CN110082467A - Detect the method and kit of exogenous N- methyl -2- pyrrole aldehyde - Google Patents
Detect the method and kit of exogenous N- methyl -2- pyrrole aldehyde Download PDFInfo
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- G—PHYSICS
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- 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
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- 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
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- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
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- G—PHYSICS
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- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/8868—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample elemental analysis, e.g. isotope dilution analysis
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Abstract
The invention discloses the methods and kit that detect exogenous N- methyl -2- pyrrole aldehyde, wherein the method for the exogenous N- methyl -2- pyrrole aldehyde of the detection includes: to extract processing to sample, to obtain extracting solution；Separating treatment is carried out to the extracting solution using gas-chromatography, so as to isolated N- methyl -2- pyrrole aldehyde；The detection of isotope ratio is carried out, to the isolated N- methyl -2- pyrrole aldehyde to obtain the testing result of isotope ratio；Determine whether the sample contains exogenous N- methyl -2- pyrrole aldehyde based on the testing result of the isotope ratio.Whether this method energy judgement sample contains exogenous N- methyl -2- pyrrole aldehyde.
Description
Technical field
The present invention relates to analytical chemistry fields, and in particular, to detect exogenous N- methyl -2- pyrrole aldehyde method and
Kit.
Background technique
Essence has the enhancing existing flavor of food and assigns the characteristic of food flavour, is widely used in food industry
In.In view of health risk brought by the perfume ingredient of part, workable essence compound is limited in the world.China
National food safety standard GB2760-2014 " food additives use standard " to the flavors and fragrances that can be made an addition in food at
It point has done and clearly to have limited, any flavors and fragrances must not be added in special provision tealeaves, coffee and pasteurization milk.
Although the technologies such as the common gas-chromatography in laboratory, liquid chromatography, gas chromatography tandem mass spectrum can measure at present
The type and content of fragrance component in food, but precise Identification fragrance is from food itself or from exogenous addition, still
It is so a difficult task.Such as perfume ingredient N- methyl -2- pyrrole aldehyde (1-Methylpyrrole-2-
Carboxaldehyde, No. CAS: 1192-58-1), it is deleted from No. 1334/2008 regulation (EC) by European Union within 2014, it must not
It is added in food.But meanwhile it is also naturally occurring a kind of fragrance component in coffee.When detection disabling essence in food
When ingredient, it is based only upon GC-MS/MS and is difficult to distinguish the source of flavor compounds.
Detection disables exogenous essence as a result, such as the method for N- methyl -2- pyrrole aldehyde has much room for improvement.
Summary of the invention
The present invention is directed at least solve one of the technical problems existing in the prior art.For this purpose, one object of the present invention
It is to propose that a kind of method for detecting exogenous N- methyl -2- pyrrole aldehyde, this method can accurately the exogenous N- of qualitative detection
Methyl -2- pyrrole aldehyde, accuracy is high, solves the problems, such as the exogenous disabling perfume ingredient detection in China.
It should be noted that the present invention is the following work based on inventor and completes:
Although the technologies such as the common gas-chromatography in laboratory, liquid chromatography, gas chromatography tandem mass spectrum can measure at present
The type and content of fragrance component in food, but can not precise Identification fragrance be from food itself or adding from exogenous
Add.Commercial synthesis product are mainly generated by processing such as petrochemical materials, before Stable isotope ratio is able to reflect millions of years
Information.During biological growth metabolism, the natural causes such as weather and environment can cause the Plantago fengdouensis of isotope in organism,
A kind of given compound can show very big difference carbon stable isotope composition is upper, so that stable isotope is with fingerprint
Feature.Therefore carbon stable isotope ratio (13C/12C is expressed as δ13C difference) can be used for identifying exogenous compounds and plant
Naturally occur compound.
Thus, according to an aspect of the present invention, the present invention provides a kind of exogenous N- methyl -2- pyrrole aldehydes of detection
Method.According to an embodiment of the invention, this method comprises: processing is extracted to sample, to obtain extracting solution；Utilize gas
Phase chromatography carries out separating treatment to the extracting solution, so as to isolated N- methyl -2- pyrrole aldehyde；To the isolated N-
Methyl -2- pyrrole aldehyde carries out the detection of isotope ratio, to obtain the testing result of isotope ratio；Based on the isotope ratio
Testing result determine whether the sample contains exogenous N- methyl -2- pyrrole aldehyde.
