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Plasma indomethacin assay by reversedphase ion pair high pressure liquid chromatography

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Abstract

A new rapid, selective and sensitive high pressure liquid Chromatographic (HPLC) assay for indomethacin in plasma is described. The method involves precipitation of proteins with perchloric acid, followed by dichloromethane extraction using flurbiprofen as an internal standard. The organic solvent was evaporated and the residue dissolved in a water-methanol (2+3) phosphate buffer mixture with an apparent pH of 6.8.

Aliquots of 100μl were injected automatically into the Chromatograph. The separation of indomethacin was achieved on a reversed phase (C18, 10μm) column with a mobile phase consisting of 65% (vol/vol) methanol in water solution of apparent pH 6.8 containing tetrabutylammonium hydrogensulfate as an ion pairing agent. Quantitation of indomethacin was performed by UV detection at 235 nm.

At a 2.0 mg.l−1 concentration of indomethacin in plasma the analytical recovery was 82.9±3.4% (n=7), the intra-day variability (CV) was 3.6% (n=7) and the inter-day variability (CV) was 11.0% (n=7). The calibration curve was linear (typical r-values>0.990) in the range of plasma concentrations as usually found during indomethacin therapy (up to 6 mg.l−1). The limit of sensitivity is 0.025 mg.l−1.

The metabolitesO-desmethylindomethacin andO-desmethyldeschlorobenzoylindomethacin did not interfere with the method.

The capacity of an analyst using this method with automated injection and peak integration is about forty samples in duplicate per day. The applicability of the method for pharmacokinetic studies is demonstrated.

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References

  1. Kwan KC, Breault GO, Umberhauer ER, McMahon FG, Duggan DE. Kinetics of indomethacin absorption, elimination and enterohepatic circulation in man. J Pharmacokin Biopharm 1976;4:255–80.

    Google Scholar 

  2. Alván G, Orme M, Bertilsson L, Ekstrand R, Palmer L. Pharmacokinetics of indomethacin. Clin Pharmacol Ther 1975;18:364–75.

    Google Scholar 

  3. Harman RE, Meisinger MAP, Davis GE, Kuehl FA. The metabolites of indomethacin, a new anti-inflammatory drug. J Pharmacol Exp Ther 1964;143:215–20.

    Google Scholar 

  4. Hucher HB, Zacchei AG, Cox SV, Brodie DA, Cantwell NHR. Studies on the absorption, distribution and excretion of indomethacin in various species. J Pharmacol Exp Ther 1966;153:237–49.

    Google Scholar 

  5. Hucher HB, Hoffman EA. Physiologic disposition and metabolic fate of indomethacin analogs. J Pharm Sci 1971;60:1049–52.

    Google Scholar 

  6. Duggan DE, Hogans AF, Kwan KC, McMahon FG. The metabolism of indomethacin in man. J Pharmacol Exp Ther 1972;181:563–75.

    Google Scholar 

  7. Champion GD, Paulus HE, Mongan E, Okun R, Pearson CM, Sarkissian E. The effect of aspirin on serum indomethacin. Clin Pharmacol Ther 1972;13:239–44.

    Google Scholar 

  8. Hvidberg E, Lausen HH, Jansen JA. Indomethacin: plasma concentrations and protein binding in man. Eur J Clin Pharmacol 1972;4:119–24.

    Google Scholar 

  9. Lindquist B, Møller Jensen K, Johansson H, Hansen T. Effect of concurrent administration of aspirin and indomethacin on serum concentrations. Clin Pharmacol Ther 1974;15:247–52.

    Google Scholar 

  10. Hare LE, Ditzler CA, Duggan DE. Radioimmunoassay of indomethacin in biological fluids. J Pharm Sci 1977;66:486–9.

    Google Scholar 

  11. Alam I, Langone JJ, Levine L. Immunoassay of indomethacin: the use of (1251) protein A to detect specific serologic binding. Prostaglandins Med 1979;2:167–75.

    Google Scholar 

  12. Ferry DG, Ferry DM, Moller PW, McQueen EG. Indomethacin estimation in plasma and serum by electron capture gas chromatography. J Chromatogr 1974;89:110–2.

    Google Scholar 

  13. Helleberg L. Determination of indomethacin in serum and urine by electron-capture gas-liquid chromatography. J Chromatogr 1976;117:167–73.

    Google Scholar 

  14. Plazonnet B, Vandenheuvel WJA. Preparation, gas chromatography and mass spectrometry of methyl and trimethylsilyl esters of indomethacin. J Chromatogr 1977;142:587–96.

    Google Scholar 

  15. Arbin A. Three alkylation methods for the determination of indomethacin in plasma by electron capture gas chromatography. J Chromatogr 1977;144:85–92.

    Google Scholar 

  16. Jensen KM. Determination of indomethacin in serum by an extractive alkylation technique and gas-liquid chromatography. J Chromatogr 1978;153:195–202.

