CN108226407A - A kind of method of quick detection rice digestion characteristics - Google Patents
A kind of method of quick detection rice digestion characteristics Download PDFInfo
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- CN108226407A CN108226407A CN201810002780.1A CN201810002780A CN108226407A CN 108226407 A CN108226407 A CN 108226407A CN 201810002780 A CN201810002780 A CN 201810002780A CN 108226407 A CN108226407 A CN 108226407A
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Abstract
The invention discloses a kind of method of quick detection rice digestion characteristics, this method is that the rice after being cooked is rapidly frozen and is immediately placed in freeze-drying instrument using liquid nitrogen to be lyophilized；Hand-ground is lyophilized polished rice and is sieved；External enzymatic liquefaction is carried out to the sample after sieving, in enzymolysis different time points sampling；It is quickly detected using blood glucose meter and and carries out data analysis.The present invention is adapted to the various rice of detection, and the digestion characteristics of rice are analyzed by simulating digestion process of the rice in human body.This have the advantage that easy to operate, quick and cost is relatively low, can rapid Estimation effectively be made to the digestion characteristics of different rice varieties, it is complicated for operation to overcome conventional rice digestion characteristics analysis method, the defects of time-consuming and cost is higher, quick analysis for rice digestion characteristics provides important technical support, also provides technical guarantee to cultivate the healthy paddy rice kind with slow digestion characteristics.
Description
Technical field
The present invention relates to a kind of methods of quick detection rice digestion characteristics, belong to food health field, more particularly to
The digestion characteristics detection and analysis method of rice.
Background technology
Rice is most important cereal crops in the world, and more than 60% population of China is using rice as staple food.Starch is rice
Main component, content be more than 90%.Starch is made of amylose and amylopectin, and the content and structure of two kinds of components are poor
It is different, it is the main reason for causing rice quality difference.Digestion characteristics of the rice in human body are current healthy food area researches
An important directions.Human body can enter the stage of a rapid digestion rice starch after edible rice, and intestinal absorption is big
Glucose is measured, blood sugar concentration is caused to steeply rise in a short time, pancreatic secretion insulin can be promoted, if instantaneous blood sugar concentration mistake
Height then can generate larger pressure to pancreas.In addition, for diabetes patient, the rice with normal stool characteristic can not be eaten
Rice.
According to the digestion characteristics of rice starch, the starch in rice can be divided into rapid digestion starch (RDS), disappeared at a slow speed
Change starch (SDS) and resistant starch (RS).The digestion rate of rice starch is reduced, can subtract its digestion in human body intestinal canal
Slowly, instantaneous blood sugar concentration is reduced, the pressure of each internal organs and human health level is improved so as to reduce hyperglycemia to human body.In addition,
The slow starch of digestion rate can be degraded into the substance beneficial to health after large intestine is entered by small intestine by coliform.
The digestion characteristics of rice measure the direct measure of generally use life rice flour or starch at present, since rice is in practical food
With when be to be entered in vivo in a manner of cooked rice and be digested, therefore, can not accurate response for the measure of raw rice flour
The digestion characteristics of cooked rice.In addition, rice digestion characteristics method for measuring method more commonly used at present is using import reagent
Box measures, but the determination step of kit is more demanding to experiment condition and measure of the glucose after treatments of the sample in terms of it is numerous
It is trivial and time-consuming, it is crucial that kit measurement cost is high, it is difficult to for the quick analysis of a large amount of samples.Therefore there is an urgent need for pass through
The existing method for measuring raw rice flour or starch is improved, a kind of digestion characteristics for cooked rice of invention measure and analysis side
Method.New method is not only required to rice in analogue body and digests process, but also need by simplifying the measure to glucose content
Cooked rice digestion characteristics accuracy and finding speed are improved, to be that the new rice variety with slow starch digestion characteristics is cultivated
Digestion with related rice based food, which measures, provides technical support.
Invention content
To overcome the shortcomings of at present in terms of rice digestion characteristics analysis, the present invention provides a kind of quick detection rice to disappear
Change the method for characteristic.
Technical solution is as follows used by this method：
A kind of method of quick detection rice digestion characteristics, this method are that the rice after being cooked is rapidly frozen using liquid nitrogen
And it is immediately placed in freeze-drying instrument and is lyophilized；Hand-ground is lyophilized polished rice and is sieved；Sample after sieving is carried out external
Enzymatic liquefaction, in enzymolysis different time points sampling；It is quickly detected using blood glucose meter and and carries out data analysis.