The method of the exogenous N- methyl -2- pyrrole aldehyde of detection of the embodiment of the present invention is carried out at separation by gas-chromatography
Reason, separates chromatographic peak and the other impurity peaks of N- methyl -2- pyrrole aldehyde, isolated N- methyl -2- pyrrole aldehyde has
Effect eliminates interference of the complex matrices to detection.On the basis of chromatographic isolation, obtained together using isotopic ratio analysis instrument
The plain ratio in position it is accurate as a result, whether judgement sample contains exogenous N- methyl -2- pyrrole aldehyde.Outside the detection of the embodiment of the present invention
The method of source property N- methyl -2- pyrrole aldehyde is easy to operate, at low cost, efficiently solves the identification of exogenous disabling perfume ingredient
Problem, especially food exogenous disable the identification problem of perfume ingredient, and the technology for being conducive to food safety Regulation department is held
Method safeguards the legitimate rights and interests of consumer.
In addition, the method for the exogenous N- methyl -2- pyrrole aldehyde of detection according to the above embodiment of the present invention, can also have
There is following additional technical characteristic:
According to an embodiment of the invention, the extraction process is Simultaneous distillation-extraction processing.
According to an embodiment of the invention, the organic phase of the Simultaneous distillation-extraction processing is methylene chloride, water phase is water.
According to an embodiment of the invention, the volume ratio of the organic phase and the water phase is 2-7:100.
According to an embodiment of the invention, the chromatographic column of the gas-chromatography separation is highly polar capillary chromatographic column, preferably
Ground is HP-INNOWAX chromatographic column, 60m × 0.25mm × 0.25 μm.
According to an embodiment of the invention, the chromatographic condition of the gas-chromatography separation are as follows: shunt mode: not shunting, the time
0.5-2min；Sampling volume: 1-3 μ L；Injector temperature: 200-300 DEG C；Flow rate of carrier gas: constant current, 0.6-1mL/min.
According to an embodiment of the invention, the gas phase of the gas-chromatography separation separates temperature program are as follows: 50 DEG C, 6min；50
DEG C/min, and 120 DEG C, 1min；2 DEG C/min, 145 DEG C, 2min；4 DEG C/min, 180 DEG C, 0min；60 DEG C/min, 240 DEG C, 1min.
According to an embodiment of the invention, the testing result based on the isotope ratio determines whether the sample contains
Exogenous N- methyl -2- pyrrole aldehyde includes: the detection that standard items are carried out with isotope ratio, to obtain isotope ratio standard
Value；The testing result of the isotope ratio is in the isotope ratio standard value range, then without exogenous N- methyl -2- pyrroles's first
Aldehyde.
According to an embodiment of the invention, the isotope ratio is detected as carbon isotope ratio δ13C detection.
According to an embodiment of the invention, the sample is coffee.
According to an embodiment of the invention, the carbon isotope ratio δ13C standard value is -35.5 ‰~-31.5 ‰.
On this basis, invention further provides a kind of exogenous N- methyl -2- pyrrole aldehyde detection kits.Root
According to the embodiment of the present invention, it is based on 200g sample, which includes: extraction reagent A: methylene chloride, 30-50mL；Extract examination
Agent B: water, 800-1200mL；And chromatographic column: highly polar capillary chromatographic column.
The kit of the embodiment of the present invention carries out Simultaneous distillation-extraction and extracts simultaneously richness from sample by extracting reagent A and B
Collect N- methyl -2- pyrrole aldehyde, recycles highly polar capillary chromatographic column to carry out gas-chromatography separating treatment, so that N- methyl -
2- pyrrole aldehyde chromatographic peak and other impurity peaks are completely separable, effectively eliminate interference of the complex matrices to detection.In N- methyl-
On the basis of 2- pyrrole aldehyde chromatographic peak and other impurity peaks are completely separable, carbon isotope ratio δ is utilized13The accurate detection of C, judgement
Whether sample contains exogenous N- methyl -2- pyrrole aldehyde.
According to an embodiment of the invention, the kit further comprises: desiccant: anhydrous sodium sulfate, 50-120mg.
According to an embodiment of the invention, the kit further comprises: standard solution: 20 μ g/mL N- methyl -2- pyrroles
Formaldehyde standard solution；And calibration solution: 20 μ g/mL normal cetanes.
According to an embodiment of the invention, the chromatographic column is HP-INNOWAX chromatographic column, 60m × 0.25mm × 0.25 μm.
