CN105806793A - Simple and effective mulberry leaf polysaccharide detection method - Google Patents
Simple and effective mulberry leaf polysaccharide detection method Download PDFInfo
- Publication number
- CN105806793A CN105806793A CN201610218597.6A CN201610218597A CN105806793A CN 105806793 A CN105806793 A CN 105806793A CN 201610218597 A CN201610218597 A CN 201610218597A CN 105806793 A CN105806793 A CN 105806793A
- Authority
- CN
- China
- Prior art keywords
- folium mori
- water
- sugar
- mulberry leaf
- extracting solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
Landscapes
- Physics & Mathematics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines Containing Plant Substances (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention discloses a simple and effective mulberry leaf polysaccharide detection method. The method specifically comprises steps as follows: mulberry leaf water-soluble carbohydrates and mulberry leaf total carbohydrates are extracted, hydrolyzed and converted into reducing sugar, the light absorption values of a mulberry leaf water-soluble carbohydrate extracting solution and a mulberry leaf total carbohydrate extracting solution in the position of 540 nm are measured with a 3,5-DNS acid (dinitrosalicylic acid) colorimetric method, the mass of the mulberry leaf water-soluble carbohydrate extracting solution and the mulberry leaf total carbohydrate extracting solution in a measurement liquid are calculated on the basis of glucose according to the glucose standard curve, the content of mulberry leaf water-soluble carbohydrates and the content of mulberry leaf total carbohydrates (on the basis of reducing sugar) are calculated, and the content of mulberry leaf polysaccharides is calculated according to the formula that the quantity of the mulberry leaf polysaccharides (%) is equal to the product obtained by multiplying the difference between the quantity of mulberry leaf total carbohydrates (%) and mulberry leaf water-soluble carbohydrates (%) by 0.9. By means of the method, the problem that the color developing condition is difficult to control due to the fact that sulfuric acid releases much heat when meeting water is solved, besides, the measurement process is not affected by monosaccharide components, the operation is simple and safe, the result is stable and reliable, and the method is suitable for detecting the content of the polysaccharides in massive mulberry leaf samples.
Description
Technical field
The invention belongs to detection field, be specifically related to a kind of simple and effective polysaccharides of Folium Mori detection method.
Background technology
Folium Mori are the leaf of Moraceae (Moraceae) Morus (Morus L.) Mulberry kind (Morus alba L.) plant, containing several functions compositions such as protein, inorganic salt, vitamin, polysaccharide, polyphenol, flavone, alkaloids, Ministry of Public Health is classified as medicine-food two-purpose type Chinese herbal medicine.
Modern study pharmacology shows, polysaccharides of Folium Mori, as one of Folium Mori active component, has significant hypoglycemic activity.At present, polysaccharides of Folium Mori detection method of content is mainly Anthrone-sulfuricacid method and phend-sulphuric acid.Anthrone-sulfuricacid method almost can measure all of carbohydrate, and phend-sulphuric acid is mainly used in the mensuration of methylated sugar, pentose and polysaccharide.When measuring polysaccharides of Folium Mori by both approaches, actually get is total sugar content, therefore accurately to detect polysaccharides of Folium Mori content and the most isolated and purified must obtain the polysaccharides of Folium Mori that purity is higher, or the total sugar result obtained is deducted reducing sugar result (reducing sugar result need to be measured by other method).But the polysaccharides of Folium Mori complex steps that isolated and purified acquisition purity is higher, poor reproducibility；And Anthrone-sulfuricacid method measures total sugar result and also can be affected by the protein that tryptophane is higher, less stable；Phend-sulphuric acid experiment condition requires strict (requirement such as Pyrogentisinic Acid is the most high), is limited by multiple condition during mensuration, and repeatability is poor.Therefore, existing detection method all can not reflect the content of polysaccharide in Folium Mori simply, accurately and efficiently.Therefore, a kind of method detecting polysaccharides of Folium Mori simple, accurate, quick of research and development is badly in need of in this field.
Summary of the invention
In view of this, it is an object of the invention to provide the detection method of a kind of polysaccharides of Folium Mori, simple to operate, safety, result is stable, reliable, is suitable for detecting a large amount of Folium Mori sample polyoses contents.
