CN106353268A - Rapid detection method of amylase content in rice - Google Patents
Rapid detection method of amylase content in rice Download PDFInfo
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- CN106353268A CN106353268A CN201610758168.8A CN201610758168A CN106353268A CN 106353268 A CN106353268 A CN 106353268A CN 201610758168 A CN201610758168 A CN 201610758168A CN 106353268 A CN106353268 A CN 106353268A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
Abstract
The invention provided a rapid detection method of amylase content in rice, which comprises the steps of 1, preparing a sample to be tested with the sample of rice and preparing 4 to 6 kinds of standard amylase powder as standard samples; 2, obtaining the sample solution and the standard solution by conducting microwaving, de-fatting and constant volume processing of the samples to be tested and the standard samples; 3, preparing the blank solution; 4, determining the absorbance of the sample solution and the standard solution at the wavelength of 720 nanometers with the spectrophotometer, wherein the spectrophotometer is zero set with the blank solution; 5, drawing the standard curve of the amylase-absorbance with the amylase content and absorbance of the standard solution and then obtaining the amylase content of the sample of rice according to the absorbance of the samples. The invention has the advantages of short processing time, good dispersing effect and improving the detection efficiency and stability of test results.
Description
Technical field
The present invention relates to the detection method of rice quality detection technique field, specifically rice grain amylose content.
Background technology
A kind of assay method of existing rice chain content of starch, it includes following steps:
1st, Rice Samples are obtained polished rice after shelling, fine grinding, take at least 10g polished rice, be ground into powder with Cyclone mill, and pass through
The screen cloth of 80 mesh～100 mesh, then adopts 85% methanol solution backflow extracting defat, places 2 days with equilibrium water after defat to powder
Get sample.
2nd, weigh 100 mg scholar's 0.5mg samples in 100ml conical flask, carefully add people's 1ml ethanol solution in sample, will
The sample being bonded in bottle wall sweeps away, and l.0mo1/l sodium hydroxide solution, in conical flask, and gently shakes up to pipette 9.0 ml, with
Afterwards mixture is heated in boiling water bath 10min with dispersing starch；Taking-up is cooled to room temperature, transfers in 100ml volumetric flask,
Plus distilled water constant volume acutely shaking mixing, obtain sample dispersion liquid.
3rd, 4 kinds of amylose standard powder are prepared, in each amylose standard powder, the mass content of amylose is different, respectively
Weigh each 100mg scholar 0.5mg of this 4 kinds of amylose standard powder to be put in different conical flasks, carefully add people in each conical flask
1ml ethanol solution, in sample, the amylose standard powder being bonded in bottle wall is swept away, and pipettes 9.0 ml l.0mo1/l hydrogen-oxygen
Change and receive solution in each conical flask, and gently shake up, subsequently mixture is heated in boiling water bath 10min with dispersing starch；Take
Go out and be cooled to room temperature, transfer to respectively in 100ml volumetric flask, plus distilled water constant volume acutely shaking mixing, obtain 4 kinds of standards
Dispersion liquid；
4th, pipette 0.09mol/l sodium hydroxide, standard scores dispersion liquid and each 5.0 ml of sample dispersion liquid, carry out following operation respectively:
Put in advance plus in the 100ml volumetric flask of people's about 50ml water, plus 1.0 ml acetic acid solutions, shake up, add the sumptuous examination of 2.0ml
Agent, adds water to scale, shakes up, and stands 10min, respectively obtains blank solution, standard solution and sample solution.
5th, use the absorbance of spectrophotometer determination sample solution, standard solution at 720nm, spectrophotometer is empty
White solution zeroing.
6th, with the amylose content of standard solution as abscissa, straight chain is drawn for vertical coordinate with corresponding absorbance and forms sediment
Powder-absorbance standard curve, the absorbance of sample solution is obtained sample solution with reference to amylose-absorbance standard curve
Amylose content, and then detect the amylose content of rice sample.
Existing this assay method have the shortcomings that detection time length and accuracy low it is therefore necessary to improve.
Content of the invention
The purpose of the present invention is to propose to a kind of method for quick of rice grain amylose content, it is time-consuming that it has detection
The high feature of short, result precision, can effectively improve efficiency, accuracy and the reliability of rice grain amylose content detection.
The purpose of the present invention can be achieved through the following technical solutions:
The method for quick of rice grain amylose content, it includes following steps:
Step 1, preparation of samples.
Rice Samples are obtained polished rice after shelling, fine grinding, polished rice is ground into all powder by 80 mesh～100 mesh sieves
Shape thing, weighs powder 100mg scholar 0.5mg as testing sample.
