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LC-MS/TOF and UHPLC-MS/MS study of in vivo fate of rifamycin isonicotinyl hydrazone formed on oral co-administration of rifampicin and isoniazid.
J Pharm Biomed Anal. 2010 Jul 08; 52(3):377-83.JP

Abstract

The formation and fate of 3-formylrifamycin isonicotinyl hydrazone (HYD) was investigated following oral co-administration of rifampicin (RIF) and isoniazid (INH) in Sprague-Dawley (SD) rats (n=5) using advanced analytical modalities. The study was carried out with 20 and 5mg/kg doses of RIF and INH, respectively. The plasma, urine and faeces samples were collected at different time points up to 48h, which were qualitatively and quantitatively evaluated for the presence of HYD after proper sample preparation. For the same, initially liquid chromatography-mass spectrometry/time-of-flight (LC-MS/TOF) method was developed in electrospray ionization (ESI) positive mode, wherein separation was achieved on a C18 column (4.6mmx250mm, 5microm), using a volatile mobile phase in a gradient mode. The presence of HYD was confirmed by accurate mass study, spiking with the standard and UV-visible spectra matching. For quantitative evaluation of HYD, a selective and sensitive ultra high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method was developed for all the three matrices. In this case, elution of HYD was achieved on a small C18 column (4.6mmx50mm, 1.8microm) using a short gradient method. The quantitation was done by selective reaction monitoring (SRM) in ESI positive mode. The validation parameters like linearity, accuracy, precision, selectivity, matrix effect, recovery and stability were assessed as per regulatory guidelines. The calibration range was established between 1 and 200ng/ml, with r(2)>0.99 in all the cases. The back calculated values for three quality control (QC) samples, and at lower limit of quantitation (LLOQ) were within 15 and 20%, respectively, of the nominal values. Similarly, the intra- and inter-day precisions were found within 15% at the four tested levels. The HYD was found to be stable for the duration of sample preparation and analysis in the controlled experimental conditions. The analysis of in vivo samples showed a significant extent of HYD in faeces, however, the interaction product was not found in plasma and urine. To verify the results, 5mg/kg oral dose of HYD standard was given to rats separately, and its presence was studied in all the three matrices. Further, in vitro plasma stability of HYD was also carried out to explain its absence in plasma and urine, which showed approximately 55% disappearance of HYD in 2h.

Authors+Show Affiliations

Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar 160 062, Punjab, India.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19692195

Citation

Prasad, Bhagwat, and Saranjit Singh. "LC-MS/TOF and UHPLC-MS/MS Study of in Vivo Fate of Rifamycin Isonicotinyl Hydrazone Formed On Oral Co-administration of Rifampicin and Isoniazid." Journal of Pharmaceutical and Biomedical Analysis, vol. 52, no. 3, 2010, pp. 377-83.
Prasad B, Singh S. LC-MS/TOF and UHPLC-MS/MS study of in vivo fate of rifamycin isonicotinyl hydrazone formed on oral co-administration of rifampicin and isoniazid. J Pharm Biomed Anal. 2010;52(3):377-83.
Prasad, B., & Singh, S. (2010). LC-MS/TOF and UHPLC-MS/MS study of in vivo fate of rifamycin isonicotinyl hydrazone formed on oral co-administration of rifampicin and isoniazid. Journal of Pharmaceutical and Biomedical Analysis, 52(3), 377-83. https://doi.org/10.1016/j.jpba.2009.07.014
Prasad B, Singh S. LC-MS/TOF and UHPLC-MS/MS Study of in Vivo Fate of Rifamycin Isonicotinyl Hydrazone Formed On Oral Co-administration of Rifampicin and Isoniazid. J Pharm Biomed Anal. 2010 Jul 8;52(3):377-83. PubMed PMID: 19692195.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - LC-MS/TOF and UHPLC-MS/MS study of in vivo fate of rifamycin isonicotinyl hydrazone formed on oral co-administration of rifampicin and isoniazid. AU - Prasad,Bhagwat, AU - Singh,Saranjit, Y1 - 2009/07/22/ PY - 2009/04/20/received PY - 2009/06/30/revised PY - 2009/07/15/accepted PY - 2009/8/21/entrez PY - 2009/8/21/pubmed PY - 2010/5/21/medline SP - 377 EP - 83 JF - Journal of pharmaceutical and biomedical analysis JO - J Pharm Biomed Anal VL - 52 IS - 3 N2 - The formation and fate of 3-formylrifamycin isonicotinyl hydrazone (HYD) was investigated following oral co-administration of rifampicin (RIF) and isoniazid (INH) in Sprague-Dawley (SD) rats (n=5) using advanced analytical modalities. The study was carried out with 20 and 5mg/kg doses of RIF and INH, respectively. The plasma, urine and faeces samples were collected at different time points up to 48h, which were qualitatively and quantitatively evaluated for the presence of HYD after proper sample preparation. For the same, initially liquid chromatography-mass spectrometry/time-of-flight (LC-MS/TOF) method was developed in electrospray ionization (ESI) positive mode, wherein separation was achieved on a C18 column (4.6mmx250mm, 5microm), using a volatile mobile phase in a gradient mode. The presence of HYD was confirmed by accurate mass study, spiking with the standard and UV-visible spectra matching. For quantitative evaluation of HYD, a selective and sensitive ultra high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method was developed for all the three matrices. In this case, elution of HYD was achieved on a small C18 column (4.6mmx50mm, 1.8microm) using a short gradient method. The quantitation was done by selective reaction monitoring (SRM) in ESI positive mode. The validation parameters like linearity, accuracy, precision, selectivity, matrix effect, recovery and stability were assessed as per regulatory guidelines. The calibration range was established between 1 and 200ng/ml, with r(2)>0.99 in all the cases. The back calculated values for three quality control (QC) samples, and at lower limit of quantitation (LLOQ) were within 15 and 20%, respectively, of the nominal values. Similarly, the intra- and inter-day precisions were found within 15% at the four tested levels. The HYD was found to be stable for the duration of sample preparation and analysis in the controlled experimental conditions. The analysis of in vivo samples showed a significant extent of HYD in faeces, however, the interaction product was not found in plasma and urine. To verify the results, 5mg/kg oral dose of HYD standard was given to rats separately, and its presence was studied in all the three matrices. Further, in vitro plasma stability of HYD was also carried out to explain its absence in plasma and urine, which showed approximately 55% disappearance of HYD in 2h. SN - 1873-264X UR - https://www.unboundmedicine.com/medline/citation/19692195/LC_MS/TOF_and_UHPLC_MS/MS_study_of_in_vivo_fate_of_rifamycin_isonicotinyl_hydrazone_formed_on_oral_co_administration_of_rifampicin_and_isoniazid_ DB - PRIME DP - Unbound Medicine ER -