For achieving the above object, the present invention utilizes saccharide monosaccharide, oligosaccharide and polysaccharide three major types, and under certain condition, oligosaccharide and polysaccharide can be hydrolyzed to the monosaccharide with reproducibility.And DNS reagent (3,5-dinitrosalicylic acid solution) it is reduced into henna amino-compound (3-amino-5-NITROSALICYLIC ACID) after reducing sugar solution altogether heat, this compound has absorption maximum at 540nm, and within the specific limits, the amount of reducing sugar is linear with light absorption value.Therefore the present invention is first with total sugar and the content of water-soluble sugar in DNS colorimetric method for determining Folium Mori, recycling formula: polysaccharides of Folium Mori (%)=[Folium Mori total sugar (%)-Folium Mori water-soluble sugar (%)] × 0.9, is calculated the content of polysaccharides of Folium Mori.To this end, provide following technical scheme:
A kind of detection method of simple effectively polysaccharides of Folium Mori, extract Folium Mori water-soluble sugar and Folium Mori total sugar respectively, then hydrolysis becomes reducing sugar, obtain Folium Mori water-soluble sugar extracting solution and Folium Mori total sugar extracting solution respectively, recycling 3, 5-dinitrosalicylic Acid Colorimetry measures Folium Mori water-soluble sugar extracting solution and Folium Mori total sugar extracting solution light absorption value at 540nm, then calculate Folium Mori water-soluble sugar extracting solution and Folium Mori total sugar extracting solution in mensuration liquid respectively according to glucose standard curve and be equivalent to the quality of glucose, it is calculated as follows Folium Mori water-soluble sugar and the content of Folium Mori total sugar again:
In formula: m2Look into water-soluble sugar in curve gained mensuration liquid and be equivalent to glucose quality, mg；
m3Look into total sugar in curve gained mensuration liquid and be equivalent to glucose quality, mg；
V0Mensuration liquid amasss, mL；
V2Water-soluble sugar extracting liquid volume, mL；
V3Total sugar extracting liquid volume, mL；
N2Water-soluble sugar extracting solution extension rate；
N3Total sugar extracting solution extension rate；
M Folium Mori sample quality, g.
And according to equation below calculating polysaccharides of Folium Mori:
Polysaccharides of Folium Mori (%)=[Folium Mori total sugar (%)-Folium Mori water-soluble sugar (%)] × 0.9.
In above-mentioned formula, total sugar refers to the summation (except starch, cellulose etc.) of monosaccharide in Folium Mori, oligosaccharide and polysaccharide, water-soluble sugar refers to the summation of monosaccharide in Folium Mori, oligosaccharide, total sugar and water-soluble sugar all content with hydrolysis as reducing sugar calculates, and 0.9 is the coefficient that reducing sugar (with glucose meter) is scaled polysaccharides of Folium Mori.
In the present invention, described 3, the compound method of 5-dinitrosalicylic acid solution is as follows: the first preparation aqueous solution that temperature is 40～45 DEG C containing sodium potassium tartrate tetrahydrate, is subsequently adding NaOH solution, sequentially adds 3,5-dinitrosalicylic acid, phenol and sodium sulfite, last constant volume makes the final concentration of 185g/L of sodium potassium tartrate tetrahydrate, the final concentration of 0.524mol/L of NaOH, 3, the final concentration of 6.3g/L of 5-dinitrosalicylic acid, the final concentration of 5.0g/L of phenol and the final concentration of 5.0g/L of sodium sulfite.
In the present invention, the extracting method of described Folium Mori water-soluble sugar extracting solution is as follows: takes new fresh mulberry leaf and dries to constant weight, pulverize, be subsequently adding distilled water water-bath and extract, be designated as extracting solution A, sequentially add acetic acid zinc solution and potassium ferrocyanide solution, centrifugal collection supernatant, obtains Folium Mori reducing sugar extracting solution, then adds hydrochloric acid in Folium Mori reducing sugar extracting solution and is hydrolyzed, regulation pH to 7.5-8.0, obtains Folium Mori water-soluble sugar extracting solution.Above-mentioned being centrifuged is not less than 6000rpm in principle, centrifugal 10min, and the supernatant collected after two times centrifugal is often no less than 60mL, and a part is used for the mensuration of reducing sugar, and another part converts for sequential hydrolysis, prepares water-soluble sugar.
