The use of high-resolution mass spectra has made it easier to measure ergocalciferol in biological material. To quantify ergocalciferol, multiple LC-MS/MS methods have been developed. Using LC-MS/MS, researchers were able to establish an accurate method for quantifying ergocalciferol in a large number of samples, which should be tested in the blood shortly after administration. The goal of this research is to improve chromatographic conditions and assess the LC-MS/MS method for reliable quantification of ergocalciferol dose form in human plasma. Methanol liquid–liquid extraction and a reverse phase C8 column were used to extract and separate ergocalciferol from human plasma, respectively. To detect and quantify ergocalciferol by LC-MS/MS, the residue following liquid-liquid extraction was dissolved using PTAD reagent along with the reconstituted solution. A quadrupole tandem mass spectrometer (LC-MS/MS) with an atmospheric pressure chemical ionisation (APCI) source was used for detection. The procedure was validated using certified reference material in accordance with FDA requirements. The LOQ for the analyte examined was 5.44 ng/mL, 65.123 ng/mL in medium-MQC, and 120.598 ng/mL in high-HQC, and the quality control samples did not exceed 20 and 15% CV, respectively. The method for determining ergocalciferol was validated, and the accuracy was found to be between 95.40 and 103.09 percent. This approach can be used to determine the amount of ergocalciferol in the systemic circulation after external intake of tablets. The LC-MS/MS test can be used to quantify ergocalciferol natural form from external ingestion and investigate the metabolism of clinical trial participants.
Author(s) Details
A. Srinivas
Department of Genetics,
Osmania University, Hyderabad, 500 007, Telangana, India.
M. Tamilselvi
Value Added Corporate Services Pvt Ltd, Besant Nagar, Chennai, 600090,
India.
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