CN101294912B - Tea polyphenol detecting test paper, standard color comparison card and usage thereof - Google Patents
Tea polyphenol detecting test paper, standard color comparison card and usage thereof Download PDFInfo
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Abstract
The invention provides tea polyphenol test paper, which is prepared by adopting transition metal, such as iron, as a color development agent, adopting tartaric acid, citric acid, oxalic acid, hydrofluoric acid and salicylsulfonic acid, or any of sylvite, sodium salt and potassium sodium tartrate thereof as a complexing agent, and adopting soluble starch, polyvinyl alcohol, agaragar, polyacrylamide gel or gelatin as a bonding agent. Meanwhile, a standard tea polyphenol concentration color card is prepared. The tea polyphenol content can be quickly and visually observed according to the contrast between the color development of a sample to be tested on the detecting test paper and the standard color card. The test paper is suitable for quickly measuring the tea polyphenol content in plants or plant products, particularly suitable for the measurement of the tea polyphenol content in tea. The test paper has the advantages of simple, convenient and rapid method, low cost and easy popularization.
Description
Technical field
The present invention relates to the effective ingredients in plant detection technique, specifically belong to a kind of tea polyphenol detecting test paper and standard color comparison card thereof and their purposes.
Background technology
Tealeaves is one of the whole world three big natural drinks, and Tea Polyphenols is the important active component of tealeaves, is the main matter of decision tealeaves local flavor and quality.
At present, mainly comprise efficient liquid chromatography determination method, atomic absorption method, UV-VIS spectrophotometry, titrimetry and electrochemical process etc. about plant polyphenol Determination on content method.Each component obtains separating and carrying out qualitative and quantitative analysis in the efficient liquid chromatography determination method mensuration material, and this method can be measured the different weight polyphenol fractions and the finger-print of plurality of active ingredients simultaneously.Atomic absorption method is to utilize basic lead acetate and Tea Polyphenols complexing to form yellow mercury oxide, by measuring excessive lead ion in the supernatant, the content of indirect determination Tea Polyphenols.Flow injection-inhibition chemoluminescence method is under alkali condition, Tea Polyphenols reduction H
2O
2, suppress luminol-H
2O
2-KIO
4The chemiluminescence of system, it suppresses luminous degree and becomes linear dependence with the content of Tea Polyphenols.Spectrophotometric method also is a common method, mainly comprises spectrophotometric colourimetry, three wavelength spectrophotometry and differential spectrophotometry.Spectrophotometric method mainly is the plant polyphenol and different substance that show color reactions in higher plant and the product, optical density by substance that show color becomes correlationship to measure polyphenol content with the concentration of polyphenol, different polyphenol and identical or different developer quality in the determinand, have characteristic absorption peak, can be by multi-wavelength and differential spectrophotometry.Chromogenic reagents such as Tea Polyphenols and potassium permanganate, diazo salt, ammonium molybdate, ferrous tartrate and associated metal ion have formed a series of Tea Polyphenols assay methods such as ferrous tartrate colourimetry (GB 8313287), diazotising-coupling branch colourimetry ammonium molybdate spectrophotometric method, permanganimetric method and potentiometric titration.Also have Tea Polyphenols under acid condition, to reduce Fe in addition
3+Be Fe
2+, Fe
2+Generate the blue look complex KFe[Fe (CN) of solubility with the potassium ferricyanide
6], by the content of colorimetric indirect determination Tea Polyphenols.A kind of image information of utilizing the image acquisition instrument to gather the Tea Polyphenols colour developing has also been described by China Agricultural University (patent No. CN 1811391A), handles based on computing machine colourity recognition system, carries out the polyphenol content method for measuring.Its advantage is to have substituted expensive spectrophotometer with image capture instrument, and the typical curve that can prestore is only measured testing sample and can be obtained its concentration.Its measuring principle is ferrous tartrate and Tea Polyphenols colour developing, directly gathers the image information of colour developing in colorimetric pool, and the colourity of testing sample and the colourity of standard model are relatively carried out the mensuration of polyphenol content.
