CN108387573A - Hydroxymethylfurfural quick detection reagent, kit and detection method in a kind of honey - Google Patents
Hydroxymethylfurfural quick detection reagent, kit and detection method in a kind of honey Download PDFInfo
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- CN108387573A CN108387573A CN201810125289.8A CN201810125289A CN108387573A CN 108387573 A CN108387573 A CN 108387573A CN 201810125289 A CN201810125289 A CN 201810125289A CN 108387573 A CN108387573 A CN 108387573A
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- reagent
- hydroxymethylfurfural
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- honey
- tacheometer
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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
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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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
- G01N21/17—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
-
- 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
- G01N21/17—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
Abstract
Hydroxymethylfurfural quick detection reagent in a kind of honey, the reagent include reagent A and reagent B, and the reagent A is hydrochloric acid solution, and the reagent B is 2 thiobarbituricacidα- solution.After technical solution above, notable benefit includes：Used kit detection hydroxymethylfurfural colour developing of the present invention is apparent, colour stable, meanwhile, the hydroxymethylfurfural standard items of 0 ~ 10mg/kg are under various concentration gradient, and absorbance is linear, coefficient R2Reach 0.9992 or more；This method is quick, and instrument is carried out at the same time 10 Air conduct measurements, and detection every time only needs 1 second, less the time required to multiple samples.
Description
Technical field
Present invention relates particularly to hydroxymethylfurfural quick detection reagent, kit and detection methods in a kind of honey, belong to
Safety of Food Quality detection technique field.
Background technology
Hydroxymethylfurfural（HMF）It is no in fresh pure honey, it is honey in storage or heating process
From fructose converting, therefore it is the one of the important signs that for distinguishing whether honey is fresh.It provides in the world, the hydroxyl in honey
Methyl furfural should not be more than 40mg/kg, also be provided in national standard GH/T 18796-2012, hydroxymethylfurfural contains in honey
40mg/kg is not to be exceeded in amount.
The method for measuring the HMF contents in honey at present mainly has liquid chromatography-mass spectrometry, high performance liquid chromatography, gas phase
Chromatography, ultraviolet spectrophotometry, derivatization spectrophotometry etc..Though wherein liquid chromatography-mass spectrometry, high performance liquid chromatography
It is so relatively accurate, but experimental procedure is complicated, time-consuming, and the precision equipment of relevant Large expensive is needed, it is unsuitable for live fast
Speed detection；And ultraviolet spectrophotometry method is simple, but due to the more accurate costliness of instrument, it is also unsuitable for field quick detection.
The in-situ check and test method of hydroxymethylfurfural is derivatization spectrophotometry and colorimetric method in common food, although behaviour
Make simply, but there is the problems such as artificial judgement aberration, accuracy is poor, accuracy of detection is low, influences the accuracy of result, and it is public
In the honey of cloth in hydroxymethylfurfural rapid detection method, reagent and product have certain toxicity, be not suitable for the market of farm produce,
The large dosage of supermarket uses.
In addition, though there are also corresponding detection reagents in market, but color developing effect is bad, the linearly related degree of colour developing
It is not high, there is a problem of that accuracy of detection is not high in instrument is examined soon.
Invention content
The purpose of the invention is to provide hydroxymethylfurfural quick detection reagent in a kind of honey for using of cooperation instrument,
Kit and corresponding detection method, can either scene complete to the quick measurement of hydroxymethylfurfural in honey, and can be right
Food security carries out real-time monitoring, has the characteristics that agents useful for same is simple, does not need complexity condition and easy to operate, quick.
In order to complete foregoing invention purpose, the present invention provides hydroxymethylfurfural quick detection reagent in a kind of honey, described
Reagent includes reagent A and reagent B, and the reagent A is hydrochloric acid solution, and the reagent B is 2- thiobarbituricacidα- solution.
Further, a concentration of 2 ~ 9.5mol/L of the reagent A, a concentration of 0.01 ~ 0.04mol/L of the reagent B.
Further, a concentration of 2mol/L of the reagent A, a concentration of 0.02mol/L of the reagent B.
The present invention also provides a kind of kits including hydroxymethylfurfural quick detection reagent in the honey.
Further, the kit includes the reagent B of reagent A described in 1 bottle of 40ml and 1 bottle of 40ml.
Further, the kit further includes sample extraction cup, 5mL centrifuge tubes and dropper.
