CN109580820A - The rapid detection method of Furnan products in a kind of dairy products - Google Patents
The rapid detection method of Furnan products in a kind of dairy products Download PDFInfo
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- CN109580820A CN109580820A CN201811562740.9A CN201811562740A CN109580820A CN 109580820 A CN109580820 A CN 109580820A CN 201811562740 A CN201811562740 A CN 201811562740A CN 109580820 A CN109580820 A CN 109580820A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G01N2030/027—Liquid chromatography
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Abstract
The present invention provides a kind of rapid detection methods of Furnan products in dairy products to be cooled to room temperature method includes the following steps: oxalic acid solution heating water bath is added in (1) in dairy products sample solution；(2) solution of trichloroacetic acid A is then added, after concussion mixing, centrifugation takes supernatant, and solution of trichloroacetic acid B is added in filter residue, after concussion mixing, centrifugation；Merge supernatant, solution of trichloroacetic acid B constant volume is added；(3) sample introduction is analyzed using high performance liquid chromatograph, selects packing material size for 2.7-3.5 μm of reverse phase C18 chromatographic column.The present invention selects 2.7-3.5 μm of packing material size of reverse phase C18 chromatographic column to complete Furnan products detection in dairy products, analyzing duration is only 12min, quantitatively it is limited to 0.01 μ g/mL, the range of linearity is 0.01-50 μ g/mL, substantially reduce analysis time, improve detection sensitivity, have the advantages that rapidly and efficiently with save solvent.
Description
Technical field
The invention belongs to the technical fields of compound test, and in particular to Furnan products is quick in a kind of dairy products
Detection method.
Background technique
In recent years, the Consciousness of food security of compatriots enhances increasingly, higher and higher for the quality requirement of dairy products.It is made in cream
The quality of product controls aspect, and Maillard reaction is a very important ring, as the reducing sugar in diary product process and
The product chaff propylhomoser of a kind of nonenzymatic browning reaction occurred between lysine, initial stage is used as monitoring Pasteur by European Union
The index of sterilization cream and UHT cream quality impairments.The Furnan products that Maillard reaction mid-term stage generates, it is also possible to as
The index of dairy products thermal damage.
So far, in dairy products Furnan products research focus primarily upon: 5 hydroxymethyl furfural, furfural,
2- furfuryl ketone and 5 methyl furfural.The existing detection method of furfural includes spectroscopic methodology and chromatography；Spectroscopic methodology has simple
The advantages such as low with expense, but this method poor selectivity, and it is also easy to produce matrix interference；Chromatography mainly include liquid chromatography and
Gas chromatography, gas chromatography sensitivity are lower than liquid chromatography, and liquid chromatography is using most furfural detection means.
Existing Furnan products liquid phase chromatography detection method includes liquid chromatography tandem UV detector method (High
Performance liquid chromatography-Ultraviolet detector, HPLC-UV), liquid chromatography tandem
Fluorescence detection (High performance liquid chromatography-Fluorescence detector, HPLC-
) and Liquid Chromatography-Tandem Mass Spectrometry detection method (High performance liquid chromatography-Mass FLD
Spectrometry, HPLC-MS): HPLC-FLD needs to perform the derivatization pretreatment to furfural substance, and process is cumbersome；HPLC-
MS is more demanding to instrument and operator, higher cost.HPLC-UV is most widely used Furnan products detection method.
It is directed to the liquid chromatography tandem UV detector method of Furnan products in existing literature, generallys use packing material size
For 5 μm of reverse phase C18 chromatographic column, a length of 25min when entirely analyzing, the range of linearity is relatively narrow, and obtains and use expensive superelevation
Effect liquid phase chromatogram instrument.
Therefore, the detection method for studying Furnan products in simple, the efficient dairy products of one kind, is that this field urgently solves
Certainly the technical issues of.
Summary of the invention
In order to solve the above-mentioned technical problem, the purpose of the present invention is to provide in a kind of dairy products Furnan products it is fast
The entire analysis duration of fast detection method, the detection method is only 12min.
Specifically, on the one hand, provide a kind of rapid detection method of Furnan products in dairy products, this method includes
Following steps:
(1) oxalic acid solution heating water bath is added in dairy products sample solution, is cooled to room temperature；
(2) solution of trichloroacetic acid A is then added to be centrifuged under revolving speed 4-5g after concussion mixing, takes supernatant, filter residue is added
Solution of trichloroacetic acid B is centrifuged under revolving speed 4-5g after concussion mixing；Merge supernatant, solution of trichloroacetic acid B constant volume is added；
(3) high performance liquid chromatograph sample introduction is used, selects packing material size for 2.7-3.5 μm of reverse phase C18 chromatographic column, flows
Xiang Weishui and acetonitrile, flow velocity 0.5-0.7mL/min, column temperature are 28-32 DEG C, after chromatography, detect Furnan products
Content.
