CN105548077A - Method for determining total starch content of rice by refractive index - Google Patents
Method for determining total starch content of rice by refractive index Download PDFInfo
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- CN105548077A CN105548077A CN201510889159.8A CN201510889159A CN105548077A CN 105548077 A CN105548077 A CN 105548077A CN 201510889159 A CN201510889159 A CN 201510889159A CN 105548077 A CN105548077 A CN 105548077A
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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
- G01N21/41—Refractivity; Phase-affecting properties, e.g. optical path length
Abstract
The invention discloses a method for determining the total starch content of rice by refractive index. The method comprises the steps of: taking a 0.65mol/L hydrochloric acid solution as an acidolysis solution, subjecting a 50ml standard sample solution, a to-be-determined sample solution and a blank solution that are prepared from the acidolysis solution to the following operations respectively: performing 100DEG C oil bath heating, adding 1mL of a 300g/L zinc sulfate solution and 1mL of a 150g/L potassium ferrocyanide solution, adding distilled water to a constant volume of 100mL, and performing filtering, thus obtaining a to-be-determined sample filtrate, a standard sample filtrate and a blank filtrate respectively; utilizing the standard sample filtrate and the blank filtrate to prepare a standard ratio solution; measuring the refractive index of the standard ratio solution and the to-be-determined sample filtrate so as to obtain the total starch content of the to-be-determined sample.
Description
Technical field
The invention belongs to field of food detection, relate to the mensuration of total starch content in rice, be specially a kind of method utilizing index determination rice total starch content.
Background technology
Starch is prevalent in all kinds of plant with polysaccharide form, especially the principal ingredient of the crop organ such as cereal kernel, stem tuber.Starch reaches 70%-80% in the ratio of rice kernel weight, and size and the rice edible quality of content of starch are closely related.Therefore, the mensuration of rice total starch content is for the attributional analysis important in inhibiting of rice.
Detect the conventional items analyzed as rice quality, the assay method of total starch content mainly contains enzyme hydrolysis method, acid-hydrolysis method and polarimetry etc.Measuring the method that total starch comparatively commonly uses in domestic and foreign literature is acid hydrolyzation method and enzyme hydrolyzation method, but these methods exist deficiency to a certain degree, as experiment complex operation, measure consuming time longer etc.Polarimetry is with the optical characteristics of starch for principle, but this method requires relatively strict in experimental implementation, and the process need testing staff measuring optical activity from the acidolysis of starch to filtrate strictly controls the time, otherwise content of starch result can be seriously on the low side.In order to meet related check mechanism and plantation, processing enterprise etc. to the requirement of rice starch determination techniques, improve detection efficiency, it is very necessary for formulating more easy, effective mensuration content of starch method.
The existing method of index determination content of starch that utilizes is mainly used in potato at present, concrete grammar is: take 20g mashed potato in small beaker, add 10mL60% perchloric acid solution, with 50mL distilled water, the whole sample liquids in beaker are transferred in 100mL volumetric flask again, in 100 DEG C of boiling water bath heating 10min, take out cooling, add zinc sulfate and each 1mL of potassium ferrocyanide, constant volume filters; Potato standard model is mixed with content of starch 8%, 10%, 12% ... 26%, obtain standard solution with said method equally; Read the index of refraction of standard solution and sample filtrate with refractometer, obtain corresponding equation with standard solution, the content of starch of potato samples can be calculated.But said method is not suitable for the mensuration of total starch content in rice.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of easy, effective method to measure total starch content in rice.The method utilizes index of refraction to measure total starch content in rice.
