CN102507479A - Simple method for measuring amylose content in rice - Google Patents
Simple method for measuring amylose content in rice Download PDFInfo
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- CN102507479A CN102507479A CN201110365254XA CN201110365254A CN102507479A CN 102507479 A CN102507479 A CN 102507479A CN 201110365254X A CN201110365254X A CN 201110365254XA CN 201110365254 A CN201110365254 A CN 201110365254A CN 102507479 A CN102507479 A CN 102507479A
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Abstract
A simple method for measuring amylose content in rice belongs to the technical field of methods for measuring the rice quality, and includes the following steps: firstly, weighing a rice standard sample and a to-be-tested sample; secondly, adding alcohol solutions in the rice standard sample and the to-be-tested sample respectively for dispersion treatment, adding and mixing sodium hydroxide solutions to the alcohol solutions after the dispersion treatment, and allowing the mixed solutions to stand at the room temperature; thirdly, performing constant volume operation for the sample solutions; fourthly, adding acetic acid, I2-KI solutions and distilled water in the solutions with constant volumes; fifthly, reading the absorbance value at the wave length of 620 nm through light splitting; and sixthly, drawing a standard curve as per the absorbance value of the standard sample and the given amylose content, and then calculating the amylose content of the to-be-tested sample corresponding to the standard curve. The method provided by the invention is simple to operate, low in cost and relatively quick.
Description
Technical field
The invention belongs to rice quality assay method technical field, be specifically related to a kind of method of simple measuring amylose content.
Background technology
Amylose is the glucose chain molecule thousands of unit head, few branch that is formed by connecting with α-1,4 glycosidic bond, and its molecular structure has very big influence to the gelatinization characteristic of starch, and amylose content is an important indicator weighing rice quality always.
The assay method of the amylose content of regulation in Ministry of Agriculture's rice matter bioassay standard (NY147-88); Concerning vast common breeding work person; Operational difficulty is arranged, be not easy to actual utilization, mainly show three aspects: one, error appears in the boiling water bath process of boiling that disappears easily: sample is placed in the volumetric flask of 100ml; Adding alkali lye disappears in boiling water bath and boils; The volumetric flask volume is relatively large, disappears once that it is on the low side to boil sample size, and between different batches because error appears in problems such as bath temperature easily; Two, boiling water bath disappear boil time-consuming: every batch of sample need disappear and boil 10min, if sample size is more,, need disappearing the time of boiling of labor, be difficult to realize the fast measuring of amylose content; Three, water wasting expense reagent: need twice constant volume in the operating process, each 100ml needs the distilled water of labor; Every volumetric flask need add 1ml acetic acid and 1.5ml Wagner's reagent; If sample size is many, reagent needs repeatedly preparation, expends reagent and has also increased the working time.For this reason, we simplify and improve the amylose content determination method on the basis of Ministry of Agriculture's standard method.
Summary of the invention
To the problem that prior art exists, the objective of the invention is to design the technical scheme of the method that a kind of simple measuring rice grain amylose content is provided, this method is easy and simple to handle, and is with low cost, quick relatively.
The method of described a kind of simple measuring rice grain amylose content is characterized in that comprising following processing step:
1) presses the standard determination method of amylose, take by weighing the rice standard model and the testing sample of amylose content distribution gradient;
2) add ethanolic solution in rice standard model and the testing sample respectively and carry out dispersion treatment, add the sodium hydroxide solution mixing after the dispersion treatment, at room temperature leave standstill then;
3) leave standstill completion after, to step 2) sample solution carry out constant volume;
4) get the constant volume solution that step 3) obtains, add acetic acid, I again
2-KI solution and distilled water, mixing leaves standstill;
5) leave standstill after, read absorbance with beam split in the 620nm wavelength;
6) absorbance of according to standard sample and known amylose content, the drawing standard curve, corresponding then typical curve calculates the amylose content of testing sample.
The method of described a kind of simple measuring rice grain amylose content, the testing sample 50mg that it is characterized in that getting the rice standard model 50mg of 100 mesh sieves in the described step 1) and cross 100 mesh sieves.
The method of described a kind of simple measuring rice grain amylose content; It is characterized in that described step 2) in to add concentration in rice standard model and the testing sample respectively be that 95% ethanol 0.5ml carries out dispersion treatment, adding concentration is the sodium hydroxide solution 4.5ml mixing of 1M after the dispersion treatment.
The method of described a kind of simple measuring rice grain amylose content is characterized in that described step 2) in leave standstill more than 16 hours under the room temperature.
The method of described a kind of simple measuring rice grain amylose content is characterized in that in the described step 3) sample solution being settled to 50ml.
