CN106769925A - A kind of method of sugar in detection feed - Google Patents
A kind of method of sugar in detection feed Download PDFInfo
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- CN106769925A CN106769925A CN201611080763.7A CN201611080763A CN106769925A CN 106769925 A CN106769925 A CN 106769925A CN 201611080763 A CN201611080763 A CN 201611080763A CN 106769925 A CN106769925 A CN 106769925A
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
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Abstract
The invention discloses a kind of method for detecting sugar in feed, comprise the steps of, A, sample are chosen：B, sample roughing, C, Specification Curve of Increasing, D, sugar content are determined, E, result are calculated, the method of sugar is using the low advantage of the convenient, simple of spectrophotometry, equipment cost in present invention detection feed, the early stage treatment that is avoided that sulfate by ion chromatography again is complicated, equipment cost is high, unfavorable popularization shortcoming, and the method is equally applicable to the analysis of polysaccharide in tea tree its hetero-organization, and polyoses content detection and analysis in other approach natural products sources.The error caused because of detection liquid intrinsic colour is also avoided that simultaneously, while the simplification detection method in the case where correct content is drawn.
Description
Technical field
The present invention relates to a kind of detection method, specifically a kind of method for detecting sugar in feed.
Background technology
Feed, is the general name of the food of owner domesticated animal, compares narrowly general feeds and generally refers to agricultural
Or the food of animal husbandry domesticated animal.Feed（Feed）Including soybean, dregs of beans, corn, fish meal, amino acid, miscellaneous dregs of rice, additive,
More than ten feedstuffs of kind such as whey powder, grease, meat meal tankage, cereal, sugar grass.
Sugar in feed is one of most important intake nutrition in animal body growth course, therefore sugar in feed contains
Amount directly determines quality of the fodder, quality of the fodder difference may cause animal body it is slow-growing, it is malnutritive situations such as, make
Into the economic loss of raiser, thus in feed sugar content detection be particularly important, detection of the prior art to sugar
In there is certain defect, such as using monosaccharide component in chromatographic process (HPLC, GC) Direct Analysis polysaccharide, use indirect method
(phend-sulphuric acid, Anthrone-sulfuricacid method) analyzes polyoses content；The former is excessively complicated because pre-treatment works, and (such as HPLC needs sample
Product derivatization, GC need pre-conversion into effumability, heat-staple derivative), what especially analysis was obtained is only monosaccharide component
Or reduced sugar situation etc., it is unfavorable for being widely used.The latter is although fairly simple, but a kind of suitable right due to being difficult to find
According to product, add the degree of accuracy it is difficult to ensure that.
The content of the invention
It is an object of the invention to provide a kind of method for detecting sugar in feed, to solve to be proposed in above-mentioned background technology
Problem.
To achieve the above object, the present invention provides following technical scheme：
The method of sugar, comprises the steps of in a kind of detection feed,
A, sample are chosen：By feed sample to be measured, in the electric heating convection oven being placed under 90 DEG C of environment, height is used after drying 1.5h
Fast pulverizer is crushed, and after crossing 20 mesh sieves, 10g is taken out from the sample after filtering, adds 80% ethanol of 500ml, and 120 take the photograph
Water-bath refluxing extraction 1h under the conditions of family name's degree, while hot suction filtration；20 times of 80% hot ethanols of filter residue weight, sonic oscillation are added in filter residue
1min, suction filtration is repeated once, and is positioned over standby under 45 DEG C of low temperature drying environment；
B, sample roughing：The ready sample obtained in step A is weighed, 50 times of the 12 of example weight degrees Celsius of aseptic distillations are added
