Abstract
Cardiovascular disease is related to the high consumption of trans fatty acid. This issue has driven SEAFAST IPB Laboratory to develop a trans fatty acid analysis method for gas chromatography based on AOCS Official Method Ce 1h-05. This research was aimed to validate the method via three steps: preparation step, procedure orientation test, and method validation . During instrument performance testing in the preparation step, linearity, limit of detection (LoD), limit of quantitation (LoQ), range, and precision parameters were validated for FAME 9t-C18:1 and C18:2 isomer mix ed standard solution. During the method validation step, specificity, precision and accuracy parameters were validated for 9t-C18:1 compound attached to food matrix, i.e. doughnut . The instrument validation result of 9t-C18:1 standard solution had fulfilled the criteria of linearity (r 2 =1, requirement r 2 > 0.990) within the range of 0.0020 to 0.1200 mg/mL; precision (RSD) 0.24%; LoD 0.0007 mg/mL and LoQ 0.0020 mg/mL. The instrument validation result for each isomer in C18:2 standard solution also met the criteria of linearity (r 2 = 0.9999, requirement r 2 > 0 . 990) with precision (RSD), LoD and LoQ respectively; 1) 9t,12t- C18:2 : 1 . 40%; 0 . 0033 mg/mL; 0.0100 mg/mL; 2) 9t,12c- C18:2 : 1.89%; 0.0015 mg/mL; 0.0045 mg/mL; 3) 9c,12t- C18:2 : 2.39%; 0 . 0015 mg/mL; 0 . 0044 mg/mL; 4) C18:2 cis: 3 . 39%; 0 . 0006 mg/mL; 0 . 0019 mg/mL. The validation result for 9t-C18:1 in 100 mg fat extract of doughnut suggested that the modified method is validated in term of specificity (Rs = 1 . 2), precision (RSD fatty acid content (mg/g) = 1.78%; RSD percentage of fatty acid content towards total fatty acid = 0.72%) and accuracy ( as percentage of recovery ) 80.54-94.25%.
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