Abstract: Wheat is a vital crop having imperative nutrients for human diet e.g., protein and certain B-vitamins (thiamin, riboflavin, niacin and pyridoxine), well-intentioned for body metabolic processes. These vitamins are concentrated in grain outer layer thus milling process affects these vitamins to a larger extent and may reduce their level. The aim of the present study is to determine the level of protein and B-vitamins in whole wheat and straight grade flours of different spring wheats. Ten spring wheat varieties were procured from different parts of the country, milled and quantified. The protein content was determined by Kjeldhal’s method and B-vitamins were analyzed through High Performance Liquid Chromatography (HPLC) using UV-VIS detector. Results revealed that the level of protein as well as the B-vitamins was high in whole wheat flours of spring wheats as compared to straight grade flour. Only 3, 15, 6 and 6% of thiamin, riboflavin, niacin and pyridoxine of whole wheat flour was detected in straight grade flour. Simple Pearson’s correlation has been studied among wheat B-vitamins and its protein content which exhibited that in whole wheat flour a positive correlation for protein content with thiamin (r = 0.54), riboflavin (r = 0.07) and niacin (r = 0.22) whereas in straight grade flour protein concentration was examined to be positively correlated with riboflavin concentration (r = 0.19) only. Conclusively, the results of the current study are useful for different stake holders for their intended uses.