In order to solve the problems of the technologies described above, the invention provides a kind of method utilizing total starch content in index determination rice, comprising the steps:
1) take, concentration as the hydrochloric acid solution of 0.65mol/L be acid hydrolysis solution;
Accurately take 5.000g rice starch standard model and add the mixing of 50mL acid hydrolysis solution, obtain standard model liquid;
The Rice Samples to be measured accurately taking milled 80 mesh sieves of 5.000g adds the mixing of 50mL acid hydrolysis solution, obtains analyte sample fluid;
Using 50mL acid hydrolysis solution as blank solution;
2), analyte sample fluid, standard model liquid and blank solution are proceeded as follows respectively successively:
1., be positioned over after in container and heat 24 ~ 26min (being preferably 25min) in 100 DEG C of oil baths (glycerine), be then cooled to room temperature;
2., by the gains (comprising sample liquid, residue) in container all pour in 100mL volumetric flask, and wash (graded washing) container with 30mL distilled water, the cleansing solution of gained is incorporated in 100mL volumetric flask;
3., in 100mL volumetric flask add 1mL300g/L solution of zinc sulfate, shake up, then add 1mL150g/L potassium ferrocyanide solution, fully shake up; Adding distil water is settled to 100mL, filters, and discards 15mL just filtrate, continues to employ remaining filtrate;
Thus obtain testing sample filtrate, standard model filtrate, blank filtrate respectively;
3) 0.0mL, 1.5mL, 2.5mL, 3.5mL, 4.0mL, 4.5mL, 5.0mL standard model filtrate, is drawn, be corresponding in turn to again and add the blank filtrate of 5.0mL, 3.5mL, 2.5mL, 1.5mL, 1.0mL, 0.5mL, 0.0mL and mix, thus corresponding configuration becomes these 7 kinds of standard proportioning solution of 0 × C, 0.3 × C, 0.5 × C, 0.7 × C, 0.8 × C, 0.9 × C, 1.0 × C; Above-mentioned 7 kinds of standard proportioning solution are named as sequence number to be successively 0 ~ sequence number be 6 standard proportioning solution;
Remarks illustrate: above-mentioned standard proportioning solution refers to that content of starch is 0 × C, 0.3 × C, 0.5 × C, 0.7 × C, 0.8 × C, 0.9 × C, 1.0 × C standard proportioning solution (sequence number is 0,1,2,3,4,5,6); Wherein, C is the total starch content of rice starch standard model, represents with percentage (%);
4), the index of refraction (Abbe refractometer such as can be utilized to measure) of measurement standard proportioning solution and testing sample filtrate, thus the final total starch content obtaining testing sample.
As the improvement utilizing the method for total starch content in index determination rice of the present invention:
Described step 4) be:
The index of refraction taking sequence number as the standard proportioning solution of 0 as blank, to be 1 ~ sequence number by sequence number be 6 standard proportioning solution carry out following operation respectively:
Using blank for the index of refraction of standard proportioning solution (sequence number is 1 ~ sequence number is 6) deduction rear as the rear index of refraction of revision, to revise the content of starch drawing standard curve corresponding to rear index of refraction and this standard proportioning solution, thus obtain linear equation; The index of refraction of testing sample filtrate is substituted into above-mentioned linear equation, the total starch content of testing sample can be obtained, represent with percentage (%).
In the present invention, room temperature generally refers to 10 ~ 30 DEG C.
The principle of the inventive method is: starch acidolysis becomes monose, the refractive properties that monose is certain because this body structure has, and available index of refraction represents.Index of refraction size and content of starch are good linear relationship.
In the present invention, rice starch standard model is the rice starch extract that starch accounts for that dry matter content reaches more than 99.5%, and namely the dry matter content of this standard model can be considered its total starch content C.The dry matter content of rice starch standard model is measured by oven drying method, dries 2-3h at standard model being placed in 105 DEG C, calculates mass change before and after drying.
Rice starch standard model of the present invention, controls to guarantee that the total starch content of rice starch standard model and C value immobilize by moisture.
The index of refraction of standard proportioning solution of the present invention and testing sample filtrate, refers to the true index of refraction of this liquid.According to the temperature corrected value of refractometer, direct reading is corrected to the reading at 20 DEG C, this reading is only the true index of refraction of solution.The direct reading of same solution at different temperatures on refractometer is different, is to form because the scale of this instrument is scribed at standard temperature 20 DEG C, therefore needs the temperature corrected value utilizing instrument to indicate that direct reading is corrected to true index of refraction.Therefore, the true index of refraction of same solution can not change with temperature.
Of the present inventionly utilize total starch content method in index determination rice, there is following technical advantage:
1, the technology of the present invention is easy and simple to handle, and agents useful for same is few and more easily obtain, and instrument is Conventional compact instrument, meets the principle of economical rationality;
2, the technology of the present invention is consuming time relatively short, does not have the process of long enzymolysis or hydrolysis, can shorten detection time, thus improves detection efficiency;
3, the influence factor in the technology of the present invention is few, and substantially do not need the humiture considering environment, temperature factor is eliminated by refractometer automatically, and humidity factor does not significantly affect implementation process after the content of starch of confirmed standard sample.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in further detail.
Fig. 1 is the implementation step schematic diagram utilizing total starch content method in index determination rice.
Fig. 2 is the canonical plotting of experiment 1.
Fig. 3 is the canonical plotting of experiment 3.
In Fig. 2 and Fig. 3, horizontal ordinate is index of refraction after the revision of standard proportioning solution, and ordinate is the known content of starch of standard proportioning solution.
