CN113640280B - Detection reagent for detecting HCN of rubber tree and preparation method thereof - Google Patents
Detection reagent for detecting HCN of rubber tree and preparation method thereof Download PDFInfo
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Abstract
The invention provides a detection reagent for detecting rubber tree HCN, which takes chloroform as a solvent, and each 20-25 mL of chloroform contains 10-15 g of 4,4' -diaminodiphenyl methane, 10-15 g of copper ethylacetoacetate, 2-5 g of sodium alginate polymer and 5-8 g of polyvinylpyrrolidone. The detection reagent prepared by the invention can be directly used for detecting the HCN content of the bark of the rubber tree, has the advantages of quick detection time, accurate result, simple operation, low cost and the like, and can be suitable for detecting the hydrogen cyanide content of a large amount of bark of the rubber tree.
Description
Technical Field
The invention relates to the technical field of biochemical detection, in particular to a detection reagent for detecting rubber tree HCN and a preparation method thereof.
Background
Natural rubber has the irreplaceable advantages of synthetic rubber in the aspects of elasticity, wear resistance, ductility and the like, and has wide application in some important industrial fields (such as aviation, aerospace, heavy-duty car and other manufacturing industries). Brazil rubber tree (Hevea brasiliensis, simply rubber tree) is the main source of natural rubber. Brazil rubber is a plant of the family Euphorbiaceae and is also a major commercial crop in tropical areas of our country. The traditional breeding cycle of Brazilian rubber tree is about 30 years, and early yield prediction is the key to shortening the breeding cycle. Dead skin (tapping panel dryness) is a major factor limiting the yield of rubber trees. According to statistics, the accumulated incidence rate of dead skin in main rubber planting areas in China is between 20% and 40%, the serious area is more than 50%, and the direct economic loss caused by dead skin of rubber trees in each year is up to 30 hundred million yuan. In addition, with the popularization of the high-yield clone and ethephon stimulated rubber production technology, the occurrence rate and the severity of dead bark of the rubber tree tend to rise year by year. Therefore, early prediction of the occurrence of dead skin and timely prevention and intervention are the most effective means for reducing the dead skin hazard at present.
Brazil rubber is a typical cyanogenic plant, and when the rubber tree develops dead skin symptoms, the trunk bark tissue releases a higher amount of HCN, i.e., HCN is associated with dead skin. Thus, early predictions of dead skin can be made by detecting HCN in bark tissue. The HCN test paper is adopted to directly detect the rubber tree leaves, the HCN content in the tree leaves is measured, the bark internal tissue of the rubber tree is different from the rubber tree leaves, various hierarchical structures exist, the color is darker, the HCN test paper is not suitable for detecting the HCN content of the bark of the rubber tree, and the detection result is inaccurate easily, so that a detection reagent suitable for detecting the HCN content of the bark of the rubber tree needs to be prepared, and an important basis is provided for early and accurate prediction of dead bark of the rubber tree.
Disclosure of Invention
Accordingly, the invention aims to provide a detection reagent for detecting the HCN of the rubber tree and a preparation method thereof, and the HCN detection reagent prepared by the invention is directly dripped to the bark of the rubber tree, so that the detection of the HCN content can be completed, and the detection reagent has the advantages of quick detection time, accurate result, simple operation, low cost and the like.
The technical scheme of the invention is realized as follows:
copper 4,4' -diaminodiphenyl methane and ethylacetoacetate can react with hydrogen cyanide to form colored compounds, and thus can be used for detecting the content of hydrogen cyanide in substances, and the chemical reaction equation is as follows:
2CuAc 2 +C 17 H 22 N 2 +2HCN→2CuCN+[C 17 H 21 N 2 ]Ac+3HAc
the bark is divided into a multilayer tissue structure such as an outer epidermis, a cambium, a phloem and the like from outside to inside, the sodium alginate polymer can enable components of the detection reagent to form a cross-linked interpenetrating network, the polyvinylpyrrolidone can remove substances such as protein, tannin and the like in the bark, the influence of nutritional components in the bark on the color development effect of the color developing agent is reduced, meanwhile, the bark has the function of a surfactant, the sodium alginate polymer and the polyvinylpyrrolidone can play a role in stabilizing and promoting the penetration of the detection reagent into the multilayer tissue structure of the bark, and the detection reagent and HCN in the bark tissue can fully act.
A detection reagent for detecting HCN of rubber tree takes chloroform as a solvent, and each 20-25 mL of chloroform contains 10-15 g of 4,4' -diaminodiphenyl methane, 10-15 g of copper ethylacetoacetate, 2-5 g of sodium alginate polymer and 5-8 g of polyvinylpyrrolidone.