Further, in step (1), the volume that oxalic acid solution is added in every g sample solution is 0.3-0.5mL, and oxalic acid is molten
The concentration of liquid is 0.12-0.16mol/L.
Further, in step (1), the temperature of heating water bath is 90-120 DEG C, time 20-30min.
Further, in step (2), the concentration of solution of trichloroacetic acid A is 35-45g/mL, the concentration of solution of trichloroacetic acid B
For 3.5-4.5g/mL.
Further, in step (2), the time of centrifugation is 12-18min.
Further, in step (3), the length of chromatographic column is 12-18cm, internal diameter 2.8-3.2mm, film thickness 2.5-
3.5μm。
Further, in step (3), sample volume is 4.5-5.5 μ L, and sample injection disc temperature is 3.8-4.2 DEG C；Ultraviolet detection wave
A length of 270-290nm.
Further, in step (3), before sample introduction, sample solution crosses 0.45 μm of nylon leaching film.
The invention has the benefit that the present invention selects the reverse phase C18 chromatographic column that packing material size is 2.7-3.5 μm to complete cream
Furnan products detect in product, and analysis duration is only 12min, are quantitatively limited to 0.01 μ g/mL, and the range of linearity is 0.01~50
μ g/mL, substantially reduces analysis time, improves detection sensitivity, has widened quantification range, has rapidly and efficiently molten with saving
The advantages of agent.The present invention can be realized using common high performance liquid chromatograph device, not need using expensive ultra high efficiency liquid phase
Chromatographic apparatus.
Detailed description of the invention
Fig. 1 is the detection chromatogram of the Furnan products of powdered milk sample in the embodiment of the present invention 1；
Fig. 2 is the detection chromatogram of the Furnan products of liquid milk sample in the embodiment of the present invention 2；
Fig. 3 is the detection chromatogram of the Furnan products of milk sample product in the embodiment of the present invention 3.
Specific embodiment
Technical solution of the present invention is illustrated to be clearer, below in conjunction with specific embodiment to skill of the invention
Art scheme is further elaborated:
In a specific embodiment, a kind of rapid detection method of Furnan products in dairy products is provided,
Method includes the following steps:
(1) oxalic acid solution heating water bath is added in dairy products sample solution, is cooled to room temperature；
(2) solution of trichloroacetic acid A is then added to be centrifuged under revolving speed 4-5g after concussion mixing, takes supernatant, filter residue is added
Solution of trichloroacetic acid B is centrifuged under revolving speed 4-5g after concussion mixing；Merge supernatant, solution of trichloroacetic acid B constant volume is added；
(3) high performance liquid chromatograph sample introduction is used, selects packing material size for 2.7-3.5 μm of reverse phase C18 chromatographic column, flows
Xiang Weishui and acetonitrile, flow velocity 0.5-0.7mL/min, column temperature are 28-32 DEG C, after chromatography, detect Furnan products
Content.
By the present invention in that being dissociated mating type furfural with oxalic acid, then solution of trichloroacetic acid is selected to remove removing protein, by gained
Solution enters liquid chromatograph analysis.The present invention selects reverse phase C18 chromatographic column of the packing material size less than 5 μm to complete four in dairy products
The detection of kind of Furnan products, when analysis a length of 12min, be quantitatively limited to 0.01 μ g/mL, the range of linearity is 0.01 μ g/mL-50 μ
G/mL substantially reduces analysis time, improves detection sensitivity.The present invention uses common high performance liquid chromatograph device
It realizes, does not need using expensive Ultra Performance Liquid Chromatography instrument device.
In step (1) of the present invention, the volume that oxalic acid solution is added in every g sample solution is 0.3-0.5mL, oxalic acid solution
Concentration is 0.12-0.16mol/L.The oxalic acid solution for selecting the same day to prepare, oxalic acid solution can reduce sample pH, in heating condition
The lower chemical bond for destroying mating type furfural, dissociate furfural compounds.
In step (1) of the present invention, the temperature of heating water bath is 90-120 DEG C, time 20-30min.