Water, at 60 DEG C, ultrasound assisted extraction 30min, by extract solution with Buchner funnel depressurize suction filtration after, filtrate decompression is concentrated into
100mL, uses 300mL95% ethanol precipitations, stirring while adding, is put in alcohol precipitation one day under 4 DEG C of cryogenic conditions, and 10000r/min is cold
Freeze centrifugation, precipitation uses absolute ethanol washing 3 times, is dried to constant weight in freeze-dryer, obtains final product refined Tea Flower polysaccharide system
Product, weigh, standby,
C, Specification Curve of Increasing：0.20g anthrones are weighed first, the 100mL75% concentrated sulfuric acids are added, and dissolving is mixed, and is made anthrone-sulphur
Acid solution, then weighs the glucose that 200mg dries to constant weight, and distilled water is settled to 100mL for mother liquor, draws 10mL mother liquor constant volumes
It is working solution to 100mL, shakes up；Accurate working solution 0,0.2,0.4,0.6,0.8,1.0mL of drawing uses water in 10mL tool plug test tubes
1mL is settled to, above-mentioned Anthrone Sulphuric acid solution 4mL is added, shaken up, after ice bath cooling, after boiling water bath boils 10min, then ice
Bath cooling 10min, absorbance is detected under 620nm wavelength；It is bent that Glucose standards sample concentration and correspondence absorbance draw standard
The linear relationship of line is y=9.5686x-0.0058, correlation r=0.9995, the range of linearity：0mg/mL~0.2mg/mL；
D, sugar content are determined：The sugar content in feed is accurately determined using the chromatography of ions, is weighed a certain amount of
Glucose, be configured to each reference substance concentration be 10ug/mL mixing reference substance, detected with chromatography of ions, claim
The sample 51.9mg for answering step B treatment to obtain is taken, adds 4mL, the trifluoroacetic acid of 2mol/L that nitrogen sealing is filled with after being dissolved,
120 DEG C of hydrolysis 6h, after hydrolysis terminates, centrifugation takes supernatant, at 40 DEG C, is dried up with Nitrogen evaporator, dissolves to blow with methyl alcohol and catches up with residual
Trifluoroacetic acid, be repeated 3 times, be finally settled to 50mL, cross 0.45um microporous barriers, carry out ion chromatography；
E, result are calculated.
As further scheme of the invention：The condition of work of the chromatography of ions is：Flow velocity：1.0mL/min, sample introduction
Amount 10uL, 30 DEG C of column temperature, ampere integrated detected, gold electrode, Dionex companies CarboPacPA10250mm × 3mm, mobile phase：
A：water；B：100mMNaOH；C：100mMNaOH/0.5MNaAc；Gradient elution：0~20min：70%A, 30%B；21~
25min：90%B → 80%B, 10%C → 20%C；25~35min：80%B → 50%B, 20%C → 50%C；36~
46min：70%A, 30%B；According to the retention time and peak area of monosaccharide solutions chromatography of ions figure after Tea Flower polysaccharide hydrolysis, meter
Calculate above-mentioned 11 monose gross mass account for refined Tea Flower polysaccharide preparation quality percentage, as Tea Flower polysaccharide contains
Amount c2 (%).
As further scheme of the invention：The specific method of the step E is：Sugar in spectrophotometry feed
Detectable concentration be multiplied by the content that conversion factor obtains sugar in final feed.
Compared with prior art, the beneficial effects of the invention are as follows：The method of sugar utilizes light splitting in present invention detection feed
Convenient, simple, the advantage that equipment cost is low of photometry detection, be avoided that the early stage treatment of sulfate by ion chromatography is complicated again,
Equipment cost is high, unfavorable popularization shortcoming, and the method is equally applicable to the analysis of polysaccharide in tea tree its hetero-organization, and other are on the way
Polyoses content detection and analysis in the natural products source of footpath.The error caused because of detection liquid intrinsic colour is also avoided that simultaneously, together
When in the case where correct content is drawn simplification detection method.
Specific embodiment
The technical scheme of this patent is described in more detail with reference to specific embodiment.
