CN112945879A - Synergistic detection method for color value of capsaicin and capsorubin - Google Patents
Synergistic detection method for color value of capsaicin and capsorubin Download PDFInfo
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Abstract
The invention provides a color value cooperative detection method of capsaicin and capsorubin, which comprises the steps of oscillating and uniformly mixing red fruit sample powder with the particle size of less than or equal to 0.4mm to be detected with an alcohol solution, standing for 0.5-2h, and carrying out centrifugal treatment to obtain a centrifugal clear liquid; diluting the centrifugal clear liquid to obtain a solution to be detected; detecting the absorbance values of the liquid to be detected at A280nm and A460 nm; obtaining the capsaicin content in the red fruit sample to be detected according to the light absorption value at A280 nm; and obtaining the color value of the capsorubin in the red fruit sample to be detected according to the light absorption value at A460 nm. The detection method can simultaneously measure the color values of the capsaicin and the capsorubin in the red fruit to be detected, has short detection time, can realize high-throughput detection, provides powerful data support for the breeding of new hybrid varieties, and improves the breeding speed of the new varieties; the method has high accuracy of detection results, adopts the alcohol solution as the extracting solution of the capsaicin and the capsorubin, is nontoxic, safe and environment-friendly, is simple and easy to implement, and is beneficial to popularization.
Description
Technical Field
The invention belongs to the technical field of vegetable and fruit quality detection, and particularly relates to a cooperative detection method for color values of capsaicin and capsorubin.
Background
Capsaicin is an alkaloid contained in pepper fruits, and the content of the capsaicin is an important factor for evaluating the quality of the pepper fruits and pepper products and also an index factor for measuring the quality of breeding materials in pepper breeding. The capsorubin is a natural pigment contained in mature capsicum fruits, has bright color, high color value, strong tinting strength, no toxicity and harmlessness, is widely applied in the food industry, is a natural colorant of food, and reflects the capsorubin content in capsicum by using the color value.
At present, the content of capsaicin is detected by adopting national standards GB/T-21266-2007 and NYT 1381-2007. The detection instruments used in the 2 methods are HPLC high performance liquid chromatographs. The high performance liquid chromatograph has the advantages of high separation efficiency, good selectivity, high detection sensitivity, accurate obtained data and the like; the method has the disadvantages of high analysis cost of each sample, high daily maintenance cost of the instrument, sample-by-sample detection for sample analysis, long time for single sample analysis and unsuitability for simultaneous preparation of a large amount of sample materials. During breeding and material screening of new varieties, the pepper fruit sample materials are generally mature and harvested at the same period, if the sample detection time is long, the detection period and complexity are increased, and the quality screening efficiency is not improved. In addition to the two methods, an enzyme-linked immunosorbent assay (an enzyme-linked immunosorbent assay for measuring the content of capsaicin in the vegetable oil and fat of NYT 3109-2017) and a spectrophotometric assay are adopted, but the two methods have a relatively limited application range.
At present, the method for quantitatively analyzing the color value of capsorubin is the most common method of the current national standard GB1886.34-2015 ultraviolet spectrophotometry, a chromophore contained in a component molecular structure of the capsorubin enables the capsorubin to have a unique light absorption area in an ultraviolet-visible light area, and a solution or crystal of the capsorubin has gorgeous red, orange or yellow under visible light. The Color value of capsorubin (Color value units) can be determined according to the characteristics of capsorubin.
However, both the HPLC method for measuring capsaicin and the spectrophotometry method for quantitatively analyzing the color value of capsorubin can only measure the color value of capsaicin or capsorubin separately.
Disclosure of Invention
The invention provides a cooperative detection method for color values of capsaicin and capsorubin, which can realize the simultaneous and rapid detection of the color values of the capsaicin and the capsorubin and improve the screening efficiency of cross breeding.