Embodiment
Embodiment 1, a kind of method utilizing total starch content in index determination rice, carry out following steps successively:
1) take, concentration as the hydrochloric acid solution of 0.65mol/L be acid hydrolysis solution;
Accurately take 5.000g rice starch standard model and add the mixing of 50mL acid hydrolysis solution, obtain standard model liquid;
The Rice Samples to be measured accurately taking milled 80 mesh sieves of 5.000g adds the mixing of 50mL acid hydrolysis solution, obtains analyte sample fluid;
Using 50mL acid hydrolysis solution as blank solution;
2), analyte sample fluid, standard model liquid and blank solution are proceeded as follows respectively successively:
1., be positioned over after in container and heat 25min in 100 DEG C of oil baths (glycerine), be then cooled to room temperature;
2., by the gains (comprising sample liquid, residue) in container all pour in 100mL volumetric flask, and wash (graded washing) container with 30mL distilled water, the cleansing solution of gained is incorporated in 100mL volumetric flask;
3., in volumetric flask add 1mL300g/L solution of zinc sulfate, shake up, then add 1mL150g/L potassium ferrocyanide solution, fully shake up; Adding distil water is settled to 100mL, filters, and discards 15mL just filtrate, continues to employ remaining filtrate;
Thus obtain testing sample filtrate, standard model filtrate, blank filtrate respectively;
3) 0.0mL, 1.5mL, 2.5mL, 3.5mL, 4.0mL, 4.5mL, 5.0mL standard model filtrate, is drawn, be corresponding in turn to again and add the blank filtrate of 5.0mL, 3.5mL, 2.5mL, 1.5mL, 1.0mL, 0.5mL, 0.0mL and mix, thus corresponding configuration becomes these 7 kinds of standard proportioning solution of 0 × C, 0.3 × C, 0.5 × C, 0.7 × C, 0.8 × C, 0.9 × C, 1.0 × C; Above-mentioned 7 kinds of standard proportioning solution are named as sequence number to be successively 0 ~ sequence number be 6 standard proportioning solution;
Remarks illustrate: above-mentioned standard proportioning solution refers to that content of starch is 0 × C, 0.3 × C, 0.5 × C, 0.7 × C, 0.8 × C, 0.9 × C, 1.0 × C standard proportioning solution (sequence number is 0,1,2,3,4,5,6); Wherein, C is the total starch content of rice starch standard model, represents with percentage (%).
4), the index of refraction of Abbe refractometer measurement standard proportioning solution and testing sample filtrate is utilized, thus the final total starch content obtaining testing sample.
Specific as follows:
The index of refraction taking sequence number as the standard proportioning solution of 0 as blank, to be 1 ~ sequence number by sequence number be 6 standard proportioning solution carry out following operation respectively:
Using blank for the index of refraction of standard proportioning solution deduction rear as the rear index of refraction of revision, to revise the content of starch drawing standard curve corresponding to rear index of refraction and this standard proportioning solution, thus obtain linear equation; The index of refraction of testing sample filtrate is substituted into above-mentioned linear equation, the total starch content of testing sample can be obtained, represent with percentage (%).
Contrast test is carried out in experiment 1, acid-hydrolysis method, polarimetry and the total starch content method in index determination rice (embodiment 1) that utilizes of the present invention with traditional.In this test, 5 rice testing samples are numbered DC1, DC2, DC3, DC4, DC5 respectively.
Method 1: acid-hydrolysis method, carries out according to the experimental procedure of the mensuration of starch " in the GB/T5009.9-2008 food ".
Method 2: polarimetry, carries out according to the experimental procedure of " the mensuration polarimetry of GB/T20378-2006 ative starch content of starch ".
Method 3: of the present inventionly utilize total starch content method in index determination rice (embodiment 1).
In the method 3 of this experiment, the total starch content of rice starch standard model is 94%, i.e. C=94%.Therefore, after the known content of starch of standard proportioning solution and corresponding revision, index of refraction is in table 1, and drawing standard curve (see Fig. 2, coefficient R
2be 0.9994), and calculating acquisition equation I is y=13353 × (a-a0)-2.29934, wherein, a is the index of refraction of testing sample, a0 is the index of refraction (a0=1.33560) of standard proportioning solution 0, and y is the content of starch (%) of testing sample.
The index of refraction a of testing sample DC1 ~ DC5 gained is respectively 1.34136,1.34095,1.34166,1.34186,1.34196, substitutes into equation I and show that content of starch is respectively 74.6%, 69.1%, 78.6%, 81.3%, 82.6%.
