CN111537460A - Method for determining content of waxy sorghum starch - Google Patents
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Abstract
The invention belongs to the technical field of starch content determination, and particularly discloses a method for determining the content of waxy sorghum starch, which comprises the following steps: (1) preparing a solution; (2) preparing a standard colorimetric solution; (3) sample treatment and sample colorimetric solution preparation; (4) and (6) loading on a machine. The method is mainly used for measuring the content of the waxy sorghum starch, and solves the problem that the inaccurate detection result of the waxy sorghum starch in the prior art can increase the production cost.
Description
Technical Field
The invention belongs to the technical field of starch content determination, and particularly discloses a method for determining the content of waxy sorghum starch.
Background
The sorghum flour is flour milled from sorghum, has the reputation of natural nutritional food, contains 65.9 to 77.4 percent of starch, 8.4 to 14.5 percent of protein and 2.4 to 10.4 percent of crude fat, and is one of favorite food. The sorghum flour is beneficial to human health when being eaten frequently, and especially has certain auxiliary medical treatment effect when being eaten frequently by people suffering from hypertension, hyperlipidemia, diabetes and the like; the glutinous sorghum starch can also be used for brewing wine and other functions.
Waxy sorghum starch has very high amylopectin content and loose structure, is suitable for rhizopus growth, and has high starch yield when sorghum liquor is prepared by using Xiaoqu, wherein non-glutinous sorghum contains certain amylose and has compact structure, the protein content is higher than that of waxy sorghum, sorghum contains pigment components such as tannin, anthocyanin and the like, the derivative phenolic compound gives special fragrance to the white liquor, and the bulkiness of the sorghum is sticky but not pasty after steaming.
In the prior art, the detection steps of the waxy sorghum starch are multiple, so that the result of detecting the waxy sorghum starch is inaccurate, and the production cost is increased for producers in production.
Disclosure of Invention
The invention aims to provide a method for measuring the content of waxy sorghum starch, which solves the problem that the inaccurate detection result of waxy sorghum starch in the prior art increases the production cost.
In order to achieve the purpose, the technical scheme of the invention is as follows: the method for measuring the content of the waxy sorghum starch comprises the following steps:
(1) solution preparation: 1mol/L sodium hydroxide: weighing 20g of sodium hydroxide, adding water to dissolve the sodium hydroxide, and fixing the volume to 500 ml; 0.1mol/LHcl hydrochloric acid solution: measuring 4.5ml of concentrated hydrochloric acid, adding the concentrated hydrochloric acid into a 500ml volumetric flask with a certain amount of water, and fixing the volume; iodine reagent: weighing 2.0g of potassium iodide, dissolving in 10ml of distilled water, adding 0.2g of iodine, and diluting with distilled water to 100ml after dissolving;
1mg/ml amylose standard solution: drying amylose in an oven at 105 ℃ for 3 hours, cooling, weighing 0.1g of amylose in a 100ml conical flask, adding 1ml of ethanol, adding 9ml of 1mol/L sodium hydroxide, slightly shaking to disperse the starch, heating the mixture in boiling water for 10 minutes to completely disperse the starch, cooling, transferring the mixture to a 100ml volumetric flask, adding water until the starch is scaled and shaking uniformly;
(2) preparing a standard colorimetric solution:
amylose standard solution colorimetric solution: absorbing 1mg/ml amylose standard solution 0.3ml \0.7ml \1.0ml \1.5ml \2.0ml \2.5ml, respectively putting into a 50ml volumetric flask, adding 30ml distilled water, adjusting the pH value to about 3.5-4.0 by using 0.1mol/LHcl hydrochloric acid solution, adding 0.5ml iodine reagent, fixing the volume to 50ml, wherein the corresponding amylose standard solution concentrations are respectively: 0.006mg/mL, 0.014mg/mL, 0.02mg/mL, 0.03mg/mL, 0.04mg/mL, 0.05mg/mL, standing for 30 minutes, and loading;
