CN108051417B - Fluorescence inspection method of edible areca-nut brine - Google Patents

Fluorescence inspection method of edible areca-nut brine Download PDF

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CN108051417B
CN108051417B CN201711346571.0A CN201711346571A CN108051417B CN 108051417 B CN108051417 B CN 108051417B CN 201711346571 A CN201711346571 A CN 201711346571A CN 108051417 B CN108051417 B CN 108051417B
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brine
fluorescence
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edible
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CN108051417A (en
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曾荣今
成奋民
陈建
陈述
龙云飞
高倩
刘思远
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Hunan University of Science and Technology
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N2021/6417—Spectrofluorimetric devices

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Abstract

The invention discloses a fluorescence inspection method of edible betel nut brine. Aiming at the defects of the current edible areca-nut brine inspection method, the invention establishes a brine fluorescence inspection method by utilizing the correlation between fluorescence and brine quality and flavor according to the fluorescence characteristics of brine. The detection method disclosed by the invention is high in sensitivity, objective and accurate in result and simple to operate, and has important application value in the aspects of detection analysis, guidance preparation and the like of edible betel nut brine.

Description

Fluorescence inspection method of edible areca-nut brine
Technical Field
The invention relates to the technical field of edible betel nut detection and analysis, in particular to a fluorescence detection method of edible betel nut brine.
Background
At present, the large betel nut processing enterprises in Hunan province reach more than 30 families, the annual output is more than 20 ten thousand tons, and the annual output value exceeds 100 million yuan. Hunan Tan is used as a source of edible betel nuts, and the betel nut processing industry becomes the leading industry in Hunan province. With the rapid development of the betel nut industry, the betel nut processing industry has become a support industry of quan, and simultaneously, the betel nut planting industry and the industrial development of betel nuts in China are greatly promoted.
The production process of edible betel nuts is complex, and comprises the steps of selecting seeds, cleaning, airing, fermenting, airing, giving fragrance, cutting the seeds, adding brine, airing and the like, wherein the brine is the main characteristic of the Hunan betel nut food. The brine is prepared by reacting calcium hydroxide, maltose and water under appropriate conditions, and then adding additives in various proportions to the mixture. The early reaction of the brine is a complex and violent exothermic reaction of maltose under high temperature and alkaline conditions, and a caramelized product with strong fluorescence is formed. The fluorescence parameters (fluorescence intensity, emission wavelength and the like) are stable, and the quality and the flavor of the brine are also stable.
At present, each areca nut enterprise has respective quality and flavor requirements on edible areca nut brine, but the quality and the flavor are mainly judged according to experience and sense, the judgment result is different from person to person, the subjectivity is strong, and no objective detection method is available. Fluorescence analysis methods have received much attention because of their high sensitivity, non-destructive and real-time in situ detection. Therefore, the establishment of an objective edible betel nut brine fluorescence inspection method has important theoretical and practical significance.
Disclosure of Invention
Aiming at the defects of the current edible areca-nut brine inspection method, the invention provides the edible areca-nut brine fluorescence inspection method which is high in sensitivity, objective and accurate in result and simple to operate.
The purpose of the invention is realized by the following modes:
a fluorescence detection method of edible areca-nut brine specifically comprises the following steps:
(1) preparing a brine test solution: accurately weighing 1.000 g of a series of brine samples respectively, adding a proper amount of distilled water for dissolving, then moving the brine samples into a 100 mL volumetric flask, carrying out constant volume to 100 mL by using the distilled water, carrying out high-speed centrifugation to obtain a series of brine clear solutions with the mass concentration (namely the mass fraction) of 1%, sucking 10mL of the brine clear solutions with the mass concentration of 1% and adding the brine clear solutions into the 100 mL volumetric flask, then adding 10mL of 0.2mol/L phosphate buffer solution (pH = 7.4), adding the distilled water for constant volume to a scale, and shaking up to obtain a series of brine test solutions with the mass concentration of 0.1%;
(2) measurement of brine fluorescence spectrum: adding 3.0 mL of 0.1% bittern test solution into 1cm thick 4 mL cuvette, respectively measuring fluorescence spectra of series bittern, setting fluorescence parameter as lambdaex=350 nm, scanning wavelength range 365-: ex =5, Em = 5;