The preparation method of described extracting solution A is as follows: taking new fresh mulberry leaf is 50 DEG C in temperature and dries to constant weight, and pulverizing, be subsequently adding the distilled water of 60 times that are equivalent to Folium Mori weight, 50 DEG C of water-baths are extracted 20min water-baths, obtained extracting solution A.
The addition of hydrochloric acid described in said hydrolyzed presses concentrated hydrochloric acid and Folium Mori reducing sugar extracting liquid volume than for 3:50；Described hydrolysis is hydrolysis 15min under 70 DEG C of water-baths.
In the present invention, the extracting method of described Folium Mori total sugar extracting solution is as follows: takes new fresh mulberry leaf and dries to constant weight, pulverize, it is subsequently adding distilled water fully to dissolve, after making total sugar complete hydrolysis with hydrochloric acid, is sequentially added into acetic acid zinc solution and potassium ferrocyanide solution, regulation pH to 7.5-8.0, filter, collect filtrate, obtain Folium Mori total sugar extracting solution.The present invention adds distilled water when fully dissolving powder of Folium Mori, first with a small amount of water by powder of Folium Mori furnishing pasty state, add the distilled water of more amount, mixing, its objective is to make that powder of Folium Mori is fully dispersed in water to be opened, beneficially Folium Mori total sugar and the extraction of Folium Mori water-soluble sugar.
Described hydrochloric acid addition is 0.5:6 (g/mL) by Folium Mori and concentrated hydrochloric acid w/v, described being hydrolyzed under boiling water bath hydrolyzes 30min, filter the most handy monolayer nylon cloth to filter, mainly due to this extracting solution contains more mucopolysaccharide, extremely complex with filter paper filtering, get rid of the filter paper constituent (carbohydrate) interference to reducing sugar test simultaneously.
The condition of described drying is 50 DEG C；The weight of described addition water is equivalent to 60 times of Folium Mori weight；In described acetic acid zinc solution, acetic acid zinc concentration is 219g/L；In described potassium ferrocyanide solution, the concentration of potassium ferrocyanide is 106g/L.Add acetic acid zinc solution and potassium ferrocyanide solution to add preferably along volumetric flask wall, so can reduce the generation of bubble, facilitate constant volume.
Most preferably, the regression equation of described glucose standard curve is y=1.5175x-0.0578 or y=1.5802x-0.063.
During said extracted, reducing sugar extracting solution the most handy 50mL volumetric flask measures, and main purpose is accurately to control the volume that reducing sugar extracting solution carries out converting, and reduces experimental error.
In the present invention, the concentrated hydrochloric acid of indication refers to the mass fraction hydrochloric acid more than 37%, oligosaccharide, polysaccharide can be hydrolyzed under concentrated hydrochloric acid effect the monosaccharide (because total sugar, water-soluble sugar, polysaccharide are all to calculate with reducing sugar) with reproducibility, and the addition of hydrochloric acid, bath temperature, water bath time be total sugar, water-soluble sugar hydrolysis be the key point of reductive monosaccharide, the condition of hydrolysis is too high, too low all can cause last measured value inaccurate.
The beneficial effects of the present invention is: the present invention utilizes total sugar and the content of water-soluble sugar in DNS colorimetric method for determining Folium Mori, recycling formula: polysaccharides of Folium Mori (%)=[Folium Mori total sugar (%)-Folium Mori water-soluble sugar (%)] × 0.9, is calculated the content of polysaccharides of Folium Mori；Relative to traditional Anthrone-sulfuricacid method and phend-sulphuric acid, DNS colorimetry avoids because sulphuric acid chance water very exothermic makes the unmanageable problem of color condition, the process that simultaneously measures is not disturbed by monosaccharide component, measurement result is closer to actual value, and total sugar, water-soluble sugar extraction process is simple, easily operation, therefore the present invention is suitable for the detection of a large amount of Folium Mori sample polyoses content, and then filter out the advantage Folium Mori kind that polyoses content is high.
Accompanying drawing explanation
In order to make the purpose of the present invention, technical scheme and beneficial effect clearer, the present invention provides drawings described below:
Fig. 1 is embodiment 1 glucose standard curve.
Fig. 2 is embodiment 2 glucose standard curve.
Detailed description of the invention
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail.The experimental technique of unreceipted actual conditions in embodiment, generally according to normal condition or according to the condition proposed by manufacturer.
