CN112147092A - Method for detecting acidic polysaccharide in water body - Google Patents
Method for detecting acidic polysaccharide in water body Download PDFInfo
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- CN112147092A CN112147092A CN202010473719.2A CN202010473719A CN112147092A CN 112147092 A CN112147092 A CN 112147092A CN 202010473719 A CN202010473719 A CN 202010473719A CN 112147092 A CN112147092 A CN 112147092A
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 160
- 229920001284 acidic polysaccharide Polymers 0.000 title claims abstract description 91
- 150000004805 acidic polysaccharides Chemical class 0.000 title claims abstract description 91
- 238000000034 method Methods 0.000 title claims abstract description 47
- 239000012086 standard solution Substances 0.000 claims abstract description 64
- 238000001514 detection method Methods 0.000 claims abstract description 48
- 238000002835 absorbance Methods 0.000 claims abstract description 36
- 238000001914 filtration Methods 0.000 claims abstract description 17
- 238000012417 linear regression Methods 0.000 claims description 22
- 239000006228 supernatant Substances 0.000 claims description 21
- 239000003086 colorant Substances 0.000 claims description 20
- 239000012528 membrane Substances 0.000 claims description 18
- 230000020477 pH reduction Effects 0.000 claims description 11
- 239000000126 substance Substances 0.000 claims description 11
- 238000000870 ultraviolet spectroscopy Methods 0.000 claims description 10
- 239000002253 acid Substances 0.000 claims description 9
- 239000000243 solution Substances 0.000 claims description 9
- 239000004417 polycarbonate Substances 0.000 claims description 7
- 229920000515 polycarbonate Polymers 0.000 claims description 7
- 229920001285 xanthan gum Polymers 0.000 claims description 7
- 235000010493 xanthan gum Nutrition 0.000 claims description 7
- 239000000230 xanthan gum Substances 0.000 claims description 7
- 229940082509 xanthan gum Drugs 0.000 claims description 7
- 239000004677 Nylon Substances 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 6
- 229920001778 nylon Polymers 0.000 claims description 6
- 235000010443 alginic acid Nutrition 0.000 claims description 5
- 239000000783 alginic acid Substances 0.000 claims description 5
- 229920000615 alginic acid Polymers 0.000 claims description 5
- 229960001126 alginic acid Drugs 0.000 claims description 5
- 150000004781 alginic acids Chemical class 0.000 claims description 5
- 239000011148 porous material Substances 0.000 claims description 4
- 238000010186 staining Methods 0.000 claims description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 3
- 229910052799 carbon Inorganic materials 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims 3
- 238000001471 micro-filtration Methods 0.000 claims 1
- 230000008569 process Effects 0.000 abstract description 8
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 6
- 238000004043 dyeing Methods 0.000 abstract description 6
- 231100000481 chemical toxicant Toxicity 0.000 abstract description 5
- 239000003440 toxic substance Substances 0.000 abstract description 5
- 238000005259 measurement Methods 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 2
- 238000004364 calculation method Methods 0.000 abstract description 2
- 238000006243 chemical reaction Methods 0.000 abstract description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical group CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 238000007796 conventional method Methods 0.000 description 6
- 230000002378 acidificating effect Effects 0.000 description 5
- 239000000047 product Substances 0.000 description 4
- 229960000583 acetic acid Drugs 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 239000012362 glacial acetic acid Substances 0.000 description 3
- 239000003002 pH adjusting agent Substances 0.000 description 3
- UJOBWOGCFQCDNV-UHFFFAOYSA-N 9H-carbazole Chemical compound C1=CC=C2C3=CC=CC=C3NC2=C1 UJOBWOGCFQCDNV-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000004177 carbon cycle Methods 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 239000012445 acidic reagent Substances 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- UBXYXCRCOKCZIT-UHFFFAOYSA-N biphenyl-3-ol Chemical group OC1=CC=CC(C=2C=CC=CC=2)=C1 UBXYXCRCOKCZIT-UHFFFAOYSA-N 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000008239 natural water Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000004804 polysaccharides Chemical class 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
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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
- 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
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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Abstract
The invention provides a method for detecting acidic polysaccharide in a water body, belonging to the field of analysis and detection. The detection method for the total acidic polysaccharide in the water body simplifies the detection process into that the two-wavelength measurement is directly carried out after dyeing, the absorbance values of the water body to be detected at the two characteristic peak wavelengths of the standard solution are measured, and then the concentration of the total acidic polysaccharide in the water body after the dyeing reaction is represented by using the absorbance ratio obtained by calculation, so that the detection accuracy is improved, the filtration is omitted, the interference of suspended matters in the water body is avoided, the detection time and the detection cost are shortened, the purpose of simply and efficiently detecting the total acidic polysaccharide in the water body is realized, the detection method provided by the application is environment-friendly and efficient, toxic chemical reagents do not need to be added, and the secondary pollution is avoided. The results of the examples show that with the detection method provided herein, the filtering operation is omitted, with a relative error as low as 2.5% compared to the conventional detection method.
