AJAC  Vol.4 No.12 , December 2013
Development and Validation of Extraction Method for the Determination of UC781 in Cervicovaginal Fluid
Abstract: Liquid chromatography plays the important and critical role in the field of clinical pharmacology of evaluating drugs in biological matrices. Studying antiretroviral drugs in the female genital tract has important implications for using drugs as a vaginal microbicide for prevention of HIV-1 sexual transmission. Accurate measurement of drug levels is extremely important in optimizing drug concentration in gel formulation. Extracting drugs from small volumes of viscous, proteinaceous substances like cervicovaginal fluid (CVF) is a practically challenging process. The proposed method was designed to introduce procedure for sample collection and drug extraction procedure for CVF matrix before the chromatographic separation. Based on this extraction method, we validated a reverse phase high performance liquid chromatography with electrospray ionization mass spectrometry assay in order to quantify UC781 in female genital tract compartment. The LC-MS method was validated based on a novel extraction technique which proved to be efficient in reducing analyte degradation with an average extraction efficiency of 72%. This method is accurate, demonstrating an average accuracy over three QC (n = 30) concentrations ranging from 99.9% to 106.1%. Average precision within-day and between-day ranged from 3.1% to 10.2% and 5.1% to 6.4%, respectively. We demonstrated that the analyte was able to maintain its stability under various conditions using this extraction method. The sample preparation, extraction, and the powerful liquid chromatography and mass spectrometry can readily be applied for accurate quantification of similar drugs in CVF.  
Cite this paper: N. Rezk, Z. Qiao and M. Jeriasy, "Development and Validation of Extraction Method for the Determination of UC781 in Cervicovaginal Fluid," American Journal of Analytical Chemistry, Vol. 4 No. 12, 2013, pp. 707-714. doi: 10.4236/ajac.2013.412085.

[1]   S. Taylor, M. Boffito and P. L. Vernazza, “Antiretroviral Therapy to Reduce the Sexual Transmission of HIV,” Journal of HIV Therapy, Vol. 8, No. 3, 2003, pp. 55-66.

[2]   T. Lalani and C. Hicks, “Does Antiretroviral Therapy Prevent HIV Transmission to Sexual Partners?” Current HIV/AIDS Reports, Vol. 10, No. 2, 2007, pp. 80-82.

[3]   J. M. Baeten and J. Overbaugh, “Measuirng the Infectiousness of Peresons with HIV-1: Opportunities for Preventing Sexual HIV-1 Transmission,” Current HIV Research, Vol. 1, No. 1, 2003, pp. 69-86.

[4]   J. A. Talameh, N. L. Rezk and A. Kashuaba, “Quantifying the HIV-1 Integrase Inhibitor Raltegravir in Female Genital Tract Secretions Using High-Performance Liquid Chromatography with Ultraviolet Detection,” Journal of Chromatography B, Vol. 878, No. 1, 2010, pp. 92-106.

[5]   J. Balzarini, M. J. Perez-Perez, S. Velazquez and A. San-Felix, “Suppression of the Breakthrough of Human Immunodeficiency Virus Type 1 (HIV-1) in Cell Culture by Thiocarboxanilide Derivatives When Used Individually or in Combination with Other HIV-1-Specific Inhibitors (i.e., TSAO Derivatives),” Proceeding of the National Academy of Sciences of the United States of America, Vol. 92, 1995, pp. 5470-5474.

[6]   J. Balzarini, W. G. Brouwer, D. C. Dao, E. M. Osika and E. De Clercq, “Identification of Novel Thiocarboxanilide Derivatives That Suppress a Variety of Drug-Resistant Mutant Human Immunodeficiency Virus Type 1 Strains at a Potency Similar to That for Wild-Type Virus,” Antimicrobial Agents and Chemotherapy, Vol. 40, No. 6, 1996, pp. 1454-1466.

[7]   R. W. J. Buckheit, T. L. Kinjerski, V. Fliakas-Boltz, J. Russell, T. L. Stup, L. A. Pallansch, et al., “Structure-Activity and Cross-Resistance Evaluations of a Series of Human Immunodeficiency Virus Type-1-Specific Compounds Related to Oxathiin Carboxanilide,” Antimicrobial Agents and Chemotherapy, Vol. 39, No. 12, 1995, pp. 2718-2727.

[8]   J. B. MacMahon, R. W. J. Buckheit, R. J. Gulakowski, M. J. Currens, D. T. Vistica, R. H. Schoemaker, et al., “Biological and Biochemical Anti-Human Immunodeficiency Virus Activity of UC 38, a New Non-Nucleoside Reverse Transcriptase Inhibitor,” Journal of Pharmacology and Expermental Therapeutics, Vol. 276, No. 1, 1996, pp. 298-305.

[9]   J. Vingerhoets, H. Azijn, E. Fransen, I. De Baere, L. Smeulders, D. Jochmans, et al., “TMC125 Displays a High Genetic Barrier to the Development of Resistance: Evidence from in Vitro Selection Experiments,” Journal of Virology, Vol. 79, No. 20, 2005, pp. 12773-12782.