In the present invention, experiment material is fresh Folium Mori, is taken from Southwest University mulberry field, dries to constant weight, pulverizes, cross 100 mesh sieves, save backup under-20 DEG C of low temperature environments at 50 DEG C.
The reagent used is as follows:
Acetic acid zinc solution is prepared: weighs 21.9g zinc acetate, adds 3mL glacial acetic acid, be dissolved in water and be diluted to 100mL, makes acetic acid zinc solution, final concentration of 219g/L.
Potassium ferrocyanide solution is prepared: weighs 10.6g potassium ferrocyanide, is dissolved in water and is diluted to 100mL, makes potassium ferrocyanide solution, final concentration of 106g/L.
Wagner's reagent is prepared: weighs 5g iodine, 10g potassium iodide, is dissolved in 100mL distilled water, makes Wagner's reagent.
DNS preparation of reagents: the preparation 500mL hydrothermal solution containing 185g sodium potassium tartrate tetrahydrate, being sequentially added into 262mL concentration wherein is the NaOH solution of 2mol/L, 6.3g 3,5-dinitrosalicylic acid, 5.0g phenol and 5.0g sodium sulfite, stirring and dissolving, transfer to after cooling, in 1000mL volumetric flask, be settled to scale with distilled water, be stored in brown bottle standby, this reagent is DNS reagent, uses after placing 7-10 days under room temperature (18～25 DEG C).
Embodiment 1
The detection method of a kind of polysaccharides of Folium Mori, comprises the following steps:
(1) Folium Mori reducing sugar and the extraction of water-soluble sugar
A, weigh powder of Folium Mori 1.00g in 250mL conical flask, add 60mL distilled water (first with a small amount of water by powder of Folium Mori furnishing pasty state), stir, 50 DEG C of water-bath 20min, be designated as extracting solution A；
B, step a gained extracting solution A (containing residue) is transferred in 100mL volumetric flask successively, by distilled water flushing conical flask 2-3 time, flushing liquor is transferred in volumetric flask in the lump, in volumetric flask, it is sequentially added into acetic acid zinc solution that 3mL concentration is 219g/L again and 3mL concentration is the potassium ferrocyanide solution of 106g/L, and with distilled water constant volume to 100mL, it is designated as extracting solution B；
C, use monolayer nylon cloth filtration step b gained extracting solution B, in filtrate collection to 50mL centrifuge tube (in view of extracting liquid volume and subsequent dose, the extracting solution in each volumetric flask need to be collected with 2 50mL centrifuge tubes)；Then centrifugal 10min under the conditions of 10000rpm, collects supernatant；Supernatant being centrifuged under the conditions of 10000rpm 10min again, collects supernatant, this supernatant is Folium Mori reducing sugar extracting solution；
D, measure 50mL step c gained Folium Mori reducing sugar extracting solution with 50mL volumetric flask, it is then transferred in 100mL volumetric flask, 3mL concentrated hydrochloric acid is added in volumetric flask, shake up, cool down rapidly with circulating water after 70 DEG C of water-bath 15min, it is subsequently poured in 250mL conical flask, regulation pH to 7.5-8.0, in the 100mL volumetric flask transferred to, by distilled water flushing conical flask 2-3 time, flushing liquor is transferred in the lump in corresponding volumetric flask, and constant volume is to 100mL, it is then transferred in 50mL centrifuge tube, is designated as Folium Mori water-soluble sugar liquid to be measured, be stored in 4 DEG C of refrigerators standby；Remaining Folium Mori reducing sugar extracting solution is stored in 4 DEG C of refrigerators, for subsequent measurements Folium Mori content of reducing sugar, is designated as Folium Mori reducing sugar liquid to be measured；
The most each sample parallel extracts three parts, obtains three parts of reducing sugar liquid to be measured and three parts of water-soluble sugar liquid to be measured, and last measurement result takes its arithmetic mean of instantaneous value.