It is as follows：
(1) preparation of samples：It weighs polished rice sample to be placed in glass culture dish, according to mass ratio 1:1.8 ratio, which adds in, steams
Culture dish is put into steamer by distilled water, and boiling is taken out after 45 minutes.The culture dish of taking-up is placed under room temperature and is cooled down 30 minutes.
It pours into liquid nitrogen to culture dish, culture dish is placed in freeze-drying instrument after the grain of rice of quick freeze boiling, 12h is lyophilized.It takes out and freezes
The grain of rice is lyophilized to fine particle using mortar hand-ground in the dry grain of rice, is more than 1mm particles using 18 mesh screens and uses 100
Mesh screen is less than 0.15mm particles.The particle for grinding and being sieved is placed in valve bag and is preserved.
(2) sodium acetate buffer is configured：It adds in 11.8mL glacial acetic acid to 850mL deionized waters, adjustment pH value to 6.0；
Add in the CaCl of 4mL 1M2Solution (dissolving 1.109g CaCl2Into 10mL deionized waters), while add in 1mL 0.49M
MgCl2Solution (adds in 995.79mg MgCl2In 6H2O to 10mL deionized waters), add in 200mg NaN3；It is settled to
1000mL；It is placed in 4 DEG C of refrigerators and preserves.
(3) digestion buffer solution configuration：Measure 400mL sodium acetate buffers.Add in 5mg pancreatin (4U/mg) and 250 μ L starch
Transglucosidase (3.26U/ μ L).Pay attention to：It is now with the current, it is preheated 30 minutes before use, being put into 37 DEG C of water-baths.
(4) sample preheats：(each sample need to carry out repeating to test twice) is weighed in sample 150mg to 50mL centrifuge tubes,
It adds in magnetic rotor and adds in 5mL deionized waters, centrifuge tube is placed in 37 DEG C of magnetic agitation water-baths with 150 revs/min
Rotating speed preheats 3 minutes.
(5) external test tube digestion reaction：After the completion of step (4), 30 points will be preheated in 20mL steps (3) using pipettor
The digestion buffer solution of clock is rapidly joined in 50mL centrifuge tubes, starts timing in the moment of addition, two repetitions are because adding in buffer solution
Time it is different, need timing respectively.
(6) sampling and examination of glucose concentration：10,20,40,60,90 and 120 minutes after digestion reaction starts respectively
It is sampled, draws digestion reaction liquid 10uL with pipettor every time, quickly drop on the test paper of blood glucose meter, wait for 5 seconds readings simultaneously
Record concentration of glucose in digestive juice, data unit mmol/L.
(7) data analysis：The digestion amount c of sample different time points is analyzed firstt, do digestion amount curve.Secondly, using enzyme
Promote kinetics First-order equation c=c0e-kt(wherein c be real time reaction object concentration, c0For initial reactant concentration, k is enzymolysis
Reaction rate constant k values, t are the real time reaction time), digestion reaction rate constants k is calculated, does the slope of reaction rate constant k
Figure.
Sample required by step (1) must quickly be ground immediately after freeze-drying is taken out, be otherwise lyophilized sample can absorb water from
And influence experiment.It must be sieved after sample grinding, sample particle size be kept between 0.15mm-1mm, to ensure that sample will not
By particle size effect, ensure the repetition stability that sample measures.
Must immediately be adjusted after glacial acetic acid is added in required by step (2) increases PH to 6.0, and should preserve before use
Under the conditions of 4 DEG C.
Digestion buffer solution required by step (3) must be now with the current, and half an hour is preheated in advance before use.
The dosage that pancreatin 0.25mg and amylo-transglucosidase 12.5ul can be added according to every 150mg samples is matched, enzyme used
The product of Sigma companies is preferably selected, to ensure the accuracy measured.
Magnetic agitation water-bath must be used required by step (4), magnetic rotor needs to be always maintained at turning in digestion process
It is dynamic, to ensure the repeatability of the consistency of digestion process and measurement result.
20ml required by step (5), which digests buffer solution, disposably to be rapidly joined.Each sample adds in digestion buffer solution
Between time difference have to be larger than 30s, otherwise can not be detected in same time sampling.
Each independent digestion reaction required by step (6) reaches sampling detection rapidly after corresponding time point, adjusts in advance
Blood glucose meter is tried, sucks 10ul digestive juices immediately after blood glucose meter gets out detection to detect concentration of glucose.