[10]   R. E. Haaland, T. Evans-Strickfaden, A. Holder, C. P. Pau, J. M. Mcnicholl, S. Chaikummaao, et al., “UC781 Microbicide Gel Retains Anti-HIV Activity in Cervicovaginal Lavag Fluids Collected Following Twice-Daialy Vaginal Application,” Antimicrobial Agents and Chemotherapy, Vol. 56, No. 7, 2012, pp. 3592-3596.

[11]   N. Rezk, “Development and Validation of LC-ESI-MS Method for Sensitive, Accurate and Rabid Determination of UC-781 in New Zealand White Rabbit Plasma,” Talanta, Vol. 85, No. 4, 2011, pp. 2074-2079.

[12]   D. H. Owen and D. F. Katz, “A Vaginal Fluide Simulant,” Contraception, Vol. 59, No. 2, 1999, pp. 91-95.

[13]   V. P. Shah, K. K. Midha, J. W. Findlay, et al., “Bioanalytical Method Validation—Revisit with a Decade of Progress,” Pharmaceutical Research, Vol. 17, No. 12, 2000, pp. 1551-1557.

[14]   US Department of Health and Human Services, Food and Drug Administration Center for Drug Evaluation and Research (CDER), Center of Veterinary Medicine (CVM), “FDA Guidance for Industry,” Pharmaceutical Research, 2001.

[15]   UNAIDA, “AIDS Epidemic Update,” Geneva, 2005.

[16]   UNAIDA, “AIDS Epidemic Update,” Geneva, 2009.

[17]   UNAIDS, “Report in the Global AIDS Epidemic,” Geneva, 2008.

[18]   “Will There Be an HIV Vaccine in the Next Decade?” Nature Medicine, Vol. 13, 2007, pp. 518-519.

[19]   L. C. Rohan and A. B. Sassi, “Vaginal Drug Delivery System for HIV Prevention,” American Association of Pharmaceutical Scientists, Vol. 11, No. 1, 2009, pp. 78-87.

[20]   B. Gulter and J. Justman, “Vaginal Microbicides and Prevention of HIV Transmission,” Lancet Infection Disease, Vol. 8, No. 11, 2008, pp. 685-697.

[21]   A. B. Moscicki, “Vaginal Microbicides: Where Are We and Where Are We Going?” Journal of Infection Chemotherapy, Vol. 14, No. 5, 2008, pp. 337-341.

[22]   S. M. Iqbal, T. B. Ball, P. Levinson, L. Maranan, W. Jaoko, C. Wachihi, et al., “Elevated Elafin/Trappin-2 the Female Genital Tract Is Associated with Protection against HIV Acquisition,” AIDS, Vol. 23, No. 13, 2009, pp. 1669-1677.

[23]   R. M. Grant, D. Hamer, T. Hope, R. Johnston, J. Lange, M. M. Lederman, et al., “Whither or Wither Microbicides?” Science, Vol. 321, No. 5888, 2008, pp. 532-534.

[24]   S. Min, A. Corbett, N. Rezk, S. Cu-Uvinc, S. Fiscus, et al., “Protease Inhibitor and Nonnucleoside Reverse Transcriptase Inhibitor Concentration in Genital Tract of HIV-1 Infected Women,” Journal of Acquired Immune Deficiency Syndrome, Vol. 37, No. 5, 2004, pp. 1577-1580.

[25]   D. Julie, R. Yeh, K. Paterson, A. Corbett, B. Jung, N. Rezk, et al., “Antiretroviral Drug Exposure in the Female Genital Tract: Implications for Oral Pre-and Post Exposure Prophylaxis,” AIDS, Vol. 21, No. 14, 2007, pp. 1899-1907.

[26]   R. Yeh, N. Rezk, A. Kashuba, et al., “Genital Tract, Cord Blood, and Amniotic Fluid Exposures of Seven Antiretroviral Drugs during and after Pregnancy in Human Immunodeficiency Virus Type 1-Infected Women,” Antimicrobial Agents and Chemotherapy, Vol. 53, No. 6, 2009, pp. 2367-2374.

[27]   A. Kawara, A. Delong, N. Rezk, et al., “Antiretroviral Drug Concentrations and HIV RNA in the Genital Tract of HIV-Infected Women Receiving Long-Term Highly Active Antiretroviral Therapy,” Clinical Infectious Diseases, Vol. 46, No. 5, 2008, pp. 719-725.

[28]   G. R. Huggins and G. Preti, “Vaginal Odors and Secretions,” Clinical Obstetrics and Gynecology, Vol. 24, No. 2, 1981, pp. 355-377.

[29]   L. L. Klein, K. R. Jonschr, M. J. Heerwagen, R. S. Gibbs and J. L. McManaman, “Reprod, Shotgun Proteomic Analysis of Vaginal Fluid from Women in Late Pregnancy,” Science, Vol. 15, No. 3, 2008, pp. 263-273.