(2) extraction of Folium Mori total sugar
A, weigh powder of Folium Mori 0.50g in 150mL tool plug triangular flask in, add 20mL distilled water (first with a small amount of water by powder of Folium Mori furnishing pasty state), stir；
B, addition 6mL concentrated hydrochloric acid, boiling water bath 30min hydrolysis in above-mentioned triangular flask；
C, 1-2 drip hydrolyzed solution on white plaque, add 1 Wagner's reagent and check whether total sugar hydrolyzes completely, if not presenting blueness, then prove to have hydrolyzed complete；
D, in above-mentioned triangular flask, add 30mL distilled water, add acetic acid zinc solution that 2mL concentration is 219g/L after shaking up and 2mL concentration is 106g/L potassium ferrocyanide solution, regulate pH to 7.5-8.0 with pH meter；
E, the liquid (containing residue) in triangular flask is transferred in 100mL volumetric flask, with distilled water flushing triangular flask 2-3 time, flushing liquor is transferred in the lump in above-mentioned volumetric flask, and constant volume is to 100mL, with monolayer nylon cloth, the extracting solution in volumetric flask is filled in 50mL centrifuge tube, being stored in 4 DEG C of refrigerators standby, this extracting solution is Folium Mori total sugar extracting solution, is designated as Folium Mori total sugar liquid to be measured.
Each sample parallel extracts three parts, obtains three parts of total sugar liquid to be measured, and last measurement result takes its arithmetic mean of instantaneous value.
(3) DNS colorimetric method for determining Folium Mori reducing sugar, water-soluble sugar, total sugar content
A, taking step (1) and (2) gained Folium Mori reducing sugar liquid to be measured, water-soluble sugar liquid to be measured, total sugar liquid to be measured in 10mL centrifuge tube respectively, 10000rpm is centrifuged 10min；
B, take each liquid 0.4mL to be measured in 10mL color comparison tube respectively, add 0.4mL distilled water and 0.6mL DNS reagent, shake up, boiling water bath 5min, it is cooled to room temperature (18～25 DEG C) with cold water immediately after taking-up, then to often pipe addition distilled water to scale (10mL), shakes up, replace liquid to be measured to do blank by same procedure operation with 0.4mL distilled water, at 540nm wavelength, measure each pipe absorbance；Each sample liquid to be measured parallel assay twice, averages.
C, drafting glucose standard curve, first preparation Glucose standards solution: accurately weigh glucose 103.6mg (drying to constant weight in advance) at 80 DEG C, after adding the dissolving of a small amount of distilled water, it is transferred in 100mL volumetric flask, it is settled to 100mL with distilled water, being stored in 4 DEG C of refrigerators standby after mixing, concentration is 1.036mg/mL；Then 9 10mL color comparison tubes are taken, it is separately added into 0.00mL, 0.08mL, 0.16mL, 0.24mL, 0.32mL, 0.40mL, 0.48mL, 0.56mL, 0.64mL glucose solution (add 0.00mL glucose solution and do blank), dilute is to 0.80mL, add 0.60mL DNS reagent, shake up, boiling water bath 5min, it is cooled to room temperature with cold water immediately after taking-up, again to often pipe addition distilled water to scale (10mL), shake up, at 540nm wavelength, measure each pipe absorbance.With glucose content (mg) as abscissa, absorbance is vertical coordinate, draws standard curve, and obtaining regression equation is y=1.5175x-0.0578, R2=0.9989 (such as Fig. 1).
D, bring sample absorbance value into standard curve y=1.5175x-0.0578, obtain each mensuration liquid and be equivalent to the milligram number of glucose, Folium Mori reducing sugar, Folium Mori water-soluble sugar, Folium Mori total sugar and the content of polysaccharides of Folium Mori can be calculated further according to formula.
In Folium Mori, reducing sugar, water-soluble sugar, total sugar and polyoses content calculate as follows:
Reducing sugar (with glucose meter) %=
Water-soluble sugar (with glucose meter) %=
Total sugar (with glucose meter) %=
Polysaccharides of Folium Mori (%)=[Folium Mori total sugar (%)-Folium Mori water-soluble sugar (%)] × 0.9
In formula: m1Look into reducing sugar in curve gained mensuration liquid and be equivalent to glucose in milligrams number, mg；
m2Look into water-soluble sugar in curve gained mensuration liquid to want when in glucose in milligrams number, mg；
m3Look into total sugar in curve gained mensuration liquid and be equivalent to glucose in milligrams number, mg；
V0Mensuration liquid amasss, V0=0.4mL；
V1Reducing sugar extracting liquid volume, V1=100mL；
V2Water-soluble sugar extracting liquid volume, V2=100mL；
V3Total sugar extracting liquid volume, V3=100mL；
N1Reducing sugar extracting solution extension rate, N1=1；
N2Water-soluble sugar extracting solution extension rate, N2=2；
N3Total sugar extracting solution extension rate, N3=1；
M Folium Mori sample quality, g.
