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अमूर्त

Determination, Fractionation and Quantification of Vitamin D in Pleurotus ostreatus (Oyster Mushroom) Using Recycling Preparative High Performance Liquid Chromatography (HPLC)

Aswir Abd Rashed1*, Rathi Devi Nair Gunasegavan1, Shahrul Razid Sarbini2

Recent studies have demonstrated that vitamin D insufficiency is common in tropical countries. Vitamin D is a fat- soluble vitamin that is obtained by sunlight exposure, diet, or supplements and it is an important factor influencing the health status of individuals. Vitamin D is naturally present in the mushroom and once exposed to a source of Ultraviolet (UV) radiation, can generate nutritionally relevant amounts of vitamin D. Hence, the purpose of this study is to determine, fractionalised and quantify the amount of vitamin D in the oyster mushroom (Pleurotus ostreatus). An amount of fine powder mushroom was weighed and incubated with 20 ml hexane and 50% potassium hydroxide mixture for 10 minutes at 37°C. The mixture solvent was then centrifuged for 45 minutes at 2000 rpm before the supernatant was stored at -20°C for a week until a white precipitate was formed. Later, it was diluted with 95% methanol and injected into the recycling preparative High-Performance Liquid Chromatography (HPLC) system. The system was operated at a flow rate of 20 ml/min with a running time of 50 minutes. The retention time for vitamin D peak was recorded at 32 minutes. The eluent was collected at this particular time before the quantification of vitamin D was performed by using Ultra High-Performance Liquid Chromatography (UHPLC) at 265 nm. It was recorded that the concentration of vitamin D in the fractionated solvent was 69.6 ± 7.25 ppm (2784 ± 290 IU). The linearity of this method was found to be within the acceptable recovery limit range (r2˃0.999). The recycling preparative HPLC has great potential as a useful tool for the determination and fractionation of vitamin D in food samples. Commercially available vitamin D3 supplements are usually derived from the wool of healthy sheep. Thus, the extraction of vitamin D from mushrooms has great potential as an alternative for vegan, vegetarian and halal conscious consumers.