Method Validation of Acrylamide in Dried Blood Spot by Liquid Chromatography-tandem Mass Spectrometry

Zenshiny Starlin, Yahdiana Harahap and Eme S. Sitepu

Background and Objective: Acrylamide (AA) is a carcinogenic substance that is easily found in working environment, food, contaminated air and tobacco smoke. This substance can be distributed rapidly through all body compartments. The aim of this study is to get the method for determining acrylamide in dried blood spot. Materials and Methods: Dried blood spot was used as the bio-sampling method and was optimized and validated by using propranolol as the internal standard. The sample was prepared using a protein precipitation technique optimized. Reversed-phase chromatography with Acquity^® UPLC BEH C18 column (1.7, 2.1× 100 mm) was used for compound separation. Results: Optimized analytical condition for this substance was eluted with the flow rate of 0.20 mL/min under a gradient of the mobile phase of 0.1% formic acid in water and acetonitrile within 3 min. Triple quadrupole mass spectrometry with electrospray ionization (ESI) in positive mode was used as quantification analysis. The Multiple Reaction Monitoring (MRM) was set at m/z 71.99>55.23 (m/z) for acrylamide and 260.2>116.2 (m/z) for propranolol. The range of concentration was linear within 2.5-100 μg mL^–1. Conclusion: All the validation parameters were fulfilled the criteria in US FDA Guideline for Bioanalytical Method Validation 2018.

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