The invention provides a color value cooperative detection method of capsaicin and capsorubin, which comprises the following steps,
oscillating and uniformly mixing the red fruit sample powder with the grain diameter of less than or equal to 0.4mm to be detected with an alcohol solution, standing for 0.5-2h, and carrying out centrifugal treatment to obtain a centrifugal clear liquid;
diluting the centrifugal clear liquid to obtain a solution to be detected;
detecting the absorbance values of the liquid to be detected at A280nm and A460 nm;
obtaining the capsaicin content in the red fruit sample to be detected according to the light absorption value at A280 nm;
and obtaining the color value of the capsorubin in the red fruit sample to be detected according to the light absorption value at the A460nm position.
Further, the mass fraction of water in the to-be-detected haw sample powder is less than or equal to 10%.
Further, the mass ratio of the mass of the to-be-detected haw sample powder to the mass of the alcoholic solution is 1: 45-65.
Further, the mass concentration of the alcohol solution is 65-75%.
Further, the alcohol solution is at least one of the following: methanol solution and ethanol solution.
Further, in the centrifugal treatment, the centrifugal temperature is 20-30 ℃, and the centrifugal time is 8-12 min.
Further, diluting the centrifugal clear liquid by adopting an alcoholic solution, wherein the mass ratio of the centrifugal clear liquid to the alcoholic solution used for dilution is 1: 3-5.
Further, in the process of uniformly mixing the mixture by oscillation, the rotating speed is 80-120rpm, the time is 35-45min, and the temperature is 45-55 ℃.
Further, the obtaining of the capsaicin content in the red fruit sample to be detected according to the light absorption value at A280nm comprises,
obtaining a regression equation according to capsaicin light absorption values of solutions of capsaicin and dihydrocapsaicin with mass concentrations of 0.2mg/L, 1mg/L, 10mg/L, 20mg/L, 40mg/L, 80mg/L and 160mg/L respectively;
substituting the light absorption value at the A280nm position into the regression equation to obtain the mass concentration of the capsaicin in the liquid to be detected;
and obtaining the capsaicin content in the red fruit sample to be detected according to the mass concentration of the capsaicin in the liquid to be detected.
Further, the obtaining of the color value of the capsorubin in the red fruit sample to be tested according to the light absorption value at A460nm comprises,
substituting the absorbance value at A460nm into equation E1％ 1cmObtaining the color value of the capsorubin in the red fruit sample to be tested, wherein E is A460 f 1000/(m p 100), and E is1％ 1cmThe color value of the capsorubin in the red fruit sample to be detected is represented, A460 represents the light absorption value of the liquid to be detected at A460nm, f represents the dilution multiple, m represents the quality of the red fruit sample powder to be detected, and p represents the drift diameter length of an enzyme label plate used when the light absorption value is detected.
One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:
the invention provides a cooperative detection method of color value of capsaicin and capsorubin, which is characterized in that red fruit sample powder to be detected is dissolved by adopting an alcoholic solution and then is subjected to centrifugal treatment, supernatant liquid subjected to centrifugal treatment is diluted to be used as liquid to be detected, light absorption values of the liquid to be detected at A280nm and A460nm are detected, and the content of capsaicin in the red fruit sample to be detected is obtained according to the light absorption value at A280 nm; and obtaining the color value of the capsorubin in the red fruit sample to be detected according to the light absorption value at A460 nm. The detection method can simultaneously measure the color values of the capsaicin and the capsorubin in the red fruit to be detected, has short detection time and high efficiency, can realize high-throughput detection, can meet the screening work of a large amount of materials of sample materials in the harvest period, and provides powerful data support for breeding new species of hybrid breeding, thereby improving the screening efficiency, being beneficial to expanding the number of hybrid combinations of breeding workers and improving the breeding speed of the new species; the method has high accuracy of detection results, adopts the alcohol solution as the extraction solution of the capsaicin and the capsorubin, is nontoxic, safe and environment-friendly, is simple and easy to implement, and is beneficial to popularization.
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In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on the drawings without creative efforts.
FIG. 1 is a process diagram of a cooperative detection method of capsaicin and capsorubin provided by the invention;
FIG. 2 is a plot of the fitted regression lines made for samples Nos. 5-28 for example 2 and comparative example 3.
Detailed Description
The present invention will be described in detail below with reference to specific embodiments and examples, and the advantages and various effects of the present invention will be more clearly apparent therefrom. It will be understood by those skilled in the art that these specific embodiments and examples are provided to illustrate the invention, and not to limit the invention.