The method of sugar, comprises the steps of in a kind of detection feed,
A, sample are chosen：By feed sample to be measured, in the electric heating convection oven being placed under 90 DEG C of environment, height is used after drying 1.5h
Fast pulverizer is crushed, and after crossing 20 mesh sieves, 10g is taken out from the sample after filtering, adds 80% ethanol of 500ml, and 120 take the photograph
Water-bath refluxing extraction 1h under the conditions of family name's degree, while hot suction filtration；20 times of 80% hot ethanols of filter residue weight, sonic oscillation are added in filter residue
1min, suction filtration is repeated once, and is positioned over standby under 45 DEG C of low temperature drying environment；
B, sample roughing：The ready sample obtained in step A is weighed, 50 times of the 12 of example weight degrees Celsius of aseptic distillations are added
Water, at 60 DEG C, ultrasound assisted extraction 30min, by extract solution with Buchner funnel depressurize suction filtration after, filtrate decompression is concentrated into
100mL, uses 300mL95% ethanol precipitations, stirring while adding, is put in alcohol precipitation one day under 4 DEG C of cryogenic conditions, and 10000r/min is cold
Freeze centrifugation, precipitation uses absolute ethanol washing 3 times, is dried to constant weight in freeze-dryer, obtains final product refined Tea Flower polysaccharide system
Product, weigh, standby,
C, Specification Curve of Increasing：0.20g anthrones are weighed first, the 100mL75% concentrated sulfuric acids are added, and dissolving is mixed, and is made anthrone-sulphur
Acid solution, then weighs the glucose that 200mg dries to constant weight, and distilled water is settled to 100mL for mother liquor, draws 10mL mother liquor constant volumes
It is working solution to 100mL, shakes up；Accurate working solution 0,0.2,0.4,0.6,0.8,1.0mL of drawing uses water in 10mL tool plug test tubes
1mL is settled to, above-mentioned Anthrone Sulphuric acid solution 4mL is added, shaken up, after ice bath cooling, after boiling water bath boils 10min, then ice
Bath cooling 10min, absorbance is detected under 620nm wavelength；It is bent that Glucose standards sample concentration and correspondence absorbance draw standard
The linear relationship of line is y=9.5686x-0.0058, correlation r=0.9995, the range of linearity：0mg/mL~0.2mg/mL；
D, sugar content are determined：The sugar content in feed is accurately determined using the chromatography of ions, is weighed a certain amount of
Glucose, be configured to each reference substance concentration be 10ug/mL mixing reference substance, detected with chromatography of ions, claim
The sample 51.9mg for answering step B treatment to obtain is taken, adds 4mL, the trifluoroacetic acid of 2mol/L that nitrogen sealing is filled with after being dissolved,
120 DEG C of hydrolysis 6h, after hydrolysis terminates, centrifugation takes supernatant, at 40 DEG C, is dried up with Nitrogen evaporator, dissolves to blow with methyl alcohol and catches up with residual
Trifluoroacetic acid, be repeated 3 times, be finally settled to 50mL, cross 0.45um microporous barriers, carry out ion chromatography；
E, result are calculated.
The condition of work of the chromatography of ions is：Flow velocity：1.0mL/min, sample size 10uL, 30 DEG C of column temperature, ampere integration inspection
Survey, gold electrode, Dionex companies CarboPacPA10250mm × 3mm, mobile phase：A：water；B：100mMNaOH；C：
100mMNaOH/0.5MNaAc；Gradient elution：0~20min：70%A, 30%B；21~25min：90%B → 80%B, 10%C
→ 20%C；25~35min：80%B → 50%B, 20%C → 50%C；36~46min：70%A, 30%B；According to Tea Flower
The retention time and peak area of monosaccharide solutions chromatography of ions figure after polysaccharide hydrolysis, the gross mass for calculating above-mentioned 11 monose account for essence
The percentage of the quality of the Tea Flower polysaccharide preparation of system, as Tea Flower polyoses content c2 (%).
The specific method of step E is：The detectable concentration of sugar is multiplied by conversion factor and obtains in spectrophotometry feed
The content of sugar in final feed.
Operation principle of the invention is：The method of sugar utilizes spectrophotometry just in present invention detection feed
The prompt, advantage that simple, equipment cost is low, is avoided that the early stage treatment of sulfate by ion chromatography is complicated, equipment cost is high, unfavorable again
The shortcoming of popularization, the method is equally applicable to the analysis of polysaccharide in tea tree its hetero-organization, and in other approach natural products sources
Polyoses content is tested and analyzed.The error caused because of detection liquid intrinsic colour is also avoided that simultaneously, while correctly contain being drawn
Simplification detection method in the case of amount.
