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Determination of betaine, a natural potential anti-inflammatory compound, using sildenafil as an internal standard in human plasma by high performance liquid chromatography–mass spectrometry

Mansee M. Pathak*, Vikas V. Vaidya, Hardik R. Shah


Betaine is a naturally occurring cationic functional group which shows a wide range of therapeutic activities. The present research work involves rapid; robust and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method developed and validated for s analysis of betaine in human plasma. This method involved a solid phase extraction and a subsequent analysis performed on an LC-MS system which contained the turbo spray ionization interface. Chromatographic separation was performed using a Gemini C18 column. Good linearity was observed over the concentration range of 10.00 ng/mL to 1203.26 ng/mL with a correlation coefficient (r² ≥ 0.9984). Sildenafil was used as the internal standard in the present method. The method involves simple isocratic chromatography conditions and mass spectrometric detection in the positive ionization mode. The intra-day and inter-day precision and accuracy of the quality control samples were within the acceptance criteria of ±15%. The total elution time was about 5 min which allows higher throughput. This validated method can be applied for analysis of real samples from a bioequivalence study involving administration of formulations containing betaine as their active therapeutic components.


Betaine; Sildenafil; LC-MS/MS; Solid phase extraction

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