Development and  Method Validation of Acrylamide in Dried Blood Spots with Propranolol as Internal Standard by High Performance Liquid Chromatography Tandem Mass Spectrometry

Zenshiny Starlin, Yahdiana Harahap, Eme Stepani Sitepu

Faculty of Pharmacy, Universitas Indonesia, Depok 16424, West Java, Indonesia

Corresponding author: Yahdiana Harahap

yahdiana03@yahoo.com

Acrylamide (AA) is a carcinogenic substance that is easily found in working environment, food, contaminated air, and tobacco smoke. This substances can be distributed rapidly through all body compartments. Therefore a highly sensitive and selective method using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) is needed. The method  was developed using propranolol as internal standard and using dried blood spots (DBS) for bio-sampling method.  This analysis was performed on Acquity® UPLC BEH C₁₈ column (1.7 mm, 2.1 mm x 100mm), eluted with 0.24 mL/min flow rate under a gradient of mobile phase of 0.1% formic acid in water and acetonitrile within 3 minutes. The DBS extraction was done by protein precipitation technique. The analysis quantification was performed  by a triple quadruple mass spectrometry with electrospray ionization (ESI) in positive ion mode. The multiple reaction monitoring (MRM) was set at 71.99 > 55.23 (m/z) for acrylamide and 260.2 > 116.2 (m/z) for propranolol. The analytical method had been fully validated according to the US FDA’s Bioanalytical Method Validation Guidance. The method was linear at concentration range of 5 – 5,000 ng/mL.

Keywords: Acrylamide, DBS, propranolol, UPLC-MS/MS, validation