Formula c is utilized in step (7)t=(cxmmol/L×10-3×180.16g/mol×25ml×10-3×0.9)/
(150mg×10-3(the c of) × 100%xFor different time points concentration of glucose) calculate each time point sample digestion amount (sample
Quality/Starting sample weight × 100% is digested)；For convenience of this experimental calculation, by enzyme kinetics Formula One into
Row variant：That is former formula c=c0e-kt；If initial reactant concentration c0≈ 1, variant are Ln (c)=- kt；Continuation variant is Ln (1-
ct)=- kt.First, digestion amount during 20 minutes early stages of analysis digestion.Secondly, different time points can be reflected by making the slope line of k
The variation tendency of digestion rate.K values are bigger, and whole digestion rate variation is gentler.In general, whole digestion rate variation is got over
Greatly, i.e., the kind that k values are smaller and 20 minutes digestion amounts are lower, more beneficial and health slower in the digestion rate of early period.
Advantages of the present invention：
The present invention has easy to operate, and cost is relatively low, digestion situation that can rapidly and efficiently in simulation rice body, so as to obtain
Obtain the digestion characteristics in rice body.The present invention provides effective technical support, Neng Gouxian for quick analysis rice digestion characteristics
Write the efficiency for improving and screening advantageous health rice.
Description of the drawings
Fig. 1 is 4 kinds of rice variety HURANG ARISO LUTA, EDAKKADAN 0-69-27,300 and E ZI 96 disappear
Change specificity analysis figure.A, treatments of the sample tracing analysis figure.B, treatments of the sample rate k value slope line analysis figures.
Fig. 2 is 4 kinds of japonica rice variety ROXANI, L 205, Zheng rice No. 5 and YR196 digestion characteristics analysis charts.A, treatments of the sample
Tracing analysis figure.B, treatments of the sample rate k value slope line analysis figures.
Specific embodiment
By following instance the invention is further described with definition and not limit.Reality in following embodiments
Proved recipe method is conventional method unless otherwise specified.The invention will be further described below in conjunction with the accompanying drawings.
Embodiment 1
Rice variety rice digestion characteristics are analyzed：
(1) preparation of samples：Selected from International Rice 4 rice variety (http://iric.irri.org/), including
HURANG ARISO LUTA(ID:IRIS_313-10966),EDAKKADAN0-69-27(ID:IRIS_313-10833),
SANBAILI(ID:) and EZI96 (ID IRIS_B264:IRIS_313-8859), it is X01, X02, X03 that above-mentioned material is numbered respectively
And X04.Paddy decladding after going out essence, weighs 1g polished rice samples and is placed in a diameter of 3 centimetres of small-size glass culture dish, press respectively
According to mass ratio 1:1.8 ratio adds in 1.8mL distilled water, culture dish is put into steamer, boiling is taken out after 45 minutes.It will take out
Culture dish be placed under room temperature and cool down 30 minutes.It pours into liquid nitrogen to culture dish, by culture dish after the grain of rice of quick freeze boiling
It is placed in freeze-drying instrument and 12h is lyophilized.The grain of rice has been lyophilized in taking-up, and the grain of rice is lyophilized to fine particle using mortar hand-ground, makes
It is less than the particle of 0.15mm with particle of 18 mesh screens more than 1mm and using 100 mesh screens.The particle that will be ground and be sieved
It is placed in valve bag and preserves.
(2) sodium acetate buffer is configured：It adds in 11.8mL glacial acetic acid to 850mL deionized waters, adjustment pH value to 6.0.
Add in the CaCl of 4mL 1M2Solution (dissolving 1.109g CaCl2Into 10mL deionized waters), while add in 1mL 0.49M
MgCl2Solution (adds in 995.79mg MgCl2In 6H2O to 10mL deionized waters), add in 200mg NaN3.It is settled to
1000mL.It is placed in 4 DEG C of refrigerators and preserves.
(3) digestion buffer solution configuration：Measure 400mL sodium acetate buffers.Add in 5mg pancreatin (4U/mg, Sigma) and 250
μ L amylo-transglucosidases (3.26U/ μ L, Sigma) preheat 30 minutes before use, being put into 37 DEG C of water-baths.
(4) sample preheats：Each number is accurately weighed respectively in three parts of sample 150mg to 50mL centrifuge tubes, is denoted as respectively
X01-1、X01-2、X01-3；X02-1、X02-2、X02-3；X03-1, X03-2, X03-3 and X04-1, X04-2, X04-3, respectively
It adds in magnetic rotor and adds in 5mL deionized waters, centrifuge tube is placed in 37 DEG C of magnetic agitation water-baths with 150 revs/min
Rotating speed preheats 3 minutes.