In the existing method of measuring Tea Polyphenols, the method precision height that has, the method that has are with strong points, but need expensive exact instrument; Though titrimetry does not need exact instrument, need the process of reagent preparation and process titration.So, set up a kind of simply, the Tea Polyphenols detection method is very important easily and efficiently.
Summary of the invention
The purpose of this invention is to provide a kind of tea polyphenol detecting test paper and standard color comparison card thereof, and a kind of this detection test paper and standard color comparison card method of polyphenol content in the working sample quickly and easily of utilizing is provided.
A kind of tea polyphenol detecting test paper provided by the invention makes by the method that comprises the steps: take by weighing bonding agent and add in the distilled water, make its dissolving, be mixed with the binder solution that concentration is 0.2-5%; Complexing agent is joined in the binder solution, stirring and dissolving, making its concentration is 0.2-1.5mol/L; Add the transition metal salt developer again, stirring and dissolving, making its concentration is 0.3-1.5mol/L, promptly gets the liquid that develops the color; With soaking 3-20 minute in the filter paper immersion colour developing liquid, to take out, drying is cut out, shaping, packing promptly gets polyphenol detection test paper;
Described bonding agent is selected from soluble starch, polyvinyl alcohol (PVA), agar, polyacrylamide gel or gelatin etc.; Complexing agent is selected from tartrate, citric acid, oxalic acid, hydrofluorite or each sylvite or sodium salt or sodium potassium tartrate tetrahydrate etc. in them; The slaine of developer chosen from Fe, copper or zinc etc.
A kind of and the matching used standard color comparison card of tea polyphenol detecting test paper provided by the invention makes by the method that comprises the steps: the Tea Polyphenols standard solution of the 3-15 kind variable concentrations of compound concentration in 0-10mg/mL; In the Tea Polyphenols standard solution of variable concentrations, develop the color respectively with the described tea polyphenol detecting test paper of claim 1,, standard color block is arranged in order, pastes by the Tea Polyphenols concentration of standard solution, make standard color comparison card according to the production standard color lump as a result that develops the color.
Described according to the production standard color lump as a result that develops the color, be that the result utilizes computing machine to carry out image scanning, Processing of Preparation obtains the Tea Polyphenols standard color block according to developing the color.Also can obtain with the method for pigmented tinting.
Described test paper can be used for the mensuration of polyphenol content in the mensuration of polyphenol content in plant or the vegetable products, particularly tealeaves.
The assay method of polyphenol content is characterized in that in a kind of tealeaves, comprises the steps: weighing tealeaves to be measured, adds the water (g/mL) of 95 ℃-100 ℃ of 25-50 times of volumes, and lixiviate made solution to be measured in 20 minutes; Get above-mentioned tea polyphenol detecting test paper and drip or dip in and get solution to be measured, in 20 seconds-10 minutes,, obtain polyphenol content with above-mentioned matching used standard color comparison card comparison.
Polyphenol content can calculate or check in according to set tabulation according to following formula.
Polyphenol content (mg/g)=(colourimetric number * V)/(W * P)
In the formula: colourimetric number (mg/mL): obtain with the colorimetric card comparison; V: the volume of extract (mL); P: for the first time Tea Polyphenols extraction ratio (%) is about 84.30% as tealeaves during with 50 times of volume floodings; W: the quality of testing sample (g).Also can calculate and make the polyphenol content table according to formula.
Compared with prior art the present invention is that primary raw material has prepared tea polyphenol detecting test paper with developer, complexing agent, bonding agent and filter paper etc., this detection test paper is applicable to the mensuration of polyphenol content in the mensuration of polyphenol content in plant or the vegetable products, particularly tealeaves.This method has characteristics such as simple, convenient, fast, can detect Tea Polyphenols intuitively, need not large-scale precision instrument, also need not the professional and technical personnel or, more be not subjected to the constraint of environment, also avoid the complicated processes of the restriction and the reagent preparation of test condition through professional training, be convenient to popular popularizing and use whenever and wherever possible, in addition, low price and being easy to carry, the holding time of test paper is long.
