Method for determination of pectin substances weight percentage in vegetal raw material

FIELD: food industry.

SUBSTANCE: invention relates to food industry, in particular to determination of pectin substances quantity in vegetal raw material. The method envisages drying plaited paper filter until constant weight, preparation of protopectin and hydratopectin, sampling. The protopectin solution acid medium is neutralised with pectin substances fraction in the solutions being analysed saponified with 40% solution of NaOH during 15 minutes. Then pectin substances are sedimented with concentrated HCl and the sediment is filtered. Then the sediments on filters are washed with cold distilled water. Filters with sediments are dried till constant weight and one calculates pectin substances fractions weight percentage from certain formulas.

EFFECT: invention enables enhanced accuracy of determination of pectin substances weight percentage in vegetal raw material, reduction of time and labour expenditures as well as specific equipment usage exclusion.

2 tbl, 2 ex

 

The invention relates to the technical biochemistry, namely to determine the amount of pectin in plant raw material.

Regardless of the nature of plant material and methods of quantitative determination of the content of pectin in it, the first step is always cooking "standardized" solutions hydrocompaction and > protopectin, the essence of which lies in a phased and full extraction of all of hydrocompaction and > protopectin from plant tissue. Thus obtained extracts hydrocompaction > protopectin and explore one of the existing methods.

There is a method of quantitative determination of pectin, which is based on the deposition of pectic acids strong mineral acids under conditions of high gravity ("bulk" method).

To 200 ml of "standardized" solution hydrocompaction or > protopectin selected for the study, add 2.5 ml of 40%NaOH solution and incubated at room temperature for 15 minutes. Formed pectic acid precipitated with 10 ml of concentrated HCl and centrifuged at 3000 rpm for 10-15 minutes in a number of graduated centrifuge tubes. In the centrifugation pectin colloids form a precipitate, the volume of which is measured in each of the wading is E. The volume of sediments in all the test tubes summarize. Then, using the calibration curve and the amount of precipitate obtained, determine the concentration of pectin [Shelukhina I.E. the Scientific basis of the technology of pectin. Frunze: Ilim, 1988, - 168 C.].

The disadvantages of this method include the above unified centrifugation parameters, which in practice does not ensure the accuracy of the analysis, as pectic acid selected from various types of raw materials, have different ability to precipitate formation in conditions of high gravity. In the precipitation yield different density, which inevitably affects occupied volume, and, in addition, part of clots pectic acids often freely placed in the liquid volume of the centrifuge tube, and not going into the sediment. In addition, the amount of rainfall in the tube is so small that the calibration tubes accurately determine the quantity that is not possible. All this inevitably leads to high errors and inaccurate test result. The disadvantages of the method should also include the need to use specific equipment (centrifuge and graduated centrifuge tubes).

There is a method of quantitative determination of pectin, which is based on the deposition of pectic acid in the form of calcium with the lei - pectates - and given their number weight ("calcium-Pechatny" method).

The beginning of the analysis is preceded by drying the necessary number of paper folded filter at a temperature of 100-105°C to constant weight.

Solution > protopectin prepare to saponification, neutralizing the acidic reaction medium, which was used during the extraction of > protopectin, sodium hydroxide.

Next is the stage of saponification of hydrocompaction or > protopectin, (according to the study "standardized" solution), which are selected based on 50 ml of the appropriate standardized solution. In the same flask was added the same volume of 0,1N. (0,4%) NaOH solution. The flask cover and leave for 8-10 hours at room temperature to conduct the saponification process carboxyl groups of pectin.

After this time the solution acidified with 50 ml of 1N. acetic acid and incubated for 30 minutes. Formed pectic acid is precipitated with 50 ml of 11.1% solution of CaCl2. Received 1 precipitation pectate calcium is filtered through a pre-dried to constant mass filters. Precipitation on the filters was washed with a 0.5% solution of CaCl2portions of 50 ml, after which precipitation repeatedly washed with cold distilled water to remove chloride ions. Completeness of washing the precipitate of pectate calcium is from ions of Cl -determined by qualitative reactions of the filtrate with AgNO3. To reduce ash residue on the filter is washed several times with hot distilled water.

Filters with the washed precipitate is carefully transferred into boxy and dried at a temperature of 100-105°C to constant weight.

