Skip to main content
Log in

High-throughput determination of theophylline and caffeine in human serum by conventional liquid chromatography-mass spectrometry

  • Original Article
  • Published:
Forensic Toxicology Aims and scope Submit manuscript

Abstract

Automated high-performance liquid chromatography/mass spectrometry (HPLC-MS) with backflush column-switching was established for ultra-fast determination of theophylline and caffeine. A 400-μl portion of serum sample diluted with ultrapure water was injected and transferred to an Oasis HLB cartridge used as a precolumn for extraction. After switching the valves, the analytes trapped in the precolumn were eluted in the backflush mode and separated with a Chromolith Performance RP-18e column (C18-bonded monolithic silica); the compounds in column effluents were then detected by atmospheric pressure chemical ionization (APCI)-MS. The present method successfully provided high-throughput determination of theophylline and caffeine within 2 min. Satisfactory linearity, reproducibility, and sensitivity could be obtained for analysis of therapeutic and toxic levels of both compounds. Because of the very simple procedure and high throughput using the conventional HPLC system, the present method seems to have high potential in the fields of forensic toxicology and emergency medicine.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Stavric B (1988) Methylxanthines: toxicity to humans. 1. Theophylline. Food Chem Toxicol 26:541–565

    Article  PubMed  CAS  Google Scholar 

  2. Stavric B (1988) Methylxanthines: toxicity to humans. 2. Caffeine. Food Chem Toxicol 26:645–662

    Article  PubMed  CAS  Google Scholar 

  3. Stavric B (1988) Methylxanthines: toxicity to humans. 3. Theobromine, paraxanthine and the combined effects of methylxanthines. Food Chem Toxicol 26:725–733

    Article  PubMed  CAS  Google Scholar 

  4. Kumazawa T, Seno H, Lee X-P, Ishii A, Watanabe-Suzuki K, Sato K, Suzuki O (1999) Extraction of methylxanthines from human body fluids by solid-phase microextraction. Anal Chim Acta 387:53–60

    Article  CAS  Google Scholar 

  5. Sheehan M, Hertel RH, Kelly CT (1977) Gaschromatographic/mass-spectrometric determination of theophylline in whole blood. Clin Chem 23:64–68

    PubMed  CAS  Google Scholar 

  6. Floberg S, Lindstrom B, Lonnerholm G (1980) Simultaneous determination of theophylline and caffeine after extractive alkylation in small volumes of plasma by gas chromatography-mass spectrometry. J Chromatogr 221:166–169

    Article  PubMed  CAS  Google Scholar 

  7. Tserng KY (1983) Gas chromatographic-mass spectrometric quantitation of theophylline and its metabolites in biological fluids. J Pharm Sci 72:526–529

    Article  PubMed  CAS  Google Scholar 

  8. Desage M, Soubeyrand J, Soun A, Brazier JL, Georges Y (1984) Automated theophylline assay using gas chromatography and a mass-selective detector. J Chromatogr 336:285–291

    Article  PubMed  CAS  Google Scholar 

  9. Bailey E, Farmer PB, Peal JA, Hotchkiss SA, Caldwell J (1987) Analytical methodology to determine stable isotopically labelled and unlabelled theophylline in human plasma using capillary gas chromatography-mass spectrometry. J Chromatogr 416:81–89

    Article  PubMed  CAS  Google Scholar 

  10. Kasuya Y, Furuta T, Shimota H (1989) Capillary gas chromatographic-mass spectrometric determination of stable isotopically labelled and unlabelled theophylline in serum and urine and of 1,3-dimethyluric acid in urine. J Chromatogr 494:101–108

    Article  PubMed  CAS  Google Scholar 

  11. Nakajima M, Sato S, Yamato S, Shimada K, Kitagawa S, Honda A, Miyamoto J, Hirano H, Suzuki M, Miyazaki H (2003) Assessment of tear concentrations on therapeutic drug monitoring. III. Determination of theophylline in tears by gas chromatography/mass spectrometry with electron ionization mode. Drug Metab Pharmacokinet 18:139–145

    Article  PubMed  CAS  Google Scholar 

  12. Saka K, Uemura K, Shintani-Ishida K, Yoshida K (2007) Acetic acid improves the sensitivity of theophylline analysis by gas chromatography-mass spectrometry. J Chromatogr B 846: 240–244

    Article  CAS  Google Scholar 

  13. Hieda Y, Kashimura S, Hara K, Kageura M (1994) Development of the method for analysis of theophylline and its related compounds using capillary high-performance liquid chromatography/fast atom bombardment-mass spectrometry (in Japanese with English abstract). Jpn J Legal Med 48:253–262

    CAS  Google Scholar 

  14. Song J, Park KU, Park HD, Yoon Y, Kim JQ (2004) Highthroughput liquid chromatography-tandem mass spectrometry assay for plasma theophylline and its metabolites. Clin Chem 50:2176–2179

    Article  PubMed  CAS  Google Scholar 

  15. Beaudry F, Lavoie J, Vachon P (2005) Development of an electrospray ionization mass spectrometric method for the quantification of theophylline in horse serum. Biomed Chromatogr 19:643–648