Description of drawings
Fig. 1: the colour developing of tea polyphenol detecting test paper in the different Tea Polyphenols concentration of 0-10mg/mL of soluble starch, sodium potassium tartrate tetrahydrate and ferric trichloride preparation and according to its standard color comparison card that is prepared into.Last figure is a standard color comparison card, and figure below is actual colour developing.
Fig. 2: the tea polyphenol detecting test paper of soluble starch, sodium potassium tartrate tetrahydrate and ferric trichloride preparation is measured the colour developing of jasmine tea and Longjing tea, with corresponding colorimetric card comparison.
Fig. 3: the tea polyphenol detecting test paper of agar powder, sodium potassium tartrate tetrahydrate and ferric trichloride preparation is measured the colour developing of Pu'er tea and Biluochun tea, with corresponding colorimetric card comparison.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.
Embodiment 1
The preparation of test paper: take by weighing soluble starch 2.5g, be dissolved in the 80mL water, be heated to water white transparency, be mixed with the starch solution of 2.5% (W/V), take by weighing sodium potassium tartrate tetrahydrate 28.2g, be dissolved in the starch solution for preparing, stirring and dissolving adds ferric trichloride 27.0g again, continue stirring and dissolving, fixed molten to 100mL, promptly get chromophoric solution, wherein the volumetric molar concentration of sodium potassium tartrate tetrahydrate and ferric trichloride is 1.0mol/L.Qualitative filter paper immersed in the chromophoric solution soaked 10 minutes, take out and place on the glass plate, promptly get tea polyphenol detecting test paper in 37 ℃ of oven dryings.
The preparation of standard color comparison card: be mixed with concentration (mg/mL) with Tea Polyphenols and be respectively 0,1.0,2.0,3.0,4.0,5.0,6.0,7.0,8.0,9.0,10.0 titer.With of the Tea Polyphenols colour developing of above-mentioned tea polyphenol detecting test paper to various criterion concentration, Tea Polyphenols colour developing with normal concentration is a benchmark, utilize computer A dobe Photoshop to edit the rgb value of different colours, compile versicolor color lump with rgb value, digital photofinishing is produced all color lumps.Colour developing test paper with the normal concentration Tea Polyphenols scans simultaneously again, both as calculated machine software carry out color comparison, the step of editor, digital photofinishing, scanning, comparison repeatedly, up to comparing out the color that is complementary with standard polyphenol concentration, in editor's color lump that digital photofinishing is produced, cut out and the Standard China Green Tea polyphenol solution color lump of color of being complementary that on test paper, develops the color, and press the colour developing of Tea Polyphenols concentration standard and arrange, paste, make Tea Polyphenols concentration standard colorimetric card, see shown in Figure 1.
The mensuration of polyphenol content in the tealeaves:
The preparation of testing sample: weighing testing sample Pilochun (a green tea), Iron Guanyin, Dragon Well tea, Pu'er, black tea and jasmine tea, add the water (g/mL) of 95 ℃-100 ℃ of 50 times of volumes respectively, lixiviate 20 minutes makes the solution to be measured of various tealeaves.
Polyphenol content is measured: get tea polyphenol detecting test paper and drip or dip in and get solution to be measured, adopted visual colorimetry and standard color comparison card comparison in 30 seconds, promptly get colourimetric number;
Every duplicate samples is measured three times, averages, and calculates the tealeaves polyphenol content by following formula.Polyphenol content (mg/g)=(colourimetric number * V)/(W * P).Wherein: colourimetric number: obtain with the colorimetric card comparison; V: the volume of extract (mL); P: extract Tea Polyphenols extraction ratio for the first time, by 84.30%; W: the quality of testing sample (g).The results are shown in Table 1 and Fig. 2, Fig. 2 shows the standard coloration card of tea polyphenol detecting test paper of soluble starch, sodium potassium tartrate tetrahydrate and ferric trichloride preparation and the colour developing result who measures jasmine tea tea and Longjing tea.The result shows: tea polyphenol detecting test paper ocular estimate result compares relative deviation less than 10% with National Standard Method, is suitable for the fast measuring of tealeaves Tea Polyphenols.