The method of described a kind of simple measuring rice grain amylose content is characterized in that drawing 0.5ml constant volume liquid in the described step 4), adds 5ml distilled water mixing, and adding concentration then successively is acetic acid 0.1ml, the I of 1M
2-KI solution 0.2ml and distilled water 4.2ml.
The method of described a kind of simple measuring rice grain amylose content is characterized in that leaving standstill in the described step 4) 20 minutes.
The present invention is simple to operate; Reasonable in design; Compared with prior art; Beneficial effect of the present invention is following: the boiling water bath of 1) well having avoided stipulating in Ministry of Agriculture's standard method disappears the process of boiling owing to reasons such as bath temperature cause occurring easily between different batches the problem of error, especially is fit to the mensuration of batch samples; 2) sample is in leaving standstill the process of alkali digestion, and operating personnel can leave the mensuration of other meter matter indexs such as carrying out glue denseness, gelatinization point, more reasonable use with arrange the time, improve assay determination efficient; 3) simplified method has been saved reagent and water resource greatly than Ministry of Agriculture standard method, and it is original 1/10 that the use of reagent and distilled water is about, all more suitable to most of breeding units and individual.
The present invention and the standard method contrast of the existing Ministry of Agriculture, its (table 1) specific as follows:
The contrast of table 1 amylose content determination simplified method and Ministry of Agriculture's standard method
Description of drawings
Fig. 1 records the correlation analysis of 21 sample amylose contents for Ministry of Agriculture's standard method and the present invention;
The linear regression relation of 4 standard model absorbances that Fig. 2 records for Ministry of Agriculture's standard method and the present invention.
Embodiment
Below in conjunction with Figure of description and Test Example the present invention is further described, the method for a kind of simple measuring amylose content of the present invention is called for short simplified method.
Test Example 1
The sample (3.4%-27.0%) of choosing 21 amylose content distribution gradient is rough through removing, mill is smart and process ground rice after, behind 60 mesh sieves and 100 mesh sieves, place the sealing bag room temperature preservation subsequent use respectively.4 rice standard models are provided by Ministry of Agriculture's rice and quality of item supervision and inspection center, and content is respectively 1.5%, 10.4%, 16.2% and 26.5%, deposit with 21 samples room temperature together behind 60 orders and 100 mesh sieves excessively respectively.
Took by weighing the sample and the standard model 100mg of 60 mesh sieves respectively; After adding 1ml concentration respectively and be 95% ethanol dispersed sample, adding 9ml concentration is the NaOH solution of 1M, after shaking up gently; Put into boiling water bath and disappear and boil 10min, take out and be settled to 100ml after being cooled to room temperature; From volumetric flask, pipette out 5ml liquid to clean in addition 100ml volumetric flask, in volumetric flask, add Wagner's reagent (the 0.2g I of 50ml water, 1ml acetic acid, 1.5ml successively
2After adding 2g KI dissolved in distilled water, be settled to 100ml, existing with join at present); Be settled to 100ml; Leave standstill 20min after shaking up, spectrophotometer 620nm wavelength reads absorbance, and the absorbance of according to standard sample and known amylose content calculate the amylose content of sample.
Took by weighing the sample and the standard model 50mg of 100 mesh sieves respectively; Put into the 50ml volumetric flask respectively, add 0.5ml concentration respectively and be 95% ethanol dispersed sample after, adding 4.5ml concentration is the NaOH solution of 1M; After shaking up gently; Fill in the volumetric flask stopper, left standstill more than the 16h under the room temperature, be settled to 50ml; From volumetric flask, pipette out in 0.5ml liquid to the clean 20ml test tube, in test tube, add 5ml distilled water mixing, adding 0.1ml concentration then successively is the acetic acid of 1M, the Wagner's reagent of 0.2ml (0.2g I
2After adding 2g KI dissolved in distilled water, be settled to 100ml, existing with join at present), 4.2ml distilled water; Leave standstill 20min after shaking up; Spectrophotometer 620nm wavelength reads absorbance, the absorbance of according to standard sample and known amylose content, drawing standard curve; Corresponding then typical curve calculates the amylose content of testing sample.
The result shows, the amylose content that two kinds of methods record is high-positive correlation, and r2=0.9908 reaches the utmost point level of signifiance, and is as shown in Figure 1, and the amylose content maximum absolute error that two kinds of methods record is less than 1.2%.
Embodiment 1
Took by weighing the standard model 100mg of 60 mesh sieves, add the ethanol dispersed sample of 1ml 95% after, add 9ml (1M) NaOH solution, after shaking up gently, put into boiling water bath and disappear and boil 10min, take out and be settled to 100ml after being cooled to room temperature; From volumetric flask, pipette out 5ml liquid to clean in addition 100ml volumetric flask; The Wagner's reagent that in volumetric flask, adds 50ml water, 1ml acetic acid, 1.5ml successively; Be settled to 100ml, leave standstill 20min after shaking up, spectrophotometer 620nm wavelength reads absorbance.