In step (2) of the present invention, the concentration of solution of trichloroacetic acid A is 35-45g/mL, and the concentration of solution of trichloroacetic acid B is
3.5-4.5g/mL.In acid condition, trichloroacetic acid can form insoluble salt with protein, and then be settled out in dairy products
Protein.
In step (2) of the present invention, the time of centrifugation is 12-18min.After centrifugation, protein precipitation is discarded, takes supernatant.
In step (3) of the present invention, the length of chromatographic column is 12-18cm, and internal diameter 2.8-3.2mm, film thickness is 2.5-3.5 μ
m.By preferred column condition, the detection of Furnan products can be realized using common high performance liquid chromatograph device.
In step (3) of the present invention, sample volume is 4.5-5.5 μ L, and sample injection disc temperature is 3.8-4.2 DEG C；Ultraviolet detection wavelength
For 270-290nm.By preferred sampling condition and Detection wavelength, quantitative limit further reduced, improve detection sensitivity.
In step (3) of the present invention, before sample introduction, sample solution crosses 0.45 μm of nylon leaching film, that may be present tiny to remove
Particle prevents liquid chromatogram pipeline blockage.
With reference to the accompanying drawings and examples, the present invention is specifically described.It will be preferably by following embodiments
Understand the present invention.
Embodiment 1
Detection method includes the following steps for Furnan products in milk powder:
(1) it weighs 2.25g powdered milk sample and adds water to 15g mixing, the addition 5mL0.15mol/L same day configures 100 DEG C of water of oxalic acid
Bath heating 25min, is cooled to room temperature；
(2) 3mL concentration 40g/mL solution of trichloroacetic acid A is then added, after shaking 10min, 4.355g is centrifuged 15min；It takes
Supernatant pours into 25mL volumetric flask, and 5mL4g/mL solution of trichloroacetic acid B is added in filter residue, after shaking 10min, 4.355g centrifugation
15min；Supernatant merges, and 4g/mL solution of trichloroacetic acid B is added and is settled to scale；
(3) it takes appropriate amount of fluid to cross 0.45 μm of nylon leaching film, is analyzed into chromatograph.Selecting packing material size is 3 μm of reverse phase C18
Chromatographic column, chromatographic column: Waters Atlantis T3 (3.0mm × 15cm, 3 μm)；Mobile phase: mobile phase A is water, Mobile phase B
For acetonitrile；Gradient elution: 0-10.0min, 5%B-95%B, 10.0-12.0min, 95%B-95%B, 12.0-12.1min,
95%B-5%B, 12.1-20.0min, 5%B-5%B；Flow velocity 0.6mL/min；Chromatogram column temperature: 30 DEG C；Sample volume: 5 μ L；Into
Sample dish temperature: 4 DEG C；Ultraviolet detection wavelength: 280nm.
Detection chromatogram in powdered milk sample as shown in Figure 1, detect 5 hydroxymethyl furfural and furfural compounds, 5- hydroxyl first
The detection limit of base furfural and furfural is respectively 57.88 μ g/100g and 12.74 μ g/100g；2- furfuryl ketone and 5- is not detected
Methyl furfural compound shows that the 2- furfuryl ketone and 5 methyl furfural compound in the milk powder are less than 0.01 μ g/ of quantitative limit
mL。
Embodiment 2
Detection method includes the following steps for Furnan products in liquid milk:
(1) 15g liquid milk sample is weighed, the addition 4.5mL0.16mol/L same day configures 90 DEG C of heating water bath 30min of oxalic acid,
It is cooled to room temperature；
(2) 3mL concentration 35g/mL solution of trichloroacetic acid A is then added, after shaking 10min, 4g is centrifuged 18min；Take supernatant
Liquid pours into 25mL volumetric flask, and 5mL3.5g/mL solution of trichloroacetic acid B is added in filter residue, and after shaking 10min, 4g is centrifuged 18min；Supernatant
Liquid merges, and 3.5g/mL solution of trichloroacetic acid B is added and is settled to scale.
(3) it takes appropriate amount of fluid to cross 0.45 μm of nylon leaching film, is analyzed into chromatograph.Selecting packing material size is 2.7 μm of reverse phase
C18 chromatographic column, chromatographic column: Waters Atlantis T3 (2.8mm × 12cm, 2.8 μm)；Mobile phase: mobile phase A is water, stream
Dynamic phase B is acetonitrile；Gradient elution: 0-10.0min, 5%B-95%B, 10.0-12.0min, 95%B-95%B, 12.0-
12.1min, 95%B-5%B, 12.1-20.0min, 5%B-5%B；Flow velocity 0.5mL/min；Chromatogram column temperature: 28 DEG C；Sample introduction
Amount: 4.5 μ L；Sample injection disc temperature: 3.8 DEG C；Ultraviolet detection wavelength: 270nm.
