DEVELOPMENT AND VALIDATION OF HPLC-UV METHOD FOR SIMULTANEOUS ANALYSIS OF ACRYLAMIDE AND GLYCIDAMIDE IN VOLUMETRIC ABSORPTIVE MICROSAMPLING

Objective: Acrylamide is a carcinogenic compound that can be found in commonly consumed foods and cigarette smoke. This metabolized by cytochrome P450 the human body to more reactive metabolite, glycidamide. study aimed optimize validate sensitive HPLC-UV method for determining acrylamide glycidamide simultaneously volumetric absorptive microsampling (VAMS) sample. Methods: Isoniazid as an internal standard was added VAMS sample containing prior protein precipitation. The analytes were separated using reversed-phase chromatography with C18 SunfireTMWaters® column (5 µm; 250 mm x 4.6 mm) ultraviolet detector. Results: optimum chromatographic condition eluted at temperature of 30 °C mobile phase 6 mmol potassium dihydrogen phosphate pH 3.5–methanol (96:4 v/v) flow rate 0.50 ml/min detected 210 nm. LLOQ obtained 1.0 µg/ml both calibration curve linear over concentration range 1.0-100.0 µg/ml. Conclusion: developed bioanalytical valid based on US FDA Guideline Bioanalytical Method Validation 2018.