Development and Validation of HPLC-Uv Method for Simultaneous Analysis of Acrylamide and Glycidamide in Volumetric Absorptive Microsampling

Abstract

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

Blood microsampling technologies: Innovations and applications in 2022

“…139 VAMS technology was also capable of detecting carcinogens, such as acrylamide and aflatoxin B 1 . 140,141 The incidence of the global COVID-19 pandemic has accelerated the use and development of decentralised blood sampling for virtual clinical trials, trials that do not require in-person visits and routine analysis. The use of TASSO-SST devices was evaluated for measuring blood protein levels in healthy subjects and non-hospitalised COVID-19 patients.…”

“…Microsampling technologies have been evaluated in biomarker research studies for the diagnosis and prognosis of various conditions and the prediction and monitoring of response to an intervention 24,41,42,132–144 . Omics‐based approaches, such as metabolomics and proteomics, are increasingly employed for the generation of large amounts of data to facilitate the rapid discovery of biomarkers.…”

“…Other studies that have implemented the VAMS technology include research for (i) the human biomonitoring of mercury concentrations for exposure assessment, 41,145 (ii) the determination of tryptophan‐related biomarkers for understanding pathologies of neurodegenerative diseases such as Alzheimer's or Parkinson's disease 138 and (iii) the measurement of endocrine and bone biomarkers 139 . VAMS technology was also capable of detecting carcinogens, such as acrylamide and aflatoxin B 1 140,141 …”

“…Microsampling technologies have been evaluated in biomarker research studies for the diagnosis and prognosis of various conditions and the prediction and monitoring of response to an intervention. 24 , 41 , 42 , 132 , 133 , 134 , 135 , 136 , 137 , 138 , 139 , 140 , 141 , 142 , 143 , 144 Omics‐based approaches, such as metabolomics and proteomics, are increasingly employed for the generation of large amounts of data to facilitate the rapid discovery of biomarkers. A study assessed the DBS and DPS technology for non‐targeted lipidomic analysis as high throughput profiling and quantification of lipids could expand knowledge of underlying disease pathologies.…”

“… 139 VAMS technology was also capable of detecting carcinogens, such as acrylamide and aflatoxin B 1 . 140 , 141 …”