Additional aspect and advantage of the invention will be set forth in part in the description, and will partially become from the following description
Obviously, or practice through the invention is recognized.
Detailed description of the invention
Above-mentioned and/or additional aspect of the invention and advantage will become from the description of the embodiment in conjunction with the following figures
Obviously and it is readily appreciated that, in which:
Fig. 1 shows the N- methyl -2- pyrrole aldehyde standard items GC/C/ of various concentration according to an embodiment of the invention
IRMS response results schematic diagram；
Fig. 2 shows N- methyl -2- pyrrole aldehyde standard under the conditions of different chromatographic columns according to an embodiment of the invention
The GC/C/IRMS response results schematic diagram of product, wherein the chromatographic column in A is DB-5MS chromatographic column；Chromatographic column in B is HP-
INNOWAX chromatographic column.
Specific embodiment
The embodiment of the present invention is described below in detail, examples of the embodiments are shown in the accompanying drawings, wherein from beginning to end
Same or similar label indicates same or similar element or element with the same or similar functions.Below with reference to attached
The embodiment of figure description is exemplary, and for explaining only the invention, and is not considered as limiting the invention.
It should be noted that term " first ", " second " are used for description purposes only, it is not understood to indicate or imply phase
To importance or implicitly indicate the quantity of indicated technical characteristic.Define " first " as a result, the feature of " second " can be with
Explicitly or implicitly include one or more of the features.Further, in the description of the present invention, unless otherwise saying
Bright, the meaning of " plurality " is two or more.
According to an aspect of the present invention, the present invention provides a kind of sides for detecting exogenous N- methyl -2- pyrrole aldehyde
Method.
The method of the exogenous N- methyl -2- pyrrole aldehyde of detection of the embodiment of the present invention is carried out at separation by gas-chromatography
Reason, so that N- methyl -2- pyrrole aldehyde chromatographic peak and other impurity peaks are completely separable, effectively eliminates complex matrices to detection
Interference.On the basis of N- methyl -2- pyrrole aldehyde chromatographic peak and other impurity peaks are completely separable, carbon isotope ratio δ is utilized13C
Accurate detection, whether judgement sample contain exogenous N- methyl -2- pyrrole aldehyde.
The method of the exogenous N- methyl -2- pyrrole aldehyde of detection of the embodiment of the present invention solves the exogenous disabling essence in China
Ingredient technical problem existing for market surpervision provides effective technological means for technical enforcement, protects consumer health, and maintenance is closed
Method food production processing enterprise interests.
The method of the exogenous N- methyl -2- pyrrole aldehyde of detection of embodiment to facilitate the understanding of the present invention, at this to this
Method is explained, according to an embodiment of the invention, this method comprises:
S100 extraction process
According to an embodiment of the invention, processing is extracted to sample, to obtain extracting solution.As a result, by extraction
Reason obtains the extracting solution containing N- methyl -2- pyrrole aldehyde from sample.
According to an embodiment of the invention, the extraction process is Simultaneous distillation-extraction processing.At Simultaneous distillation-extraction
Reason, N- methyl -2- pyrrole aldehyde are concentrated, the Determination of Trace Volatile ingredient N- methyl -2- pyrrole aldehyde in sample can be isolated
Come, to improve the accuracy of detection.
According to an embodiment of the invention, the organic phase of the Simultaneous distillation-extraction processing is methylene chloride, water phase is water.One
Aspect, methylene chloride polarity is close with N- methyl -2- pyrrole aldehyde, keeps extraction efficiency higher；On the other hand, methylene chloride boiling point
It is low, it carries out volatile when rotary evaporation concentration.According to an embodiment of the invention, the volume ratio of the organic phase and the water phase
For 2-7:100.The ratio not only can guarantee good extraction efficiency, but also the dosage of the effectively save organic reagent of energy as a result,.
The separation of S200 gas-chromatography
According to an embodiment of the invention, carrying out separating treatment, isolated N- to the extracting solution using gas-chromatography
Methyl -2- pyrrole aldehyde chromatographic peak.It is separated by gas-chromatography, so that N- methyl -2- pyrrole aldehyde chromatographic peak and other impurities
Peak is completely separable, effectively eliminates interference of the complex matrices to detection in sample, detects convenient for subsequent isotope method.