Detection chromatogram as shown in Fig. 2, detect two kinds of compounds of 5 hydroxymethyl furfural and furfural in liquid milk sample,
The detection limit of 5 hydroxymethyl furfural and furfural compounds is respectively 51.95 μ g/100g and 5.64 μ g/100g；2- furan is not detected
Furfuryl ketone and 5 methyl furfural compound show 2- furfuryl ketone and 5 methyl furfural chemical combination in the ultra-high-temperature sterilized milk
Object is less than 0.01 μ g/mL of quantitative limit.
Embodiment 3
Detection method includes the following steps for Furnan products in pasteurization milk:
(1) 15g pasteurization milk sample is weighed, the addition 7.5mL0.12mol/L same day configures 120 DEG C of heating water baths of oxalic acid
20min is cooled to room temperature；
(2) 3mL concentration 45g/mL solution of trichloroacetic acid A is then added, after shaking 12min, 5g is centrifuged 18min；Take supernatant
Liquid pours into 25mL volumetric flask, and 5mL4.5g/mL solution of trichloroacetic acid B is added in filter residue, and after shaking 12min, 5g is centrifuged 18min；Supernatant
Liquid merges, and 4.5g/mL solution of trichloroacetic acid B is added and is settled to scale.
(3) it takes appropriate amount of fluid to cross 0.45 μm of nylon leaching film, is analyzed into chromatograph.Selecting packing material size is 3.5 μm of reverse phase
C18 chromatographic column, chromatographic column: WatersAtlantisT3 (3.2mm × 18cm, 3.2 μm)；Mobile phase: mobile phase A is water, flowing
Phase B is acetonitrile；Gradient elution: 0-10.0min, 5%B-95%B, 10.0-12.0min, 95%B-95%B, 12.0-
12.1min, 95%B-5%B, 12.1-20.0min, 5%B-5%B；Flow velocity 0.7mL/min；Chromatogram column temperature: 32 DEG C；Sample introduction
Amount: 5.5 μ L；Sample injection disc temperature: 4.2 DEG C；Ultraviolet detection wavelength: 290nm.
Detection chromatogram in pasteurization milk sample as shown in figure 3, detect 5 hydroxymethyl furfural and two kinds of furfural changes
The detection limit of conjunction object, 5 hydroxymethyl furfural and furfural compounds is respectively 6.69 μ g/100g and 3.87 μ g/100g；It is not detected
2- furfuryl ketone and 5 methyl furfural compound show 2- furfuryl ketone and 5 methyl furfural in the pasteurization milk
It closes object and is less than 0.01 μ g/mL of quantitative limit.
Finally, it should be noted that the above various embodiments is only to help to understand technical solution of the present invention and core concept,
Rather than its limitations；Although present invention has been described in detail with reference to the aforementioned embodiments, the ordinary skill people of this field
Member it is understood that it is still possible to modify the technical solutions described in the foregoing embodiments, or to part of or
Person's all technical features are equivalently replaced, and these modify or replace the protection scope for also falling into the claims in the present invention
It is interior.
Claims (8)
1. the rapid detection method of Furnan products in a kind of dairy products, which is characterized in that method includes the following steps:
(1) oxalic acid solution heating water bath is added in dairy products sample solution, is cooled to room temperature；
(2) solution of trichloroacetic acid A is then added to be centrifuged under revolving speed 4-5g after concussion mixing, takes supernatant, trichlorine is added in filter residue
Acetic acid solution B is centrifuged under revolving speed 4-5g after concussion mixing；Merge supernatant, solution of trichloroacetic acid B constant volume is added；
(3) high performance liquid chromatograph sample introduction is used, selects packing material size for 2.7-3.5 μm of reverse phase C18 chromatographic column, mobile phase is
Water and acetonitrile, flow velocity 0.5-0.7mL/min, column temperature are 28-32 DEG C, after chromatography, detect containing for Furnan products
Amount.
2. the rapid detection method of Furnan products in dairy products according to claim 1, which is characterized in that step
(1) in, the volume that oxalic acid solution is added in every g sample solution is 0.3-0.5mL, and the concentration of oxalic acid solution is 0.12-
0.16mol/L。
3. the rapid detection method of Furnan products in dairy products according to claim 1, which is characterized in that step
(1) in, the temperature of heating water bath is 90-120 DEG C, time 20-30min.