    Google Scholar 

  17. Sibeon RG, Baty JD, Baber N, Chan K, Orme ML. Quantitative gas-liquid Chromatographic method for the determination of indomethacin in biological fluids. J Chromatogr 1978;153:189–94.

    Google Scholar 

  18. Evans MA. GLC microdetermination of indomethacin in plasma. J Pharm Sci 1980;69:219–20.

    Google Scholar 

  19. Palmér L, Bertilsson L, Alván G, Orme M, Sjöqvist F, Holmstedt B. Indomethacin: quantitative determination in plasma by mass fragmentography including pilot pharmacokinetics in man. In: Robinson HJ, Vane JR, eds, Prostaglandin synthetase inhibitors. New York: Raven Press, 1973:963.

    Google Scholar 

  20. Søndergaard I, Steiness E. Determination of indomethacin in plasma and urine by direct quantitative thin-layer chromatography. J Chromatogr 1979;162:485–8.

    Google Scholar 

  21. Van der Meer MJ, Hundt HKL. Estimation of serum indomethacin at therapeutic levels by means of thin-layer chromatography and spectrophotometry. J Chromatogr 1980;181:282–5.

    Google Scholar 

  22. Bayne WF, East T, Dye D. High-pressure liquid Chromatographic method with postcolumn, in-line hydrolysis and fluorometric-detection for indomethacin in biological fluids. J Pharm Sci 1981;70:458–9.

    Google Scholar 

  23. Bernstein MS, Evans MA. High-performance liquid chromatography — fluorescense analysis for indomethacin and metabolites in biological fluids. J Chromatogr 1982;229:179–87.

    Google Scholar 

  24. Skellern GG, Salole EG. A high-speed liquid Chromatographic analysis of indomethacin in plasma. J Chromatogr 1975;114:483–5.

    Google Scholar 

  25. Soldin SJ, Gero T. A liquid-chromatographic analysis for indomethacin in serum. Clin Chem 1979;25:589–91.

    Google Scholar 

  26. Terweij-Groen CP, Heemstra S, Kraak JC. Rapid determination of indomethacin and salicylic acid in serum by means of reversed-phase liquid chromatography. J Chromatogr 1980;181:385–97.

    Google Scholar 

  27. Kazmi S, Ali A, Plakogiannis FM. High-pressure liquid Chromatographic determination of indomethacin in plasma after ointment application and oral administration. Drug Dev Ind Pharmacy 1981;7:359–65.

    Google Scholar 

  28. Shinek JL, Rao NGS, Wahba Khalil SK. High performance liquid Chromatographic analysis of tolmetin, indomethacin and sulindac in plasma. J Liquid Chromatogr 1981;4:1987–2013.

    Google Scholar 

  29. Jun HW, Suryakusuma H. Improved and rapid liquid Chromatographic determination of indomethacin in serum. Analytical Letters 1982;15(B13):1063–71.

    Google Scholar 

  30. Cooper JK, McKay G, Hawes EM, Midha KK. High-performance liquid Chromatographic assay of indomethacin and its application in pharmacokinetics in healthy volunteers. J Chromatogr 1982;233:289–96.

    Google Scholar 

  31. Greizerstein HB, McLaughlin IG. The rapid determination of indomethacin in 50μl blood samples. J Liq Chromatogr 1982;5:337–43.

    Google Scholar 

  32. Jonkman JHG. Ionpaar-extractie als isoleringsmethode bij de analyse van geneesmiddelen en metabolieten in lichaamsvloeistoffen. I. Inleiding en algemene principes. Pharm Weekbl 1975;110:649–55.

    Google Scholar 

  33. Jonkman JHG. Ionpaar-extractie als isoleringsmethode bij de analyse van geneesmiddelen en metabolieten in lichaamsvloeistoffen. II. Praktische uitvoering en toepassingen. Pharm Weekbl 1975:110:673–89.

    Google Scholar 

  34. Smith RV, Stewart JT. Textbook of biopharmaceutic analysis: a description of methods for the determination of drugs in biological fluids. Philadelphia: Lea & Febiger, 1981:44.

    Google Scholar 

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Authors and Affiliations

  1. Drug Monitoring Unit, Laboratory for Drug Analysis, P.O. Box 147, 9400, AC Assen, The Netherlands

    J. H. G. Jonkman, W. J. V. Van Der Boon, R. Schoenmaker & A. Holtkamp

Authors
  1. J. H. G. Jonkman
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  2. W. J. V. Van Der Boon
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  3. R. Schoenmaker
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  4. A. Holtkamp
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In honour of Professor Faber on the occasion of his 25-years' professorate.

Correspondence.

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Jonkman, J.H.G., Van Der Boon, W.J.V., Schoenmaker, R. et al. Plasma indomethacin assay by reversedphase ion pair high pressure liquid chromatography. Pharmaceutisch Weekblad Scientific Edition 5, 313–318 (1983). https://doi.org/10.1007/BF02074861

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  • DOI: https://doi.org/10.1007/BF02074861

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