(3) analysis of fluorescence intensity and emission wavelength of brine: and respectively taking the fluorescence intensity values at the maximum emission wavelengths according to the fluorescence spectra to obtain a fluorescence intensity graph corresponding to the series of brines, and respectively taking the emission wavelengths at the maximum fluorescence intensities to obtain a maximum emission wavelength graph corresponding to the series of brines.
Further, the series of brines in the steps (1), (2) and (3) include brines prepared at different reaction temperatures, brines prepared at different reaction times, brines prepared at different material ratios, brines prepared in different feeding modes, and brines of different batches under the same condition.
The fluorescence detection method of edible areca-nut brine is applied to the detection and analysis of the edible areca-nut brine.
The invention has the beneficial effects that:
according to the fluorescence characteristics of the brine, the correlation between fluorescence and the quality and flavor of the brine is utilized to establish a fluorescence detection method of the brine. The method has the advantages of high sensitivity, objective and accurate result and simple operation. Has important application value in the aspects of detection and preparation of edible areca-nut brine and the like.
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FIG. 1 shows fluorescence spectra of brines prepared at different reaction temperatures: the mass concentration of the sample was 0.1%, and the fluorescence parameter was set to λex=350 nm, scanning wavelength range 365-: ex =5, Em = 5.
FIG. 2 is a graph showing the influence of reaction temperature on the fluorescence intensity of brine: the mass concentration of the sample was 0.1%, and the fluorescence parameter was set to λex=350 nm，λem=440 nm, slit width: ex =5, Em = 5.
FIG. 3 is a graph of the effect of reaction temperature on the maximum emission wavelength of brine: the mass concentration of the sample was 0.1%. Fluorescence parameter set to λex=350 nm, slit width: ex =5, Em = 5.
FIG. 4 is a graph of fluorescence spectra of brines with different reaction times: the mass concentration of the sample was 0.1%, and the fluorescence parameter was set to λex=350 nm, slit width: ex =5, Em = 5.
FIG. 5 is a graph showing the effect of different reaction times on the fluorescence intensity of brine: the mass concentration of the sample was 0.1%, and the fluorescence parameter was set to λex=350 nm，λem=440 nm, slit width: ex =5, Em = 5.
FIG. 6 is a graph of the effect of different reaction times on the maximum emission wavelength of brine: the mass concentration of the sample was 0.1%, and the fluorescence parameter was set to λex=350 nm, slit width: ex =5, Em = 5.
FIG. 7 is a graph of the maximum emission wavelength of different batches of orthohalide prepared in duplicate: the mass concentration of the sample was 0.1%. Fluorescence parameterIs set to lambdaex=350 nm, slit width: ex =5, Em = 5.
Fig. 8 is a plot of fluorescence intensity (fig. 8A) and maximum emission wavelength (fig. 8B) for different batches of brine prepared under the same conditions: the mass concentration of the sample was 0.1%, and the fluorescence parameter was set to λex=350 nm, slit width: ex =5, Em = 5.
Detailed Description
The invention is described in further detail below with reference to the figures and the embodiments.
Example 1
A fluorescence detection method of edible areca-nut brine specifically comprises the following steps:
(1) preparing a brine test solution: accurately weighing 1.000 g of brine samples prepared at different reaction temperatures respectively, adding a proper amount of distilled water for dissolving, then transferring into a 100 mL volumetric flask, using distilled water for constant volume to 100 mL, and then carrying out high-speed centrifugation to obtain a series of 1% brine clear solutions. Sucking 10mL of 1% brine clear liquid, adding the brine clear liquid into a 100 mL volumetric flask, adding 10mL of 0.2mol/L phosphate buffer solution (pH = 7.4), adding distilled water to a constant volume to a scale, and shaking up to obtain 0.1% brine test solution prepared at different reaction temperatures;
(2) measurement of brine fluorescence spectrum: 3.0 mL of 0.1% brine test solution is added into a 4 mL cuvette with the thickness of 1cm, and fluorescence spectra of the brine prepared at different reaction temperatures are respectively measured. (FIG. 1). Fluorescence parameter set to λex=350 nm, scanning wavelength range 365-: ex =5, Em = 5. As can be seen from FIG. 1, the fluorescence spectra of the brines with different reaction temperatures are very different.
