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Pharmaceutisch Weekblad

, Volume 11, Issue 6, pp 232–235 | Cite as

Optimization of thein vitro glucuronidation of ibuprofen using factorial design

  • Nico R. Niemeijer
  • Jan H. de Boer
  • Thijs K. Gerding
  • Rokus A. de Zeeuw
Short Communications

Abstract

Ibuprofen was chosen as a test compound to perform a multivariate design in order to determine the highest yield of thein vitro glucuronidation reaction in relation to the concentration of reacting and activating substances. Preliminary studies with a univariate design indicated that the concentration of Mg2+ had no significant effect on the glucuronidation and that Triton X-100 could be omitted as it appeared to inhibit the glucuronidation. In a 33 factorial design the influence of concentrations of ibuprofen, UDP glucuronic acid and enzyme, respectively, on the yield of ibuprofen glucuronide was established. It was concluded that the highest amount of ibuprofen glucuronide formed (within the limits of this design) was achieved with an ibuprofen concentration of 486 μM, a UDP-glucuronic acid concentration of 3 mM and an enzyme concentration of 3.57 mg/ml. Using this methodology it is possible to optimize the glucuronidation yield in a more rational way, which can be useful in the upscaling of enzymatic reactions.

Keywords

Enzyme activation Glucuronidase Ibuprofen Optimization Multivariate design Univariate design 

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References

  1. 1.
    Upton RA, Buskin JN, Williams RL, Holford NHG, Riegelman S. Negligible excretion of unchanged ketoprofen, naproxen and probenicid in urine. J Pharm Sci 1980;69:1254–57.PubMedGoogle Scholar
  2. 2.
    Wilson BK, Thompson JA. Glucuronidation of propranolol by dog liver microsomes. Drug Metab Dispos 1984;12:161–4.PubMedGoogle Scholar
  3. 3.
    Thompson JA, Hull JE, Norris KJ. Glucuronidation of propranolol and 4′-hydroxypropranolol. Drug Metab Dispos 1981;9:466–71.PubMedGoogle Scholar
  4. 4.
    Del Villar E, Sanchez E, Tephly TR. Morfine metabolism. II. Studies on morphine glucuronyltransferase activity in intestinal microsomes of rats. Drug Metab Dispos 1974;2:370–4.PubMedGoogle Scholar
  5. 5.
    Gerding TK, Drenth BFH, De Zeeuw RA. Separation of N-0437 enantiomers by RP-HPLC after pre-column derivatization with D(+)-glucuronic acid. J HRC&CC 1987;10:523–5.Google Scholar
  6. 6.
    Pallante S, Lyle MA, Fenselau C. Synthesis and characterization of glucuronides of 5′-hydroxy-d 9-tetrahydrocannobinol and 11-hydroxy-d 9-tetrahydrocannabinol. Drug Metab Dispos 1978;6:389–95.PubMedGoogle Scholar
  7. 7.
    Lehman JP, Fenselau C. Synthesis of quaternary ammonium-linked glucuronides by rabbit hepatic microsomal UDP-glucuronyl-transferase and analysis by fast-atom bombardment mass spectrometry. Drug Metab Dispos 1982;10:446–9.PubMedGoogle Scholar
  8. 8.
    Box GEP, Hunter WG, Hunter JS. Statistics for experiments. Chichester: John Wiley, 1978.Google Scholar
  9. 9.
    Davies OL. The design and analysis of industrial experiments. 2nd ed. London: Longman Group, 1979.Google Scholar
  10. 10.
    Inoue T, Suzuki S, Niwaguchi T. The metabolism of l-phenyl-2-(N-methyl-N-benzylamino)propane(benzphetamine)in vitro in the rat. Xenobiotica 1983;13:241–9.PubMedGoogle Scholar
  11. 11.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem 1951;193:265–75.PubMedGoogle Scholar

Copyright information

© Royal Dutch Association for Advancement of Pharmacy 1989

Authors and Affiliations

  • Nico R. Niemeijer
    • 1
  • Jan H. de Boer
    • 1
  • Thijs K. Gerding
    • 1
  • Rokus A. de Zeeuw
    • 1
  1. 1.Department of Analytical Chemistry and ToxicologyUniversity of GroningenAW Groningenthe Netherlands

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