It is calculated Folium Mori reducing sugar, water-soluble sugar, total sugar result as shown in table 1:
Folium Mori reducing sugar, water-soluble sugar, total sugar measurement result in table 1, embodiment 1
Note: each sample parallel extracts three groups, often group parallel assay twice, the arithmetic mean of instantaneous value of twice mensuration is meansigma methods in group, and in three groups of groups, the arithmetic mean of instantaneous value of meansigma methods is meansigma methods between group, lower same.
According to the measurement result in table 1, can in the hope of embodiment 1 in the content of polysaccharides of Folium Mori be:
Polysaccharides of Folium Mori (%)=(24.12%-13.19%) × 0.9=9.84%
Embodiment 2
Related reagent preparation, Folium Mori reducing sugar, water-soluble sugar, total sugar extracting method with embodiment 1.
Glucose standards solution is prepared: prepare Glucose standards solution, final concentration of 1.056mg/mL according to the method for embodiment 1.
Glucose standard curve makes: take 11 10mL color comparison tubes, it is separately added into 0.00mL, 0.08mL, 0.16mL, 0.24mL, 0.32mL, 0.40mL, 0.48mL, 0.56mL, 0.64mL, 0.72mL, 0.80mL glucose solution (add 0.00mL glucose solution and do blank), dilute is to 0.80mL, add 0.60mL DNS reagent, shake up, boiling water bath 5min, it is cooled to room temperature with cold water immediately after taking-up, again to often pipe addition distilled water to scale (10mL), shake up, at 540nm wavelength, measure each pipe absorbance.With glucose content (mg) as abscissa, absorbance is vertical coordinate, draws standard curve, and obtaining regression equation is y=1.5802x-0.0631, R2=0.9997 (such as Fig. 2).
In sample sugar content mensuration and calculate with embodiment 1.
Folium Mori reducing sugar in embodiment 2, water-soluble sugar, total sugar measurement result are shown in Table 2:
Folium Mori reducing sugar, water-soluble sugar, total sugar measurement result in table 2 embodiment 2
According to the measurement result in table 2, can in the hope of embodiment 2 in the content of polysaccharides of Folium Mori be:
Polysaccharides of Folium Mori (%)=(34.56%-17.24%) × 0.9=15.59%
Embodiment 1 and embodiment 2 result show: utilizing DNS colorimetric method for determining Folium Mori reducing sugar, water-soluble sugar and total sugar, collimation is good, and can be obtained the content of polysaccharides of Folium Mori by calculating.
Finally illustrate is, preferred embodiment above is only in order to illustrate technical scheme and unrestricted, although the present invention being described in detail by above preferred embodiment, but skilled artisan would appreciate that, in the form and details it can be made various change, without departing from claims of the present invention limited range.
Claims (9)
1. the detection method of a simple effectively polysaccharides of Folium Mori, it is characterised in that: extract Folium Mori water-soluble sugar and Folium Mori total sugar respectively, so
Rear hydrolysis becomes reducing sugar, obtains Folium Mori water-soluble sugar extracting solution and Folium Mori total sugar extracting solution respectively, recycles 3,5-dinitro
Salicylic acid colorimetric method for determining Folium Mori water-soluble sugar extracting solution and Folium Mori total sugar extracting solution light absorption value at 540nm, then according to Portugal
Grape Standard for Sugars curve calculates Folium Mori water-soluble sugar extracting solution and Folium Mori total sugar extracting solution in mensuration liquid respectively and is equivalent to the weight of glucose
Amount, is calculated as follows Folium Mori water-soluble sugar and the content of Folium Mori total sugar:
In formula: m2Look into water-soluble sugar in curve gained mensuration liquid and be equivalent to glucose quality, mg；
m3Look into total sugar in curve gained mensuration liquid and be equivalent to glucose quality, mg；
V0Mensuration liquid amasss, mL；
V2Water-soluble sugar extracting liquid volume, mL；
V3Total sugar extracting liquid volume, mL；
N2Water-soluble sugar extracting solution extension rate；
N3Total sugar extracting solution extension rate；
M Folium Mori sample quality, g.