    Article  PubMed  CAS  Google Scholar 

  16. Hori Y, Fujisawa M, Shimada K, Hirose Y, Yoshioka T (2006) Method for screening and quantitative determination of serum levels of salicylic acid, acetaminophen, theophylline, phenobarbital, bromvalerylurea, pentobarbital, and amobarbital using liquid chromatography/electrospray mass spectrometry. Biol Pharm Bull 29:7–13

    Article  PubMed  CAS  Google Scholar 

  17. Arinobu T, Hattori H, Seno H, Ishii A, Suzuki O (2002) Comparison of SSI with APCI as an interface of HPLC-mass spectrometry for analysis of a drug and its metabolites. J Am Soc Mass Spectrom 13:204–208

    Article  PubMed  CAS  Google Scholar 

  18. Arinobu T, Hattori H, Iwai M, Ishii A, Kumazawa T, Suzuki O, Seno H (2002) Liquid chromatographic-mass spectrometric determination of haloperidol and its metabolites in human plasma and urine. J Chromatogr B 776:107–113

    Article  CAS  Google Scholar 

  19. Arinobu T, Hattori H, Ishii A, Kumazawa T, Lee X-P, Suzuki O, Seno H (2003) Comparison of sonic spray ionization with atmospheric pressure chemical ionization as interface of liquid chromatography-mass spectrometry for the analysis of some local anesthetics. Chromtographia 57:301–307

    Article  CAS  Google Scholar 

  20. Arinobu T, Hattori H, Ishii A, Kumazawa T, Lee X-P, Kojima S, Suzuki O, Seno H (2003) Rapid analysis of pentazocine in human plasma by liquid chromatography/mass spectrometry with sonic spray ionization. Anal Chim Acta 492:249–252

    Article  CAS  Google Scholar 

  21. Lee X-P, Kumazawa T, Sato J, Shoji Y, Hasegawa C, Karibe C, Arinobu T, Seno H, Sato K (2003) Simple method for the determination of benzodiazepines in human body fluids by high-performance liquid chromatography-mass spectrometry. Anal Chim Acta 492:223–231

    Article  CAS  Google Scholar 

  22. Hattori H, Arinobu T, Iwai M, Suzuki O, Seno H (2003) Liquid chromatographic/mass spectrometric determination of morphine, codeine and cocaine in human sera using a new internal surface reversed phase column. Jpn J Forensic Toxicol 21:47–54

    CAS  Google Scholar 

  23. Aoki R, Arinobu T, Kumazawa T, Hattori H, Noguchi H (2007) An automated on-line method for simultaneous analysis of phenothiazines in human serum by high-performance liquid chromatography/sonic spray ionization mass spectrometry using backflush column switching. Forensic Toxicol 25:8–15

    Article  CAS  Google Scholar 

  24. Katagi M, Tatsuno M, Nishikawa M, Tsuchihashi H (1999) On-line solid-phase extraction liquid chromatographycontinuous flow frit fast atom bombardment mass spectrometric and tandem mass spectrometric determination of hydrolysis products of nerve agents alkyl methylphosphonic acids by pbromophenacyl derivatization. J Chromatogr A 833:169–179

    Article  PubMed  CAS  Google Scholar 

  25. Masucci JA, Caldwell GW, Jones WJ, Juzwin SJ, Sasso PJ, Evangelisto M (2001) The use of on-line and off-line chromatographic extraction techniques coupled with mass spectrometry for support of in vivo and in vitro assays in drug discovery. Curr Top Med Chem 1:463–471

    Article  PubMed  CAS  Google Scholar 

  26. Katagi M, Nishikawa M, Tatsuno M, Miki A, Tsuchihashi H (2001) Column-switching high-performance liquid chromatography-electrospray ionization mass spectrometry for identification of heroin metabolites in human urine. J Chromatogr B 751:177–185

    Article  CAS  Google Scholar 

  27. Miki A, Tatsuno M, Katagi M, Nishikawa M, Tsuchihashi H (2002) Simultaneous determination of eleven benzodiazepine hypnotics and eleven relevant metabolites in urine by columnswitching liquid chromatography-mass spectrometry. J Anal Toxicol 26:87–93

    PubMed  CAS  Google Scholar 

  28. Miki A, Katagi M, Tsuchihashi H (2003) Determination of methamphetamine and its metabolites incorporated in hair by column-switching liquid chromatography-mass spectrometry. J Anal Toxicol 27:95–102

    PubMed  CAS  Google Scholar 

  29. Friedrich G, Rose T, Rissler K (2003) Determination of testosterone metabolites in human hepatocytes I. Development of an on-line sample preparation liquid chromatography technique and mass spectroscopic detection of 6β-hydroxytestosterone. J Chromatogr B 784:49–61

    Article  CAS  Google Scholar 

  30. Uges DRA (2004) Hospital toxicology. In: Moffat AC, Osselton MD, Widdop B (eds) Clarke’s analysis of drugs and poisons. Pharmaceutical, London, pp 3–36

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Tetsuya Arinobu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Arinobu, T., Hattori, H., Kumazawa, T. et al. High-throughput determination of theophylline and caffeine in human serum by conventional liquid chromatography-mass spectrometry. Forensic Toxicol 27, 1–6 (2009). https://doi.org/10.1007/s11419-008-0058-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11419-008-0058-6

Keywords

Navigation