Further describing, the preparation method of the sodium alginate polymer comprises the following steps:
step Sa: adding sodium alginate into a sodium periodate solution with the mass concentration of 0.25-0.30%, and stirring for 1-2 hours in a dark place to obtain a reaction solution;
step Sb: adding 5-8% calcium chloride aqueous solution into the reaction solution, stirring for 50-60 min, adding 4-6% sodium polyacrylate solution, and performing ultrasonic emulsification at 25-35 ℃ for 5-10 min to obtain emulsion;
step Sc: centrifuging the emulsion at 3000-4000 rpm for 20-30 min, and spray drying the precipitate at 75-85 ℃ for 25-35 min to obtain the sodium alginate polymer.
Further illustrating that the mass ratio of the sodium alginate to the sodium periodate to the calcium chloride to the sodium polyacrylate is 0.02-0.025: 0.2 to 0.3: 55-65: 0.5 to 0.8.
Further described, a method for preparing a detection reagent for detecting rubber tree HCN, comprising the steps of:
(1) Adding 4,4' -diaminodiphenyl methane and copper acetoacetate into chloroform, and stirring for 10-15 min to obtain a mixed solution;
(2) Taking the mixed solution, adding polyvinylpyrrolidone, and carrying out ultrasonic treatment for 25-30 min at an ultrasonic frequency of 2.5-3 MHz to obtain a primary solution;
(3) Adding sodium alginate polymer into the primary liquid, shearing at 50-55 deg.c and shearing speed of 5000-8000 rpm for 12-18 min, and cooling to obtain the detection reagent.
Further, the application method of the detection reagent for detecting the HCN of the rubber tree is characterized in that the detection reagent is dripped on the bark of the rubber tree and is kept stand for 5-10 min, so that the detection of the HCN of the rubber tree is completed.
Compared with the prior art, the invention has the beneficial effects that: the invention can realize the detection of the HCN content of the bark of the rubber tree by the scientifically prepared HCN detection reagent, and has clear detection result and high prediction accuracy.
According to the invention, substances such as protein, tannin and the like in bark can be removed by adopting the sodium alginate polymer and the polyvinylpyrrolidone, the influence of nutrient components in bark on the color development effect of the color developing agent is reduced, the polyvinylpyrrolidone can improve the compatibility among components of the detection reagent and improve the dispersibility of liquid, the function of stabilizing the detection reagent is achieved, the sodium alginate polymer and the polyvinylpyrrolidone cooperate with each other, the detection reagent is promoted to quickly and uniformly permeate into bark tissues, the detection reagent and HCN in bark are promoted to fully act, the color development effect is more obvious, and the result is more accurate.
Drawings
FIG. 1 is a schematic view of the bark of the present invention divided into an interior and an exterior
Detailed Description
In order to better understand the technical content of the present invention, the following provides specific examples to further illustrate the present invention.
The experimental methods used in the embodiment of the invention are conventional methods unless otherwise specified.
Materials, reagents, and the like used in the examples of the present invention are commercially available unless otherwise specified.
Example 1
A detection reagent for detecting the HCN of the rubber tree takes chloroform as a solvent, and each 22mL of chloroform contains 12g of 4,4' -diaminodiphenylmethane, 12g of copper acetoacetate, 3g of sodium alginate polymer and 5g of polyvinylpyrrolidone.
The preparation method of the sodium alginate polymer comprises the following steps:
step Sa: the mass ratio of the sodium alginate to the sodium periodate solution to the calcium chloride aqueous solution to the sodium polyacrylate solution is 0.022:0.2:60:0.6, taking 0.022g of sodium alginate, adding 0.2g of sodium periodate solution with the mass concentration of 0.28%, and stirring for 2 hours in a dark place to obtain a reaction solution;
step Sb: 60g of calcium chloride water solution with the mass concentration of 6% is added into the reaction solution, the mixture is stirred for 55min, 0.6g of sodium polyacrylate solution with the mass concentration of 5% is added, and the mixture is subjected to ultrasonic emulsification at 30 ℃ for 8min to obtain emulsion;
step Sc: centrifuging the emulsion at 3500rpm for 25min, collecting precipitate, and spray drying at 80deg.C for 30min to obtain sodium alginate polymer.