4. the rapid detection method of Furnan products in dairy products according to claim 1, which is characterized in that step
(2) in, the concentration of solution of trichloroacetic acid A is 35-45g/mL, and the concentration of solution of trichloroacetic acid B is 3.5-4.5g/mL.
5. the rapid detection method of Furnan products in dairy products according to claim 1, which is characterized in that step
(2) in, the time of centrifugation is 12-18min.
6. the rapid detection method of Furnan products in dairy products according to claim 1, which is characterized in that step
(3) in, the length of chromatographic column is 12-18cm, and internal diameter 2.8-3.2mm, film thickness is 2.5-3.5 μm.
7. the rapid detection method of Furnan products in dairy products according to claim 1, which is characterized in that step
(3) in, sample volume is 4.5-5.5 μ L, and sample injection disc temperature is 3.8-4.2 DEG C；Ultraviolet detection wavelength is 270-290nm.
8. the rapid detection method of Furnan products in dairy products according to claim 1, which is characterized in that step
(3) in, before sample introduction, sample solution crosses 0.45 μm of nylon leaching film.
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Cited By (9)
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| CN111024907A (en) * | 2019-12-26 | 2020-04-17 | 光明乳业股份有限公司 | Method for rapidly evaluating content of furfural substances in liquid dairy product based on storage temperature and humidity of liquid dairy product |
| CN111024906A (en) * | 2019-12-26 | 2020-04-17 | 光明乳业股份有限公司 | Method for rapidly evaluating content of furfural substances in solid dairy product based on storage temperature and humidity of solid dairy product |
| CN111048159A (en) * | 2019-12-30 | 2020-04-21 | 光明乳业股份有限公司 | Evaluation method of liquid milk heat treatment process |
| CN111983158A (en) * | 2020-08-20 | 2020-11-24 | 光明乳业股份有限公司 | Method for rapidly identifying doping amount of reconstituted milk in raw milk |
| CN112415153A (en) * | 2020-11-04 | 2021-02-26 | 薛伟锋 | Method for predicting residual shelf life of infant milk powder |
| CN112649274A (en) * | 2020-11-27 | 2021-04-13 | 光明乳业股份有限公司 | Extraction method of furfural compounds |
| CN112684125A (en) * | 2020-12-08 | 2021-04-20 | 光明乳业股份有限公司 | Method for rapidly evaluating content of furfural substances in prepared milk after heat treatment |
| CN112986437A (en) * | 2020-12-30 | 2021-06-18 | 光明乳业股份有限公司 | Purification detection method for furfural compounds in coffee dairy products |
| CN115389642A (en) * | 2021-05-25 | 2022-11-25 | 中粮营养健康研究院有限公司 | Method for judging Maillard reaction degree in dairy product |
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Cited By (9)
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| CN111024907A (en) * | 2019-12-26 | 2020-04-17 | 光明乳业股份有限公司 | Method for rapidly evaluating content of furfural substances in liquid dairy product based on storage temperature and humidity of liquid dairy product |
| CN111024906A (en) * | 2019-12-26 | 2020-04-17 | 光明乳业股份有限公司 | Method for rapidly evaluating content of furfural substances in solid dairy product based on storage temperature and humidity of solid dairy product |
| CN111048159A (en) * | 2019-12-30 | 2020-04-21 | 光明乳业股份有限公司 | Evaluation method of liquid milk heat treatment process |
| CN111983158A (en) * | 2020-08-20 | 2020-11-24 | 光明乳业股份有限公司 | Method for rapidly identifying doping amount of reconstituted milk in raw milk |
| CN112415153A (en) * | 2020-11-04 | 2021-02-26 | 薛伟锋 | Method for predicting residual shelf life of infant milk powder |
| CN112649274A (en) * | 2020-11-27 | 2021-04-13 | 光明乳业股份有限公司 | Extraction method of furfural compounds |
| CN112684125A (en) * | 2020-12-08 | 2021-04-20 | 光明乳业股份有限公司 | Method for rapidly evaluating content of furfural substances in prepared milk after heat treatment |
| CN112986437A (en) * | 2020-12-30 | 2021-06-18 | 光明乳业股份有限公司 | Purification detection method for furfural compounds in coffee dairy products |
| CN115389642A (en) * | 2021-05-25 | 2022-11-25 | 中粮营养健康研究院有限公司 | Method for judging Maillard reaction degree in dairy product |
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