Determination of Vitamin D3 Content in High, Low and Zero Fat Food Using High Performance Liquid Chromatography

Abstract
Background: Vitamin D, has a significant role in bone metabolism and helps calcium absorption in the body. There are only few vitamin D assay methods available for zero fat food.
Aims of Study: (1) To develop an accurate and sensitive LC method for the quantification of vitamin D3 in food by optimization of each step of the analytical method: Extraction, sample preparation, separation and detection. (2) To validate the developed method. (3) To apply the method to quantify the total vitamin D3 in food from several species.
Material and Methods: In this study, a rapid, simple, and economical reversed phase liquid chromatographic method was described for the determination of vitamin D3, in some high, low and zero fat samples (milk products, cereal and chewing gum samples). The isolation of fat soluble vitamins includes a saponification step and an extraction step with petroleum ether and diethyl ether. The vitamin D3 content of samples was determined by reversed phase liquid chromatog-raphy. Ultra violet-Visible (UV-VIS) detector and C18 column were used for this purpose.
Results: The linearity of standard curves of vitamin D3 were 10-200 mg/ml expressed of the correlation coefficient r2 = 0.9992). The detection (LOD) and quantification (LOQ) limits were 5.09 and 15.42mg/ml, respectively. The accuracy was 101.37±4.37.
Conclusion: The described reversed-phase HPLC method is favorable compared with other published HPLC-UV methods (20 and 21) because of its stability-indicating nature, short run time and wide analytical range with outstanding linearity, accuracy and precision. The proposed method allows the determination of vitamin D3 in a single chromatographic run and is suitable for the analysis of the stability of vitamin D3. The obtained results from the assay of vitamin D3 in commer-cial nutrition supplements confirmed that the method is appropriate for the routine analysis of various food samples.