-7085(94)00127-N CrossRefGoogle Scholar
  10. 10.
    Bergqvist Y, Funding L, Kaneko A, Krysen B, Leek T (1998) J Chromatogr B Biomed Sci Appl 719:141–149. doi: 10.1016/S0378-4347(98)00382-X CrossRefGoogle Scholar
  11. 11.
    Bergqvist Y, Hopstadius C (2000) J Chromatogr B Analyt Technol Biomed Life Sci 741:189–193. doi: 10.1016/S0378-4347(00)00082-7 CrossRefGoogle Scholar
  12. 12.
    Lejeune D, Souletie I, Houze S, Bricon TL, Bras JL, Gourmel B, Houze P (2007) J Pharm Biomed Anal 43:1106–1115. doi: 10.1016/j.jpba.2006.09.036 CrossRefGoogle Scholar
  13. 13.
    Ebeshi BU, Obodozie OO, Bolaji OO, Ogunbona FA (2005) Afr J Biotechnol 4:856–861Google Scholar
  14. 14.
    Hoskins JM, Shenfield GM, Gross AS (1997) J Chromatogr B Biomed Sci Appl 696:81–87. doi: 10.1016/S0378-4347(97)00225-9 CrossRefGoogle Scholar
  15. 15.
    Taylor RB, Alexander C, Nathwani D, Zimbler N (1996) J Liq Chromatogr Relat Technol 19:1317–1328. doi: 10.1080/10826079608006320 CrossRefGoogle Scholar
  16. 16.
    Kusaka M, Setiabudy R, Chiba K, Ishizaki T (1996) Am J Trop Med Hyg 54:189–196Google Scholar
  17. 17.
    Wanwimolruk S, Pratt EL (1995) J Liq Chromatogr 18:4097–4105. doi: 10.1080/10826079508013747 CrossRefGoogle Scholar
  18. 18.
    Kolawole JA, Taylor RB, Moody RR (1995) J Chromatogr B Biomed Appl 674:149–154. doi: 10.1016/0378-4347(95)00293-3 CrossRefGoogle Scholar
  19. 19.
    Chaulet FJ, Grelaud G, Patrick MB, Mounier C, Brazier LJ (1994) J Pharm Biomed Anal 12:111–117. doi: 10.1016/0731-7085(94)80018-9 CrossRefGoogle Scholar
  20. 20.
    Taylor RB, Behrens R, Moody RR (1990) J Chromatogr A 527:490–497Google Scholar
  21. 21.
    Leveque NL, Charman WN, Chiu FCK (2006) J Chromatogr B Analyt Technol Biomed Life Sci 830:314–321. doi: 10.1016/j.jchromb.2005.11.004 CrossRefGoogle Scholar
  22. 22.
    Guidance for industry: bioanalytical method validation, U.S. Department of health and human services, food and drug administration centre for drug evaluation and research (CDER), centre for veterinary medicine (CVM) 2001Google Scholar

Copyright information

© Vieweg+Teubner | GWV Fachverlage GmbH 2009

Authors and Affiliations

  • Satish G. Pingale
    • 1
    • 2
  • Ketan K. Nerurkar
    • 2
  • Ajit M. Padgaonkar
    • 2
  • Uttam D. Pawar
    • 2
  • Kiran V. Mangaonkar
    • 1
  1. 1.Department of Chemistry, Mithibai College of ArtsChaun Institute of Science & Amrutben Jivanlal College of Commerce and EconomicsMumbaiIndia
  2. 2.Drug Monitoring Research InstituteNavi MumbaiIndia

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