Table 1 the inventive method and National Standard Method (GB 8313287) tealeaves polyphenol content is measured relatively
Embodiment 2
1. the preparation of test paper: take by weighing agar powder 0.2g, be dissolved in the 80mL water, be heated to water white transparency, be mixed with the starch solution of 0.2% (W/V), take by weighing sodium potassium tartrate tetrahydrate 28.2g, be dissolved in the starch solution for preparing, stirring and dissolving adds ferric trichloride 27.0g again, continue stirring and dissolving, fixed molten to 100mL, promptly get chromophoric solution, wherein the volumetric molar concentration of sodium potassium tartrate tetrahydrate and ferric trichloride is 1.0mol/L.Qualitative filter paper immersed in the chromophoric solution soaked 5 minutes, take out and place on the glass plate, promptly get tea polyphenol detecting test paper in shady and cool dry place airing.
2. the preparation of standard color comparison card: Tea Polyphenols is mixed with concentration (mg/mL) and is respectively 0,1.0,2.0,3.0,4.0,5.0,6.0,7.0,8.0,9.0,10.0 titer.Measuring test paper with above-mentioned Tea Polyphenols is benchmark to the Tea Polyphenols colour developing of various criterion concentration, makes standard color comparison card with the method for drawing pigment toning.
3. the preparation of testing sample and assay: the method by embodiment 1 is carried out.The result shows: the tea polyphenol detecting test paper visual observation is compared relative deviation less than 10% with National Standard Method, is suitable for the fast measuring of tealeaves Tea Polyphenols.
Embodiment 3
1. the preparation of test paper: take by weighing soluble starch 2.0g, be dissolved in the 80mL water, be heated to water white transparency, be mixed with the starch solution of 2% (W/V), take by weighing sodium oxalate 4.02g, be dissolved in the starch solution for preparing, stirring and dissolving adds ferric trichloride 13.5g again, continue stirring and dissolving, fixed molten to 100mL, promptly get chromophoric solution, wherein the volumetric molar concentration of sodium oxalate and ferric trichloride is respectively 0.3mol/L and 0.5mol/L.Qualitative filter paper immersed in the chromophoric solution soaked 5 minutes, take out and place on the glass plate, promptly get tea polyphenol detecting test paper in 37 ℃ of oven dryings.
2. the preparation of standard color comparison card: colour developing is a benchmark to the Tea Polyphenols of various criterion concentration with above-mentioned tea polyphenol detecting test paper, prepares its respective standard colorimetric card, and method for making is with embodiment 1.
Embodiment 4
1. the preparation of test paper: take by weighing soluble starch 4.0g, be dissolved in the 80mL water, be heated to water white transparency, be mixed with the starch solution of 4% (W/V), take by weighing sodium oxalate 2.68g, be dissolved in the starch solution for preparing, stirring and dissolving adds ferric nitrate 12.12g again, continue stirring and dissolving, fixed molten to 100mL, promptly get chromophoric solution, wherein the volumetric molar concentration of sodium oxalate and ferric nitrate is respectively 0.2mol/L and 0.3mol/L.Qualitative filter paper immersed in the chromophoric solution soaked 5 minutes, take out and place on the glass plate, promptly get tea polyphenol detecting test paper in 37 ℃ of oven dryings.
2. the preparation of standard color comparison card: be mixed with Tea Polyphenols concentration (mg/mL) and be respectively 0,0.5,1.5,2.5,3.5,4.5,5.5 titer.Colour developing is a benchmark to the Tea Polyphenols of various criterion concentration with above-mentioned tea polyphenol detecting test paper, preparation respective standard colorimetric card, and method for making is seen embodiment 1.
3. testing sample black tea and Biluochun tea are prepared into liquid to be measured by the method for embodiment 1.