Description
Technical Field
The invention relates to the field of analysis and detection, in particular to a method for detecting acidic polysaccharide in a water body.
Background
The acidic polysaccharide has strong biological activity, has the functions of reducing blood fat, regulating immunity, resisting blood coagulation, protecting liver and the like, and is the most widely applied bioactive polysaccharide in the fields of life science and medicine. Wherein, in aquatic ecology field, the acidic polysaccharide in the nature water has high stickness, is the main binder of large-scale gel in the water, influences the carbon cycle process in the water, and its content has important effect to the carbon cycle in the water, and the influence to climate change simultaneously does not have a small amount of variation.
At present, in the prior art, the content of acidic polysaccharide in a water body is mainly detected by adopting a single-wavelength ultraviolet-visible spectrum method, after the method utilizes alcian blue for dyeing under an acidic condition, suspended matters in the water body are removed by filtering to reduce interference, and then a single-wavelength ultraviolet-visible spectrophotometer is used for determination. But inevitably brings about other disturbances in the filtration process, such as adsorption of the staining substances by the filtration membrane, adsorption of the filtration apparatus and the vessel, and the like. Although errors can be controlled by the same water sample and the same measuring steps, the increase of the operation steps inevitably brings interference, increases the uncertainty and workload of detection, and cannot realize quick and convenient detection. In addition, the carbazole method and the m-hydroxybiphenyl method for detecting high-content acidic polysaccharide require toxic reagents, generate secondary pollution and are not suitable for natural water bodies with low content of acidic polysaccharide. Therefore, it is needed to provide a method for detecting acidic polysaccharides in water with reduced interference during detection, simple operation and high efficiency.
Disclosure of Invention
The detection method provided by the invention simplifies the detection process into dyeing-acidification-two-wavelength measurement, avoids the interference caused by the filtration process, shortens the detection time and cost, is environment-friendly and efficient, does not need to add toxic chemical reagents, and avoids secondary pollution.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a method for detecting total acidic polysaccharide in a water body, which comprises the following steps:
(1) preparing a standard solution with gradient concentration, wherein a standard substance in the standard solution comprises one of xanthan gum, alginic acid and uronic acid;
(2) pre-treating the standard solution in the step (1), wherein the pre-treating comprises the following steps: mixing the standard solution with a coloring agent, and then sequentially carrying out acidification treatment and standing to obtain a supernatant;
(3) determining two characteristic peak wavelengths lambda of the supernatant in the step (2) by using an ultraviolet-visible spectrophotometer1And λ2And measuring said supernatant at said two wavelengths λ1And λ2The absorbance of the standard solution is calculated to obtain the standard solution at two wavelengths lambda1And λ2The absorbance ratio of (a); establishing a standard curve by taking the absorbance ratio as a horizontal coordinate and the concentration of the corresponding standard solution as a vertical coordinate to obtain a linear regression equation;
(4) adopting the same method as the step (2) to pretreat the water body to be detected, and measuring the two wavelengths lambda of the water body to be detected1And λ2And (4) substituting the absorbance ratio into the linear regression equation in the step (3) to obtain the concentration of the total acidic polysaccharide in the water body.
Preferably, the coloring agent in the step (2) is acid alcian blue with the mass concentration of 0.02%.
Preferably, the volume ratio of the coloring agent to the standard solution is 1 (3-15).
Preferably, the acidification treatment in the step (2) is to adjust the pH of a solution obtained by mixing the standard solution and the coloring agent to 2-5.
