I want to calculate the concentration of vitamin in blood plasma. I followed a procedure from an article. taken 1 ml of plasma, added 5 ml of hexane (to extract the compound), centrifuged. two phase formed, taken 4 ml of the top organic layer, dried under purging with nitrogen gas. Added 2 ml methanol and injected 10 ul in HPLC. Compared with the calibration and found 25 ug/ml.
Now, please help me to do the back calculation to find how much vitamin present in the plasma. since I dried and added methanol. How could i account all those?
There are several reliable immune assays including ELISA to quantify Vitamin D in plasma and serum. What exactly are the vitamins you want to measure? Sorry i can not advise you about MS/ LCMS and HPLC. It might be necessary to consider plasma binding proteins and fibrin that may interfere with your organic extraction..perhaps you can try de-fibrination of the plasma before extraction....just guessing ?
Thanks for your reply Dr. Simon. I'm using Vitamin E. Consider I'm doing in vitro studies, with simulated gastro-intestinal conditions. Instead of plasma, i'm taking 1 ml of digesta and doing the same procedure, mentioned above. I can't go for assays. Now how to calculate the amount of vitamin present in the digesta..
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