(3) sample treatment and sample colorimetric solution preparation: randomly extracting 200g of sample, uniformly mixing, putting 50g of uniformly mixed sample into an electric skin grinding machine for grinding, wherein the surface of the grinded sorghum is white and smooth, a small amount of impurities are in the concave part, pouring out the sample, smashing the sample by using a smashing machine, weighing 3g of smashed sample, drying at 105 ℃ for 3 hours, cooling, weighing to obtain W, putting into a filter paper bag, and putting into a Soxhlet fat extractor for extracting and degreasing; taking out the filter paper bag, drying the starch in an oven at 105 ℃ for 3 hours, cooling and weighing to obtain a sample mass W1; weighing 0.1000g of a degreased sample into a 100ml conical flask, adding 1ml of ethanol, adding 9ml of 1mol/L sodium hydroxide, slightly shaking to disperse starch, heating the mixture in boiling water for 10 minutes to completely disperse the starch, cooling, transferring the mixture into a 100ml volumetric flask, adding water until the scales are scaled, and shaking uniformly to obtain a sample solution; absorbing 5.0ml of sample liquid into a 50ml volumetric flask, adding 30ml of distilled water, adjusting the pH value to be about 3.5-4.0 by using 0.1mol/LHcl hydrochloric acid solution, adding 0.5ml of iodine reagent into the sample, fixing the volume to 50ml, standing for 30 minutes, and loading on a machine;
(4) and (3) loading: measuring absorbance values of standard colorimetric solutions and preparing a wavelength curve, using distilled water as a blank, using a 1ml cuvette to measure amylose absorbance values A at the wavelengths of 620nm and 479nm, preparing a dual-wavelength amylose standard curve by using A620-479, and finding out the amylose standard curve to obtain the concentration Y1 of amylose in a sample so as to calculate the content of the amylose in the dehydrated and degreased sample; the amylose content is Y1 multiplied by 10 multiplied by W1/W multiplied by 100%; amylopectin content ═ 1-amylose content.
Further, in the step (3), a skin grinding machine is used for grinding skin for 4 times, and the time for grinding skin for each time is 5 min.
Further, the sample crushed by the crusher in the step (3) is 60-80 meshes.
Further, the degreasing method in the step (3) comprises the following steps: 105ml of methanol was added to the flask, and defatting was performed under heating and reflux for 4 to 6 hours.
Further, the drying temperature of the sample and the starch crushed in the step (3) are both 105 ℃, and the drying time is both 3 hours.
The technical principle and the beneficial effects of the technical scheme are as follows:
(1) the scheme adopts absorbance to calculate, so that the data is more accurate;
(2) in the scheme, the color reaction after the iodine is added in a dispersing way can more intuitively explain the purity of the sample;
(3) in the scheme, the amylopectin content is indirectly obtained by measuring the amylose content, so that the inaccurate detection data result caused by color development error in the process of directly detecting the amylopectin is avoided.
Drawings
FIG. 1 is a graph of amylose standards in the present invention.
Detailed Description
The following is further detailed by way of specific embodiments:
the laboratory apparatus used in this example: electric skin grinding machine (Jiliang JX95-3), miniature high-speed universal sample crusher, electronic balance, electric heating blowing drying oven, Soxhlet fat extractor, constant temperature water bath, PH meter, spectrophotometer, condenser
Reagents used in this example: amylose standard samples (potato amylose pure samples of agricultural product quality and safety institute of agricultural science of Heilongjiang province), amylopectin standard samples (rice amylopectin pure samples of agricultural product quality and safety institute of Heilongjiang province), methanol, sodium hydroxide, hydrochloric acid, iodine, potassium iodide and absolute ethyl alcohol.