And according to equation below calculating polysaccharides of Folium Mori:
Polysaccharides of Folium Mori (%)=[Folium Mori total sugar (%)-Folium Mori water-soluble sugar (%)] × 0.9.
The detection method of a kind of simple effectively polysaccharides of Folium Mori, it is characterised in that described 3,5-dinitro
The compound method of salicylic acid solution is as follows: the first preparation aqueous solution that temperature is 40～45 DEG C containing sodium potassium tartrate tetrahydrate, then adds
Entering NaOH solution, sequentially add 3,5-dinitrosalicylic acid, phenol and sodium sulfite, last constant volume makes Soluble tartar.
The final concentration of 0.524mol/L of the final concentration of 185g/L of sodium, NaOH, 3, the 5-final concentration of 6.3g/L of dinitrosalicylic acid, benzene
The final concentration of 5.0g/L of phenol and the final concentration of 5.0g/L of sodium sulfite.
A kind of detection method of simple effectively polysaccharides of Folium Mori, it is characterised in that: described Folium Mori water-soluble sugar
The extracting method of extracting solution is as follows: takes new fresh mulberry leaf and dries to constant weight, pulverize, is subsequently adding distilled water water-bath and extracts, be designated as carrying
Take liquid A, sequentially add acetic acid zinc solution and potassium ferrocyanide solution, centrifugal collection supernatant, obtain Folium Mori reducing sugar extracting solution,
Then in Folium Mori reducing sugar extracting solution, add hydrochloric acid be hydrolyzed, regulate pH to 7.5～8.0, obtain Folium Mori water-soluble sugar extracting solution.
A kind of detection method of simple effectively polysaccharides of Folium Mori, it is characterised in that: described extracting solution A's
Preparation method is as follows: takes new fresh mulberry leaf and is 50 DEG C in temperature and dries to constant weight, pulverizes, be subsequently adding and be equivalent to Folium Mori weight
The distilled water of 60 times, 50 DEG C of water-baths are extracted 20min, are obtained extracting solution A.
A kind of detection method of simple effectively polysaccharides of Folium Mori, it is characterised in that: the addition of described hydrochloric acid
By concentrated hydrochloric acid and Folium Mori reducing sugar extracting liquid volume than for 3:50；Described being hydrolyzed under 70 DEG C of water-baths hydrolyzes 15min.
A kind of detection method of simple effectively polysaccharides of Folium Mori, it is characterised in that: described Folium Mori total sugar extracts
The extracting method of liquid is as follows: takes new fresh mulberry leaf and dries to constant weight, pulverizes, be subsequently adding distilled water and fully dissolve, make always with hydrochloric acid
After sugar complete hydrolysis, being sequentially added into acetic acid zinc solution and potassium ferrocyanide solution to zinc acetate and potassium ferrocyanide, regulation pH is extremely
7.5-8.0, filters, and collects filtrate, obtains Folium Mori total sugar extracting solution.
A kind of detection method of simple effectively polysaccharides of Folium Mori, it is characterised in that: described hydrochloric acid addition is pressed
Folium Mori and concentrated hydrochloric acid w/v are 0.5:6 (g/mL), described in be hydrolyzed under boiling water bath hydrolyze 30min.
8. according to the detection method of simple effectively polysaccharides of Folium Mori a kind of described in any one of claim 3～7, it is characterised in that: described acetic acid
In zinc solution, acetic acid zinc concentration is 219g/L；In described potassium ferrocyanide solution, the concentration of potassium ferrocyanide is 106g/L.