Preferably, the standing time in the step (2) is 20-120 min.
Preferably, the water body to be detected is a water body with the concentration of the soluble organic carbon less than 10 mg/L.
The invention also provides a detection method of the dissolved acidic polysaccharide in the water body, which comprises the following steps: firstly, filtering a water body to be detected by using a microporous filter membrane to obtain a pretreated water body; the water body is pretreated by using the total acidic polysaccharide in the water body according to the technical scheme, and the concentration of the obtained acidic polysaccharide is the concentration of the dissolved acidic polysaccharide in the water body.
Preferably, the material of the microporous filter membrane comprises one of nylon, nylon mixed material and polycarbonate membrane.
Preferably, the pore diameter of the microporous filter membrane is 0.2-0.5 μm.
The invention also provides a detection method of the granular acidic polysaccharide in the water body, which comprises the following steps: dividing the water body to be detected into two parts, and detecting one part of the water body to be detected by using the method for detecting the total acidic polysaccharides in the water body according to the technical scheme to obtain the concentration of the total acidic polysaccharides in the water body; detecting the other water body to be detected by using the dissolved acidic polysaccharide in the water body by using the technical scheme to obtain the concentration of the dissolved acidic polysaccharide in the water body; and subtracting the concentration of the dissolved acidic polysaccharide in the water body from the concentration of the total acidic polysaccharide in the water body to obtain the concentration of the granular acidic polysaccharide in the water body.
The invention provides a method for detecting total acidic polysaccharide in a water body, which comprises the following steps: firstly, preparing a standard solution with gradient concentration, then mixing the standard solution with a coloring agent, sequentially carrying out acidification treatment and standing pretreatment, and determining two characteristic peak wavelengths lambda of the standard solution by using an ultraviolet-visible spectrophotometer1And λ2And at two wavelengths λ1And λ2The absorbance of the standard solution at two wavelengths lambda1And λ2The absorbance ratio is an abscissa, the concentration of the corresponding standard solution is an ordinate, a standard curve is established, and a linear regression equation is obtained; the same method is used for obtaining the water body to be measured at two wavelengths lambda1And λ2Substituting the absorbance ratio into the linear regression equation to finally obtain the concentration of the total acidic polysaccharide in the water body. The detection method for the total acidic polysaccharide in the water body simplifies the detection process into that the total acidic polysaccharide in the water body is directly measured by two wavelengths after dyeing under an acidic condition, the absorbance values of the water body to be detected at the wavelengths of two characteristic peaks of a standard solution are measured, and the concentration of the total acidic polysaccharide in the water body after dyeing reaction is represented by the light absorption ratio obtained by calculation, so that the detection accuracy is improved, the filtration is omitted, the interference of suspended matters in the water body is avoided, the detection time and the detection cost are shortened, and the purpose of simply and efficiently detecting the total acidic polysaccharide in the water body is realizedThe method is environment-friendly and efficient, does not need to add toxic chemical reagents, and avoids secondary pollution. The results of the examples show that with the detection method provided herein, the filtering operation is omitted, with a relative error as low as 2.5% compared to the conventional detection method.
Drawings
FIG. 1 is a graph of a standard curve obtained in example 1 of the present invention and a linear regression equation;
FIG. 2 is a graph of a standard curve obtained in example 5 of the present invention and a linear regression equation.
Detailed Description
The invention provides a method for detecting total acidic polysaccharide in a water body, which comprises the following steps:
(1) preparing a standard solution with gradient concentration, wherein a standard substance in the standard solution comprises one of xanthan gum, alginic acid and uronic acid;
(2) pre-treating the standard solution in the step (1), wherein the pre-treating comprises the following steps: mixing the standard solution with a coloring agent, and then sequentially carrying out acidification treatment and standing to obtain a supernatant;
(3) determining two characteristic peak wavelengths lambda of the supernatant in the step (2) by using an ultraviolet-visible spectrophotometer1And λ2And measuring said supernatant at said two wavelengths λ1And λ2The absorbance of the standard solution is calculated to obtain the standard solution at two wavelengths lambda1And λ2The absorbance ratio of (a); establishing a standard curve by taking the absorbance ratio as a horizontal coordinate and the concentration of the corresponding standard solution as a vertical coordinate to obtain a linear regression equation;
(4) adopting the same method as the step (2) to pretreat the water body to be detected, and measuring the two wavelengths lambda of the water body to be detected1And λ2And (4) substituting the absorbance ratio into the linear regression equation in the step (3) to obtain the concentration of the total acidic polysaccharide in the water body.