The content of starch of table 1, standard proportioning solution and index of refraction
Remarks illustrate: for " standard proportioning solution 1 ", the computing formula of known content of starch % is: 0.3 × C=0.3 × 94%=28.2%.In like manner, content of starch 0.5 × C, the 0.7 × C of standard proportioning solution 2 ~ 5,0.8 × C, 0.9 × C, 1.0 × C calculate 47.0%, 65.8%, 75.2%, 84.6%, 94.0% respectively.
Table 2, three kinds of methods measure the content of starch Comparative result of rice testing sample
In this experimental technique 3, the related coefficient of typical curve is 0.9994, is greater than 0.998, illustrates that the method is effectively feasible.Rice testing sample measures by method 1,2,3 respectively and obtains content of starch (see table 2), method 1 result of all samples and the relative differences of method 3 result, in 0.1% to 1.6%, meet the regulation " relative differences is no more than 10% " in standard GB/T5009.9-2008; Method 2 result of all samples and the absolute phase difference of method 3 result, in 0.1% to 1.5%, meet the regulation " absolute phase difference is no more than 2% " in standard GB/T20378-2006.Prove thus, of the present inventionly utilize total starch content method in index determination rice to be accurately.
Test 2, utilize total starch content method in index determination rice (embodiment 1) with of the present invention, testing staff A repeats sample DC1 in five test experience 1, and testing staff B repeats five times and detects DC1; Calculation equation is with experiment 1.
Five indexs of refraction of testing staff A, sample DC1 are respectively 1.34136,1.34138,1.34135,1.34131,1.34129, substitute into equation I and show that content of starch is respectively 74.6%, 74.9%, 74.5%, 73.9%, 73.7%.
Five indexs of refraction of testing staff B, sample DC1 are respectively 1.34126,1.34131,1.34132,1.34128,1.34133, substitute into equation I and show that content of starch is respectively 73.3%, 73.9%, 74.1%, 73.5%, 74.2%.
The relative standard deviation RSD of testing staff A, B five replication results is respectively 0.7% and 0.5%, illustrates that testing staff utilizes the repeatability of the inventive method mensuration same sample better.The RSD of ten reproducible results is 0.7%, meets " RSD is less than 2.0% " this requirement of generally acknowledging, illustrates that the repeatability of the inventive method, accuracy are all more satisfactory.
Test 3, utilize total starch content method in index determination rice (embodiment 1) with of the present invention, 5 testing samples again in test experience 1.In this experiment, rice starch standard model is due to the change of water percentage, and its total starch content is 90%, i.e. C=90%.
After the known content of starch of standard proportioning solution and corresponding revision, index of refraction is in table 3, and drawing standard curve (see Fig. 3, coefficient R
2be 0.9995), and calculating acquisition equation II is y=13432 × (a-a0)-2.65909, wherein, a is the index of refraction of testing sample, a0 is the index of refraction (a0=1.33565) of standard proportioning solution 0, and y is the content of starch (%) of testing sample.
The content of starch of table 3 standard proportioning solution and index of refraction
In this experiment, the index of refraction a of testing sample DC1 ~ DC5 gained is respectively 1.34143,1.34098,1.34171,1.34193,1.34198, substitutes into equation II and show that content of starch is respectively 75.0%, 68.9%, 78.7%, 81.7%, 82.4%.5 sample experiment 1 methods 2 (accuracy of the industry confessed testing result is high) record content of starch and are all less than 2.0% with the absolute phase difference recording content of starch with this experiment, meet the regulation of GB/T20378-2006 " absolute phase difference is no more than 2% ", namely when starch standard model content of starch changes, also can obtain effective equation, thus obtain result accurately.Demonstrate the inventive method thus and have good versatility.
Generally speaking, of the present inventionly total starch content method in index determination rice is utilized to be a kind of easy, effectively and accurately method.
Contrast experiment 1-1, make hydrochloric acid solution into perchloric acid solution, volumetric molar concentration is constant, and volume is also constant; All the other are equal to embodiment 1.5 testing samples in test experience 1 are carried out by the method.In this experiment, the total starch content of rice starch standard model is 94%, i.e. C=94%.
Contrast experiment 1-2, make the concentration of hydrochloric acid solution into 0.55mol/L by 0.65mol/L, volume is constant; All the other are equal to embodiment 1.5 testing samples in test experience 1 are carried out by the method.In this experiment, the total starch content of rice starch standard model is 94%, i.e. C=94%.
Contrast experiment 1-3, make the concentration of hydrochloric acid solution into 0.75mol/L by 0.65mol/L, volume is constant; All the other are equal to embodiment 1.5 testing samples in test experience 1 are carried out by the method.In this experiment, the total starch content of rice starch standard model is 94%, i.e. C=94%.
The starch result of contrast experiment 1-1,1-2,1-3 gained and with the comparing in table 4 of experiment 1 method 2 acquired results.
