AAPS PharmSciTech

, Volume 13, Issue 1, pp 202–210 | Cite as

Sensitive and Rapid HPLC Quantification of Tenofovir from Hyaluronic Acid-Based Nanomedicine

Article

Abstract

The purpose of this study was to develop and validate a rapid, sensitive, and specific reversed-phase high-performance liquid chromatography method for the quantitative determination of native tenofovir (TNF) for various applications. Different analytical performance parameters such as linearity, precision, accuracy, limit of quantification (LOQ), limit of detection (LOD), and robustness were determined according to International Conference on Harmonization (ICH) guidelines. A Bridge™ C18 column (150 × 4.6 mm, 5 μm) was used as stationary phase. The retention time of TNF was 1.54 ± 0.03 min (n = 6). The assay was linear over the concentration range of 0.1–10 μg/mL. The proposed method was sensitive with LOD and LOQ values equal to 50 and 100 ng/mL, respectively. The method was accurate with percent mean recovery from 95.41% to 102.90% and precise as percent RSD (relative standard deviation) values for intra-day, and inter-day precision were less than 2%. This method was utilized for the estimation of molar absorptivity of TNF at 259 nm (ε 259 = 12,518 L/mol/cm), calculated from linear regression analysis. The method was applied for determination of percentage of encapsulation efficiency ( 22.93 ± 0.04%), drug loading (12.25 ± 1.03%), in vitro drug release profile in the presence of enzyme (43% release in the first 3 h) and purification analysis of hyaluronic acid-based nanomedicine.

KEY WORDS

high-performance liquid chromatography hyaluronic acid molar absorptivity nanomedicine tenofovir 

Notes

ACKNOWLEDGMENTS

The project described was supported by Grant Number R01AI087304 from the National Institute of Allergy And Infectious Diseases. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Allergy And Infectious Diseases or the National Institutes of Health. We gratefully acknowledge Mr. Jack Liu (Zhenjiang Dong Yuan Biotech Co., Ltd., Jiangsu, China) for providing samples of hyaluronic acid.