A preparation method of a detection reagent for detecting rubber tree HCN comprises the following steps:
(1) Adding 4,4' -diaminodiphenyl methane and copper acetoacetate into chloroform, and stirring for 12min to obtain a mixed solution;
(2) Adding polyvinylpyrrolidone into the mixed solution, and performing ultrasonic treatment at frequency of 2.8MHz for 28min to obtain primary solution;
(3) Adding sodium alginate polymer into the primary liquid, shearing at 55 ℃ with the shearing speed of 6500rpm for 15min, and cooling to obtain the detection reagent.
A using method of a detection reagent for detecting the HCN of the rubber tree comprises the steps of dripping the detection reagent onto the bark of the rubber tree, and standing for 8min to finish the detection of the HCN of the rubber tree.
Example 2
A detection reagent for detecting the HCN of the rubber tree takes chloroform as a solvent, and each 20mL of chloroform contains 10g of 4,4' -diaminodiphenylmethane, 10g of copper acetoacetate, 2g of sodium alginate polymer and 5g of polyvinylpyrrolidone.
The preparation method of the sodium alginate polymer comprises the following steps:
step Sa: the mass ratio of the sodium alginate to the sodium periodate solution to the calcium chloride aqueous solution to the sodium polyacrylate solution is 0.025:0.3:65:0.8 g of sodium alginate is taken, 0.3g of sodium periodate solution with the mass concentration of 0.28% is added, and the mixture is stirred for 2 hours in a dark place to obtain a reaction solution;
step Sb: adding 65g of calcium chloride aqueous solution with the mass concentration of 6% into the reaction solution, stirring for 55min, adding 0.8g of sodium polyacrylate solution with the mass concentration of 5%, and performing ultrasonic emulsification at 30 ℃ for 8min to obtain emulsion;
step Sc: centrifuging the emulsion at 3500rpm for 25min, collecting precipitate, and spray drying at 80deg.C for 30min to obtain sodium alginate polymer.
Example 3
A detection reagent for detecting the HCN of the rubber tree takes chloroform as a solvent, and each 25mL of chloroform contains 15g of 4,4' -diaminodiphenylmethane, 15g of copper acetoacetate, 5g of sodium alginate polymer and 8g of polyvinylpyrrolidone.
The preparation method of the sodium alginate polymer comprises the following steps:
step Sa: the mass ratio of the sodium alginate to the sodium periodate solution to the calcium chloride aqueous solution to the sodium polyacrylate solution is 0.020:0.2:55:0.5 g of sodium alginate is taken, 0.3g of sodium periodate solution with the mass concentration of 0.30% is added, and the mixture is stirred for 2 hours in a dark place to obtain a reaction solution;
step Sb: adding 55g of calcium chloride water solution with the mass concentration of 8% into the reaction solution, stirring for 60min, adding 0.5g of sodium polyacrylate solution with the mass concentration of 5%, and performing ultrasonic emulsification at 35 ℃ for 8min to obtain emulsion;
step Sc: centrifuging the emulsion at 4000rpm for 25min, collecting precipitate, and spray drying at 85deg.C for 30min to obtain sodium alginate polymer.
A method for preparing a detection reagent for detecting HCN of rubber tree of examples 2 and 3, comprising the steps of:
(1) Adding 4,4' -diaminodiphenyl methane and copper acetoacetate into chloroform, and stirring for 12min to obtain a mixed solution;
(2) Adding polyvinylpyrrolidone into the mixed solution, and performing ultrasonic treatment at frequency of 3MHz for 30min to obtain primary solution;
(3) Adding sodium alginate polymer into the primary liquid, shearing at 50 ℃, shearing speed 8000rpm for 18min, and cooling to obtain the detection reagent.
A method for using a detection reagent for detecting the HCN of the rubber tree comprises the steps of dripping the detection reagent onto the bark of the rubber tree, and standing for 10min to finish the detection of the HCN of the rubber tree.
Comparative example 1
A detection reagent for detecting HCN of rubber tree according to embodiment 1, which is different in that: sodium alginate polymer 0.5g and polyvinylpyrrolidone 2g.
Comparative example 2
A detection reagent for detecting HCN of rubber tree according to embodiment 1, which is different in that: 5g of 4,4' -diaminodiphenylmethane, 8g of copper acetoacetate and 15mL of chloroform.
Comparative example 3
A detection reagent for detecting HCN of rubber tree according to embodiment 1, which is different in that: 18g of 4,4' -diaminodiphenylmethane, 20g of copper acetoacetate, 30mL of chloroform, 10g of sodium alginate polymer and 12g of polyvinylpyrrolidone.
