A research group from the Analytical Chemistry and Chemical Engineering Department of Alcalá University has developed a procedure for estimating the content of 7S and 11S globulins in soybeans by Perfusion reversed-phase high-performance liquid chromatography (RP-HPLC). This invention represents a breakthrough for quality control analysis of these products.
The group is seeking for commercial agreements with technical assistance or license agreements patent.
New and innovative aspects
This invention represents a breakthrough allowing the possibility of estimate separately the contents of the mentioned globulins that have different characteristics and different functional properties, so that this estimation would allow to select properly the most suitable variety of soy bean for the development of a particular food according to the functional properties that are more necessary for such elaboration.
Moreover, this method is very fast because the chromatographic conditions used allow to obtain the soybean chromatogram in less than three minutes.
Main advantages of its use
- The procedure is accessible to most analysis laboratories because it can be carried out a basic instrumentation.
- The procedure is based on a chromatographic separation using a perfusion column and it is capable of separating the chromatographic peaks of the soybean proteins corresponding to 11S globulin and to the fraction (7S globulin + serum proteins) in soybean samples.
- The area measurement of the chromatographic peaks corresponding to 11S globulin and to the fraction (7S globulin + serum) coupled with the total area measurement of the peaks and the value of the total protein content of soybeans, allows to estimate the content of each of the majority globulins in the soybean.
The chromatograms for soybeans have been obtained with a high performance liquid chromatograph equipped with a degassing system, a quaternary pump, a thermostatted column compartment, an autosampler and a variable wavelength UV absorption detector that allows to perform mobile phase composition gradients. The column used has a perfusion filling in reversed phase mode.
The separation is performed with a perfusion reversed-phase column with dimensions of 50 x 4.6 mm internal diameter packed with particles of crosslinked polystyrene with divinylbenzene with a size of 10 μm. The chromatographic method consists of a linear binary gradient: from 5 to 25% of mobile phase (MP) B in 1.7 min and from 25 to 45% MP B in 1.3 min followed by a reverse gradient from 45 to 5% MP B in 1 min to equilibrate the column with the initial conditions between the analysis. The mobile phases are prepared with water-trifluoroacetic acid 0.1% (v/v) (mobile phase A) and acetonitrile-trifluoroacetic acid 0.1% (v/v) (mobile phase B). The mobile phase flow is 3 mL/min, temperature 60 °C and UV detection is performed at 254 nm. The mobile phases are filtered with PVDF filters (0.45 μm) and they are continuously degassed by vacuum. The injection volume is 20 μL.
The preparation of the soybeans solutions is performed by dissolving the appropriate amount of milled sample in a volume of suitable Milli-Q water followed by sonication at 25ºC during 5 min and centrifugation (3362g) during 10 min.
- To select properly the most suitable variety of soy bean for the development of a particular food according to the functional properties that are more necessary for such elaboration.
Intellectual property status
Patent granted with date 8/10/2009
Current development status
Already on the market
Desired business relationship
Commercial agreement with technical assistance