REFERENCES

  1. 1.
    Checa A, Oliver R, Hernández-Cassou S, Saurina J. Determination of HIV drugs in biological matrices: a review. Anal Chim Acta. 2009;647(1):1–13.PubMedCrossRefGoogle Scholar
  2. 2.
    Zidan AS, Spinks C, Fortunak J, Habib M, Khan MA. Near-infrared investigations of novel anti-HIV tenofovir liposomes. AAPS J. 2010;12(2):202–14.PubMedCrossRefGoogle Scholar
  3. 3.
    Choi SU, Bui T, Ho RJ. pH-Dependent interactions of indinavir and lipids in nanoparticles and their ibility to entrap a solute. J Pharm Sci. 2008;97(2):931–43.PubMedCrossRefGoogle Scholar
  4. 4.
    Rohan LC, Sassi AB. Vaginal drug delivery systems for HIV prevention. AAPS J. 2009;11(1):78–87.PubMedCrossRefGoogle Scholar
  5. 5.
    Ndesendo VM, Pillay V, Choonara YE, Buchmann E, Bayever DN, Meyer LC. A review of current intravaginal drug delivery approaches employed for the prophylaxis of HIV/AIDS and prevention of sexually transmitted infections. AAPS PharmSciTech. 2008;9(2):505–20.PubMedCrossRefGoogle Scholar
  6. 6.
    D'Cruz OJ, Uckun FM. Clinical development of microbicides for the prevention of HIV infection. Curr Pharm Des. 2004;10(3):315–36.PubMedCrossRefGoogle Scholar
  7. 7.
    Abdool Karim Q, Abdool Karim SS, Frohlich JA, Grobler AC, Baxter C, Mansoor LE, et al. Effectiveness and safety of tenofovir gel, an antiretroviral microbicide, for the prevention of HIV infection in women. Science. 2010;329(5996):1168–74.PubMedCrossRefGoogle Scholar
  8. 8.
    Rohan LC, Moncla BJ, Kunjara Na Ayudhya RP, Cost M, Huang Y, et al. In vitro and ex vivo testing of tenofovir shows it is effective as an HIV-1 microbicide. PLoS One. 2010;5(2):1–13.CrossRefGoogle Scholar
  9. 9.
    Alukda D, Sturgis T, Youan BB. Formulation of tenofovir-loaded functionalized solid lipid nanoparticles intended for HIV prevention. J Pharm Sci. 2011;100(8):3345–56. doi: 10.1002/jps.22529.PubMedCrossRefGoogle Scholar
  10. 10.
    Mayer KH, Maslankowski LA, Gai F, El-Sadr WM, Justman J, Kwiecien A, et al. Safety and tolerability of tenofovir vaginal gel in abstinent and sexually active HIV-infected and uninfected women. AIDS. 2006;20(4):543–51.PubMedCrossRefGoogle Scholar
  11. 11.
    Rosen RK, Morrow KM, Carballo-Dieguez A, Mantell JE, Hoffman S, Gai F, et al. Acceptability of tenofovir gel as a vaginal microbicide among women in a phase I trial: a mixed-methods study. J Womens Health (Larchmt). 2008;17(3):383–92.CrossRefGoogle Scholar
  12. 12.
    Saxena BB, Han YA, Fu D, Rathnam P, Singh M, Laurence J, Lerner S. Sustained release of microbicides by newly engineered vaginal rings. AIDS. 2009;23(8):917–22.PubMedCrossRefGoogle Scholar
  13. 13.
    Kandagal PB, Manjunatha DH, Seetharamappa J, Kalanur SS. RP-HPLC method for the determination of tenofovir in pharmaceutical formulations and spiked human plasma. Anal Lett. 2008;41(4):561–70.CrossRefGoogle Scholar
  14. 14.
    Sentenac S, Fernandez C, Thuillier A, Lechat P, Aymard G. Sensitive determination of tenofovir in human plasma samples using reversed-phase liquid chromatography. J Chromatogr B Analyt Technol Biomed Life Sci. 2003;793(2):317–24.PubMedCrossRefGoogle Scholar
  15. 15.
    Jullien V, Tréluyer JM, Pons G, Rey E. Determination of tenofovir in human plasma by high-performance liquid chromatography with spectrofluorimetric detection. J Chromatogr B Analyt Technol Biomed Life Sci. 2003;785(2):377–81.PubMedCrossRefGoogle Scholar
  16. 16.
    Takahashi M, Kudaka Y, Okumura N, Hirano A, Banno K, Kaneda T. Determination of plasma tenofovir concentrations using a conventional LC-MS method. Biol Pharm Bull. 2007;30(9):1784–6.PubMedCrossRefGoogle Scholar
  17. 17.
    Yadav M, Mishra T, Singhal P, Goswami S, Shrivastav PS. Rapid and specific liquid chromatographic tandem mass spectrometric determination of tenofovir in human plasma and its fragmentation study. J Chromatogr Sci. 2009;47(2):140–8.PubMedGoogle Scholar
  18. 18.
    Delahunty T, Bushman L, Robbins B, Fletcher CV. The simultaneous assay of tenofovir and emtricitabine in plasma using LC/MS/MS and isotopically labeled internal standards. J Chromatogr B Analyt Technol Biomed Life Sci. 2009;877(20–21):1907–14.PubMedGoogle Scholar
  19. 19.
    Bennetto-Hood C, Long MC, Acosta EP. Development of a sensitive and specific liquid chromatography/mass spectrometry method for the determination of tenofovir in human plasma. Rapid Commun Mass Spectrom. 2007;21(13):2087–94.PubMedCrossRefGoogle Scholar
  20. 20.
    Raju NA. Simultaneous estimation of tenofovir disproxil fumerate, emtricitabine & efavirenz in tablet dosage forms by isocratic-RP-HPLC. J Pharm Res. 2009;2(6):1103–6.Google Scholar
  21. 21.