The invention prepares a standard solution with gradient concentration, and a standard substance in the standard solution comprises one of xanthan gum, alginic acid and uronic acid. The range and gradient value of the gradient concentration are not particularly limited in the present invention, and the concentration range is within the range that is conventional in the art. In the embodiment of the invention, the preparation range of the gradient concentration is 0-10 mg/L; the gradient concentrations were 0mg/L, 2mg/L, 4mg/L, 6mg/L, 8mg/L and 10 mg/L. The method for preparing the standard solution is not particularly limited in the present invention, and the standard solution may be prepared by a method known in the art. In the present invention, the solvent of the standard solution is preferably pure water. The sources of xanthan gum, alginic acid and uronic acid are not particularly limited in the present invention, and commercially available products well known to those skilled in the art may be used.
After obtaining the standard solution, the invention carries out pretreatment on the standard solution, wherein the pretreatment comprises the following steps: and mixing the standard solution with a coloring agent, and then sequentially carrying out acidification treatment and standing to obtain a supernatant. In the invention, in the acidification treatment process, the mixed liquid of the standard substance and the coloring agent is adjusted to an acidic environment by the acidic reagent, so that the coloring agent and the acidic groups in the standard substance form salt bonds, and the standard substance and the coloring agent are promoted to react fully, so that the mixed liquid after the standard substance and the coloring agent react has a characteristic absorption peak when being scanned by an ultraviolet-visible spectrometer.
In the present invention, the coloring agent is preferably acid alcian blue having a mass concentration of 0.02%. The source of the acid alcian blue is not particularly limited in the present invention, and a commercially available product well known to those skilled in the art may be used.
In the invention, the volume ratio of the coloring agent to the standard solution is preferably 1 (3-15), and more preferably 1 (5-10).
In the invention, the acidification treatment is preferably to adjust the pH of a solution obtained by mixing the standard solution and the coloring agent to be 2-5, and the pH is more preferably 2.5. According to the invention, the pH value of the solution obtained by mixing the standard solution and the coloring agent is controlled within the range, so that the phenomenon that the coloring agent and the standard substance are not reacted fully when the pH value is too high or too low can be avoided. The pH adjuster used in the acidification treatment of the present invention is not particularly limited, and any pH adjuster known to those skilled in the art may be used. In the present invention, the pH adjusting agent used in the acidification treatment is preferably glacial acetic acid.
In the invention, the standing time is preferably 20-120 min, and more preferably 30-60 min.
After obtaining the supernatant, the invention uses an ultraviolet-visible spectrophotometer to determine the wavelength lambda of two characteristic peaks of the supernatant1And λ2And measuring said supernatant at said two wavelengths λ1And λ2The absorbance of the standard solution is calculated to obtain the standard solution at two wavelengths lambda1And λ2The absorbance ratio of (b). The ultraviolet-visible spectrophotometer of the present invention is not particularly limited, and a commercially available product known to those skilled in the art may be used. In the present invention, the determination of two characteristic peak wavelengths λ of the supernatant is carried out1And λ2Preferably, the supernatant is scanned in the full wavelength range by using an ultraviolet-visible spectrophotometer to obtain an ultraviolet-visible light spectrogram of the standard solution, then a characteristic absorption peak of the standard substance is selected from the spectrogram, and the wavelength lambda of two characteristic peaks is determined1And λ1。
Obtaining a standard solution at two wavelengths lambda1And λ2After the absorbance ratio is obtained, the invention establishes a standard curve by taking the absorbance ratio as a horizontal coordinate and the concentration of the corresponding standard solution as a vertical coordinate to obtain a linear regression equation. The method for obtaining the linear regression equation is not particularly limited in the present invention, and the linear regression equation can be obtained by a method well known in the art.