Throughout the specification, unless otherwise specifically noted, terms used herein should be understood as having meanings as commonly used in the art. Accordingly, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. If there is a conflict, the present specification will control.
Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or can be prepared by an existing method.
It is noted that, in this document, relational terms such as "first" and "second," and the like, may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions.
In order to solve the technical problems, the embodiment of the invention provides the following general ideas:
the invention provides a color value cooperative detection method of capsaicin and capsorubin, which is combined with figure 1 and comprises the following steps,
s1, oscillating and uniformly mixing the red fruit sample powder with the grain size of less than or equal to 0.4mm and the alcoholic solution, standing for 0.5-2h, and centrifuging to obtain a centrifugal clear solution;
s2, diluting the centrifugal clear liquid to obtain a liquid to be detected;
s3, detecting the absorbance values of the liquid to be detected at A280nm and A460 nm;
s4, obtaining the capsaicin content in the red fruit sample to be detected according to the light absorption value at the A280nm position; and obtaining the color value of the capsorubin in the red fruit sample to be detected according to the light absorption value at the A460nm position.
The invention provides a cooperative detection method of color value of capsaicin and capsorubin, which is characterized in that red fruit sample powder to be detected is dissolved by adopting an alcoholic solution and then is subjected to centrifugal treatment, supernatant liquid subjected to centrifugal treatment is diluted to be used as liquid to be detected, light absorption values of the liquid to be detected at A280nm and A460nm are detected, and the content of capsaicin in the red fruit sample to be detected is obtained according to the light absorption value at A280 nm; and obtaining the color value of the capsorubin in the red fruit sample to be detected according to the light absorption value at A460 nm. The detection method can simultaneously measure the color values of the capsaicin and the capsorubin in the red fruit to be detected, has short detection time and high efficiency, can realize high-throughput detection, can meet the screening work of a large amount of materials of sample materials in the harvest period, and provides powerful data support for breeding new species of hybrid breeding, thereby improving the screening efficiency, being beneficial to expanding the number of hybrid combinations of breeding workers and improving the breeding speed of the new species; the method is simple and easy to implement, and is favorable for popularization. The red fruit sample to be tested can be hot pepper.
In the invention, the unit of the capsaicin content is mg/g or g/kg, and the unit mass of the capsaicin in the red fruit sample powder to be detected is expressed. The grain size of the red fruit sample powder to be detected is controlled, so that the mixing of the red fruit sample powder and an alcohol solution can be accelerated, and capsaicin and capsorubin can be extracted.
As an implementation manner of the embodiment of the invention, the mass fraction of water in the to-be-detected haw sample powder is less than or equal to 10%.
The water content in the red fruit sample powder to be detected is controlled to improve the quality of the sample, and the fruit is mildewed due to the overhigh water content.
As an implementation manner of the embodiment of the present invention, a mass ratio of the to-be-measured haw sample powder to the alcoholic solution is 1: 45-65. The alcohol solution is mixed with the red fruit sample powder to be tested to extract capsaicin and capsorubin, so as to prepare the liquid to be tested containing the capsaicin and the capsorubin.
As an implementation manner of the embodiment of the invention, the mass concentration of the alcoholic solution is 65-75%.
As an implementation manner of the embodiment of the present invention, the alcohol solution is any one of the following: methanol solution and ethanol solution.
As an implementation mode of the embodiment of the invention, in the centrifugal treatment, the centrifugal temperature is 20-30 ℃, and the centrifugal time is 8-12 min. Controlling the adverse effect of overhigh centrifugal temperature, and the adverse effect of overlow centrifugal temperature on extracting the capsaicin and the capsorubin; the short centrifugation time, incomplete extraction of capsaicin and capsorubin, too long centrifugation time and increased energy consumption.
As an implementation mode of the embodiment of the invention, the centrifugal clear liquid is diluted by adopting an alcoholic solution, and the mass ratio of the centrifugal clear liquid to the alcoholic solution used for dilution is 1: 3-5. The dilution is to make the concentration of the diluted solution corresponding to the absorbance of the solution within the effective range of the standard curve.