Comparative example 4
A detection reagent for detecting HCN of rubber tree according to embodiment 1, which is different in that: the preparation method of the detection reagent is different, chloroform is taken, 4' -diaminodiphenyl methane and copper ethylacetoacetate are added, and the mixture is stirred for 10 to 15 minutes to obtain a mixed solution; adding sodium alginate polymer into the mixed solution, performing thermal shearing, and cooling to obtain a primary solution; and adding polyvinylpyrrolidone into the primary liquid, and performing ultrasonic treatment to obtain the detection reagent.
1. Experimental procedure
Selecting barks of 5 rubber trees H1-H5 without dead skin and 5 rubber trees T1-T5 with different degrees of dead skin, wherein T1 represents the dead skin length of a cutting surface of the rubber tree below 2cm, T2 represents the dead skin length of the cutting surface between 2cm and 1/4 of the secant length, T3 represents the dead skin length of the cutting surface between 1/4 and 1/2 of the secant length, T4 represents the dead skin length of the cutting surface between 1/2 and 3/4 of the secant length, T5 represents the dead skin length of the cutting surface between 3/4 and the full line dead skin, dividing the barks into inner parts and outer parts according to FIG. 1, and respectively performing a color reaction experiment, wherein the experimental results are shown in Table 1:
note that: + represents color development, -represents no color development
As is clear from the above table, the color development part is the bark inside part, the barks of H1-H5 are not developed, the barks of T1-T5 are developed in bluish green to different degrees, the rubber trees of examples 1-3 which die in different degrees are developed in different colors, the rubber tree of comparative example 1 is developed in T4 and T5 only, the rubber tree of comparative example 2 is developed in T5 only, none of comparative example 3 is developed in T3-T5 of comparative example 4.
In conclusion, the invention has the advantages of quick detection of the HCN content of the bark of the rubber tree, accurate result, simple operation, low cost and the like, and can be applied to the field detection of the production of the rubber tree by controlling the proportion of the 4,4' -diaminodiphenylmethane, the copper acetoacetate, the chloroform, the sodium alginate polymer and the polyvinylpyrrolidone.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.
Claims (6)
1. A detection reagent for detecting HCN in a rubber tree, characterized by: chloroform is used as a solvent, and each 20-25 mL of chloroform contains 10-15 g of 4,4' -diaminodiphenylmethane, 10-15 g of copper ethylacetoacetate, 2-5 g of sodium alginate polymer and 5-8 g of polyvinylpyrrolidone;
the preparation method of the sodium alginate polymer comprises the following steps:
step S1: adding sodium alginate into a sodium periodate solution with the mass concentration of 0.25-0.30%, and stirring in a dark place for 1-2h to obtain a reaction solution;
step S2: adding a calcium chloride aqueous solution with the mass concentration of 5-8% into the reaction solution, stirring for 50-60 min, adding a sodium polyacrylate solution with the mass concentration of 4-6%, and performing ultrasonic emulsification at 25-35 ℃ for 5-10 min to obtain an emulsion;
step S3: and centrifuging the emulsion at a rotating speed of 3000-4000 rpm for 20-30 min to obtain a precipitate, and spray-drying the precipitate at 75-85 ℃ for 25-35 min to obtain the sodium alginate polymer.
2. A test reagent for detecting HCN in rubber trees according to claim 1, characterized in that: the mass ratio of the sodium alginate to the sodium periodate solution to the calcium chloride aqueous solution to the sodium polyacrylate solution is 0.02-0.025: 0.2-0.3: 55-65: 0.5 to 0.8.
3. The method for preparing a detection reagent for detecting HCN of rubber tree according to claim 1 or 2, characterized in that: the method comprises the following steps:
(1) Adding 4,4' -diaminodiphenyl methane and copper acetoacetate into chloroform, and stirring for 10-15 min to obtain a mixed solution;
(2) Adding polyvinylpyrrolidone into the mixed solution, and performing ultrasonic treatment to obtain a primary solution;
(3) And adding sodium alginate polymer into the primary liquid, carrying out thermal shearing, and cooling to obtain the detection reagent.
4. A method for preparing a detection reagent for detecting HCN in rubber trees according to claim 3, characterized in that: the ultrasonic frequency of the ultrasonic treatment is 2.5-3 MHz, and the time is 25-30 min.
5. A method for preparing a detection reagent for detecting HCN in rubber trees according to claim 3, characterized in that: the thermal shearing is carried out at 50-55 ℃ with the shearing speed of 5000-8000 rpm for 12-18 min.
6. The method for using the detection reagent for detecting the HCN of the rubber tree according to any one of claims 1 to 5, which is characterized in that: and (3) dropwise adding the detection reagent onto the bark of the rubber tree, and standing for 5-10 min to finish detection of the HCN of the rubber tree.
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