The present invention also provides a kind of detection method of hydroxymethylfurfural in honey using the kit, the detection sides
Method includes multi-parameter biochemistry tacheometer standard curve establishment step S1 and multi-parameter biochemistry tacheometer detecting step S2；
The multi-parameter biochemistry tacheometer standard curve establishment step S1 includes,
Hydroxymethylfurfural analytical standard product are diluted to form standard dilutions by S11；
Reagent A and reagent B are added into the standard dilutions by S12, complete chromogenic reaction；
S13 takes 2mL distilled water in clean dried cuvette, is put into the multi-parameter biochemistry tacheometer channel, completes instrument
School zero；
The standard dilutions after the completion of reaction are poured into cuvette, are put into the inspection of the multi-parameter biochemistry tacheometer by S14
Channel is surveyed, the spectral value of standard dilutions described in various concentration is measured, generates standard curve, is calculated using the standard curve
The concentration of the standard dilutions, and the linear dependence of the standard curve is assessed, if the standard curve is linear
Dependence among equations coefficients R2Between 0.999 to 1, then it is assumed that the reagent linear dependence is suitable, and enters the multi-parameter and give birth to
Change tacheometer detecting step S2；
The multi-parameter biochemistry tacheometer detecting step includes,
S21 prepares honey detection sample；
S22 detects addition reagent A and reagent B in sample to the honey, completes chromogenic reaction；
S23 takes distilled water in clean dried cuvette, is put into the multi-parameter biochemistry tacheometer channel, completes instrument school
Zero；
Solution after the completion of reaction is poured into cuvette by S24, is put into the corresponding sense channel of the multi-parameter biochemistry tacheometer,
Detection is completed, and calculates using standard curve the concentration of the hydroxymethylfurfural of the sample.
Further, the detection optical source wavelength of the multi-parameter biochemistry tacheometer is 420nm.
After technical solution above, notable benefit includes：1. used kit of the present invention detects hydroxymethylfurfural
Colour developing is apparent, colour stable, meanwhile, for the hydroxymethylfurfural standard items of 0 ~ 10mg/kg under various concentration gradient, absorbance is in line
Property, coefficient R2Reach 0.999 or more；2. kit can coordinate instrument to detect upload automatically, easy to operate, it is not required to people
Work interpretation, it is as a result more acurrate, it is convenient for Site Detection and real-time oversight management；3. this method is quick, instrument is carried out at the same time 10 and leads to
Road detects, and detection every time only needs 1 second, less the time required to multiple samples.This method is suitable in field quick detection honey
Hydroxymethyl furfural content can make the quality level of honey product quick judgement, and recall rate is high, to safeguard the economy of people
Interests and the edible safety for ensureing people.
Specific implementation mode
Embodiment 1：
Hydroxymethylfurfural quick detection kit in detection honey used in the present invention, wherein reagent portion is by reagent A and examination
Agent B compositions, wherein reagent A are 1 bottle of the hydrochloric acid solution of the 2mol/L of 40mL；Reagent B is thio bar of the 2- of the 0.02mol/L of 40mL
Than 1 bottle of appropriate acid solution；Testing tool part includes：Sample extraction cup：5；5mL centrifuge tubes：5；Dropper：1 packet；Specification：
1 part.
It is that above-mentioned quick detection kit establishes standard curve, including walks as follows below with multi-parameter biochemistry tacheometer
Suddenly：
Step 1, the preparation of sample weigh the 5- hydroxyl first that 0.1013g is purchased from Shanghai Aladdin biotech inc
Base furfural analytical standard product, are diluted to 0,0.25,0.5, Isosorbide-5-Nitrae, and 8,10mg/L, 2mL is settled in 5mL centrifuge tubes；
0.4mL reagent As, 0.4mL reagent B, then 30 ~ 60 DEG C of water-baths 20 are added into standard dilutions for step 2, colour developing
Minute；
Step 3, instrument school zero take 2mL distilled water in clean dried cuvette, are put into multi-parameter biochemistry tacheometer channel,
" hydroxymethylfurfural " is selected on " detection project " column, then clicks " instrument school zero ", instrument automatic adjustment wavelength to 420nm,
Complete instrument calibration；
Step 4, standard curve are established, the solution after the completion of reaction are poured into cuvette, it is corresponding to be put into multi-parameter biochemistry tacheometer
Sense channel, click " standard curve foundations ", instrument automatically adjusts wavelength to 420nm, generates standard curve, and using giving birth to
At standard curve measure the standard dilutions in sense channel, the content value of the hydroxymethylfurfural calculated is 0,
0.285,0.488,0.991,3.951,7.950,10.07mg/L, gained linear equation coefficient R2It is 0.9998.