As an implementation mode of the embodiment of the invention, in the oscillation and uniform mixing, the rotating speed is 80-120rpm, the time is 35-45min, and the temperature is 45-55 ℃. In the present invention, rpm represents a rotation speed per minute. The stirring can shorten the mixing time of the red fruit sample powder to be detected and the alcohol solution, and accelerate the extraction of the capsaicin and the capsorubin.
As an implementation manner of the embodiment of the present invention, the obtaining the capsaicin content in the red fruit sample to be tested according to the light absorption value at a280nm includes,
s401, obtaining a regression equation according to capsaicin light absorption values of solutions of capsaicin and dihydrocapsaicin with mass concentrations of 0.2mg/L, 1mg/L, 10mg/L, 20mg/L, 40mg/L, 80mg/L and 160mg/L respectively;
s402, substituting the light absorption value at the A280nm into the regression equation to obtain the mass concentration of capsaicin in the liquid to be detected;
and S403, obtaining the capsaicin content in the red fruit sample to be detected according to the mass concentration of the capsaicin in the liquid to be detected.
Capsaicin and dihydrocapsaicin are standard chemical reagents, two curves are drawn by taking the concentration of capsaicin and dihydrocapsaicin solutions as vertical coordinates (Y) and the light absorption value of the capsaicin and the dihydrocapsaicin solutions as horizontal coordinates (X) respectively and are regressed respectively to obtain two linear equations, and then the two linear equations are combined into 1. For example, capsaicin taggant Y68.08A 280R20.999 dihydrocapsaicin standard Y-29.18A 280R20.999, the final formula is R of Y97.26 × a28020.999. For example, the linear regression equation may be Y-97.26 × a280, and the absorbance at a280nm is substituted into the formula Y-97.26 × a280 to obtain the mass concentration of capsaicin in the liquid to be measured; wherein A280 represents the light absorption value of the liquid to be measured at A280nm, and Y represents the mass concentration of capsaicin in the liquid to be measured, and the unit is mg/ml.
R2 of formula Y of 97.26 × a280 is 0.999, which has a high degree of fit and a small error. The method comprises the following steps of obtaining the capsaicin quality fraction in a red fruit sample to be detected according to the mass concentration of the capsaicin in the liquid to be detected, and obtaining the capsaicin quality fraction through the following method: substituting the concentration of the capsaicin in the solution to be detected into a formula W ═ Y × V × f/(m × 1000) to obtain the mass fraction of the capsaicin in the red fruit sample to be detected, wherein W is the mass fraction of the capsaicin and has the unit of mg/g; v is the volume of the alcohol solution used for mixing, ml; f is the dilution multiple of 4; m is the mass of the red fruit sample powder to be measured, and the unit is g.
As an implementation manner of the embodiment of the present invention, the obtaining the color value of capsorubin in the red fruit sample to be tested according to the absorbance value at a460nm includes,
substituting the absorbance value at A460nm into equation E1％ 1cmObtaining the color value of the capsorubin in the red fruit sample to be tested, wherein E is A460 f 1000/(m p 100), and E is1％ 1cmThe color value of the capsorubin in the red fruit sample to be detected is represented, A460 represents the light absorption value of the liquid to be detected at A460nm, f represents the dilution multiple, m represents the mass of the red fruit sample powder to be detected, the unit is g, and p represents the drift diameter length of the elisa plate used for detecting the light absorption value, and the unit is cm. The formula for calculating the color number is the light absorption value of 1cm transmitted by the solution with the mass concentration of 1%.
The method for detecting the color value of capsaicin and capsorubin in a synergistic manner according to the present invention will be described in detail with reference to examples, comparative examples and experimental data.