Took by weighing the standard model 50mg of 100 mesh sieves, add 0.5ml concentration and be 95% ethanol dispersed sample after, adding 4.5ml concentration is the NaOH solution of 1M, after shaking up gently, has filled in the volumetric flask stopper, leaves standstill more than the 16h under the room temperature, is settled to 50ml; From volumetric flask, pipette out in 0.5ml liquid to the clean 20ml test tube, in test tube, add 5ml water mixing earlier, adding to 0.1ml concentration then successively is the acetic acid of 1M, the Wagner's reagent of 0.2ml (0.2g I
2After adding 2g KI dissolved in distilled water, be settled to 100ml, existing with join at present) and 4.2ml distilled water, leaving standstill 20min after shaking up, spectrophotometer 620nm wavelength reads absorbance.
The result shows, the absorbance of the standard model that two kinds of methods record becomes high-positive correlation, r
2=0.9997, as shown in Figure 2, and the absorbance that leaves standstill the above sample of 16h will be higher than to disappear in the boiling water bath and boils the absorbance of 10min.
Embodiment 2
Took by weighing the sample and the standard model 50mg of 100 mesh sieves, all samples is prepared 2 groups, add 0.5ml concentration and be 95% ethanol dispersed sample after; Adding 4.5ml concentration is the NaOH solution of 1M, after shaking up gently, has filled in the volumetric flask stopper; Leave standstill under the room temperature, middle every at a distance from the digestion situation of 0.5h observation sample in alkali lye, if be lower than 16h; Then the sample of part high amylose content still has the graininess sample in the volumetric flask bottom, leaves standstill more than the 16h (comprising 16h), and then sample is transparent fully; This moment, wherein one group volumetric flask was settled to 50ml, constant volume when another group continues to leave standstill to 120h; From the volumetric flask that constant volume is crossed, pipette out 0.5ml liquid to the clean tube of 20ml; The Wagner's reagent and the 4.2ml distilled water that in test tube, add 5ml distilled water, 0.1ml acetic acid, 0.2ml successively; Leave standstill 20min after shaking up, spectrophotometer 620nm wavelength reads absorbance.
The result shows; The measured value that leaves standstill the sample amylose content of 120h leaves standstill basically all having reduced of 16h; Have only 1 sample to increase to 3.8%, but on the whole, the measured value of amylose content change little (like table 2) by original 3.4%; Maximum difference is 1.1%, the experimental error requirement of the person's screening material that meets the breeding work.Can explain thus, measure for batch samples, if in 1 day, can not accomplish; The good stopper of sample plug is at room temperature placed; As long as supporting respective standard sample when measuring, promptly testing sample and standard model at room temperature standing time consistent, still can be used for mensuration after 2-3 days.
Table 2 simplified method leave standstill 16 hours with leave standstill the comparison that recorded amylose content in 5 days
Claims (7)
1. the method for a simple measuring rice grain amylose content is characterized in that comprising following processing step:
1) presses the standard determination method of amylose, take by weighing the rice standard model and the testing sample of amylose content distribution gradient;
2) add ethanolic solution in rice standard model and the testing sample respectively and carry out dispersion treatment, add the sodium hydroxide solution mixing after the dispersion treatment, at room temperature leave standstill then;
3) leave standstill completion after, to step 2) sample solution carry out constant volume;
4) get the constant volume solution that step 3) obtains, add acetic acid, I again
2-KI solution and distilled water, mixing leaves standstill;
5) leave standstill after, read absorbance with beam split in the 620nm wavelength;
6) absorbance of according to standard sample and known amylose content, the drawing standard curve, corresponding then typical curve calculates the amylose content of testing sample.
2. the method for a kind of simple measuring rice grain amylose content as claimed in claim 1, the testing sample 50mg that it is characterized in that getting the rice standard model 50mg of 100 mesh sieves in the described step 1) and cross 100 mesh sieves.
3. the method for a kind of simple measuring rice grain amylose content as claimed in claim 1; It is characterized in that described step 2) in to add concentration in rice standard model and the testing sample respectively be that 95% ethanol 0.5ml carries out dispersion treatment, adding concentration is the sodium hydroxide solution 4.5ml mixing of 1M after the dispersion treatment.
4. the method for a kind of simple measuring rice grain amylose content as claimed in claim 1 is characterized in that described step 2) in leave standstill more than 16 hours under the room temperature.