Wherein, specification is needed, chromatographic isolation processing and the detection of isotope ratio can be online progress, can also
Independently to carry out.Chromatographic isolation is utilized, keeps N- methyl -2- pyrrole aldehyde chromatographic peak and other impurity peaks completely separable,
Isolated N- methyl -2- pyrrole aldehyde chromatographic peak is directly entered online isotope ratio detection device and is detected, and passes through N- first
The position of base -2- pyrrole aldehyde chromatographic peak can judge N- methyl -2- pyrroles from Multiple components isotope ratio testing result
The result of formaldehyde.If chromatographic isolation processing and isotope ratio detection be not it is online, can directly pass through N- methyl -2- pyrrole
N- methyl -2- pyrrole aldehyde is collected in the peak position that goes out for coughing up formaldehyde chromatographic peak, then carries out subsequent isotope ratio detection, obtains N- first
The testing result of the isotope ratio of base -2- pyrrole aldehyde.
According to an embodiment of the invention, the chromatographic column of the gas-chromatography separation is highly polar capillary chromatographic column.It is based on
The chemical property of N- methyl -2- pyrrole aldehyde, highly polar capillary chromatograph can be effectively by N- methyl -2- pyrrole aldehyde from complexity
Matrix in separate.Inventor's discovery uses HP-INNOWAX chromatographic column, 60m × 0.25mm × 0.25 μm, N- methyl -2-
Pyrrole aldehyde can be kept completely separate with other impurity peaks, and the Accurate Determining of carbon isotope ratio may be implemented.
According to an embodiment of the invention, the chromatographic condition of the gas-chromatography separation are as follows: shunt mode: not shunting, the time
0.5-2min, it is preferable that be 1min；Sampling volume: 1-3 μ l, it is preferable that be 1 μ l；Injector temperature: 200-300 DEG C, preferably
Ground is 250 DEG C；Flow rate of carrier gas: constant current, 0.6-1mL/min, it is preferable that be 0.8mL/min.Gas-chromatography separating effect as a result,
It is good, so that N- methyl -2- pyrrole aldehyde chromatographic peak and other impurity peaks are completely separable, be conducive to improve subsequent exogenous N- first
The accuracy of the result of base -2- pyrrole aldehyde detection.
According to an embodiment of the invention, the gas phase of the gas-chromatography separation separates temperature program are as follows: 50 DEG C, 6min；50
DEG C/min, and 120 DEG C, 1min；2 DEG C/min, 145 DEG C, 2min；4 DEG C/min, 180 DEG C, 0min；60 DEG C/min, 240 DEG C, 1min.
The temperature program can realize efficiently separating for object and other compounds, raising detection efficiency with the very short time.
The detection of S300 isotope ratio
According to an embodiment of the invention, the detection of isotope ratio is carried out to the isolated N- methyl -2- pyrrole aldehyde, with
Just the testing result of isotope ratio is obtained.It is conventional since the additive amount of exogenous N- methyl -2- pyrrole aldehyde is usually extremely micro
The detection accuracy of detection method is difficult to accurately distinguish with the presence or absence of exogenous N- methyl -2- pyrrole aldehyde, and inventor is based on not
With the compound in source its stable isotope ratio can discrepant principle, utilize the detection of stable isotope ratio to realize synthesis
With the differentiation of natural food additives, solves the detection problem of exogenous N- methyl -2- pyrrole aldehyde.
According to an embodiment of the invention, the testing result based on the isotope ratio determines whether the sample contains
Exogenous N- methyl -2- pyrrole aldehyde includes: the detection that standard items are carried out with isotope ratio, to obtain isotope ratio standard
Value；The testing result of the isotope ratio is in the isotope ratio standard value range, then without exogenous N- methyl -2- pyrroles's first
Aldehyde.Wherein, it should be noted that standard items are the corresponding standard items of sample, for example, sample is coffee, then standard items are same product
Kind is free of the coffee of exogenous N- methyl -2- pyrrole aldehyde, so that the isotope ratio based on standard items determines whether containing exogenous
N- methyl -2- pyrrole aldehyde.
According to an embodiment of the invention, the isotope ratio is detected as carbon isotope ratio δ13C detection.Due to N- methyl-
2- pyrrole aldehyde is mainly made of C and H, δ13The measurement of H is easier to that isotope fractionation occurs, so inventor selects carbon isotope
Than being detected.
S400 is qualitatively judged
According to an embodiment of the invention, the testing result based on the isotope ratio determines whether the sample contains external source
Property N- methyl -2- pyrrole aldehyde.