(5) external test tube digestion reaction：After the completion of step (4), 30 points will be preheated in 20mL steps (3) using pipettor
The digestion buffer solution of clock is quickly separately added into 50mL centrifuge tubes, is firstly added X01-1 and is started to be it from 0 timing, waits for 30s
X01-2 is added in afterwards and sets up another timer starts for it from 0 timing, and so on.
(6) sampling and examination of glucose concentration：10,20,40,60,90 and 120 minutes after digestion reaction starts respectively
It is sampled, each digestion reaction liquid 10ul is quickly dropped on the test paper of blood glucose meter, is waited for 5 seconds readings and is recorded in digestive juice
Concentration of glucose, data unit mmol/L.
(7) data analysis：According to above-mentioned 3 reproducible results, immediate two repetitions of selection, after calculating average value is used for
It is continuous to calculate；Utilize formula ct=(cxmmol/L×10-3×180.16g/mol×25ml×10-3×0.9)/(150mg×10-3)
× 100% (cxFor different time points concentration of glucose) calculate each time point sample digestion amount (sample digested quality/rise
Beginning example weight × 100%) and do using time (minute is abscissa), digestion amount is the digestion amount curve (Fig. 1 a) of ordinate；
If initial reactant concentration c0≈ 1 utilizes formula Ln (1-ct)=- kt (t is the real time reaction time), is calculated k values and makees k
It is worth slope figure (Fig. 1 b), acquires X01k=-0.008799479；X02k=-0.008069202；X03k=-0.007532601；
X04k=-0.006968963.According to interpretation of result, in this 4 samples, the whole digestion rate of X03 and X03 are slower.
Embodiment 2
Japonica rice variety rice digestion characteristics are analyzed：
(1) preparation of samples：Selected from International Rice 4 japonica rice variety (http://iric.irri.org/), including
ROXANI(ID:IRIS_313-8216),L205(IRIS_313-8192),ZD5(ID:) and YR196 (ID IRIS_B240:
IRIS_B055), it is G01, G02, G03 and G04 that above-mentioned material is numbered respectively.Paddy decladding after going out essence, weighs 1g polished rice respectively
Sample is placed in a diameter of 3 centimetres of small-size glass culture dish, according to mass ratio 1:1.8 ratio adds in 1.8mL distilled water, will
Culture dish is put into steamer, and boiling is taken out after 45 minutes.The culture dish of taking-up is placed under room temperature and is cooled down 30 minutes.Pour into liquid nitrogen
Culture dish is placed in freeze-drying instrument into culture dish, after the grain of rice of quick freeze boiling, 12h is lyophilized.Rice has been lyophilized in taking-up
Grain is lyophilized the grain of rice to fine particle using mortar hand-ground, using particle of 18 mesh screens more than 1mm and uses 100 mesh
It is sieved through particle of the filter less than 0.15mm.The particle for grinding and being sieved is placed in valve bag and is preserved.
(2) sodium acetate buffer is configured：It adds in 11.8mL glacial acetic acid to 850mL deionized waters, adjustment pH value to 6.0.
Add in the CaCl of 4mL 1M2Solution (dissolving 1.109gCaCl2Into 10mL deionized waters), while add in 1mL 0.49M MgCl2
Solution (adds in 995.79mg MgCl2In 6H2O to 10mL deionized waters), add in 200mg NaN3.It is settled to 1000mL.It puts
It is preserved in 4 DEG C of refrigerators.
(3) digestion buffer solution configuration：Measure 400mL sodium acetate buffers.Add in 5mg pancreatin (4U/mg, Sigma) and 250
μ L amylo-transglucosidases (3.26U/ μ L, Sigma) preheat 30 minutes before use, being put into 37 DEG C of water-baths.
(4) sample preheats：Each number is weighed respectively in three parts of sample 150mg to 50mL centrifuge tubes, is denoted as G01- respectively
1、G01-2、G01-3；G02-1、G02-2、G02-3；G03-1, G03-2, G03-3 and G04-1, G04-2, G04-3, are separately added into
Magnetic rotor simultaneously adds in 5mL deionized waters, and centrifuge tube is placed in 37 DEG C of magnetic agitation water-baths with 150 revs/min of rotating speed
Preheating 3 minutes.
(5) external test tube digestion reaction：After the completion of step (4), 30 points will be preheated in 20mL steps (3) using pipettor
The digestion buffer solution of clock is quickly separately added into 50mL centrifuge tubes, is firstly added G01-1 and is started to be it from 0 timing, waits for 30s
G01-2 is added in afterwards and sets up another timer starts for it from 0 timing, and so on.