5. the method for a kind of simple measuring rice grain amylose content as claimed in claim 1 is characterized in that in the described step 3) sample solution being settled to 50ml.
6. the method for a kind of simple measuring rice grain amylose content as claimed in claim 1 is characterized in that drawing 0.5ml constant volume liquid in the described step 4), adds 5ml distilled water mixing, and adding concentration then successively is acetic acid 0.1ml, the I of 1M
2-KI solution 0.2ml and distilled water 4.2ml.
7. the method for a kind of simple measuring rice grain amylose content as claimed in claim 1 is characterized in that leaving standstill in the described step 4) 20 minutes.
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Cited By (11)
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|---|---|---|---|---|
| CN104006998A (en) * | 2014-06-11 | 2014-08-27 | 云南农业大学 | Method for separating different components of starch of rice and determining contents thereof |
| CN104215633A (en) * | 2014-09-11 | 2014-12-17 | 中国水稻研究所 | Paper chip detection method for rapidly determining content of amylose in paddy rice and application system adopted in paper chip detection system for rapidly determining content of amylose in paddy rice |
| CN104833671A (en) * | 2015-01-09 | 2015-08-12 | 中国水稻研究所 | Measurement method of absolute amylase content of rice |
| CN105548077A (en) * | 2015-12-07 | 2016-05-04 | 中国水稻研究所 | Method for determining total starch content of rice by refractive index |
| CN106053617A (en) * | 2016-04-14 | 2016-10-26 | 中国水稻研究所 | Gel permeation chromatographic method for simplified determination of relative molecular weight of amylose and amylopectin of paddy rice |
| CN106353268A (en) * | 2016-08-30 | 2017-01-25 | 广东省粮食科学研究所 | Rapid detection method of amylase content in rice |
| CN108918449A (en) * | 2018-07-13 | 2018-11-30 | 河南工业大学 | A kind of paddy xanthochromia degree detection method based on UV-VIS spectrophotometry |
| 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 |
| CN110927152A (en) * | 2019-12-05 | 2020-03-27 | 中国科学院西北高原生物研究所 | Method for rapidly detecting amylose and amylopectin in highland barley |
| CN113358598A (en) * | 2021-05-26 | 2021-09-07 | 四川农业大学 | Method for rapidly determining amylose content of wheat crops based on iodine dyeing method |
| CN115684055A (en) * | 2022-11-04 | 2023-02-03 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | Method for measuring amylose content in sweet potato starch |
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Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104006998A (en) * | 2014-06-11 | 2014-08-27 | 云南农业大学 | Method for separating different components of starch of rice and determining contents thereof |
| CN104215633A (en) * | 2014-09-11 | 2014-12-17 | 中国水稻研究所 | Paper chip detection method for rapidly determining content of amylose in paddy rice and application system adopted in paper chip detection system for rapidly determining content of amylose in paddy rice |
| CN104833671B (en) * | 2015-01-09 | 2017-07-21 | 中国水稻研究所 | A kind of assay method of the absolute amylose content of rice |
| CN104833671A (en) * | 2015-01-09 | 2015-08-12 | 中国水稻研究所 | Measurement method of absolute amylase content of rice |
| CN105548077A (en) * | 2015-12-07 | 2016-05-04 | 中国水稻研究所 | Method for determining total starch content of rice by refractive index |
| CN105548077B (en) * | 2015-12-07 | 2018-05-15 | 中国水稻研究所 | Utilize the method for total starch content in index determination rice |
| CN106053617A (en) * | 2016-04-14 | 2016-10-26 | 中国水稻研究所 | Gel permeation chromatographic method for simplified determination of relative molecular weight of amylose and amylopectin of paddy rice |
| CN106053617B (en) * | 2016-04-14 | 2018-06-15 | 中国水稻研究所 | A kind of simplified gel permeation chromatography for measuring amylose in rice and amylopectin relative molecular weight |
| CN106353268A (en) * | 2016-08-30 | 2017-01-25 | 广东省粮食科学研究所 | Rapid detection method of amylase content in rice |
| CN108918449A (en) * | 2018-07-13 | 2018-11-30 | 河南工业大学 | A kind of paddy xanthochromia degree detection method based on UV-VIS spectrophotometry |
| 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 |
| CN110927152A (en) * | 2019-12-05 | 2020-03-27 | 中国科学院西北高原生物研究所 | Method for rapidly detecting amylose and amylopectin in highland barley |
| CN113358598A (en) * | 2021-05-26 | 2021-09-07 | 四川农业大学 | Method for rapidly determining amylose content of wheat crops based on iodine dyeing method |
| CN115684055A (en) * | 2022-11-04 | 2023-02-03 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | Method for measuring amylose content in sweet potato starch |
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Application publication date: 20120620 |