Perfume ingredient is added in food often to improve flavor, and exogenous N- methyl -2- pyrroles's first whether is added in coffee
Aldehyde is the emphasis of detection.According to an embodiment of the invention, the sample is coffee.According to an embodiment of the invention, in coffee
The carbon isotope ratio δ of source property N- methyl -2- pyrrole aldehyde13C standard value is -35.5 ‰~-31.5 ‰.That is, if institute
The carbon isotope ratio δ of sample13The value of C is then free of exogenous N- methyl -2- pyrroles's first in -35.5 ‰~-31.5 ‰ ranges
Aldehyde, and if containing exogenous N- methyl -2- pyrrole aldehyde not in -35.5 ‰~-31.5 ‰ ranges.
On this basis, invention further provides a kind of exogenous N- methyl -2- pyrrole aldehyde detection kits.Root
According to the embodiment of the present invention, it is based on 200g sample, which includes: extraction reagent A: methylene chloride, 30-50mL；Extract examination
Agent B: water, preferably ultrapure water, 800-1200mL；And chromatographic column: highly polar capillary chromatographic column.
The kit of the embodiment of the present invention carries out Simultaneous distillation-extraction and extracts simultaneously richness from sample by extracting reagent A and B
Collect N- methyl -2- pyrrole aldehyde, recycles highly polar capillary chromatographic column to carry out gas-chromatography separation, so that N- methyl -2- pyrrole
It coughs up that formaldehyde chromatographic peak is completely separable with other impurity peaks, effectively eliminates interference of the complex matrices to detection.In N- methyl -2- pyrrole
Cough up formaldehyde chromatographic peak and other impurity peaks it is completely separable on the basis of, utilize carbon isotope ratio δ13The accurate detection of C, judgement sample
Whether exogenous N- methyl -2- pyrrole aldehyde is contained.
According to an embodiment of the invention, the kit further comprises: desiccant: anhydrous sodium sulfate, 50-120mg.By
This, the moisture in desiccant absorption extraction liquid prevents interference of the moisture to detection.
According to an embodiment of the invention, the kit further comprises: standard solution: 20 μ g/mL N- methyl -2- pyrroles
Formaldehyde standard solution；And calibration solution: 20 μ g/mL normal cetanes.Since N- methyl -2- pyrrole aldehyde is low pole chemical combination
Object, so correcting fluid selects normal cetane similar with N- methyl -2- pyrrole aldehyde polarity.According to an embodiment of the invention,
The chromatographic column is HP-INNOWAX chromatographic column, 60m × 0.25mm × 0.25 μm.Good separating effect as a result,.
Below with reference to specific embodiment, the present invention will be described, it should be noted that these embodiments are only explanation
Property, and be not considered as limiting the invention.
The solution of the present invention is explained below in conjunction with embodiment.It will be understood to those of skill in the art that following
Embodiment is merely to illustrate the present invention, and should not be taken as limiting the scope of the invention.Particular technique or item are not specified in embodiment
Part, it described technology or conditions or is carried out according to the literature in the art according to product description.Agents useful for same or instrument
Production firm person is not specified in device, and being can be with conventional products that are commercially available, such as can purchase from Sigma company.
Embodiment 1
Endogenic in natural coffee beans and commercial synthesis N- methyl -2- pyrrole aldehyde δ is measured in the present embodiment respectively13C value, specific as follows:
(1) extraction process: (a) by the coffee bean sample comminution of no any exogenous N- methyl -2- pyrrole aldehyde addition.
(b) 200g sample is weighed, is placed in 1L boiling flask, be added and extract reagent B: ultrapure water is connected to Likens-
The side of Nickerson SDE method will extract reagent A: methylene chloride is added in 250mL boiling flask, even
It is connected to the other side of Likens-Nickerson SDE method, two sides boiling flask is heated by electric boiling plate.
(c) after heating extraction 180 minutes, with desiccant dryness extract.Then, using the rotary evaporation for being set as 35 DEG C
Extract is concentrated to 1mL by device.
Distillation sequence while the N- methyl -2- pyrrole aldehyde standard items of commercial synthesis use same.
