The analysis of phosphorylated compounds by LC are often problematic because these compounds have high tendency to interact with metal ions and cause issues such as analyte loss, peak tailing, and inconsistent results. Recently, a new hybrid surface technology (HST) has been developed. It provides a layer over the metal surface to mitigate the analyte metal interaction. This work investigates the effect of this hybrid surface on the UPLC-MS/MS analysis of B group vitamins, including the phosphorylated vitamers. Comparison studies were carried out on two UPLC-MS setups, one comprises conventional stainless-steal instrument and column, and the other one comprises instrument and column that incorporate hybrid surface technology. Improvements in peak area, peak shape, and sensitivity were observed for all B vitamins using the UPLC-MS setup incorporating the hybrid surface technology. The enhance performance of B-group vitamin analysis using the HST will be discussed.