If the mass of the precipitate obtained from the difference between the weight of buxa with the dried filter and the precipitate pectin and weight buxa with the dried filter to filter sediment does not exceed 0.03 g, the calculation of the content of individual fractions of pectin in plant tissue exercise by the formula (1):

where X is the content of the corresponding fractions of pectin in the studied material, % of wet weight;

mf+n- weight buxa with a filter and dried to constant weight sediment, g;

mf- weight buxa with the dried filter to filter, g;

Vbulb- volume of the volumetric flask with "standardized" solution, ml;

0,9235 - the conversion factor for pochtovuyu acid;

100 is the conversion factor to percentage;

msamplethe weight of the investigated material, g;

Vthe extras- volume standardized solution that you want to take for the experiment.

If the mass of the precipitate obtained from the difference between the weight of buxa with the dried filter and the precipitate pectin and weight budget the sa with the dried filter to filter sediment exceeds 0.03 g, the experiment should be repeated with a smaller number of standardized solution, the volume of which is determined by the formula (2):

[Methods of quantitative determination of pectic substances in plant material. Methodical instructions for practical exercises / Donchenko L.V., V.V. Kondratenko, Kondratenko T.U. - Krasnodar, 2005, to 30 C.].

The disadvantages of the described method include high accuracy, 0.3%, which may be associated with the transition in sediment pectate calcium repectively impurities, as well as the high complexity and a greater duration of the analysis process (often there is a need to alter the analysis because of the excess weight pectate calcium 0.03 g).

The technical result of the invention is to improve the accuracy of the results of determining the mass fraction of pectin in plant material, reducing labor costs and time required for analysis, and excluding the use of specific equipment.

The technical result is achieved by the fact that in the known method, including drying of paper folded filters to constant weight, the preparation of "standardized" solutions > protopectin and hydrocompaction, sampling, neutralization of the acidic solution > protopectin, amylene the fractions of pectin substances in the samples, the precipitation of the pectin substances, filtration, precipitation, precipitation washing on the filter with distilled water, drying filters with precipitation to constant weight, the calculation of the mass fraction of pectin fractions according to the proposed method, the saponification of pectin in the analyzed solutions spend 40%NaOH solution for 15 minutes, pectin is precipitated with concentrated HCl, and the mass fraction of pectin substances are calculated according to formulas (3) and (4):

for hydrocompaction

where ωse- mass fraction of particles of hydrocompaction, %;

m1- the weight of the portion of the studied raw material taken for analysis, g;

m2- weight of sediment, g;

0.8 - conversion factor volume "standardized" solution hydrocompaction taken for research, the total volume of "standardized" solution hydrocompaction;

100 is the conversion factor to percentage;

for > protopectin

where ωnn- mass fraction of particles of > protopectin, %;

0,4 - coefficient conversion amount "standardized" solution > protopectin taken for research, the total volume of "standardized" solution > protopectin.

Application of 40%NaOH solution instead of 0,1N. NaOH solution for saponification pectin reduces the La times with 8-10 hours to 15 minutes.

The use of concentrated HCl to provide fast and efficient neutralization of alkaline saponified solutions (instead of preceding the deposition of 11.1%solution of CaCl2, acidification of solutions of 50 ml of 1N. acetic acid with a 30-minute exposure) and total deposition saponified fractions of pectin, which increases the validity of the definition.

The method is implemented as follows. Dried necessary number of paper folded filters to constant weight, to prepare a "standardized" solutions > protopectin and hydrocompaction, take samples, neutralize the acidic solution > protopectin, researched solutions omelet 40%NaOH solution for 15 minutes, then precipitated pectin concentrated HCl, precipitation was filtered, washed with distilled water from chlorine ions, then the filters with precipitation dried to constant weight, calculated mass fractions of fractions of pectin by the formulas (3) and (4):

for hydrocompaction

for > protopectin

Examples of the implementation of the proposed method

Example 1. Prepare the required number of paper folded filters, drying them in the tube in a drying Cabinet at 105°C to constant mass.

Definition provocat three replications.

Take a sample of 10 g of dry beet pulp and cook "standardized" solutions hydrocompaction and > protopectin.

After that from the total number of the solutions prepared fractions of pectin substances selected sample volume of 200 ml and the sample solution > protopectin neutralize the acidic environment titration with 0,1N. the NaOH solution in the presence of phenolphthalein indicator (until a steady slightly pink color).