(2) isolate and purify: using the silent winged generation of match, your 1310 GC gas chromatograph of company's T race is separated, chromatographic column
For HP-INNOWAX capillary column (Agilent Technologies,Germany)(60m x 0.250mm x
0.25 μm), using following GC condition: not shunt mode sample introduction, 250 DEG C of injector temperature, 50 DEG C of initial oven temperature (keeps 6
Minute), 120 DEG C (being kept for 1 minute) are warming up to the heating rate of 50 DEG C/min, are warming up to 145 with the heating rate of 2 DEG C/min
DEG C (keep 2 minutes), is warming up to 180 DEG C with the heating rate of 4 DEG C/min, is then warming up to the heating rate of 60 DEG C/min
240 DEG C (being kept for 2 minutes), carrier gas used is helium, flow 0.8mL/min.
(3) carbon isotope ratio measures: using DELTA V Advantage Stable isotope ratio mass spectrograph to through gas phase
Isolated N- methyl -2- pyrrole aldehyde carries out δ13C is measured, and is corrected in continuous mode using calibration solution A.
Natural coffee beans sample and commercial synthesis N- methyl -2- pyrrole aldehyde δ13C measurement result such as table 1.
As shown in Table 1, endogenous N- methyl -2- pyrrole aldehyde δ in above-mentioned natural coffee beans sample13C measurement result-
It is the δ of endogenous N- methyl -2- pyrrole aldehyde in coffee in 35.5 ‰~-31.5 ‰ ranges13C range.Above-mentioned commercial synthesis N-
The δ of methyl -2- pyrrole aldehyde solution13C measurement result is in -27.9 ‰~-23.9 ‰ ranges.According to one-way analysis of variance knot
Fruit (P < 0.001), natural coffee beans sample and commercial synthesis N- methyl -2- pyrrole aldehyde δ13C measurement result is entirely different, utilizes
Carbon isotope ratio δ13The accurate detection of C, it can be determined that whether sample contains exogenous N- methyl -2- pyrrole aldehyde.
Embodiment 2
Utilize the external source of the method detection coffee samples of the exogenous N- methyl -2- pyrrole aldehyde of detection of the embodiment of the present invention
Property N- methyl -2- pyrrole aldehyde, specific as follows:
(1) extraction process:
(a) 200g coffee samples are weighed, are placed in 1L boiling flask, is added and extracts reagent B, be connected to Likens-
The side of Nickerson SDE method.
(b) reagent A will be extracted to be added in 250ml boiling flask, is connected to Likens-Nickerson while distills extraction
Take the other side of device.Two sides boiling flask is heated by electric boiling plate.
(c) after heating extraction 180 minutes, with desiccant dryness extract.Then, using the rotary evaporation for being set as 35 DEG C
Extract is concentrated to 1mL by device.
(2) according to the method for embodiment 1, gas-chromatography separation is carried out to extracting solution, then carries out carbon isotope ratio δ13C's
Measurement；
By the δ of institute's sample13C is the same as endogenous δ13C compares, and can identify in the food whether is N- methyl -2- pyrrole aldehyde
For exogenous components, coffee samples N- methyl -2- pyrrole aldehyde δ13C measurement result is as shown in table 2.
Table 2
| Coffee samples | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 |
| δ13C(‰) | -32.88 | -31.69 | -31.26 | -32.02 | -30.60 | -30.62 | -31.68 | -32.71 | -30.22 | -31.22 | -27.19 |
As shown in Table 2, the N- methyl -2- pyrrole aldehyde δ of preceding ten coffee samples13C measurement result is in endogenous result
Within the scope of, and the measurement result of the 11st coffee samples deviates considerably from, and can be accredited as containing exogenous components.
Embodiment 3
δ is carried out to various concentration N- methyl -2- pyrrole aldehyde in the present embodiment13C measurement, specific as follows:
1) the N- methyl -2- pyrrole aldehyde standard items (1-50ppm) of various concentration are subjected to gas phase as described in Example 1
The measurement of carbon isotope ratio is carried out after chromatographic isolation.
2) response results of the N- methyl -2- pyrrole aldehyde standard items of surveyed various concentration are as shown in Figure 1, the results showed that mesh
Mark object is all oxidized to CO2, the rate of recovery is good, shows the N- methyl -2- pyrroles of the various concentration extracted from food samples
The Accurate Determining of carbon isotope ratio may be implemented in formaldehyde.
