Abstract

Cholesterol analysis by derivatization technique is a time consuming, costly, and complex process while analyzing cholesterol without derivation is a simple, and quick method.Researchers analyzed cholesterol using both derivatization and non-derivatization techniques successfully. The objective of this study was to investigate the effect of derivatization in cholesterol analysis particularly on bakery goods.The retention time of non-derivatized cholesterol (11.62 min) and non-derivatized α-tocopherol standard (11.60 min) was very close in HP-5 capillary GC column andthey eluted together while injected as mixed standard. As a result, cholesterol content determined by non-derivatized technique could be overestimated due to the presence of α-tocopherol inbakery products. The peak resolution (Rs) between derivatized cholesterol and derivatized α-tocopherol standard using the appliedgradient GC condition was 3.1 which is well separated (>1.5) based on AOAC guidelines. The derivatized gas chromatographic cholesterol analysis method was verified by limit of detection (LOD; 0.03 mg/100 g), limit of quantification (LOQ; 0.08 mg/100 g), linearity (R2; 0.999),precision (repeatability: relative standard deviation (RSD) 1.5 %; reproducibility: RSD 1.9 %), and accuracy (102.1 % recovery). The verified cholesterol analysis method was subsequently applied to determine cholesterol content in selected bakery items, yielding a range of 2.76 ± 0.06 mg/100 g (chrysanthemum bread) to 114.26 ± 4.72 mg/100 g (castella).

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