In the example, substantially as shown in figure 1, a method for determining the starch content of waxy sorghum comprises the steps of:
solution preparation:
1mol/L sodium hydroxide: 20g of sodium hydroxide is weighed, dissolved in water and added to 500ml of volume, and 0.1mol/LHcl hydrochloric acid solution: measuring 4.5ml of concentrated hydrochloric acid, adding the concentrated hydrochloric acid into a 500ml volumetric flask with a certain amount of water, and fixing the volume; iodine reagent: weighing 2.0g of potassium iodide, dissolving in 10ml of distilled water, adding 0.2g of iodine, and diluting with distilled water to 100ml after dissolving;
1mg/ml amylose standard solution (weighed according to purity): drying amylose in an oven at 105 ℃ for 3 hours, cooling, weighing 0.1g of amylose in a 100ml conical flask, carefully adding 1ml of ethanol, adding 9ml of 1mol/L sodium hydroxide, slightly shaking to disperse the starch, heating the mixture in boiling water for 10 minutes to completely disperse the starch, cooling, transferring the mixture to a 100ml volumetric flask, adding water until the starch is scaled and shaking uniformly; iodine reagent: 2.0g of potassium iodide was weighed, dissolved in 10ml of distilled water, and then 0.2g of iodine was added, and the solution was diluted with distilled water to 100ml (the solution was prepared immediately before use).
1mg/ml amylose standard solution (weighed according to purity): will be provided withStraight barDrying chain starch in an oven at 105 deg.C for 3 hr, cooling, weighing 0.1g amylopectin into 100ml conical flask, carefully adding 1ml ethanol, adding 9ml1mol/L sodium hydroxide, shaking gently to disperse starch, heating the mixture in boiling water for 10 min to completely disperse, cooling, transferring to 100ml volumetric flask, adding water to scale, and shaking.
Preparing a standard colorimetric solution:
amylose standard solution colorimetric solution: amylose standard solution colorimetric solution: absorbing 1mg/ml amylose standard solution 0.3ml \0.7ml \1.0ml \1.5ml \2.0ml \2.5ml, respectively putting into a 50ml volumetric flask, adding 30ml distilled water, adjusting the pH value to about 3.5-4.0 by using 0.1mol/LHcl hydrochloric acid solution, adding 0.5ml iodine reagent, fixing the volume to 50ml, wherein the corresponding amylose standard solution concentrations are respectively: 0.006mg/mL, 0.014mg/mL, 0.02mg/mL, 0.03mg/mL, 0.04mg/mL, 0.05mg/mL, standing for 30 minutes, and loading;
absorbing 1mg/ml amylose standard solution 0.3ml \0.7ml \1.0ml \1.5ml \2.0ml \2.5ml, respectively putting into a 50ml volumetric flask, adding 30ml distilled water, adjusting the pH value to about 3.5-4.0 by using 0.1mol/LHcl hydrochloric acid solution, adding 0.5ml iodine reagent, fixing the volume to 50ml, wherein the corresponding amylose standard solution concentrations are respectively: 0.006mg/mL, 0.014mg/mL, 0.02mg/mL, 0.03mg/mL, 0.04mg/mL, 0.05mg/mL, and allowed to stand for 30 minutes.
