SU1635132A1 - Method for determination of biological infection of grapes and wine materials - Google Patents

Abstract

The invention relates to the food industry, in particular to the winemaking and canning industries, and concerns the identification of raw materials affected by molds. The purpose of the invention is to improve the accuracy of determining the viability of molds. This is achieved by selecting a substance that characterizes the presence of such a lesion. Rutin is used as such substance.

Description

The invention relates to the food industry, in particular to the winemaking and canning industries, and concerns the identification of raw materials affected by molds.

The purpose of the invention is to improve the accuracy of determining the viability of molds.

The essence of the method is as follows.

The grapes are processed, the pulp is pressed, the sesosmotek and the first pressure are separated and rutin is introduced into the average sample of the wort, incubated followed by determination of the change in rutin concentration. The average wort sample is taken in an amount of 50 cm3, 5 mg of rutin is administered and divided in half by 25 cm3. One part of the sample is sterilized at 100 ° C for 10 minutes, and the other is incubated at 37 ° C for 30 minutes.

As a result of the incubation, rutin is cleaved to rutinase disaccharide and aglycone under the action of rutinase, a specific mold enzyme. Samples are filtered through a membrane filter and rutin concentration is determined by high performance liquid chromatography. By reducing the rutin concentration above 10%, the raw material is recognized as affected by mold. The absence of differences in the concentration of rutin in the control and test samples, the value of not exceeding 10%, serves as the basis for the recognition of raw materials healthy.

The method is carried out as follows.

Example 1. Fet - ska varieties of grapes are sorted into healthy and diseased with molds, processed in a crusher-ridge separator, pulp is pressed, the gravity wort and the first pressure are separated and an average sample is taken according to GOST 14137-74. Average samples from each batch of processed grapes in three

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Replications in an amount of 50 cm each are placed in cones into which 5 mg of rutin was administered. Samples were divided in half by 25 cm, one half was sterilized by boiling at 100 ° C for 10 minutes, and the other half of each sample was left unchanged. Sterilized control samples are placed together with the subjects in a thermostat and incubated at 37 ° C for 0.5 h. A 5 cm sample is taken from each sample, filtered through a 0.45 µm membrane filter, and the concentration of Milichrom chromatograph is determined. on a column of 62x2 mm, with a silabor Cia sorbent. A 15 µm membrane filtered sample was injected into the chromatograph and eluted with a mixture of acetonitrile and water in a ratio of 20:30 (v / v) at pH 2.8 adjusted with phosphoric acid at a rate of 30 µl / min of the flow of the mobile phase. Detection is carried out at 360 nm with identification of the rutin peak by comparing the reduced sample retention time with the reduced retention time of the pure rutin solution analyzed under the same conditions. The rutin concentration is determined by a calibration schedule. I compared the concentrations of rutin in the control and the studied company to judge the incidence of raw materials with molds.

In tab. Table 1 shows the data characterizing the rutin concentration in the control and test samples of healthy and affected Fetka ska grapes.

In samples of wort obtained from a healthy raw material and in a sterilized sample, the concentration of rutin did not change, the deviations did not exceed the error of the analysis method. During the sterilization process, inactivation of the mold enzymes occurs and the Putin introduced into the sample is not subjected to splitting, its content does not change. In a sample of grape-affected grapes, the concentration of rutin decreased, it was destroyed by deupvia of enzymes present in the grape-affected grapes.

PRI mme R 2. Grapes of the Riesling, Rhenish, Pino, Traminer, Aligote, Amur varieties are processed on a scraper-ridge separator, the grape is not sorted for healthy and moldy years. take the average sample according to GOST 14137-74. Medium samples of each grade of processed grapes in an amount of 50 cm are placed in cones, into each of which 5 mg of rutin is added. Samples are divided in half by 25 cm3, one half is sterilized by boiling for 10

min, and the other half of each sample is left unchanged. Sterilized control samples are placed together with the subjects in a thermostat and incubated with

37 ° C for 0.5 h. From each sample, 5 cm3 of sample is taken, filtered through a 0.45 µm membrane filter, and the rutin concentration on a Milichrom chromatograph on a 62x2 mm column with Silaborb С1b sorbent is determined. A 15 µl membrane filtered sample is injected into the chromatograph and eluted with a mixture of acetonitrile and water in a ratio of 20:80 by volume at pH 2.8, adjusted with phosphoric acid, with

5, the flow rate of the mobile phase is 30 μL / min. Detection is carried out at 360 nm with identification of the rutin peak by comparing the reduced sample retention time with the reduced retention time of the pure rutin solution analyzed under the same conditions. The rutin concentration is determined by a calibration schedule. Decrease of rutin concentration in control and

The 5 samples studied after 0.5 hours, more than 10%, makes it possible to recognize the raw material as affected by mold. If the decrease in the rutin concentration does not exceed 10%, then the rutin concentration is determined.

0 every hour, but not more than 4.5 hours in total. In the absence of differences in the content of rutin in the control and test samples after 4.5 hours, raw materials were considered healthy.

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In tab. 2 shows the data characterizing the concentration of rutin in the samples of grapes in relation to the control.

0 In samples of wort Riesling, Rhenski and Aligote after 0.5 h of incubation, a noticeable decrease in the routine occurs (relative to the control), which allows us to state the fact of infestation of these batches

5 grapes mold. Pinot wort sortoecorse is affected by mold, which is confirmed by 3.5 h of incubation.

No changes in concentration were detected in the Traminer and Amur wort samples.

0 rutin (with respect to the control) during the whole incubation time, which allows to establish the fact of the absence of damage to the raw material by mold, raw health. The examples presented show

5 the possibility of using the determination of rutinase activity in a mold affected raw material intended for processing. Rutinase is a specific mold enzyme whose activity is found in the digestion of

rutin added to the sample, and rutin can serve as an indicator for determining molds in the feedstock.

The proposed method (unlike the well-known one) provides: specificity of determination, greater accuracy of determination with varying degrees of material damage; the ability to monitor the progress of the pasteurization process, bentonite processing and other technological operations associated with the removal of enzymes from the processed material.

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Claims (1)

Claims The method for determining the bio-opacity of grapes and wine materials by the presence of the product of the metabolism of fungi, wherein rutin is introduced into the sample of the test product, followed by dividing it into two parts, one of which is sterilized, the concentration the rutin in these parts and compare them with each other, and the determination of the presence of a product of metabolism is carried out on the basis of the comparison result. Table 1 table 2