Related coefficient (the R of table 4, contrast experiment 1-1,1-2,1-3 gained
2), starch result and absolute phase difference
Remarks: absolute phase difference is contrast experiment 1-1,1-2,1-3 starch result and the difference of testing 1 method 2 starch result.
From table 4, the coefficient R of contrast experiment 1-1,1-2,1-3 standard curve
2be respectively 0.9969,0.9917,0.9932, all do not reach 0.998.And, in contrast experiment 1-1,1-2,1-3 all samples content of starch with experiment 1 method 2 gained content of starch, both absolute phase difference are all more than 2.0%, and namely above-mentioned contrast and experiment does not meet the regulation of GB/T20378-2006 " absolute phase difference is no more than 2% ".Prove thus, change hydrochloric acid solution in the present invention in contrast experiment and concentration is impracticable, therefore in the present invention, use 0.65mol/L hydrochloric acid solution to be reasonable and important.
Contrast experiment 2, make DEG C oil bath of 100 in embodiment 1 into 100 DEG C of boiling water baths, all the other are equal to embodiment 1.5 testing samples in test experience 1 are carried out by the method.In this experiment, the total starch content of rice starch standard model is 94%, i.e. C=94%.
In this experiment, the related coefficient of typical curve is 0.9972, does not reach 0.998.The content of starch of 5 samples is respectively 69.5%, 62.2%, 73.3%, 77.1%, 78.3%, is respectively 4.4%, 7.2%, 5.4%, 2.7%, 4.6%, all more than 2.0% with the absolute phase difference of experiment 1 method 2 acquired results.Therefore 100 DEG C of oil baths are used to be reasonable and important in the present invention.
Finally, it is also to be noted that what enumerate above is only several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be had.All distortion that those of ordinary skill in the art can directly derive from content disclosed by the invention or associate, all should think protection scope of the present invention.
Claims (2)
1. utilize the method for total starch content in index determination rice, it is characterized in that comprising the steps:
1) take, concentration as the hydrochloric acid solution of 0.65mol/L be acid hydrolysis solution;
Accurately take 5.000g rice starch standard model and add the mixing of 50mL acid hydrolysis solution, obtain standard model liquid;
The Rice Samples to be measured accurately taking milled 80 mesh sieves of 5.000g adds the mixing of 50mL acid hydrolysis solution, obtains analyte sample fluid;
Using 50mL acid hydrolysis solution as blank solution;
2), analyte sample fluid, standard model liquid and blank solution are proceeded as follows respectively successively:
1., be positioned over after in container and heat 24 ~ 26min in 100 DEG C of oil baths, be then cooled to room temperature;
2., by the gains in container all pour in 100mL volumetric flask, and with 30mL distilled water washing container, the cleansing solution of gained is incorporated in 100mL volumetric flask;
3., in 100mL volumetric flask add 1mL300g/L solution of zinc sulfate, shake up, then add 1mL150g/L potassium ferrocyanide solution, fully shake up; Adding distil water is settled to 100mL, filters, and discards 15mL just filtrate, continues to employ remaining filtrate;
Thus obtain testing sample filtrate, standard model filtrate, blank filtrate respectively;
3) 0.0mL, 1.5mL, 2.5mL, 3.5mL, 4.0mL, 4.5mL, 5.0mL standard model filtrate, is drawn, be corresponding in turn to again and add the blank filtrate of 5.0mL, 3.5mL, 2.5mL, 1.5mL, 1.0mL, 0.5mL, 0.0mL and mix, thus corresponding configuration becomes these 7 kinds of standard proportioning solution of 0 × C, 0.3 × C, 0.5 × C, 0.7 × C, 0.8 × C, 0.9 × C, 1.0 × C; Above-mentioned 7 kinds of standard proportioning solution are named as sequence number to be successively 0 ~ sequence number be 6 standard proportioning solution;
4), the index of refraction of measurement standard proportioning solution and testing sample filtrate, thus the final total starch content obtaining testing sample.
2. the method utilizing total starch content in index determination rice according to claim 1, is characterized in that:
Described step 4) be:
The index of refraction taking sequence number as the standard proportioning solution of 0 as blank, to be 1 ~ sequence number by sequence number be 6 standard proportioning solution carry out following operation respectively:
Using blank for the index of refraction of standard proportioning solution deduction rear as the rear index of refraction of revision, to revise the content of starch drawing standard curve corresponding to rear index of refraction and this standard proportioning solution, thus obtain linear equation; The index of refraction of testing sample filtrate is substituted into above-mentioned linear equation, the total starch content of testing sample can be obtained, represent with percentage (%).
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