9. the detection method of a kind of simple effectively polysaccharides of Folium Mori described in claim 1, it is characterised in that: described glucose standard curve
Regression equation is y=1.5175x-0.0578 or y=1.5802x-0.0631.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610218597.6A CN105806793B (en) | 2016-04-08 | 2016-04-08 | A kind of simple and effective mulberry leaf polysaccharide detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610218597.6A CN105806793B (en) | 2016-04-08 | 2016-04-08 | A kind of simple and effective mulberry leaf polysaccharide detection method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105806793A true CN105806793A (en) | 2016-07-27 |
| CN105806793B CN105806793B (en) | 2019-04-02 |
Family
ID=56459828
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610218597.6A Expired - Fee Related CN105806793B (en) | 2016-04-08 | 2016-04-08 | A kind of simple and effective mulberry leaf polysaccharide detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105806793B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110530806A (en) * | 2019-08-22 | 2019-12-03 | 中国林业科学研究院林产化学工业研究所 | A kind of method of poly-pentose and content of cellulose in quick measurement broadleaf raw material |
| CN112683830A (en) * | 2021-03-16 | 2021-04-20 | 南京林业大学 | Simple galactomannan degradation product quantification method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103884667A (en) * | 2014-04-18 | 2014-06-25 | 湖北工业大学 | Method for measuring polysaccharide content of medlar extracting solution |
-
2016
- 2016-04-08 CN CN201610218597.6A patent/CN105806793B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103884667A (en) * | 2014-04-18 | 2014-06-25 | 湖北工业大学 | Method for measuring polysaccharide content of medlar extracting solution |
Non-Patent Citations (4)
| Title |
|---|
| 全艳玲等: "黄伞多糖提取优化", 《中国酿造》 * |
| 吴雪辉等: "微波提取油茶饼粕中多糖的工艺研究", 《食品工业科技》 * |
| 杨浩等: "3,5-二硝基水杨酸比色法测定淫羊藿多糖的含量", 《天津化工》 * |
| 苏亚玲等: "灵芝菌生物活性多糖与总糖的测定", 《食品科学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110530806A (en) * | 2019-08-22 | 2019-12-03 | 中国林业科学研究院林产化学工业研究所 | A kind of method of poly-pentose and content of cellulose in quick measurement broadleaf raw material |
| CN110530806B (en) * | 2019-08-22 | 2022-03-29 | 中国林业科学研究院林产化学工业研究所 | Method for rapidly determining content of pentosan and cellulose in broad-leaved wood raw material |
| CN112683830A (en) * | 2021-03-16 | 2021-04-20 | 南京林业大学 | Simple galactomannan degradation product quantification method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105806793B (en) | 2019-04-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Xi et al. | Determination of tea polysaccharides in Camellia sinensis by a modified phenol-sulfuric acid method | |
| CN103623059B (en) | A kind of pharmaceutical composition and application thereof, containing its medicament | |
| CN105806793A (en) | Simple and effective mulberry leaf polysaccharide detection method | |
| CN104374762A (en) | Laser Raman spectrum detection method for quickly analyzing content of basic flavine in soybean hull | |
| CN1979133A (en) | Method for detecting leechee ploysaccharose content | |
| CN105911154A (en) | Method for determination of chlorogenic acid, galuteolin and total flavone content of honeysuckle | |
| CN111419743A (en) | Preparation method and application of vitis amurensis extract | |
| CN103655844B (en) | The preparation method of Changshan grapefruit peel pomace extract and the preparation containing this extract | |
| CN103884667A (en) | Method for measuring polysaccharide content of medlar extracting solution | |
| CN108084291A (en) | The method that polysaccharide is extracted in dendrobium candidum | |
| CN103222628A (en) | Method for extracting dietary fibers | |
| CN101644682A (en) | Fast detecting method of rhenium | |
| CN103698291B (en) | Rapid test method for adulteration of Chinese date honey | |
| CN103616339B (en) | Detection method for preparation containing ganoderan | |
| CN110530806B (en) | Method for rapidly determining content of pentosan and cellulose in broad-leaved wood raw material | |
| CN102384907A (en) | Method by utilize vanillin-sulfuric acid colorimetry to measure glabridin content | |
| CN102033051B (en) | Continuous flow measuring method for pectin content in plant | |
| CN101339137B (en) | Method for determining saponin content of SmilaxscobinicaulisC.H.Wringh or its formulation | |
| CN110174366A (en) | The extraction detection method of glycine betaine in a kind of fructus lycii | |
| CN106248599A (en) | A kind of detection method of fungus polysaccharide | |
| CN103908513B (en) | The extracting method of shinyleaf yellowhorn fruit shell total saponins | |
| CN100408999C (en) | Determination method of balsum pear saponin | |
| CN108318600A (en) | A kind of detection method of food additives content | |
| CN103335871A (en) | Test sample preparation method of tobacco pectin content detection | |
| CN107290244A (en) | A kind of quantitative analysis method of low cellulosic plant chemical composition |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190402 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |