Carla Aiolfi Guimaraes, Farid Menaa, Bouzid Menaa, Ivo Lebrun, Joyce S. Quenca-Guillen, Aline Vivian Vatti Auada, Lucildes P. Mercuri, Paula Ferreira, Maria Inês Rocha Miritello Santoro

Abstract: The development of facile and rapid quantification of biologically active biomolecules such as isotretitoin in therapeutic drugs contained in many generic formu- lations is necessary for determining their efficiency and their quality to improve the human health care. Isotretritoin finds its applications in the maintenance of epithelial tissues. Different processes to date such as normal phase HPLC, or gas chromatrography am- ong others are able to separate and quantify isote- troin. However, the extraction is quite complex and in the case of HPLC, the analysis requires long retention times. In such context, an isocratic reversed- phase high-performance liquid chromatography (HP- LC) technique coupled with an UV-vis detector is described here for easy separation and quantification of 13-cis-retinoic (isotretinoin) from soft gelatin capsule formulations. The isotretinoin was extracted from three different commercial drug samples with tetrahydrofuran (THF) solvent by a procedure that can be completed in less than 10 minutes. Subsequent separation and quantification were accomplished in less than 5 minutes under isocratic reversed-phase conditions on a Lichrospher RP18 column and a mobile phase consisting of 0.01% TFA/acetonitrile (15/85, v/v) at a flow rate of 1.0 mL/min. Isotretoin was detected for the three samples via its UV-vis absorbance at 342 nm. The method was validated and the results showed good linearity, precision and accuracy for sensitive and selective quantitative determination of isotretinoin in the different pharmaceutical formulations. We found that the average isotretinoin content in two of the three commercial pro- ducts fell outside the 90-110% United States Pha- rmacopeia specifications. Consequently, the facile extraction and the precise method for the biomole- cule quantification open up tremendous possibilities in improving the quality control of drugs which can exist as different generic brands.

Keywords: 13-Cis Retinoic Acid; Reversed-Phase Chromatography; Isotretitoin Extraction; Isotretinoin Quantification; Pharmaceuticals Formulation

Cite this paper: nullGuimaraes, C. , Menaa, F. , Menaa, B. , Lebrun, I. , Quenca-Guillen, J. , Auada, A. , Mercuri, L. , Ferreira, P. and Santoro, M. (2010) Determination of isotretinoin in pharmaceutical formulations by reversed-phase HPLC. Journal of Biomedical Science and Engineering, 3, 454-458. doi: 10.4236/jbise.2010.35063.

References

[1]   Gatti, R., Gioia, M.G. and Cavrini, V. (2000) Analysis and stability study of retinoids in pharmaceuticals by fluorescence. Journal of Pharmaceutical and Biomedical Analysis, 23(1), 147-159.

[2]   Taylor, P.W. and Keenan, M.H.J. (2006) Pharmaceutical quality of generic isotretinoin products, compared with Roaccutane. Current Medical Research and Opinion, 22(3), 603-615.

[3]   Ioele, G., Cione, E., Risoli, A., Genchi G. and Ragno, G. (2005) Accelerated photostability study of tretinoin and isotretinoin in liposomes formulations. International Jour- nal of Pharmaceutics, 60(1-2), 251-260.

[4]   Wetterich, U. and Mutschler, E. (1995) Quality of cefotaxine sodium preparations. Arzneimittel-Forschung Drug Research, 45(1), 74-80.

[5]   Lambert, P.A. and Conway, B.R. (2003) Pharmaceutical quality of ceftriaxone generic products compared with Rocephin. Journal Chemotherapy, 15(4), 357-368.

[6]   Lammer, E.J., Chen, D.T., Hoar, R.M., Agnish, N.D., Benke, P.J. and Braun, J.T. (1985) Retinoic acid em- bryopathy. New England Journal of Medicine, 313(14), 837-841.

[7]   Willhite, C.C., Wier, P.J. and Berry, D.L. (1989) Dose response and structure-activity considerations in retinoid- induced dysmor phogênesis. Critical Reviews in Toxicology, 20(2), 689-695.

[8]   (2007) United States pharmacopeia. XXX, Pharma- copeia Convention, Maryland, 2428-2429.

[9]   Lima, E.M., Diniz, D.G. and Antoniosi-Filho, N.R. (2005) Development of a gas chromatography method for the determination of isotretinoin and its degradation products in pharmaceuticals. Journal of Pharmaceutical of Bio- medical Analysis, 38(4), 678-685.

[10]   Tashtoush, M.B. Jacobson, E.L. and Jacobson, M.K. (2007) A rapid HPLC method for simultaneous deter- mination of tretinoin and isotretinoin in dermatological formulations. Journal of Pharmaceutical and Biomedical Analysis, 43(3), 859-864.

[11]   Tan, X., Meltzer, N. and Lindenbaum, S. (1992) Solid- state stability studies of 13-cis-retinoic acid and all- transretinoic acid using microcalorimetry and HPLC analysis. Pharmaceutical Research, 9(9), 1203-1208.

[12]   Allwood, M.C. and Plane, J.H. (1984) The degradation of vitamin A exposed to ultraviolet radiation. International Journal of Pharmaceutics, 19(2), 207-213.

[13]   Crank, G. and Pardijanto, M.S. (1995) Photooxidations and photosensitized oxidations of vitamin A and its palmitate ester. Journal of Photochemistry and Photobio- logy A: Chemistry, 85(1-2), 93-100.

[14]   Bempong, D.K., Honigberg, I.L. and Meltzer, N.M. (1993) Separation of 13-cis and all-trans retinoic acid and their photodegradable products using capillary zone electrophoresis and micellar electrokinetic chromato- graphy (MEC). Journal of Pharmaceutical and Biomedi- cal Analysis, 11(9), 829-833.