Haemolysis Effect Estimation of Doxycycline Hyclate 150 mg Delay Release Tablet in Bio-analysis (Human Plasma) by Liquid Chromatography-Tandem Mass Spectrometry
- *Corresponding Author:
- Paresh Mistry
Accutest Research Laboratories (I) Pvt. Ltd
S.G. Highway, Ahmedabad-380054, Gujarat, India
E-mail: [email protected]
Received Date: May 07, 2013; Accepted Date: August 22, 2013; Published Date: August 31, 2013
Citation: Mistry P, Menon S (2013) Haemolysis Effect Estimation of Doxycycline Hyclate 150 mg Delay Release Tablet in Bio-analysis (Human Plasma) by Liquid Chromatography-Tandem Mass Spectrometry. J Bioequiv Availab 5:215-223. doi: 10.4172/jbb.1000161
Copyright: © 2013 Mistry P, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
A fast, simple and particular method for estimation of Doxycycline in healthy human plasma was validated using Minocycline as IS. The analyte and IS were extracted from plasma using SPE. The compound was estranged on a RP column with an isocratic mobile phase consisting of 0.1% formic acid in water and acetonitrile (12:88, v/v), and detected by tandem mass spectrometry in positive ion mode. The ion transition recorded in several reaction monitoring mode were m/z 294.1→225.1 for Doxycycline and m/z 286.1→217.1 for IS. Linearity in plasma was observed over the concentration range 0.3-30 ng/mL for Doxycycline. The mean recovery for Doxycycline was 83.7%, with a lower limit of quantification of 0.3 ng/mL. The coefficient of variation of the assay was less than 6.8%, and accuracy of 96.1% to 102.2%. The validated method was applied to bioequivalence study of 150 mg Doxycycline Hyclate tablet in healthy human volunteers. The validated method was used to expose study samples of bioequivalence study of 150 mg Doxycycline Hyclate delay release tablet in 36 healthy human volunteers. Total 50 samples from individual volunteers identified as Haemolyzed, which were analyze initial and repeat again to cross check the method reproducibity for Haeamolysis effect and compared which found acceptable range. Bioequivalence was prove for test and reference using validated method to experimental samples (Figure 1).