Then to selected and prepared for research samples "standardized" solutions hydrocompaction > protopectin and add 2.5 ml of 40%NaOH solution and incubated at room temperature for 15 minutes, amylea pectic substances. The indicator dyes solutions in a rich crimson color.

After a specified time formed pectic acid precipitated with 10 ml of concentrated HCl. The formed precipitation was filtered through dried to constant weight paper folded filters and repeatedly washed with cold distilled water to remove chloride ions. Complete cleaning sludge from chlorine ions is determined by qualitative reactions of wash water with AgNO3.

After filtration the filters with precipitation neatly stack in buxy and dried in a drying Cabinet at 105°C to constant mass.

After this is about defining the weight of precipitation, subtracting from the mass of the dried filters with precipitation mass of the dried filters to filter, and the mass proportion of pectic substances in the investigated raw materials are determined using formulas (3) and (4). The results of the calculations are entered in the table 1.

1.;.

2.;.

3.;.

Table 1
A series of experimentsFraction of pectinThe precipitation, gThe volume fraction of pectin, %
1Hydrocompaction0,10911,37
> Protopectin0,448611,22
2Hydrocompaction0,10521,36
> Protopectin0,4178of 11.45
3Hydrocompaction 0,10611,33
> Protopectin0,461511,54

Example 2. Prepare the required number of paper folded filters, drying them in the tube in a drying Cabinet at 105°C to constant mass.

The determination is carried out in three replications.

Take a portion of 25 g of raw fruits wild pears and cook "standardized" solutions hydrocompaction and > protopectin.

After that from the total number of the solutions prepared fractions of pectin substances selected sample volume of 200 ml and the sample solution > protopectin neutralize the acidic environment titration with 0,1N. the NaOH solution in the presence of phenolphthalein indicator (until a steady slightly pink color).

Then to selected and prepared for research samples "standardized" solutions hydrocompaction > protopectin and add 2.5 ml of 40%NaOH solution and incubated at room temperature for 15 minutes, amylea pectic substances. The indicator dyes solutions in a rich crimson color.

After a specified time formed pectic acid precipitated with 10 ml of concentrated HCl. The formed precipitation was filtered through dried to constant weight paper skladka the s filters and repeatedly washed with cold distilled water to remove chloride ions.

Complete cleaning sludge from chlorine ions is determined by qualitative reactions of wash water with AgNO3.

After filtration the filters with precipitation neatly stack in buxy and dried in a drying Cabinet at 105°C to constant mass.

Then determine the mass of precipitation by subtracting from the mass of the dried filters with precipitation mass of the dried filters to filter, and the mass proportion of pectic substances in the investigated raw materials are determined using formulas (3) and (4). The calculation results are recorded in table 2.

1.;.

2.;.

3.;.

Table 2
A series of experimentsFraction of pectinThe precipitation, gThe volume fraction of pectin, %
1Hydrocompaction0,09600,48
> Protopectin0,15211,52
2 Hydrocompaction0,09200,46
> Protopectin0,14751,48
3Hydrocompaction0,09710,49
> Protopectin0,15401,54

The technical result is achieved only when the combination of the described process parameters, which allows to obtain accurate data at high precision parallel tests.

A positive effect in the implementation of the developed method is the ease of implementation and cost savings of time and labor on all stages of the process.

The method of determining the mass fraction of pectin in plant material, including drying of paper folded filters to constant weight, the preparation of standard solutions > protopectin and hydrocompaction, sampling, neutralization of the acidic solution > protopectin, saponification fractions of pectin substances in the samples, the deposition of pectin substances, filtration, precipitation, precipitation washing on the filter with distilled water, drying filters with precipitation to constant weight, the calculation of the mass fraction of actinomy substances fractions, characterized in that the saponification of pectin in the analyzed solutions spend 40%NaOH solution for 15 min, pectin besieging concentrated HCl, and the mass fraction of pectin substances are calculated according to formulas
for hydrocompaction

where ωse- mass fraction of particles of hydrocompaction, %;
m1- the weight of the portion of the studied raw material taken for analysis, g;
m2- weight of sediment, g;
0.8 - conversion factor volume "standardized" solution hydrocompaction taken for research, the total volume of "standardized" solution hydrocompaction;
100 is the conversion factor to percentage;
for > protopectin

where ωPP- mass fraction of particles of > protopectin, %;
0,4 - coefficient conversion amount "standardized" solution > protopectin taken for research, the total volume of "standardized" solution > protopectin.



 

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