After the linear regression equation is obtained, the water body to be detected is pretreated by adopting the same pretreatment method as the standard solution, and the supernatant of the water body to be detected is obtained. In the invention, the water body to be detected is preferably water body with the concentration of the soluble organic carbon less than 10 mg/L. In the present invention, the volume of the water to be measured is preferably the same as the volume of the standard solution.
After the supernatant of the water body to be detected is obtained, the supernatant of the water body to be detected is detected at two wavelengths lambda1And λ2Substituting the absorbance ratio into the linear regression equation to obtain the concentration of the total acidic polysaccharide in the water body. In the invention, the total acidic polysaccharide in the water body comprises the water body in solutionThe solution acidic polysaccharide and the granular acidic polysaccharide in the water body.
The method for detecting the total acidic polysaccharide in the water body simplifies the detection process into dyeing-acidification-two-wavelength measurement, avoids the interference caused by the filtration process, shortens the detection time and cost, is environment-friendly and efficient, does not need to add toxic chemical reagents, and avoids secondary pollution
The invention also provides a detection method of the dissolved acidic polysaccharide in the water body, which comprises the following steps: firstly, filtering a water body to be detected by using a microporous filter membrane to obtain a pretreated water body; the method for detecting the total acidic polysaccharide in the water body in the technical scheme is used for detecting the pretreated water body, and the concentration of the obtained acidic polysaccharide is the concentration of the dissolved acidic polysaccharide in the water body.
In the present invention, the material of the microporous filter membrane preferably includes one of nylon, nylon mixed material and polycarbonate membrane, and more preferably polycarbonate membrane. The source of the microporous filter membrane is not particularly limited in the present invention, and commercially available products known to those skilled in the art may be used.
In the invention, the pore diameter of the microporous filter membrane is preferably 0.2-0.5 μm, and more preferably 0.4 μm.
The invention also provides a detection method of the granular acidic polysaccharide in the water body, which comprises the following steps: dividing the water body to be detected into two parts, and detecting one part of the water body to be detected by using the method for detecting the total acidic polysaccharides in the water body in the technical scheme to obtain the concentration of the total acidic polysaccharides in the water body; detecting the other water body to be detected by using the method for detecting the dissolved acidic polysaccharide in the water body in the technical scheme to obtain the concentration of the dissolved acidic polysaccharide in the water body; and subtracting the concentration of the dissolved acidic polysaccharide in the water body from the concentration of the total acidic polysaccharide in the water body to obtain the concentration of the granular acidic polysaccharide in the water body.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Taking lake water A of the spring dish-shaped lake as a water body to be detected, and specifically detecting the lake water A in the spring dish-shaped lake in the following steps:
detection of total acidic polysaccharides in lake water A
(1) Dissolving xanthan gum into pure water to prepare gradient concentration standard solutions with the concentrations of 0mg/L, 2mg/L, 4mg/L, 6mg/L, 8mg/L and 10mg/L respectively;
(2) pre-treating the standard solution in the step (1), wherein the pre-treating comprises the following steps: respectively taking 5mL of standard solution with each concentration, respectively adding 1mL of coloring agent, acidifying with glacial acetic acid until the pH value of the solution is 2.5, shaking up, standing for 30min, and respectively taking supernatant of the standard solution with each concentration;
(3) scanning the supernatant in the step (2) in the full wavelength range by using an ultraviolet-visible spectrophotometer to determine two characteristic peak wavelengths lambda1And λ2340nm and 620nm, respectively, and measuring the two wavelengths lambda of the supernatant1And λ2The absorbance of the standard solution is calculated to obtain the standard solution at two wavelengths lambda1And λ2The absorbance ratio a (from λ)1Absorbance ratio of (A) to (B)2Absorbance of (d) obtained); establishing a standard curve by taking the absorbance ratio a as a horizontal coordinate and the concentration of the corresponding standard solution as a vertical coordinate to obtain a linear regression equation;
FIG. 1 shows the standard curve obtained in example 1 and a linear regression equation, wherein the linear regression equation is that y is 479.5a-500.3 (R)20.986), the absorbance ratio a is an abscissa, and the concentration y of the corresponding standard solution is an ordinate;
(4) and (3) pretreating the lake water A by adopting the same method as the step (2), measuring the absorbance ratio of the lake water A at the wavelengths of 340nm and 620nm, and substituting the measured absorbance ratio into the linear regression equation in the step (3) to obtain the concentration of the total acidic polysaccharides in the lake water A.
