Submit Manuscript

Journal of Biological Sciences

Year: 2008 | Volume: 8 | Issue: 6 | Page No.: 1110-1114
Research Article

Optimization of Rat Liver Microsomal Stability Assay Using HPLC

S.K. Mondal
Department of Pharmaceutical Technology, Jadavpur University, Kolkata-700 032, India
U.K. Mazumder
Department of Pharmaceutical Technology, Jadavpur University, Kolkata-700 032, India
N.B. Mondal
Steroid and Terpenoid Chemistry Division, Indian Institute of Chemical Biology, 4 Raja SC Mullick Road, Kolkata-700 032, India
S. Banerjee
Steroid and Terpenoid Chemistry Division, Indian Institute of Chemical Biology, 4 Raja SC Mullick Road, Kolkata-700 032, India
Microsomal stability of atenolol, propranolol HCl, verapamil HCl, imipramine HCl, midazolam HCl andrographolide and 14-Deoxyandrographolide was performed and quantitation was done by HPLC. The results with a single batch microsome showed good day-to-day reproducibility and variation in percent stability was within ± 5.71% of the average for all the compounds for the entire study period of 2 years. Microsome was found to be stable at -80 °C store for at least upto 2 years. Two batches of microsomes from each of 3 vendors were also used for stability study where similar type of animal, microsome preparation method and assay protocol were used. The findings demonstrated variations for both batch to batch and vendor to vendor were within 10%. Results indicated that variability in stability data can be minimized by selecting a fixed type of animal, keeping similar method of microsome preparation and following same assay protocol.
PDF Fulltext XML References Citation

Comments

p.giriraj Reply

dear sir,
its a very good study,i would like to learn more from this study.i also like to do my project work in the same study. so please send me contact information to the crosponding author and more deatials regarding this
thank you
p.giriraj

Susanta Kumar ondal Reply

Dear Giriraj,
Thanks for your comments. I am glad that the paper is useful to you. you may contact me for futher information.
Best wishes,
Susanta

Susanta Kumar Mondal Reply

Thanks for your comments

Best regards
Susanta

Leave a Comment

Your email address will not be published. Required fields are marked *

Article Trend

Total views 5521

References

  1. De-Waterbeemd, H., D.A. Smith, K. Beaumont and D.K. Walker, 2001. Property-based design: Optimization of drug absorption and pharmacokinetics. J. Med. Chem., 44: 1313-1333.
    CrossRefPubMedDirect Link
  2. Di, L., E.H. Kerns, Y. Hong, T.A. Kleintop, O.J. McConnell and D.M. Huryn, 2003. Optimization of a higher throughput microsomal stability screening assay for profiling drug discovery candidates. J. Biomol. Screen., 8: 453-462.
    CrossRefPubMedDirect Link
  3. Giuliano, C., M. Jairaj, C.M. Zafiu and R. Laufer, 2005. Direct determination of unbound intrinsic drug clearance in the microsomal stability assay. Drug Metab. Dispos., 33: 1319-1324.
    CrossRefPubMedDirect Link
  4. Hasegawa, A., Y. Kawaguchi, H. Nakasa, H. Nakamura, S. Ohmori, I. Ishii and M. Kitada, 2002. Effects of Kampo extracts on drug metabolism in rat liver microsomes: Rhei Rhizoma extract and Glycyrrhizae Radix extract inhibit drug oxidation. Jap. J. Pharmacol., 89: 164-170.
    CrossRefPubMedDirect Link
  5. Ito, K. and J.B. Houston, 2004. Comparison of the use of liver models for predicting drug clearance using in vitro kinetic data from hepatic microsomes and isolated hepatocytes. Pharm. Res., 21: 785-792.
    CrossRefPubMedDirect Link
  6. Jenkins, K.M., R. Angeles, M.T. Quintos, R. Xu, D.B. Kassel and R.A. Rourick, 2004. Automated high throughput ADME assays for metabolic stability and cytochrome P450 inhibition profiling of combinatorial libraries. J. Pharm. Biomed. Anal., 34: 989-1004.
    CrossRefPubMedDirect Link
  7. Li, A.P., 2004. In vitro approaches to evaluate ADMET drug properties. Curr. Top. Med. Chem., 4: 701-706.
    CrossRefPubMedDirect Link
  8. Lin, J.H. and A.D. Rodrigues, 2001. In vitro models for early studies of drug metabolism. In: Pharmacokinetic optimization in Drug research: Biological, Physicochemical and computational strategies. Testa, B., H. Van De Waterbeemd, G. Folkers and R. Guy (Eds.). Verlag Helvetica Chimica Acta, Switzerland, ISBN-10: 3-906390-22-5 pp: 217-243.
  9. Linget, J.M. and P.D. Vignaud, 1999. Automation of metabolic stability studies in microsomes, cytosol and plasma using a 215 Gilson liquid handler. J. Pharm Biomed. Anal., 19: 893-901.
    CrossRefPubMedDirect Link
  10. Lowry, O.H., N.J. Rosebrough, A.L. Farr and R.J. Randall, 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem., 193: 265-275.
    CrossRefPubMedDirect Link
  11. Matsuda, T., M. Kuroyanagi, S. Sugiyama, K. Umehara, A. Ueno and K. Nishi, 1994. Cell Differentiation-inducing diterpenes from Andrographis paniculata nees. Chem. Pharm. Bull., 42: 1216-1225.
    PubMedDirect Link
  12. Sinko, P.J., 1999. Drug selection in early drug development: Screening for acceptable pharmacokinetic properties using combined in vitro and computational approaches. Curr. Opin. Drug Disc. Dev., 2: 42-48.
    Direct Link
  13. Trubetskoy, O.V., J.R. Gibson and B.D. Marks, 2005. Highly miniaturized formats for in vitro drug metabolism assays using vivid® fluorescent substrates and recombinant human cytochrome P450 enzymes. J. Biomol. Screen., 10: 56-66.
    CrossRefPubMedDirect Link
  14. Xu, R., C. Nemes, K.M. Jenkins, R.A. Rourick, D.B. Kassel and C.Z.C. Liu, 2002. Application of parallel liquid chromatography/mass spectrometry for high throughput microsomal stability screening of compound libraries. J. Am. Soc. Mass Spectrom. 13: 155-165.
    CrossRefPubMedDirect Link

Keywords