4. assay: adopt the tea polyphenol detecting test paper ocular estimate to measure, the results are shown in Figure 3.
Embodiment 5
1. the preparation of test paper: take by weighing soluble starch 1.5g, be dissolved in the 80mL water, be heated to water white transparency, be mixed with the starch solution of 1.5% (W/V), take by weighing sodium fluoride 4.2g, be dissolved in the starch solution for preparing, stirring and dissolving adds ferric trichloride 40.5g again, continue stirring and dissolving, fixed molten to 100mL, promptly get chromophoric solution, wherein the volumetric molar concentration of sodium fluoride and ferric trichloride is respectively 1.0mol/L and 1.5mol/L.Qualitative filter paper immersed in the chromophoric solution soaked 5 minutes, take out and place on the glass plate, promptly get tea polyphenol detecting test paper in 37 ℃ of oven dryings.
2. the preparation of standard color comparison card: colour developing is a benchmark to the Tea Polyphenols of various criterion concentration with above-mentioned tea polyphenol detecting test paper, preparation respective standard colorimetric card, and method for making is with embodiment 1.
Embodiment 6
1. the preparation of test paper: take by weighing soluble starch 2.5g, be dissolved in the 80mL water, be heated to water white transparency, be mixed with the starch solution of 2.5% (W/V), take by weighing sodium fluoride 4.2g, be dissolved in the starch solution for preparing, stirring and dissolving adds ferric nitrate 40.4g again, continue stirring and dissolving, fixed molten to 100mL, promptly get chromophoric solution, wherein the volumetric molar concentration of sodium fluoride and ferric nitrate is 1.0mol/L.Qualitative filter paper immersed in the chromophoric solution soaked 10 minutes, take out and place on the glass plate, promptly get tea polyphenol detecting test paper in 37 ℃ of oven dryings.
2. the preparation of standard color comparison card: colour developing is a benchmark to the Tea Polyphenols of various criterion concentration with above-mentioned tea polyphenol detecting test paper, preparation respective standard colorimetric card, and method for making is with embodiment 1.
Claims (5)
1. a tea polyphenol detecting test paper is characterized in that, makes by the method that comprises the steps: take by weighing bonding agent and add in the distilled water, make its dissolving, be mixed with the binder solution that concentration is 0.2-2.5%; Complexing agent is joined in the binder solution, stirring and dissolving, making its concentration is 0.3-1.0mol/L; Add the transition metal salt developer again, stirring and dissolving, making its concentration is 0.3-1.0mol/L, promptly gets the liquid that develops the color; With soaking 3-20 minute in the filter paper immersion colour developing liquid, to take out, drying is cut out, shaping, packing promptly gets polyphenol detection test paper;
Described bonding agent is selected from soluble starch, polyvinyl alcohol (PVA), agar, polyacrylamide gel or gelatin; Complexing agent is selected from tartrate, citric acid, oxalic acid, hydrofluorite, sulfosalicylic acid or each sylvite or sodium salt or sodium potassium tartrate tetrahydrate in them; Developer is selected from ferric trichloride or ferric nitrate.
2. a standard color comparison card is characterized in that, makes by the method that comprises the steps: the Tea Polyphenols standard solution of the 3-15 kind variable concentrations of compound concentration in 0-10mg/ml; In the Tea Polyphenols standard solution of variable concentrations, develop the color respectively with the described tea polyphenol detecting test paper of claim 1,, standard color block is arranged in order, pastes by the Tea Polyphenols concentration of standard solution, make standard color comparison card according to the production standard color lump as a result that develops the color.
3. according to the described standard color comparison card of claim 2, it is characterized in that, described according to the production standard color lump as a result that develops the color, be that the result utilizes computing machine to carry out image scanning, Processing of Preparation obtains the Tea Polyphenols standard color block according to developing the color.
4. the mensuration that is used for plant or vegetable products polyphenol content according to the described test paper of claim 1.
5. the mensuration that is used for the tealeaves polyphenol content according to the described test paper of claim 1.
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