Embodiment 4
Influence of the different gas chromatographic columns to N- methyl -2- pyrrole aldehyde separating effect is investigated in the present embodiment, specifically such as
Under:
DB-5MS (30m × 0.25mm × 0.25 μm) and HP- is respectively adopted in N- methyl -2- pyrrole aldehyde standard items
INNOWAX (60m × 0.25mm × 0.25 μm) capillary chromatographic column carries out carbon isotope ratio value after carrying out gas-chromatography separation
Measurement.As a result as shown in Fig. 2, in DB-5MS, N- methyl -2- pyrrole aldehyde overlaps with other impurity peaks, Wu Fayou
Effect separation；And under conditions of HP-INNOWAX (60m × 0.25mm × 0.25 μm) chromatographic column, N- methyl -2- pyrrole aldehyde with
Other impurity peaks are kept completely separate, and the Accurate Determining of carbon isotope ratio may be implemented.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or spy described in conjunction with this embodiment or example
Point is included at least one embodiment or example of the invention.In the present specification, schematic expression of the above terms are not
Centainly refer to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can be any
One or more embodiment or examples in can be combined in any suitable manner.
Although an embodiment of the present invention has been shown and described, it will be understood by those skilled in the art that: not
A variety of change, modification, replacement and modification can be carried out to these embodiments in the case where being detached from the principle of the present invention and objective, this
The range of invention is defined by the claims and their equivalents.
Claims (10)
1. a kind of method for detecting exogenous N- methyl -2- pyrrole aldehyde characterized by comprising
Processing is extracted to sample, to obtain extracting solution；
Separating treatment is carried out to the extracting solution using gas-chromatography, so as to isolated N- methyl -2- pyrrole aldehyde；
The detection of isotope ratio is carried out, to the isolated N- methyl -2- pyrrole aldehyde to obtain the detection knot of isotope ratio
Fruit；And
Determine whether the sample contains exogenous N- methyl -2- pyrrole aldehyde based on the testing result of the isotope ratio.
2. the method according to claim 1, wherein the extraction process be Simultaneous distillation-extraction processing,
Optionally, the organic phase of the Simultaneous distillation-extraction processing is methylene chloride, and water phase is water,
Optionally, the volume ratio of the organic phase and the water phase is 2-7:100.
3. the method according to claim 1, wherein the chromatographic column of gas-chromatography separation is highly polar capillary
Pipe chromatographic column, it is preferable that be HP-INNOWAX chromatographic column, 60m × 0.25mm × 0.25 μm.
4. the method according to claim 1, wherein the chromatographic condition of gas-chromatography separation are as follows:
Shunt mode: not shunting, time 0.5-2min；
Sampling volume: 1-3 μ l；
Injector temperature: 200-300 DEG C；
Flow rate of carrier gas: constant current, 0.6-1mL/min.
5. the method according to claim 1, wherein the gas phase of gas-chromatography separation separates temperature program
Are as follows: 50 DEG C, 6min；50 DEG C/min, 120 DEG C, 1min；2 DEG C/min, 145 DEG C, 2min；4 DEG C/min, 180 DEG C, 0min；60℃/
Min, 240 DEG C, 1min.
6. the method according to claim 1, wherein the testing result based on the isotope ratio determines institute
It states sample and whether contains exogenous N- methyl -2- pyrrole aldehyde and include:
Standard items are carried out with the detection of isotope ratio, to obtain isotope ratio standard value；
The testing result of the isotope ratio is in the isotope ratio standard value range, then without exogenous N- methyl -2- pyrroles
Formaldehyde.
7. the method according to claim 1, wherein the isotope ratio is detected as carbon isotope ratio δ13C inspection
It surveys.
8. the method according to the description of claim 7 is characterized in that the sample be coffee,
Optionally, the carbon isotope ratio δ13C standard value is -35.5 ‰~-31.5 ‰.
9. a kind of exogenous N- methyl -2- pyrrole aldehyde detection kit, which is characterized in that be based on 200g sample, comprising:
Extract reagent A: methylene chloride, 30-50mL；
Extract reagent B: water, 800-1200mL；And
Chromatographic column: highly polar capillary chromatographic column.
10. kit according to claim 9, which is characterized in that further comprise:
Desiccant: anhydrous sodium sulfate, 50-120mg,
Optionally, further comprise:
Standard solution: 20 μ g/mL N- methyl -2- pyrrole aldehyde standard solution；And
Calibration solution: 20 μ g/mL normal cetanes,
Optionally, the chromatographic column is HP-INNOWAX chromatographic column, 60m × 0.25mm × 0.25 μm.
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