Analytical Procedural Validation of Policosanol Compounds

Currently, the widely accepted method for determining policosanol contents is gas chromatography-mass spectrometry (GC–MS), but measurement method validation is lacking. The validation is essential to improve measurement efficiency for greater accuracy and precision. The aim of the study purposed to validate policosanol analysis by GC–MS using silylation derivatization to improve measurement efficiency. The linearity (calibration curve performance); the lower limit of detection; and the lower limit of quantification, accuracy, precision, stability, and robustness of policosanol extracted from standard policosanol solutions and defatted rice bran were evaluated. The percentage of recovery of each policosanol concentration of 50, 100, and 150 mg/L ranged from 98.90 to 102.02%, 99.28 to 101.37%, and 99.67 to 101.37%, respectively, which is under the AOAC acceptance criteria. The accuracy of method by relative standard deviations (RSD) showed that intra-day recovery percentage and inter-day precision analysis followed the AOAC acceptance criteria. Limit of detection (LOD) and limit of quantitation (LOQ) of policosanol were 0.6–0.84 mg/L and 2.09–2.82 mg/L, respectively. Policosanol derivatized with N , O -bis(trimethylsilyl)-trifluoroacetamide was not stable under storage, with variation in excess of 15%. Policosanol standard stock solution stored at 4 °C for more than 1 week was not recommended for further use. The ruggedness or robustness of the method by changing the tested parameters including derivative temperature, light exposure, storage temperature solvent type, reaction time, analysis time period, and ultrasonic wave exposure was examined. The ruggedness test showed that changes in factors related to policosanol analysis had no significant impact ( p