(3) Analysis of fluorescence intensity and emission wavelength of brine: according to the fluorescence spectrum, the fluorescence intensity values at the maximum emission wavelengths are respectively taken to obtain a relation graph (figure 2) of the influence of the reaction temperature on the fluorescence intensity of the brine, and the emission wavelengths at the maximum fluorescence intensities are respectively taken to obtain a relation graph (figure 3) of the influence of the reaction temperature on the maximum emission wavelengths of the brine. As can be seen from FIGS. 2 and 3, the reaction temperature has a significant effect on both the fluorescence intensity and the maximum emission wavelength of the brine, and in general, the fluorescence intensity and the maximum emission wavelength increase with the increase of the temperature, and the fluorescence intensity and the fluorescence emission wavelength can be used as the detection indexes of the brine.
Example 2
A fluorescence detection method of edible areca-nut brine specifically comprises the following steps:
(1) preparing a brine test solution: accurately weighing 1.000 g of brine samples prepared in different reaction times respectively, adding a proper amount of distilled water for dissolving, then transferring into a 100 mL volumetric flask, carrying out constant volume to 100 mL by using the distilled water, carrying out high-speed centrifugation to obtain a series of 1% brine clear liquid, sucking 10mL of 1% brine clear liquid, adding into the 100 mL volumetric flask, adding 10mL of 0.2mol/L phosphate buffer solution (pH = 7.4), adding the distilled water for constant volume to a scale, and shaking uniformly to obtain 0.1% brine test solutions prepared in different reaction times;
(2) measurement of brine fluorescence spectrum: 3.0 mL of 0.1% brine test solution was added to a 1cm thick 4 mL cuvette and the fluorescence spectra were measured separately to obtain fluorescence spectra of the brines prepared at different reaction times (FIG. 4). Fluorescence parameter set to λex=350 nm, scanning wavelength range 365-: ex =5, Em = 5. As can be seen from fig. 4, the fluorescence spectra of the brines with different reaction times are greatly different.
(3) Analysis of fluorescence intensity and emission wavelength of brine: according to the fluorescence spectrum, the fluorescence intensity values at the maximum emission wavelengths are respectively taken to obtain a relationship graph (figure 5) of the influence of different reaction times on the fluorescence intensity of the brine, and the emission wavelengths at the maximum fluorescence intensities are respectively taken to obtain a relationship graph (figure 6) of the influence of different reaction times on the maximum emission wavelengths of the brine. As can be seen from fig. 5 and 6, the reaction time has a large influence on both the fluorescence intensity and the maximum emission wavelength of the brine, and generally speaking, the longer the reaction time is, the higher the fluorescence intensity and the maximum emission wavelength are, and further, the fluorescence intensity and the fluorescence emission wavelength can be used as the detection index of the brine.
Example 3
A fluorescence detection method of edible areca-nut brine specifically comprises the following steps:
(1) preparing a brine test solution: accurately weighing 1.000 g of brine samples of different batches prepared under the same conditions respectively, adding a proper amount of distilled water for dissolving, then transferring into a 100 mL volumetric flask, using distilled water for constant volume to 100 mL, and then carrying out high-speed centrifugation to obtain a series of 1% brine clear solutions. Sucking 10mL of 1% brine clear liquid, adding the brine clear liquid into a 100 mL volumetric flask, adding 10mL of 0.2mol/L phosphate buffer solution (pH = 7.4), adding distilled water to a constant volume to a scale, and shaking up to obtain different batches of 0.1% brine test solutions prepared under the same condition;
(2) measurement of brine fluorescence spectrum: 3.0 mL of 0.1% brine test solution was added to a 1cm thick 4 mL cuvette, and the fluorescence spectra of the series of brines were measured, respectively, to obtain fluorescence spectra of brines of different batches prepared under the same conditions (FIG. 7). Fluorescence parameter set to λex=350 nm, scanning wavelength range 365-: ex =5, Em = 5. As can be seen from fig. 7, the fluorescence spectra of different batches of brine prepared under the same conditions are very close.
(3) Analysis of fluorescence intensity and emission wavelength of brine: according to the fluorescence spectrum, the fluorescence intensity values at the maximum emission wavelengths are respectively taken to obtain fluorescence intensity maps (fig. 8A) of different batches of brine prepared under the same condition, and the emission wavelengths at the maximum fluorescence intensities are respectively taken to obtain the maximum emission wavelength maps (fig. 8B) of different batches of brine prepared under the same condition. As can be seen from fig. 8A and 8B, the fluorescence intensity and the maximum emission wavelength of the brines prepared under the same conditions are substantially the same, which further illustrates that the fluorescence intensity and the fluorescence emission wavelength can be used as the brine detection indexes.
The above examples show that the fluorescence detection method of the present invention is very reliable for detecting brine.
The above-described embodiments are intended to illustrate rather than to limit the invention, and any modifications and variations of the present invention are within the spirit and scope of the claims.