    Rezk NL, Crutchley RD, Kashuba AD. Simultaneous quantification of emtricitabine and tenofovir in human plasma using high-performance liquid chromatography after solid phase extraction. J Chromatogr B Analyt Technol Biomed Life Sci. 2005;822(1–2):201–8.PubMedGoogle Scholar
  22. 22.
    Sun D, Wang H, Wang B, Guo R. Development and validation of a sensitive LC-MS/MS method for the determination of adefovir in human serum and urine. J Pharm Biomed Anal. 2006;42(3):372–8.PubMedCrossRefGoogle Scholar
  23. 23.
    Huidobro AL, Rupérez FJ, Barbas C. LC methods for acyclovir and related impurities determination. J Pharm Biomed Anal. 2005;37(4):687–94.PubMedCrossRefGoogle Scholar
  24. 24.
    Choi KY, Chung H, Min KH, Yoon HY, Kim K, Park JH, Kwon IC, Jeong SY. Self-assembled hyaluronic acid nanoparticles for active tumor targeting. Biomaterials. 2010;31(1):106–14.PubMedCrossRefGoogle Scholar
  25. 25.
    Al-Ghananeem AM, Malkawi AH, Muammer YM, Balko JM, Black EP, Mourad W, et al. Intratumoral delivery of Paclitaxel in solid tumor from biodegradable hyaluronan nanoparticles formulations. AAPS PharmSciTech. 2009;10(2):410–7.PubMedCrossRefGoogle Scholar
  26. 26.
    Carole E, Schanté GZ, Herlin C, Vandamme TF. Chemical modifications of hyaluronic acid for the synthesis of derivatives for a broad range of biomedical applications. Carbohyd Polym. 2011;85(3):469–89.CrossRefGoogle Scholar
  27. 27.
    Swyer GI. The hyaluronidase content of semen. Biochem J. 1947;41(3):409–13.Google Scholar
  28. 28.
    Nirogi RVS, Kandikere VN, Shukla M, Mudigonda K, Maurya S, Boosi R, et al. Sensitive and selective liquid chromatography–tandem mass spectrometry method for the quantification of azithromycin in human plasma. Anal Chim Acta. 2005;553(1–2):1–8.CrossRefGoogle Scholar
  29. 29.
    Khan A, Khan MI, Iqbal Z, Shah Y, Ahmad L, Watson DG. An optimized and validated RP-HPLC/UV detection method for simultaneous determination of all-trans-retinol (vitamin A) and alpha-tocopherol (vitamin E) in human serum: comparison of different particulate reversed-phase HPLC columns. J Chromatogr B Analyt Technol Biomed Life Sci. 2010;878(25):2339–47.PubMedCrossRefGoogle Scholar
  30. 30.
    Xu M, Ju W, Xia X, Tan H, Chen M, Zhang J, et al. Determination of rimantadine in rat plasma by liquid chromatography/electrospray mass spectrometry and its application in a pharmacokinetic study. J Chromatogr B Analyt Technol Biomed Life Sci. 2008;864(1–2):123–8.PubMedGoogle Scholar
  31. 31.
    Liebhafsky HA, Pfeiffer HG. Beer's law in analytical chemistry. J Chem Educ. 1953;30(9):450–2.CrossRefGoogle Scholar
  32. 32.
    ICH Guideline. Q2 (R1). Validation of analytical procedures: text and methodology. Geneva Switzerland: ICH Secretariat; 2005.Google Scholar
  33. 33.
    Pelillo M, Cuvelier ME, Biguzzi B, Gallina Toschi T, Berset C, Lercker G. Calculation of the molar absorptivity of polyphenols by using liquid chromatography with diode array detection: the case of carnosic acid. J Chromatogr A. 2004;1023(2):225–9.PubMedCrossRefGoogle Scholar
  34. 34.
    Hu Z, Xia X, Tang L (inventors). Process for synthesizing oil and surfactant-free hyaluronic acid nanoparticles and microparticles. United States patent 7,601,704 B2. 2009 October 13.Google Scholar
  35. 35.
    Hackley V, Ferraris C. The use of nomenclature in dispersion science and technology, NIST Recommended Practice Guide. NIST SP. 2001; 960-3; 72; SPGoogle Scholar
  36. 36.
    Romero-Pérez A, García-García E, Zavaleta-Mancera A, Ramírez-Bribiesca JE, Revilla-Vázquez A, Hernández-Calva LM, et al. Designing and evaluation of sodium selenite nanoparticles in vitro to improve selenium absorption in ruminants. Vet Res Commun. 2010;34(1):71–9.PubMedCrossRefGoogle Scholar
  37. 37.
    Owen DH, Katz DF. A review of the physical and chemical properties of human semen and the formulation of a semen simulant. J Androl. 2005;26(4):459–69.PubMedCrossRefGoogle Scholar
  38. 38.
    Owen DH, Katz DF. A vaginal fluid simulant. Contraception. 1999;59(2):91–5.PubMedCrossRefGoogle Scholar
  39. 39.
    Patel S, Baghel US, Rajesh P, Prabhakar D, Engla G, Nagar PN. Spectrophotometric method development and validation for simultaneous estimation of tenofovir disoproxil fumarate and emtricitabine in bulk drug and tablet dosage form. Int J Pharm Clin Res. 2009;1:28–30.Google Scholar
  40. 40.
    O'Neil MJ, Smith A, Heckelman PE. The Merck index. 13th ed. New Jersey: Merck research laboratories; 2001.Google Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2011

Authors and Affiliations

  1. 1.Laboratory of Future Nanomedicines and Theoretical Chronopharmaceutics, Division of Pharmaceutical Sciences, School of PharmacyUniversity of Missouri-Kansas CityKansas CityUSA

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