Claims (3)
1. it is a kind of detect feed in sugar method, it is characterised in that comprise the steps of,
A, sample are chosen：By feed sample to be measured, in the electric heating convection oven being placed under 90 DEG C of environment, height is used after drying 1.5h
Fast pulverizer is crushed, and after crossing 20 mesh sieves, 10g is taken out from the sample after filtering, adds 80% ethanol of 500ml, and 120 take the photograph
Water-bath refluxing extraction 1h under the conditions of family name's degree, while hot suction filtration；20 times of 80% hot ethanols of filter residue weight, sonic oscillation are added in filter residue
1min, suction filtration is repeated once, and is positioned over standby under 45 DEG C of low temperature drying environment；
B, sample roughing：The ready sample obtained in step A is weighed, 50 times of the 12 of example weight degrees Celsius of aseptic distillations are added
Water, at 60 DEG C, ultrasound assisted extraction 30min, by extract solution with Buchner funnel depressurize suction filtration after, filtrate decompression is concentrated into
100mL, uses 300mL95% ethanol precipitations, stirring while adding, is put in alcohol precipitation one day under 4 DEG C of cryogenic conditions, and 10000r/min is cold
Freeze centrifugation, precipitation uses absolute ethanol washing 3 times, is dried to constant weight in freeze-dryer, obtains final product refined Tea Flower polysaccharide system
Product, weigh, standby,
C, Specification Curve of Increasing：0.20g anthrones are weighed first, the 100mL75% concentrated sulfuric acids are added, and dissolving is mixed, and is made anthrone-sulphur
Acid solution, then weighs the glucose that 200mg dries to constant weight, and distilled water is settled to 100mL for mother liquor, draws 10mL mother liquor constant volumes
It is working solution to 100mL, shakes up；Accurate working solution 0,0.2,0.4,0.6,0.8,1.0mL of drawing uses water in 10mL tool plug test tubes
1mL is settled to, above-mentioned Anthrone Sulphuric acid solution 4mL is added, shaken up, after ice bath cooling, after boiling water bath boils 10min, then ice
Bath cooling 10min, absorbance is detected under 620nm wavelength；It is bent that Glucose standards sample concentration and correspondence absorbance draw standard
The linear relationship of line is y=9.5686x-0.0058, correlation r=0.9995, the range of linearity：0mg/mL~0.2mg/mL；
D, sugar content are determined：The sugar content in feed is accurately determined using the chromatography of ions, is weighed a certain amount of
Glucose, be configured to each reference substance concentration be 10ug/mL mixing reference substance, detected with chromatography of ions, claim
The sample 51.9mg for answering step B treatment to obtain is taken, adds 4mL, the trifluoroacetic acid of 2mol/L that nitrogen sealing is filled with after being dissolved,
120 DEG C of hydrolysis 6h, after hydrolysis terminates, centrifugation takes supernatant, at 40 DEG C, is dried up with Nitrogen evaporator, dissolves to blow with methyl alcohol and catches up with residual
Trifluoroacetic acid, be repeated 3 times, be finally settled to 50mL, cross 0.45um microporous barriers, carry out ion chromatography；
E, result are calculated.
2. it is according to claim 1 it is a kind of detect feed in sugar method, it is characterised in that the chromatography of ions
Condition of work is：Flow velocity：1.0mL/min, sample size 10uL, 30 DEG C of column temperature, ampere integrated detected, gold electrode, Dionex companies
CarboPacPA10250mm × 3mm, mobile phase：A：water；B：100mMNaOH；C：100mMNaOH/0.5MNaAc；Gradient is washed
It is de-：0~20min：70%A, 30%B；21~25min：90%B → 80%B, 10%C → 20%C；25~35min：80%B →
50%B, 20%C → 50%C；36~46min：70%A, 30%B；According to monosaccharide solutions chromatography of ions after Tea Flower polysaccharide hydrolysis
The retention time and peak area of figure, the gross mass for calculating above-mentioned 11 monose account for the quality of refined Tea Flower polysaccharide preparation
Percentage, as Tea Flower polyoses content c2 (%).
3. it is according to claim 1 it is a kind of detect feed in sugar method, it is characterised in that the step E's is specific
Method is：The detectable concentration of sugar is multiplied by conversion factor and obtains containing for sugar in final feed in spectrophotometry feed
Amount.
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| CN201611080763.7A CN106769925A (en) | 2016-11-30 | 2016-11-30 | A kind of method of sugar in detection feed |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103558166A (en) * | 2013-10-25 | 2014-02-05 | 贵州信邦制药股份有限公司 | Method for measuring content of polysaccharides in glossy privet fruit and astragalus membranaceus healthy energy-strengthening preparation |
| CN104730009A (en) * | 2015-02-27 | 2015-06-24 | 浙江经贸职业技术学院 | Method for measuring polysaccharide content in tea flower |
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2016
- 2016-11-30 CN CN201611080763.7A patent/CN106769925A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103558166A (en) * | 2013-10-25 | 2014-02-05 | 贵州信邦制药股份有限公司 | Method for measuring content of polysaccharides in glossy privet fruit and astragalus membranaceus healthy energy-strengthening preparation |
| CN104730009A (en) * | 2015-02-27 | 2015-06-24 | 浙江经贸职业技术学院 | Method for measuring polysaccharide content in tea flower |
Non-Patent Citations (1)
| Title |
|---|
| 张星海等: "茶树花多糖的修正系数蒽酮-硫酸检测新方法研究", 《茶叶科学》 * |
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Application publication date: 20170531 |