Prepare 4 to 6 kinds of amylose standard powder, in each amylose standard powder, the mass content of amylose is different, point
Another name takes each amylose standard powder 100mg scholar 0.5mg as standard sample.
Respectively testing sample and standard sample are put in different beakers.
The preparation of step 2, sample solution and standard solution.
Step 2.1, is sequentially added into 95% ethanol solution 1 ml and 1.0 mol/l hydroxides in each beaker of step 1
Sodium solution 9.0 ml, is placed in heating 10s～20s in microwave environment, takes out and be cooled to room temperature, transfer to 100ml capacity after shaking up
In bottle, plus distilled water constant volume shaking up, respectively obtain sample dispersion liquid.
Step 2.2, respectively the sample dispersion liquid 10ml～50ml of removing step 2.1 be put in separatory funnel, to the leakage of point liquid
Add 5ml～20ml petroleum ether, convolution shake 1min～5min in bucket, place into after collecting lower floor's aqueous phase in separatory funnel and add
5ml～20ml petroleum ether, convolution shake 1min, collects lower floor's aqueous phase, respectively obtains defat sample.
Step 2.3, during respectively defat sample 5.00 ml of removing step 2.2 is in the 100 ml volumetric flasks to volumetric flask according to
After secondary addition distilled water 50ml, 1.0 mol/l acetic acid solution 1 ml and iodine reagent 2.0ml, add distilled water constant volume, shake up, quiet
Put 10 min, sample solution and multiple standard solution are obtained respectively.
Step 3, the preparation of blank solution.
Prepare 0.09mol/l sodium hydroxide solution as blank sample, pipette blank sample 5.00 ml in 100 ml capacity
In bottle, after sequentially adding distilled water 50ml, 1.0 mol/l acetic acid solution 1 ml and iodine reagent 2.0ml in volumetric flask, add and steam
Distilled water constant volume, shakes up, and stands 10 min, obtains blank solution.
Step 4, using the absorbance of spectrophotometer difference determination sample solution, each standard solution at 720nm, light splitting
Photometer blank solution returns to zero.
Step 5, with the amylose content of standard solution as abscissa and corresponding absorbance be vertical coordinate draw straight chain
Starch-absorbance standard curve, the absorbance of sample solution is obtained testing sample with reference to amylose-absorbance standard curve
Amylose content, thus detecting the amylose content of Rice Samples.
Prioritization scheme, in step 2.1, the frequency of microwave is 2000mhz～3000mhz.
Prioritization scheme further, in step 1, amylose standard powder is contained by potato amylose and amylopectin quality
The waxy rice flour of amount more than 99% is uniformly mixed.
The present invention has substantive distinguishing features following outstanding and significantly improves:
1st, make starch fast and effectively gelatinizing, the dispersion in rice in the present invention by the microwave treatment of step 2.1, there is place
Reason takes the feature short, dispersion effect is good, effectively accelerates detection efficiency, significantly improves the stability of testing result.
2nd, in the present invention, ungrease treatment is carried out to sample dispersion liquid by step 2.2, due to Oryza glutinosa sample in sample dispersion liquid
Product are uniformly dispersed, and make defatting step have the characteristics that efficiency high, time-consuming short, effect are good.
3rd, the present invention have the characteristics that to process quick, can have strong operability, can mass disposal Rice Samples, during sample pre-treatments
Between short, thoroughly, data variation degree is less, favorable reproducibility for starch gelatinization.
Brief description
Fig. 1 is the standard curve of the detection method drafting of the present invention.
Fig. 2 is the standard curve drawn by the detection method of prior art.
Specific embodiment
The invention will be further described below.
Embodiment
The method for quick of rice grain amylose content, it includes following steps:
Step 1, preparation of samples.
Rice Samples are obtained polished rice after shelling, fine grinding, polished rice is ground into all powder by 80 mesh～100 mesh sieves
Shape thing, weighs powder 100mg scholar 0.5mg as testing sample.
Prepare 4 kinds of amylose standard powder, each amylose standard powder is by potato amylose and amylopectin quality
The waxy rice flour of content more than 99% is uniformly mixed, and its amylose mass content is respectively 1.5%, 10.4%, 16.2% He
26.5%, weigh each amylose standard powder 100mg scholar 0.5mg respectively as standard sample.
Respectively testing sample and standard sample are put in different beakers.
The preparation of step 2, sample solution and standard solution.
