Yeni nesil antidiyabetiklerin biyolojik sıvılardan HPLC ile yanyana analizi

To the best of our knowledge a fully automated on-line SPE-LC-UV method for the simultaneous determination of Sitagliptin, Saxagliptin, Vildagliptin and Metformin in human urine samples allowing the repetitive direct injection of biological fluid was developed and validated for the first time in this study. Type 2 diabetes is one of the most common diseases in the world. For the treatment of this disease gliptin drugs act by inhibiting the destruction of incretin hormones. They are either used alone in tablets or in combination with metformin. As their amount of excretion is high in urine the developed method was applied to the urine samples. For the chromatographic separation a Gemini C18 analytical column with a mobile phase mixture of 10 mM o-phosphoric acid-methanol in gradient elution and for the fractionation of urine samples LiChrospher® ADS SPE-column was used through the study. Fractionation, transfer, elution and separation of the spiked urine samples were achieved in just 9.6 min runtime. The developed method was validated according to ICH Guidelines. The calibration curves were found to be linear in a wide the range between 10 ng/mL and 100 μg/mL with satisfactorily regression coefficients. LOD and LOQ values were found to be satisfactory as the urine excretion taken into account. Accuracy, inter-day and intra-day precision were studied on low, medium and high level of concentrations by using Excel software to perform ANOVA test. The developed method could be applied in clinical laboratories in terms of its practical use, easily found detection and reliable results.