Second, detection of dissolved acidic polysaccharide in lake water A
Firstly, filtering lake water A by utilizing a polycarbonate microporous filter membrane with the aperture of 0.4 mu m to obtain a pretreated water body; and detecting the pretreated water body by using the detection method of the total acidic polysaccharides in the lake water A, wherein the concentration of the obtained acidic polysaccharides is the concentration of the dissolved acidic polysaccharides in the lake water A.
Example 2
Taking the lake water B of the disk lake in winter as the water body to be detected, detecting the concentration of the total acidic polysaccharides and the concentration of the dissolved acidic polysaccharides in the lake water B according to the method in the embodiment 1, wherein the specific experimental results are shown in Table 1.
Example 3
Taking lake water C in the Bay of Aixi lake in autumn as the water body to be detected, detecting the concentration of total acidic polysaccharides and the concentration of dissolved acidic polysaccharides in the lake water C according to the method in the embodiment 1, and the specific experimental results are shown in Table 1.
Example 4
Taking lake water D of the summer Yanghu as a water body to be detected, detecting the concentration of total acidic polysaccharide and the concentration of dissolved acidic polysaccharide in the lake water D according to the method in the embodiment 1, wherein the specific experimental results are shown in Table 1;
example 5
Taking river water E in midstream of Xinjiang in spring as water to be detected, detecting the concentration of total acidic polysaccharides and the concentration of dissolved acidic polysaccharides in the lake water E according to the method in the embodiment 1, wherein 0.5ml of coloring agent is added, and the specific experimental results are shown in Table 1.
FIG. 2 is a graph of the standard curve obtained in example 5 and a linear regression equation with y being 188.0a' -195.5 (R)20.987), the absorbance ratio a' is plotted on the abscissa and the concentration y of the corresponding standard solution is plotted on the ordinate.
Example 6
Taking river water F at the downstream of Xinjiang in spring as water to be detected, detecting the concentration of total acidic polysaccharides and the concentration of dissolved acidic polysaccharides in the river water F according to the method in the embodiment 5, wherein the specific experimental results are shown in Table 1.
Comparative example
The lake water A in the example 1 is used as a water body to be detected, the concentration of the total acidic polysaccharides and the concentration of the dissolved acidic polysaccharides in the lake water A in the example 1 are determined according to a conventional method, and the specific detection process is as follows:
determination of total acidic polysaccharides in lake water A by conventional method
(1) Dissolving xanthan gum into pure water to prepare gradient concentration standard solutions with the concentrations of 0mg/L, 2mg/L, 4mg/L, 6mg/L, 8mg/L and 10mg/L respectively;
(2) pre-treating the standard solution in the step (1), wherein the pre-treating comprises the following steps: respectively taking 5ml of standard solution with each concentration, respectively adding 1ml of coloring agent, acidifying with glacial acetic acid until the pH value of the solution is 2.5, shaking up, standing for 30min, and filtering with a polycarbonate filter membrane with the pore diameter of 0.4 μm to obtain filtrate of the standard solution with each concentration;
(3) scanning the filtrate obtained in the step (2) in a full wavelength range by using an ultraviolet-visible spectrophotometer, and measuring the absorbance of the filtrate at the wavelength of 610 nm; establishing a standard curve by taking the absorbance as a horizontal coordinate and the concentration of the corresponding standard solution as a vertical coordinate to obtain a linear regression equation;
(4) and (3) pretreating the lake water A by adopting the same method as the step (2), measuring the absorbance of the lake water A at the wavelength of 610nm, substituting the absorbance into the linear regression equation in the step (3) to obtain the concentration of the total acidic polysaccharides in the lake water A, wherein the specific experimental results are shown in Table 1.
Secondly, measuring the dissolved acidic polysaccharide in the lake water A by a conventional method
Firstly, filtering lake water A by utilizing a polycarbonate filter membrane with the aperture of 0.4 mu m to obtain a pretreated water body; detecting the pretreated water body by using the conventional detection method of the total acidic polysaccharides in the lake water A, wherein the concentration of the obtained acidic polysaccharides is the concentration of the dissolved acidic polysaccharides in the lake water A;
the lake water B, the lake water C, the lake water D, the river water E and the river water F in the embodiments 2-6 are respectively used as water bodies to be detected, the concentrations of the total acidic polysaccharides and the concentrations of the dissolved acidic polysaccharides in the lake water B, the lake water C, the lake water D, the river water E and the river water F in the embodiments 2-6 are respectively determined according to the conventional steps, and specific experimental results are shown in Table 1.
