A research group from the Analytical Chemistry Department of Alcalá University has developed an ultrafast procedure for detection of whey protein of cow's milk by Perfusion reversed-phase high-performance liquid chromatography in dairy-type products of soybean. This invention represents a breakthrough for routine quality control analysis of these products.
The group is seeking for commercial agreements with technical assistance or license agreements patent.
New and innovative aspects
Although the whey protein of cow's milk and soybean proteins have been characterized and quantitatively determined separately using different techniques, until recently it there has not been separated simultaneously both types of proteins, nor in as little time (a few minutes).
Main advantages of its use
- This method can be performed with basic instrumentation and therefore, it is accessible to the majority of the analytical laboratories.
-The gradient used allows the simultaneous separation of the soybean protein and the whey protein of cow's milk.
- It is a simple method because it does not require a complex previous treatment of the sample or patterns.
- It allows the ultrafast separation and determination of these two types of proteins, making it ideal for industrial application.
- It allows the detection of adulteration due to the addition of whey proteins of cow's milk to dairy-type commercial products made from soy protein isolate.
- It allows simultaneously and separately to quantify soybean proteins and whey proteins of cow's milk.
- It allows to determine the content of soybean protein and whey proteins from cow's milk in samples with other additives containing nitrogen and for which existing methods for the determination of total nitrogen, do not allow to specify the protein content of the sample.
The analytical method which allows the simultaneous determination of soybean proteins and whey protein of cow's milk by perfusion reversed-phase high-performance liquid chromatography (HPLC) is carried out in a high pressure liquid chromatograph allowing to perform gradients of mobile phase composition and it has coupled a detector for ultraviolet-visible absorption (UV-Vis) of variable wavelength. The chromatographic column used is a column with a filling of perfusion in reversed phase mode.
The method consists of a linear and binary gradient in three steps, whose volume (Gradient time x Flow) is 15 mL. For a flow of 3mL/min (linear flow rate of 1058 cm/hr), the gradient is designed as follows: 5 to 25% of mobile phase (MP) B in 1.70 min, from 25 to 43% MP in 0.30 min and from 34 to 41% of MP B in 3 min. This gradient is followed by another linear and inverse gradient from 41 to 5% MP B in 1 min and 1 min at 5% of MP B to rebalance the column in the initial conditions. It is possible the use of this gradient to other flows maintaining constant the volume of the gradient. The mobile phases used are: MP A, grade HPLC water with 0.10% (v/v) of trifluoroacetic acid; MP B, HPLC grade acetonitrile with 0.10% (v/v) of trifluoroacetic acid. The mobile phases are filtered through nylon membranes of 0.45 _m and they are degassed before and/or during their use.
The separation is carried out at a temperature of the column constant and equal to 60ºC and the detection is performed at a wavelength of 254 nm.
- Routine quality control analysis of these products
- Quality control and food safety.
Intellectual property status
Patent granted with date 14/11/2000
Current development status
Already on the market
Desired business relationship
Commercial agreement with technical assistance