Extraction, identification and quantification of water buffalo milk proteins using a combination of reverse phase high-performance liquid chromatography coupled to diode-array detector (RP-HPLC-DAD) and near-infrared spectroscopy (NIRS) measurements

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Vinicius da Silva Botelho Duarte GomesRafael Mezzomo

Abstract

The development of novel methods to estimate the number of protein genetic variants of caseins (CN) and whey proteins in buffalo’s milk is necessary to optimize the technological and nutritional properties of dairy derivative products from this species. The literature has shown that buffalo’s milk CN and whey proteins can be successfully quantified using RP-HPLC-DAD. However, pure standards of genetic variants of CN and whey proteins in buffalo’s milk are not commercially available. This condition makes necessary their isolation and purification by preparative chromatography, which is difficult and requires specialized equipment. Thus, we proposed a protocol to quantify different casein genetic variants and major whey proteins in the milk of water buffaloes, using the combination of measurements derived from RP-HPLC-DAD and NIRS techniques. Our data showed estimates of casein variants and whey proteins similar to those reported by the classical method RP-HPLC-DAD that uses purified proteins and calibration curves.

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