Step 2.1, is sequentially added into 95% ethanol solution 1 ml and 1.0 mol/l hydroxides in each beaker of step 1
Sodium solution 9.0 ml, is placed in after shaking up in the environment that microwave frequency is 2500mhz and heats 15s, take out and be cooled to room temperature, transfer to
In 100ml volumetric flask, plus distilled water constant volume shaking up, respectively obtain sample dispersion liquid.
Step 2.2, respectively the sample dispersion liquid 20ml of removing step 2.1 be put in separatory funnel, into separatory funnel plus
Enter 10ml petroleum ether, convolution shake 1min～5min, after collecting lower floor's aqueous phase, place into addition 10ml petroleum ether in separatory funnel,
Convolution shake 1min, collects lower floor aqueous phase, respectively obtains defat sample, and now the volume of defat sample is substantially or 20ml.
Step 2.3, during respectively defat sample 5.00 ml of removing step 2.2 is in the 100 ml volumetric flasks to volumetric flask according to
After secondary addition distilled water 50ml, 1.0 mol/l acetic acid solution 1 ml and iodine reagent 2.0ml, add distilled water constant volume, shake up, quiet
Put 10 min, sample solution and multiple standard solution are obtained respectively.
The compound method of iodine reagent is: weigh 2.000g ± 0.005g potassium iodide, plus appropriate distilled water to make saturation molten
After liquid, add 0.200g ± 0.001g iodine, iodine after all dissolving quantitatively moves to solution in 100ml brown volumetric flask, plus steam
Distilled water, to scale, shakes up, now with the current, keeps in Dark Place.
Step 3, the preparation of blank solution.
Prepare 0.09mol/l sodium hydroxide solution as blank sample, pipette blank sample 5.00 ml in 100 ml capacity
In bottle, after sequentially adding distilled water 50ml, 1.0 mol/l acetic acid solution 1 ml and iodine reagent 2.0ml in volumetric flask, add and steam
Distilled water constant volume, shakes up, and stands 10 min, obtains blank solution.
Step 4, using the absorbance of spectrophotometer difference determination sample solution, each standard solution at 720nm, light splitting
Photometer blank solution returns to zero.
Step 5, with the amylose content of standard solution as abscissa and corresponding absorbance be vertical coordinate draw straight chain
Starch-absorbance standard curve (see figure 1), the absorbance of sample solution is obtained with reference to amylose-absorbance standard curve
The amylose content of testing sample, detects the amylose content of Rice Samples.
In the present embodiment, rice sample has prepared 4 kinds altogether, is respectively labeled as sample 1 to sample 4, every kind of rice sample is carried out
3 parts of parallel assays, in this 4 kinds of rice samples being detected by the method for the present invention, amylose content is shown in Table 1.
Carry out the contrast test of the present invention, contrast test adopts the detection method described in background technology, to having a competition simultaneously
Test detection rice sample be the same sample of the present embodiment 1 to sample 4, in contrast test step 3 using and the present embodiment phase
4 kinds of same amylose standard powder, are shown in Fig. 2 by amylose-absorbance standard curve that testing result is drawn, and contrast test is examined
The amylose content testing result of the 4 kinds of samples surveyed is shown in Table 1.
Fig. 1 and Fig. 2 is visible for contrast, and the dependency of the standard curve that the present invention draws is higher, illustrates that the standard of the present invention is bent
Line reliability is higher, and the rice sample amylose content accuracy being calculated with the standard curve of the present invention is higher；Can by table 1
See, the standard deviation of 4 kinds of rice sample amylose contents of present invention detection is respectively less than the testing result of existing method, and this
The bright coefficient of variation is respectively less than the coefficient of variation of existing method, illustrates that the testing result discreteness of the present invention is little, and accuracy is high.