After having standard curve above, so that it may to utilize standard curve above, quick detection kit and above-mentioned
Multi-parameter biochemistry tacheometer employed in calibration process, examines the content value of hydroxymethylfurfural included in sample
It surveys, detection process includes the following steps：
Step 1, the preparation of sample weigh in 1.0g commercially available honey to sample extraction cup, and 10mL distilled water is added, uses glass
Stick stirs and evenly mixs, and stands to be checked；
Step 2, colour developing, takes 2.00mL samples to be tested in 5mL centrifuge tubes, be then added into centrifuge tube 0.4mL reagent As,
0.4mL reagent B, then 30 ~ 50 DEG C of water-baths 20 minutes；
Step 3, instrument school zero take 2mL distilled water in clean dried cuvette, are put into multi-parameter biochemistry tacheometer channel
It is interior, " hydroxymethylfurfural " is selected on " detection project " column, then clicks " instrument school zero ", instrument automatically adjusts wavelength extremely
420nm, calibration are completed.
Solution after the completion of reaction is poured into cuvette by step 4, pattern detection, is put into the corresponding sense channel of instrument, point
It hits " pattern detection ", instrument shows that the content value 48.23mg/kg of hydroxymethylfurfural, result are the doubtful positive.
Next the rate of recovery for verifying hydroxymethylfurfural quick detection kit in the honey, includes the following steps：
Step 1 weighs in 4 parts of 1.0g commercially available honey to sample extraction cup, and hydroxymethylfurfural standard items are added thereto respectively
0,15,30,50mg/L, stand one day.
Step 2 is separately added into 10mL distilled water into sample described in step 1, is stirred and evenly mixed with glass bar, stands
It is to be checked.
Step 3, colour developing, takes 2.00mL samples to be tested in 5mL centrifuge tubes, 0.4mL is then added into centrifuge tube
Reagent A, 0.4mL reagent B, then 30 ~ 60 DEG C of water-baths 20 minutes.
Step 4, instrument school zero take 2mL distilled water in clean dried cuvette, are put into more used by the calibration of front
In parameter biochemistry tacheometer channel, " hydroxymethylfurfural " is selected on " detection project " column, then clicks " instrument school zero ", instrument
Device automatically adjusts wavelength to 420nm, and calibration is completed.
Solution after the completion of reaction is poured into cuvette by step 5, is put into the corresponding sense channel of instrument, clicks " sample inspection
Survey ", instrument show hydroxymethylfurfural content value, convert the rate of recovery be 86.80%, 96.20%, 104.28%.
Embodiment 2：
The concentration of reagent A in reagent portion in embodiment 1 is adjusted to 9.5mol/L, the concentration of reagent B is adjusted to
The concentration of 0.04mol/L, the standard dilution of preparation are respectively 0,0.4,0.8,1.6,2.4,3.2mg/L, other to be basically unchanged,
The content value of the hydroxymethylfurfural of apparatus measures standard dilution is 0,0.784,1.088,1.700,2.304,2.912mg/L,
Gained linear equation coefficient R2It is 0.999.
Embodiment 3：
The concentration of reagent A in reagent portion in embodiment 2 is adjusted to 5.1mol/L, the concentration of reagent B is adjusted to
0.01mol/L, other to be basically unchanged, the content value of the hydroxymethylfurfural of apparatus measures standard dilution is 0,0.783,1.089,
1.710,2.314,2.915mg/L, gained linear equation coefficient R2It is 0.9992.
From data above as can be seen that the concentration range of reagent A is a concentration of the 0.01 of 2 ~ 9.5mol/L and reagent B
When ~ 0.04mol/L, the color developing effect and linear dependence of reagent are all relatively good, all can serve as detection reagent use.Its
In, a concentration of 0.02mol/L of a concentration of 2mol/L and reagent B of reagent A, linear dependence is best.Generally, linear equation
Coefficient R2Think that the linearly related degree of the reagent is suitable when between 0.999 to 1, is suitble to detection.
