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Application of a rapid and selective method for the simultaneous determination of protease inhibitors, lopinavir and ritonavir in human plasma by UPLC-ESI-MS/MS for bioequivalence study in Indian subjects.
J Pharm Biomed Anal. 2009 May 01; 49(4):1115-22.JP

Abstract

A high throughput and rugged ultra performance liquid chromatography tandem mass spectrometry (UPLC-ESI-MS/MS) method is developed and validated for the selective determination of protease inhibitors -- lopinavir (LPV) and ritonavir (RTV) in human plasma. Plasma samples were prepared by solid phase extraction of the analytes and their deuterated analogs as internal standard (IS) using Waters Oasis HLB cartridges. The chromatographic separation was achieved in a run time of 1.2 min on Waters Acquity UPLC BEH C18 column (50 mm x 2.1 mm, 1.7 microm) under isocratic conditions. The mobile phase consisted of 10 mM ammonium formate, pH 4.0 adjusted with formic acid and methanol (10:90, v/v). The protonated precursor --> product ion transitions for lopinavir, ritonavir, d(8)-lopinavir and d(6)-ritonavir were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring (MRM) and positive ion mode. A linear dynamic range of 2.9-1452 ng/mL and 29.6-14379 ng/mL was established for ritonavir and lopinavir respectively using 0.1 mL human plasma. The mean relative recovery of lopinavir (96.6%), ritonavir (97.5%), d(8)-lopinavir (85.5%) and d(6)-ritonavir (86.3%) from spiked plasma samples was consistent and reproducible. The method was successfully applied to a bioequivalence study of [200(lopinavir)+50(ritonavir)]mg tablet formulation in 36 healthy human subjects under fasting conditions.

Authors+Show Affiliations

Bioanalytical Research Department, Veeda Clinical Research, Ambawadi, Ahmedabad, India.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Randomized Controlled Trial

Language

eng

PubMed ID

19282124

Citation

Yadav, Manish, et al. "Application of a Rapid and Selective Method for the Simultaneous Determination of Protease Inhibitors, Lopinavir and Ritonavir in Human Plasma By UPLC-ESI-MS/MS for Bioequivalence Study in Indian Subjects." Journal of Pharmaceutical and Biomedical Analysis, vol. 49, no. 4, 2009, pp. 1115-22.
Yadav M, Rao R, Kurani H, et al. Application of a rapid and selective method for the simultaneous determination of protease inhibitors, lopinavir and ritonavir in human plasma by UPLC-ESI-MS/MS for bioequivalence study in Indian subjects. J Pharm Biomed Anal. 2009;49(4):1115-22.
Yadav, M., Rao, R., Kurani, H., Singhal, P., Goswami, S., & Shrivastav, P. S. (2009). Application of a rapid and selective method for the simultaneous determination of protease inhibitors, lopinavir and ritonavir in human plasma by UPLC-ESI-MS/MS for bioequivalence study in Indian subjects. Journal of Pharmaceutical and Biomedical Analysis, 49(4), 1115-22. https://doi.org/10.1016/j.jpba.2009.02.010
Yadav M, et al. Application of a Rapid and Selective Method for the Simultaneous Determination of Protease Inhibitors, Lopinavir and Ritonavir in Human Plasma By UPLC-ESI-MS/MS for Bioequivalence Study in Indian Subjects. J Pharm Biomed Anal. 2009 May 1;49(4):1115-22. PubMed PMID: 19282124.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Application of a rapid and selective method for the simultaneous determination of protease inhibitors, lopinavir and ritonavir in human plasma by UPLC-ESI-MS/MS for bioequivalence study in Indian subjects. AU - Yadav,Manish, AU - Rao,Rajasekhar, AU - Kurani,Hemal, AU - Singhal,Puran, AU - Goswami,Sailendra, AU - Shrivastav,Pranav S, Y1 - 2009/02/20/ PY - 2008/11/26/received PY - 2009/01/22/revised PY - 2009/02/10/accepted PY - 2009/3/14/entrez PY - 2009/3/14/pubmed PY - 2009/7/17/medline SP - 1115 EP - 22 JF - Journal of pharmaceutical and biomedical analysis JO - J Pharm Biomed Anal VL - 49 IS - 4 N2 - A high throughput and rugged ultra performance liquid chromatography tandem mass spectrometry (UPLC-ESI-MS/MS) method is developed and validated for the selective determination of protease inhibitors -- lopinavir (LPV) and ritonavir (RTV) in human plasma. Plasma samples were prepared by solid phase extraction of the analytes and their deuterated analogs as internal standard (IS) using Waters Oasis HLB cartridges. The chromatographic separation was achieved in a run time of 1.2 min on Waters Acquity UPLC BEH C18 column (50 mm x 2.1 mm, 1.7 microm) under isocratic conditions. The mobile phase consisted of 10 mM ammonium formate, pH 4.0 adjusted with formic acid and methanol (10:90, v/v). The protonated precursor --> product ion transitions for lopinavir, ritonavir, d(8)-lopinavir and d(6)-ritonavir were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring (MRM) and positive ion mode. A linear dynamic range of 2.9-1452 ng/mL and 29.6-14379 ng/mL was established for ritonavir and lopinavir respectively using 0.1 mL human plasma. The mean relative recovery of lopinavir (96.6%), ritonavir (97.5%), d(8)-lopinavir (85.5%) and d(6)-ritonavir (86.3%) from spiked plasma samples was consistent and reproducible. The method was successfully applied to a bioequivalence study of [200(lopinavir)+50(ritonavir)]mg tablet formulation in 36 healthy human subjects under fasting conditions. SN - 1873-264X UR - https://www.unboundmedicine.com/medline/citation/19282124/Application_of_a_rapid_and_selective_method_for_the_simultaneous_determination_of_protease_inhibitors_lopinavir_and_ritonavir_in_human_plasma_by_UPLC_ESI_MS/MS_for_bioequivalence_study_in_Indian_subjects_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0731-7085(09)00115-0 DB - PRIME DP - Unbound Medicine ER -