Claims (3)

1. A fluorescence detection method of edible areca-nut brine is characterized by comprising the following steps:
(1) preparing a brine test solution: accurately weighing 1.000 g of a series of brine samples respectively, adding a proper amount of distilled water for dissolving, then transferring into a 100 mL volumetric flask, fixing the volume to 100 mL by using the distilled water, and then carrying out high-speed centrifugation to obtain a series of brine clear liquids with the mass concentration of 1%; sucking 10mL of 1% brine clear liquid, adding the brine clear liquid into a 100 mL volumetric flask, adding 10mL of 0.2mol/L phosphate buffer solution, adding distilled water to a constant volume to a scale, and shaking up to obtain a series of brine test solutions with the mass concentration of 0.1%;
(2) measurement of brine fluorescence spectrum: adding 3.0 mL of 0.1% bittern test solution into 1cm thick 4 mL cuvette, respectively measuring fluorescence spectra of series bittern, setting fluorescence parameter as lambdaex=350 nm, scanning wavelength range 365-: ex =5, Em = 5;
(3) analysis of fluorescence intensity and emission wavelength of brine: and respectively taking the fluorescence intensity values at the maximum emission wavelengths according to the fluorescence spectra to obtain a fluorescence intensity graph corresponding to the series of brines, and respectively taking the emission wavelengths at the maximum fluorescence intensities to obtain a maximum emission wavelength graph corresponding to the series of brines.
2. The fluorescence detection method of edible areca-nut brine according to claim 1, which is characterized in that: the series of brine in the steps (1), (2) and (3) comprises brine prepared at different reaction temperatures, brine prepared at different reaction times, brine prepared at different material ratios, brine prepared in different feeding modes and brine of different batches under the same condition.
3. Use of the fluorescence detection method of edible areca brine according to claim 1 or 2 in the detection and analysis of edible areca brine.
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103743842A (en) * 2014-01-24 2014-04-23 广西大学 Method for constructing and using Guilin brine HPLC (High Performance Liquid Chromatography) fingerprint spectrums
CN104483338A (en) * 2014-12-23 2015-04-01 山东省第四地质矿产勘查院 Method for measuring bromine in brine through filter paper method sample preparation and X-ray fluorescent spectrometry

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104807777A (en) * 2015-04-28 2015-07-29 中国食品发酵工业研究院 Rapid detection method for areca-nut water content based on near infrared spectrum analysis technology

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103743842A (en) * 2014-01-24 2014-04-23 广西大学 Method for constructing and using Guilin brine HPLC (High Performance Liquid Chromatography) fingerprint spectrums
CN104483338A (en) * 2014-12-23 2015-04-01 山东省第四地质矿产勘查院 Method for measuring bromine in brine through filter paper method sample preparation and X-ray fluorescent spectrometry

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