It is sharp up to the researched and developed life of instrument science and technology Nantong Co., Ltd that multi-parameter biochemistry tacheometer in above-mentioned experiment is all made of intelligence
The II type multi-parameter biochemistry tacheometers of DPR- of production.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (8)
1. hydroxymethylfurfural quick detection reagent in a kind of honey, which is characterized in that the reagent includes reagent A and reagent B, institute
It is hydrochloric acid solution to state reagent A, and the reagent B is 2- thiobarbituricacidα- solution.
2. hydroxymethylfurfural quick detection reagent in honey as described in claim 1, which is characterized in that the reagent A it is dense
Degree is 2 ~ 9.5mol/L, a concentration of 0.01 ~ 0.04mol/L of the reagent B.
3. hydroxymethylfurfural quick detection reagent in honey as claimed in claim 2, which is characterized in that the reagent A it is dense
Degree is 2mol/L, a concentration of 0.02mol/L of the reagent B.
4. a kind of includes the examination of hydroxymethylfurfural quick detection reagent in honey as described in any one of claims 1 to 3
Agent box.
5. kit as claimed in claim 4, which is characterized in that the kit includes reagent A described in 1 bottle of 40ml and 1 bottle
The reagent B of 40ml.
6. kit as claimed in claim 5, which is characterized in that the kit further includes sample extraction cup, 5mL centrifuge tubes
And dropper.
7. the detection method of hydroxymethylfurfural in a kind of honey using the kit described in any one of claim 4 to 6,
It is characterized in that, the detection method includes, multi-parameter biochemistry tacheometer standard curve establishment step S1 and the biochemical speed of multi-parameter
Survey instrument detecting step S2；
The multi-parameter biochemistry tacheometer standard curve establishment step S1 includes,
Hydroxymethylfurfural analytical standard product are diluted to form standard dilutions by S11；
Reagent A and reagent B are added into the standard dilutions by S12, complete chromogenic reaction；
S13 takes 2mL distilled water in clean dried cuvette, is put into the multi-parameter biochemistry tacheometer channel, completes instrument
School zero；
The standard dilutions after the completion of reaction are poured into cuvette, are put into the inspection of the multi-parameter biochemistry tacheometer by S14
Channel is surveyed, the spectral value of standard dilutions described in various concentration is measured, generates standard curve, is calculated using the standard curve
The concentration of the standard dilutions, and the linear dependence of the standard curve is assessed, if the standard curve is linear
Dependence among equations coefficients R2Between 0.999 to 1, then it is assumed that the reagent linear dependence is suitable, and enters the multi-parameter and give birth to
Change tacheometer detecting step S2；
The multi-parameter biochemistry tacheometer detecting step includes,
S21 prepares honey detection sample；
S22 detects addition reagent A and reagent B in sample to the honey, completes chromogenic reaction；
S23 takes distilled water in clean dried cuvette, is put into the multi-parameter biochemistry tacheometer channel, completes instrument school
Zero；
Solution after the completion of reaction is poured into cuvette by S24, is put into the corresponding sense channel of the multi-parameter biochemistry tacheometer,
Detection is completed, and calculates using the standard curve of the generation concentration of the hydroxymethylfurfural of the sample.
8. utilized the detection method as described in claim 7, which is characterized in that the detection light source of the multi-parameter biochemistry tacheometer
Wavelength is 420nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810125289.8A CN108387573A (en) | 2018-02-08 | 2018-02-08 | Hydroxymethylfurfural quick detection reagent, kit and detection method in a kind of honey |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810125289.8A CN108387573A (en) | 2018-02-08 | 2018-02-08 | Hydroxymethylfurfural quick detection reagent, kit and detection method in a kind of honey |
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| CN201810125289.8A Pending CN108387573A (en) | 2018-02-08 | 2018-02-08 | Hydroxymethylfurfural quick detection reagent, kit and detection method in a kind of honey |
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Cited By (3)
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| CN109709096A (en) * | 2019-01-23 | 2019-05-03 | 云南万兴隆集团蜂业有限公司 | A kind of honey device for fast detecting |
| CN110658146A (en) * | 2019-11-12 | 2020-01-07 | 浙江华康药业股份有限公司 | Method for rapidly determining content of hydroxymethylfurfural in high fructose corn syrup |
| CN110749590A (en) * | 2019-10-26 | 2020-02-04 | 中国科学院新疆理化技术研究所 | Colorimetric reagent for rapidly detecting sucrose on site and preparation method thereof |
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Application publication date: 20180810 |