The test material is derived from germplasm resources which are collected and stored by agricultural courtyard of Sichuan province and have high pungency, separation and purification for many years, have high pungency and high color value content, and are 100 parts (shown in Table 1) in total, and belong to annual hot peppers, wherein 10 parts of conical peppers are contained; 99 parts of long pepper, which is subdivided into 7 parts of horn pepper, 11 parts of linear pepper and 72 parts of horn pepper according to the shape of the fruit. The experiment is developed in a new experiment base of horticulture research institute of academy of agricultural sciences of Sichuan province, 100 parts of materials are sown and grown in a multi-span greenhouse in 1 month of 2020, a hole plate matrix mode is adopted, the specification of the hole plate is 50 holes, 2 seeds are planted in each hole, after the seeds are covered by the matrix, a plastic film is used for covering the seeds for heat preservation and moisture preservation, and after the seeds germinate and cotyledon grows out, the film is uncovered. And (3) field planting in the greenhouse in 2020 and 4 months, applying 50kg of compound fertilizer, 80kg of phosphate fertilizer and 100kg of organic fertilizer to every 667 square meters before field planting, wherein the field application is adopted, the width of the chamber is 80cm, and the width of the ditch is 60 cm. And (3) carrying out double-row field planting on the compartment surface, wherein 1 plant is planted in each hole, the plant spacing is 40cm, and 20 plants are planted in each material. Harvesting 100 parts of materials in total after 8 months, wherein the materials are numbered as 1, 2 and 3 … … 100, and dividing the red fruit sample of each part of planting material into 2 parts, which are respectively named as a and b; the fruit at the 4 th to 6 th nodes of the main branches of the plant fruiting bodies is selected as a sample for each material.
TABLE 1
Example 1
Preparing haw powder:
weighing the fresh samples (10 paprika red fruits) with the number of 1-4 and named as a, then filling the fresh samples into a kraft paper bag according to the number, drying the fresh samples to constant mass at 65 ℃, weighing the fresh samples, crushing the peel into powder by using a small crusher, and sieving the powder by using a 40-mesh sieve to obtain the powder for detecting the color value of capsorubin and capsaicin. The powders were numbered and 0.10000 was weighed out on a ten thousandth balance (Siderris, Germany) into 10ml centrifuge tubes and the experiment was repeated 3 times.
Preparing a solution to be detected:
adding 6ml of 70% ethanol (Sigma-Aldrich for HPLC, 99.9% or more) into the 4 centrifuge tubes, and shaking in a constant temperature shaker at 50 deg.C at 100rpm for 40 min; the mixture was taken out from the shaker, loaded onto a centrifuge rack and allowed to stand in the dark for 1 hour, and then the centrifuge tube samples were put into a centrifuge in batches at 25 ℃ and 8000g for 10 minutes to obtain 4 parts by volume of 6ml (powder mass: ethanol 1:60) of supernatant.
And respectively adding ethanol into 4 parts of supernatant to dilute, wherein the mass ratio of the supernatant to the ethanol is 1:4, and shaking and uniformly mixing to obtain 4 parts of solution to be detected. The 4 solutions to be tested were spotted in 200. mu.l in order on a 96-well UV microplate (Greiner UV-Star UV Special type, Germany) and the plate was set on a full-band reader (Saimer Feishale multiskanGO). Before loading the 96-well plate into the instrument, ensuring the computer to be normal, defining the name of the micropore in the corresponding software Skanit of the computer, setting the wave sections of A280nm and A460nm, clicking the option of accurately reading the light absorption value, and obtaining the light absorption values of the liquid to be detected at A280nm and A460nm after the light absorption value of the whole 96-well plate is read for less than 60 s. The absorbance values for a280nm, numbered 1 and 2, were 0.1374 and 0.1460, respectively, and the absorbance values for a460nm were 0.3172 and 0.2499, respectively.
Calculating the content of capsaicin:
the obtained absorbance values at the positions of 4 a280nm are respectively substituted into a standard curve calculation equation Y of 97.26 × a280, and capsaicin concentrations of 4 solutions to be measured are obtained, wherein the capsaicin concentrations of the solutions to be measured with numbers of 1 and 2 are 13.36352 and 14.19996 respectively. Wherein Y is the concentration mg/ml of capsaicin in the sample.
The capsaicin contents in 4 samples were obtained by substituting the capsaicin concentrations in 4 solutions to be tested into the formula W ═ Y × V × f/(m × 1000), as shown in table 2. W is the capsaicin content, and the unit is mg/g; v is 70% ethanol solution; f is the dilution multiple of 4; and m is the sample weighing mass.