TABLE 1 comparison of results determined in different waters using the two-wavelength method of the present application with conventional methods
The relative error is calculated from (two wavelength determination concentration-conventional method determination concentration)/conventional method determination concentration.
The embodiment, the comparative example and the table 1 show that the detection method for the acidic polysaccharide in the water body simplifies the detection process into direct two-wavelength measurement after dyeing under an acidic condition, realizes the purposes of omitting filtration, avoiding the interference of suspended matters in the water body, shortening the detection time and cost, and realizing the purpose of detecting the total acidic polysaccharide in the water body with simple operation and high efficiency, and compared with the conventional detection method, the detection method provided by the invention has the advantages that the relative error is as low as 2.5%, the detection method is environment-friendly and high-efficiency, toxic chemical reagents are not required to be added, and the secondary pollution is avoided.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A method for detecting total acidic polysaccharides in a water body comprises the following steps:
(1) preparing a standard solution with gradient concentration, wherein a standard substance in the standard solution comprises one of xanthan gum, alginic acid and uronic acid;
(2) pre-treating the standard solution in the step (1), wherein the pre-treating comprises the following steps: mixing the standard solution with a coloring agent, and then sequentially carrying out acidification treatment and standing to obtain a supernatant;
(3) determining two characteristic peak wavelengths lambda of the supernatant in the step (2) by using an ultraviolet-visible spectrophotometer1And λ2And measuring said supernatant at said two wavelengths λ1And λ2The absorbance of the standard solution is calculated to obtain the standard solution at two wavelengths lambda1And λ2The absorbance ratio of (a); establishing a standard curve by taking the absorbance ratio as a horizontal coordinate and the concentration of the corresponding standard solution as a vertical coordinate to obtain a linear regression equation;
(4) adopting the same method as the step (2) to pretreat the water body to be detected, and measuring the two wavelengths lambda of the water body to be detected1And λ2And (4) substituting the absorbance ratio into the linear regression equation in the step (3) to obtain the concentration of the total acidic polysaccharide in the water body.
2. The detection method according to claim 1, wherein the staining agent in the step (2) is acid alcian blue with a mass concentration of 0.02%.
3. The detection method according to claim 2, wherein the volume ratio of the staining agent to the standard solution is 1 (3-15).
4. The detection method according to claim 1, wherein the acidification treatment in the step (2) is to adjust the pH of the solution obtained by mixing the standard solution and the staining agent to 2-5.
5. The detection method according to claim 1, wherein the standing time in the step (2) is 20-120 min.
6. The detection method according to claim 1, wherein the water to be detected is a water body with a concentration of soluble organic carbon less than 10 mg/L.
7. A method for detecting dissolved acidic polysaccharide in a water body comprises the following steps: firstly, filtering a water body to be detected by using a microporous filter membrane to obtain a pretreated water body; the method for detecting the pretreated water body is characterized in that the concentration of the obtained acidic polysaccharide is the concentration of the dissolved acidic polysaccharide in the water body by using the detection method of any one of claims 1 to 6.
8. The method of claim 7, wherein the material of the microporous filter membrane comprises one of nylon, nylon mixed material and polycarbonate membrane.
9. The detection method according to claim 7 or 8, wherein the pore size of the microfiltration membrane is 0.2 to 0.5 μm.
10. A method for detecting granular acidic polysaccharide in a water body comprises the following steps: dividing the water body to be detected into two parts, and detecting one part of the water body to be detected by using the detection method of any one of claims 1 to 6 to obtain the concentration of total acidic polysaccharides in the water body; detecting another water body to be detected by using the detection method of any one of claims 7 to 9 to obtain the concentration of the dissolved acidic polysaccharide in the water body; and subtracting the concentration of the dissolved acidic polysaccharide in the water body from the concentration of the total acidic polysaccharide in the water body to obtain the concentration of the granular acidic polysaccharide in the water body.
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