Sample treatment and sample colorimetric solution preparation:
randomly extracting 200g of samples, uniformly mixing, putting 50g of uniformly mixed samples into an electric skin grinding machine for skin grinding for 4 times (5 minutes each time), wherein the surface of the ground sorghum is white and smooth, a small amount of impurities exist in concave parts, pouring out the samples, smashing the samples by using a miniature high-speed universal sample smashing machine, sieving the samples by using a 60-80-mesh sieve (superfine sieve), weighing 3g of smashed samples, putting the smashed samples into an electrothermal blowing drying box at 105 ℃ for drying for 3 hours, cooling, weighing to obtain W, putting the W into a filter paper bag, and putting the filter paper bag into a Soxhlet fat extractor for extraction and degreasing. The method comprises the following steps: adding 105ml of methanol into a flask, heating, refluxing and degreasing for 4-6 hours, taking out the filter paper bag, drying in the air, drying the starch in an oven at 105 ℃ for 3 hours, cooling, and weighing to obtain the sample mass W1. Weighing 0.1000g of degreased sample into a 100ml conical flask, carefully adding 1ml of ethanol, adding 9ml of 1mol/L sodium hydroxide, gently shaking to disperse the starch, heating the mixture in boiling water for 10 minutes to completely disperse the starch, cooling, transferring the mixture into a 100ml volumetric flask, and adding water until the scale marks are shaken up. Sucking 5.0ml of the sample liquid into a 50ml volumetric flask, adding 30ml of distilled water, adjusting the pH value to be about 3.5-4.0 by using 0.1mol/LHcl hydrochloric acid solution, adding 0.5ml of iodine reagent into the sample, fixing the volume to 50ml, standing for 30 minutes, and loading on the machine.
Firstly, processing amylose on a machine:
determination of standard colorimetric solution absorbance value and preparation of wavelength curve
Using distilled water as a blank (the same as the standard colorimetric solution except that amylose was not added), the amylose content in the dehydrated and defatted sample was calculated by measuring the absorbance value A of amylose at wavelengths 620nm and 479nm using a 1ml cuvette, preparing a standard curve for dual-wavelength amylose from A620 to 479, and finding out the standard curve for amylose to obtain the concentration Y1 of amylose in the sample.
Amylose content (in sample dry weight) Y1 × 10 × W1/W × 100%.
(1) In the formula, 10 is: (((Y1X 50)/5) X100)/0.1; since the unit of Y1 is mg/mL, when mg is converted to g, it needs to be divided by 1000 to obtain Y1X 10.
Amylopectin content (in dry sample weight) 1-amylose content.
Description of the drawings: in this method, the lower limit of amylose measurement is 1%, and the upper limit of amylopectin measurement is 99%.
(II) amylose standard curve detection data
As shown in table 1:
TABLE 1
Correction equation: abs 22.10941 × concentration +0.00815
Correlation coefficient: r-0.99987
As can be seen from table 1, for 6 standard samples, 3 parallel sample determinations were performed, with a deviation SD of 0.0005, 0.0003, 0.0005, 0.0006, 0.0021, 0.0074, and a mean deviation of 0.0019; the standard deviation% SD is 0.38, 0.10,0.10,0.09,0.24,0.67, the average standard deviation is 0.26, which shows that the precision of the measured data is good. And the correlation coefficient R is 0.99987, the linearity is very good, and the measurement result meets the requirement in the linear range. As can be seen from Table 1, as the concentration of the standard sample increases, the absorbance of the Abs increases, and the deviation SD and% SD standard increase, showing a positive correlation.
The foregoing is merely an example of the present invention and common general knowledge of known specific structures and features of the embodiments is not described herein in any greater detail. It should be noted that, for those skilled in the art, without departing from the structure of the present invention, several changes and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent.