And (3) calculating the color value of capsorubin:
substituting the obtained 4 absorbance values at A460nm into standard curve calculation equation E1％ 1cmObtaining the color value of the capsorubin of 4 parts of liquid to be tested, wherein A460 is the absorbance value of a sample at 460 nm; f is the dilution multiple; m is the sample weighing mass; p is the length of the drift diameter of the 96-well enzyme label plate of 0.58 cm.
Example 2
Example 2 the same as example 1 except that the remaining sample numbers 5 to 100 were used as the test sample and the extraction solution was methanol solution instead of ethanol solution, in example 2 with reference to example 1. The results of the measurements are shown in Table 3.
Comparative example 1
Comparative example 1 provides a method for measuring the capsaicin content, which is detected by the capsaicin content method provided by the national standard GB/T-21266-2007, the samples are named as b and are numbered 1 and 2, and the results are shown in Table 2.
Comparative example 2
Comparative example 2 provides a method for measuring the color value of capsorubin, which uses an extract of capsorubin from acetone, and a spectrophotometer to test the same, using the samples designated as b and numbered 3 and 4. The results are shown in Table 2.
Comparative example 3
Comparative example 3 provides a method for measuring the capsaicin content, which is used for detecting the capsaicin content provided by the national standard GB/T-21266-2007, and the used sample is the sample named as the sample with the number of 5-28 in example 2. The results are shown in Table 3.
TABLE 2
TABLE 3
TABLE 3
According to the contents in tables 2 and 3, the color value synergic detection method of capsaicin and capsorubin provided by the invention has the advantages that the content of capsaicin in the samples No. 1-2 detected in the example 1, namely the sample No. 1, is respectively 0.753 and 0.829mg/g, the pungency grades are all 6 grades, the detection results of the comparative example 1 on the samples No. 1-2 by adopting the national standard are 0.690 and 0.895mg/g, the pungency grades are all 6 grades, and the content of capsaicin detected by the method provided by the example 1 is close to the detection result of the national standard, and the pungency grade is consistent with the national standard. The color values of the capsorubin of the samples No. 3 to No. 4 measured by the method are respectively 20.735 +/-2.451 and 14.016 +/-0.763, the color values of the capsorubin of the samples No. 3 to No. 4 measured by the comparative example 2 are respectively 22.535 +/-3.036 and 14.222 +/-0.551, and the variance analysis shows that the color values of the capsorubin of the samples No. 3 to No. 4 measured by the example 1 and the color values of the capsorubin of the samples No. 3 to No. 4 measured by the example 2 have no significant difference, which shows that the color values of the capsorubin measured by the method and the extract of the capsorubin of acetone have no significant difference by a standard method of.
Table 3 illustrates the capsaicin contents of the fruit samples obtained in example 2 (inventive method) and comparative example 3 (national standard method) of sample Nos. 5 to 28, respectively. The capsaicin content values of the 24 fruit samples in the comparative example 3 are used as independent variables, the capsaicin content values of the 20 fruit samples in the example are used as variables, a regression relationship is established, a regression curve is fitted, a regression equation y is obtained, the regression equation y is 0.9869x, and a coefficient R is determined20.9788, the variation degree of the capsaicin content obtained by the method is only 2.12% (the value is obtained from 1-0.9788, see figure 2), the determination coefficient is 0.9788 and is close to 1, the method provided by the invention is close to the capsaicin content detected by a national standard method, the accuracy is high, the variance analysis has no obvious difference, and the method can replace the national standard detection method.
