In the Egyptian market, the most commonly used milkfat is buffalo's milkfat (BMF) as it is used by many consumers. Due to its high value and high cost, BMF is prone to adulteration with other cheap vegetable oils (VO). The common VO that are added to BMF are Shortening (SHO) and Hydrogenated Palm Kernel Oil (HPKO). Therefore, the current study was conducted to propose a two-steps protocol that can be used to detect adulteration of BMF with VO in general, and with SHO and HPKO in particular. SHO and HPKO were added, separately, to BMF by using different proportions (10, 20, 30, 40, and 50%). Reichert Meissl value (RM) and sterol fractions were used (first step) to detect the addition of VO to BMF. The addition of SHO and HPKO resulted in a significant decrease in the RM values when added with a proportion of 30% or more. The β-sitosterol values showed a significant increase when SHO or HPKO was added (10% or more). Fatty acids profile was used (second step) to determine the type of VO added to BMF. It was found that the addition of HPKO to BMF resulted in a significant increase in the levels of lauric acid (C12:0), palmitic acid (C16:0), and saturated fatty acids. While the addition of SHO to BMF resulted in a significant decrease in myristic acid (C14:0) and an elevation in palmitic acid (C16:0). Three different BMF samples from the local market were tested by using this two-steps protocol, and it succeeded to detect the addition of VO to BMF and to identify the type of VO that had been used. The obtained results suggested that the determination of β-sitosterol and fatty acids composition could be used for ascertaining the quality of milk fat.
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