Claims (5)
1. The method for measuring the content of the waxy sorghum starch is characterized by comprising the following steps of:
(1) solution preparation: 1mol/L sodium hydroxide: weighing 20g of sodium hydroxide, adding water to dissolve the sodium hydroxide, and fixing the volume to 500 ml; 0.1mol/LHcl hydrochloric acid solution: measuring 4.5ml of concentrated hydrochloric acid, adding the concentrated hydrochloric acid into a 500ml volumetric flask with a certain amount of water, and fixing the volume; iodine reagent: weighing 2.0g of potassium iodide, dissolving in 10ml of distilled water, adding 0.2g of iodine, and diluting with distilled water to 100ml after dissolving;
1mg/ml amylose standard solution: drying amylose in an oven at 105 ℃ for 3 hours, cooling, weighing 0.1g of amylose in a 100ml conical flask, adding 1ml of ethanol, adding 9ml of 1mol/L sodium hydroxide, slightly shaking to disperse the starch, heating the mixture in boiling water for 10 minutes to completely disperse the starch, cooling, transferring the mixture to a 100ml volumetric flask, adding water until the starch is scaled and shaking uniformly;
(2) preparing a standard colorimetric solution:
amylose standard solution colorimetric solution: absorbing 1mg/ml amylose standard solution 0.3ml \0.7ml \1.0ml \1.5ml \2.0ml \2.5ml, respectively putting into a 50ml volumetric flask, adding 30ml distilled water, adjusting the pH value to about 3.5-4.0 by using 0.1mol/LHcl hydrochloric acid solution, adding 0.5ml iodine reagent, fixing the volume to 50ml, wherein the corresponding amylose standard solution concentrations are respectively: 0.006mg/mL, 0.014mg/mL, 0.02mg/mL, 0.03mg/mL, 0.04mg/mL, 0.05mg/mL, standing for 30 minutes, and loading;
(3) sample treatment and sample colorimetric solution preparation: randomly extracting 200g of sample, uniformly mixing, putting 50g of uniformly mixed sample into an electric skin grinding machine for grinding, wherein the surface of the grinded sorghum is white and smooth, a small amount of impurities are in the concave part, pouring out the sample, smashing the sample by using a smashing machine, weighing 3g of smashed sample, drying at 105 ℃ for 3 hours, cooling, weighing to obtain W, putting into a filter paper bag, and putting into a Soxhlet fat extractor for extracting and degreasing; taking out the filter paper bag, drying the starch in an oven at 105 ℃ for 3 hours, cooling and weighing to obtain a sample mass W1; weighing 0.1000g of a degreased sample into a 100ml conical flask, adding 1ml of ethanol, adding 9ml of 1mol/L sodium hydroxide, slightly shaking to disperse starch, heating the mixture in boiling water for 10 minutes to completely disperse the starch, cooling, transferring the mixture into a 100ml volumetric flask, adding water until the scales are scaled, and shaking uniformly to obtain a sample solution; absorbing 5.0ml of sample liquid into a 50ml volumetric flask, adding 30ml of distilled water, adjusting the pH value to be about 3.5-4.0 by using 0.1mol/LHcl hydrochloric acid solution, adding 0.5ml of iodine reagent into the sample, fixing the volume to 50ml, standing for 30 minutes, and loading on a machine;
(4) and (3) loading: measuring absorbance values of standard colorimetric solutions and preparing a wavelength curve, using distilled water as a blank, using a 1ml cuvette to measure amylose absorbance values A at the wavelengths of 620nm and 479nm, preparing a dual-wavelength amylose standard curve by using A620-479, and finding out the amylose standard curve to obtain the concentration Y1 of amylose in a sample so as to calculate the content of the amylose in the dehydrated and degreased sample; the amylose content is Y1 multiplied by 10 multiplied by W1/W multiplied by 100%; amylopectin content ═ 1-amylose content.
2. The method for measuring the content of waxy sorghum starch according to claim 1, wherein in the step (3), the skin is ground 4 times by using a skin grinding machine, and the time for grinding the skin is 5min each time.
3. The method for measuring the content of waxy sorghum starch according to claim 1, wherein the sample after being pulverized by the pulverizer in the step (3) is 60-80 mesh.
4. The method for measuring the content of waxy sorghum starch according to claim 1, wherein the degreasing method in the step (3) is: 105ml of methanol was added to the flask, and defatting was performed under heating and reflux for 4 to 6 hours.
5. The method for measuring the content of waxy sorghum starch according to claim 1, wherein the drying temperature and the drying time of the sample and starch after pulverization in the step (3) are both 105 ℃ and 3 hours.
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2020
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| RU2644754C1 (en) * | 2016-12-21 | 2018-02-13 | Федеральное государственное бюджетное научное учреждение "Федеральный научный центр пищевых систем им. В.М. Горбатова" РАН | Method for determining amylose content in starch |
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