Dissolving to-be-detected haw sample powder by adopting an alcoholic solution, carrying out centrifugal treatment, diluting supernatant liquid subjected to centrifugal treatment to be used as to-be-detected liquid, detecting light absorption values of the to-be-detected liquid at A280nm and A460nm, and obtaining the capsaicin content in the to-be-detected haw sample according to the light absorption value at A280 nm; and obtaining the color value of the capsorubin in the red fruit sample to be detected according to the light absorption value at A460 nm. The detection method prepares a sample, the extracting solution microplate reader can simultaneously output two groups of light absorption value data of capsaicin and color value once, can simultaneously measure the color values of the capsaicin and the capsorubin in the red fruit to be detected, has short detection time and high efficiency, and can realize high-throughput detection, for example, the execution detection period of 100 samples is 1 week, the comparison example 3 is that 20 samples only detect the content of the capsaicin, and the execution detection period is 2 weeks. Therefore, the method provided by the invention can meet the screening work of a large amount of materials of sample materials in the harvest period, and provide powerful data support for breeding new species of hybrid breeding, thereby improving the screening efficiency, facilitating the breeding workers to expand the hybrid combination quantity and improving the breeding speed of the new species; the method has high accuracy of detection results, adopts the alcohol solution as the extracting solution of the capsaicin and the capsorubin, is nontoxic, safe and environment-friendly, is simple and easy to implement, and is beneficial to popularization.
Finally, it is also to be noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (10)
1. A color value cooperative detection method of capsaicin and capsorubin is characterized by comprising the following steps,
oscillating and uniformly mixing the red fruit sample powder with the grain diameter of less than or equal to 0.4mm to be detected with an alcohol solution, standing for 0.5-2h, and centrifuging to obtain a centrifugal clear liquid;
diluting the centrifugal clear liquid to obtain a solution to be detected;
detecting the absorbance values of the liquid to be detected at A280nm and A460 nm;
obtaining the capsaicin content in the red fruit sample to be detected according to the light absorption value at A280 nm;
and obtaining the color value of the capsorubin in the red fruit sample to be detected according to the light absorption value at the A460nm position.
2. The method for cooperatively detecting the color values of capsaicin and capsorubin according to claim 1, wherein the mass fraction of water in the powder of the red fruit sample to be detected is less than or equal to 10%.
3. The method for cooperatively detecting color values of capsaicin and capsorubin according to claim 1, wherein the mass ratio of the mass of the red fruit sample powder to be detected to the mass of the alcoholic solution is 1: 45-65.
4. The method for detecting color values of capsaicin and capsorubin cooperatively according to claim 3, wherein the mass concentration of the alcohol solution is 65-75%.
5. The method for detecting color values of capsaicin and capsorubin cooperatively according to claim 1, wherein the alcohol solution is at least one of: methanol solution and ethanol solution.
6. The method for cooperatively detecting color values of capsaicin and capsorubin according to claim 1, wherein the centrifugation temperature is 20-28 ℃ and the centrifugation time is 8-12 min.
7. The method for synergistically detecting color values of capsaicin and capsorubin according to claim 1, wherein the centrifuged clear liquid is diluted by an alcohol solution, and the mass ratio of the centrifuged clear liquid to the diluted alcohol solution is 1: 3-5.
8. The method for cooperatively detecting the color values of capsaicin and capsorubin according to claim 1, wherein the rotating speed is 80-120rpm, the time is 35-45min, and the temperature is 45-55 ℃ in the process of uniformly oscillating and mixing.
9. The method as claimed in claim 1, wherein the obtaining of the content of capsaicin in the red fruit sample to be tested according to the absorbance at A280nm comprises,
obtaining a regression equation according to capsaicin light absorption values of solutions of capsaicin and dihydrocapsaicin with mass concentrations of 0.2mg/L, 1mg/L, 10mg/L, 20mg/L, 40mg/L, 80mg/L and 160mg/L respectively;
substituting the light absorption value at the A280nm position into the regression equation to obtain the mass concentration of the capsaicin in the liquid to be detected;
and obtaining the capsaicin content in the red fruit sample to be detected according to the mass concentration of the capsaicin in the liquid to be detected.
10. The method as claimed in claim 1, wherein the obtaining of the color value of capsorubin in the red fruit sample to be tested according to the absorbance at A460nm comprises,
substituting the absorbance value at A460nm into equation E1％ 1cmObtaining the color value of the capsorubin in the red fruit sample to be tested, wherein E is A460 f 1000/(m p 100), and E is1％ 1cmThe color value of the capsorubin in the red fruit sample to be detected is represented, A460 represents the light absorption value of the liquid to be detected at A460nm, f represents the dilution multiple, m represents the quality of the red fruit sample powder to be detected, and p represents the drift diameter length of an enzyme label plate used when the light absorption value is detected.
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