(6) sampling and examination of glucose concentration：10,20,40,60,90,120 minutes after digestion reaction starts respectively
It is sampled, each digestion reaction liquid 10ul is quickly dropped on the test paper of blood glucose meter, is waited for 5 seconds readings and is recorded in digestive juice
Concentration of glucose, data unit mmol/L.
(7) data analysis：According to above-mentioned 3 reproducible results, immediate two repetitions of selection, after calculating average value is used for
It is continuous to calculate；Utilize formula ct=(cxmmol/L×10-3×180.16g/mol×25ml×10-3×0.9)/(150mg×10-3)
× 100% (cxFor different time points concentration of glucose) calculate each time point sample digestion amount (sample digested quality/rise
Beginning example weight × 100%) and do using time (minute is abscissa), digestion amount is the digestion amount curve (Fig. 2 a) of ordinate；
If initial reactant concentration c0≈ 1 utilizes formula Ln (1-ct)=- kt (t is the real time reaction time), is calculated k values and makees k
It is worth slope figure (Fig. 2 b), acquires G01k=-0.008473803；G02k=-0.007873653；G03k=-0.006054867；
G04k=-0.005820867.According to interpretation of result, in this 4 samples, the whole digestion rate of G01 and G02 are slower.
Claims (2)
- A kind of 1. method of quick detection rice digestion characteristics, it is characterised in that be rapidly frozen to the rice after cooked using liquid nitrogen And it is immediately placed in freeze-drying instrument and is lyophilized；Hand-ground is lyophilized polished rice and is sieved；Sample after sieving is carried out external Enzymatic liquefaction, in enzymolysis different time points sampling；It is quickly detected using blood glucose meter and and carries out data analysis.
- A kind of 2. method of quick detection rice digestion characteristics, which is characterized in that include the following steps：(1) preparation of samples：It weighs polished rice sample to be placed in glass culture dish, according to mass ratio 1:1.8 ratio adds in distilled water, Culture dish is put into steamer, is taken out after sample boiling, is cooled down 30 minutes under room temperature；It pours into liquid nitrogen to culture dish, it is rapid cold Culture dish is placed in vacuum freeze-drying 12h in freeze drier immediately after freezing the grain of rice of boiling；The freeze-drying grain of rice is taken out, uses mortar hand The dynamic grinding freeze-drying grain of rice is less than using particle of 18 mesh screens more than 1mm and to fine particle using 100 mesh screens The particle of 0.15mm；(2) sodium acetate buffer is configured：It adds in 11.8mL glacial acetic acid to 850mL deionized waters, adjustment pH value to 6.0；It adds in The CaCl of 4mL 1M2Solution, while add in 1mL 0.49M MgCl2Solution adds in 200mg NaN3；It is settled to 1000mL；(3) digestion buffer solution configuration：The prepared sodium acetate buffers of 400mL are measured, add in 5mg pancreatin (4U/mg) and 250 μ L Amylo-transglucosidase (3.26U/ μ L)；It is preheated 30 minutes before use, being put into 37 DEG C of water-baths；(4) sample preheats：It accurately weighs in sample 150mg to 50mL centrifuge tubes, add in magnetic rotor and adds in 5mL deionizations Centrifuge tube is placed in 37 DEG C of magnetic agitation water-baths and is preheated 3 minutes with 150 revs/min of rotating speed by water；(5) external test tube digestion reaction：After the completion of step (4), disappeared using 30 minutes warmed-up in pipettor aspiration step (3) Change buffer solution 20mL to rapidly join in 50mL centrifuge tubes, start timing, timing respectively between difference repeats in the moment of addition；(6) sampling and examination of glucose concentration：10,20,40,60,90 and 120 minutes after digestion reaction starts take respectively Sample is drawn digestion reaction liquid 10ul, is quickly dropped on the test paper of blood glucose meter every time, reads and to record glucose in digestive juice dense Degree, data unit mmol/L；(7) data analysis：The digestion amount c of sample different time points is analyzed firstt, do digestion amount curve.Secondly, it is anti-using enzymatic Answer dynamics First-order equation c=c0e-kt, wherein c be real time reaction object concentration, c0For initial reactant concentration, k is enzyme digestion reaction Rate constants k value, t are the real time reaction time, calculate digestion reaction rate constants k, do the slope figure of reaction rate constant k.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111733210A (en) * | 2020-06-15 | 2020-10-02 | 扬州大学 | Method for rapidly determining rice enzymolysis quantity in high flux manner |
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| CN102329853A (en) * | 2010-07-12 | 2012-01-25 | 中国农业机械化科学研究院 | Method for determining isolated soy protein digestibility in vitro |
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