Claims (3)
1. the method for quick of rice grain amylose content is it is characterised in that include following steps:
Step 1, preparation of samples；
Rice Samples are obtained polished rice after shelling, fine grinding, polished rice is ground into all powderies by 80 mesh～100 mesh sieves
Thing, weighs powder 100mg scholar 0.5mg as testing sample；
Prepare 4 to 6 kinds of amylose standard powder, in each amylose standard powder, the mass content of amylose is different, claims respectively
Take each amylose standard powder 100mg scholar 0.5mg as standard sample；
Respectively testing sample and standard sample are put in different beakers；
The preparation of step 2, sample solution and standard solution；
Step 2.1, is sequentially added into 95% ethanol solution 1 ml in each beaker of step 1 and 1.0 mol/l sodium hydroxide are molten
Liquid 9.0 ml, is placed in heating 10s～20s in microwave environment, takes out and be cooled to room temperature, transfer in 100ml volumetric flask after shaking up,
Plus distilled water constant volume shaking up, respectively obtain sample dispersion liquid；
Step 2.2, respectively the sample dispersion liquid 10ml～50ml of removing step 2.1 be put in separatory funnel, into separatory funnel
Add 5ml～20ml petroleum ether, convolution shake 1min～5min, after collecting lower floor's aqueous phase, place into addition 5ml in separatory funnel
～20ml petroleum ether, convolution shake 1min, collects lower floor's aqueous phase, respectively obtains defat sample；
Step 2.3, during respectively defat sample 5.00 ml of removing step 2.2 is in the 100 ml volumetric flasks to volumetric flask successively plus
After entering distilled water 50ml, 1.0 mol/l acetic acid solution 1 ml and iodine reagent 2.0ml, add distilled water constant volume, shake up, stand 10
Min, is obtained sample solution and multiple standard solution respectively；
Step 3, the preparation of blank solution；
Prepare 0.09mol/l sodium hydroxide solution as blank sample, pipette blank sample 5.00 ml in 100 ml volumetric flasks
In, after sequentially adding distilled water 50ml, 1.0 mol/l acetic acid solution 1 ml and iodine reagent 2.0ml in volumetric flask, add distillation
Water constant volume, shakes up, and stands 10 min, obtains blank solution；
Step 4, using the absorbance of spectrophotometer difference determination sample solution, each standard solution at 720nm, spectrophotometric
Meter blank solution returns to zero；
Step 5, with the amylose content of standard solution as abscissa and corresponding absorbance for vertical coordinate draw amylose-
Absorbance standard curve, the absorbance of sample solution is obtained the straight of testing sample with reference to amylose-absorbance standard curve
Chain content of starch, thus detect the amylose content of Rice Samples.
2. rice grain amylose content according to claim 1 method for quick it is characterised in that: step 2.1 institute
In the microwave environment stated, the frequency of microwave is 2000mhz～3000mhz.
3. rice grain amylose content according to claim 1 and 2 method for quick it is characterised in that: in step 1
Amylose standard powder uniformly mixes system by the waxy rice flour of potato amylose and amylopectin mass content more than 99%
Become.
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Cited By (6)
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| CN108901826A (en) * | 2018-07-03 | 2018-11-30 | 安徽荃银高科种业股份有限公司 | A kind of method of fast accurate breeding High quality and diseases resistance rice varieties |
| CN108956910A (en) * | 2018-07-25 | 2018-12-07 | 江西省农业科学院水稻研究所 | A kind of screening technique of noodle rice kind |
| CN110261332A (en) * | 2019-05-30 | 2019-09-20 | 扬州大学 | A kind of method of simple grain rice measurement content of amylose in rice |
| CN110333329A (en) * | 2019-05-08 | 2019-10-15 | 重庆瑞钛科技有限公司 | The rapid detection method of amylopectin content in wine brewing grain |
| CN111122470A (en) * | 2019-12-27 | 2020-05-08 | 海南大学 | Method for detecting amylose content of single-grain wheat |
| CN111537460A (en) * | 2020-05-22 | 2020-08-14 | 四川绵竹剑南春酒厂有限公司 | Method for determining content of waxy sorghum starch |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108901826A (en) * | 2018-07-03 | 2018-11-30 | 安徽荃银高科种业股份有限公司 | A kind of method of fast accurate breeding High quality and diseases resistance rice varieties |
| CN108901826B (en) * | 2018-07-03 | 2020-03-31 | 安徽荃银高科种业股份有限公司 | Method for quickly and accurately breeding high-quality disease-resistant rice variety |
| CN108956910A (en) * | 2018-07-25 | 2018-12-07 | 江西省农业科学院水稻研究所 | A kind of screening technique of noodle rice kind |
| CN108956910B (en) * | 2018-07-25 | 2019-05-14 | 江西省农业科学院水稻研究所 | A kind of screening technique of noodle rice kind |
| CN110333329A (en) * | 2019-05-08 | 2019-10-15 | 重庆瑞钛科技有限公司 | The rapid detection method of amylopectin content in wine brewing grain |
| CN110261332A (en) * | 2019-05-30 | 2019-09-20 | 扬州大学 | A kind of method of simple grain rice measurement content of amylose in rice |
| CN111122470A (en) * | 2019-12-27 | 2020-05-08 | 海南大学 | Method for detecting amylose content of single-grain wheat |
| CN111537460A (en) * | 2020-05-22 | 2020-08-14 | 四川绵竹剑南春酒厂有限公司 | Method for determining content of waxy sorghum starch |
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