CN105510504A - Method for measuring imidazole, musk xylene and sesamol residue quantity in edible essence - Google Patents
Method for measuring imidazole, musk xylene and sesamol residue quantity in edible essence Download PDFInfo
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Abstract
The invention discloses a method for measuring the imidazole, musk xylene and sesamol residue quantity in edible essence and namely discloses a method for quantitatively analyzing the imidazole, musk xylene and sesamol residue quantity in edible essence. The method is a method for measuring the imidazole, musk xylene and sesamol residue quantity in edible essence based on a gas chromatography-tandem mass spectrometer. The method includes the steps of preparation of an internal standard solution, a standard work solution and a sample solution, gas chromatography-tandem mass spectrometer analysis, calculation of a measurement result, and the like. The method is easy and convenient to operate, sensitive in response and accurate in qualitative and quantitative analysis, and interference caused when sample matrixes are complex is effectively reduced.
Description
Technical field
The present invention relates to a kind of method measuring imidazoles, musk xylene and sesamol residual quantity in flavoring essence based on gas chromatography-tandem mass spectrometer, belong to food additives physical and chemical index detection technique field.
Background technology
Imidazoles (molecular structural formula is as follows), i.e. 1,3-ribavirin, be a five-ring heterocycles aromaticity organic compound, chemical formula is C
3h
4n
2, molecular weight is 68.08, No. CAS is 288-32-4, and it is also an alkaloid, and in white or ligth yellow crystalline solid, soluble in water, ethanol, ether, chloroform, pyridine, be slightly soluble in benzene, is atomicly dissolved in sherwood oil, has acidity, also have alkalescence.Imidazoles is plane five-membered cyclic compound, there is permanent dipole between its two nitrogen-atoms, and polarity is very strong, and dipole moment is 3.61D, and intermolecularly there is hydrogen bond association, result in imidazoles and has unusual high boiling point (256 DEG C).There are 6 electron conjugated large π keys in molecule, therefore there is typical aromaticity.The nitrogen-atoms be connected with σ key with hydrogen provides pair of electrons, and in ring, all the other four atoms respectively provide an electronics Cheng Jian.Toxicology data shows that this material is poisonous, is 18.80mg/kg to its mouse oral LD50, and injection LD50 is 610mg/kg, its toxicity and means of defence similar to ethylenediamine.In addition, this material may have harm to environment, should pay special attention to water body.Comprehensive literature result, in general, imidazoles is not directly used as food additives and is used, and very large may be introduced into use as accessory substance.
Musk xylene (molecular structural formula is as follows), also known as 2,4,6-trinitro--3,3-dimethyl-5-tert-butyl benzene, by m-xylene nitration reaction and obtaining again after isobutylene alkylation, it is single compound, is different from the multicomponent mixture that Moschus belongs to natural.Chemical formula is C
12h
15n
3o
6molecular weight is 297.27, No. CAS is 81-15-2, presents slight yellow powdery or acicular crystal, the thicker musk odor of tool, fusing point is 112.5-114.5 DEG C, and boiling point is 200-202 DEG C, and the solubleness in 95% ethanol is 7g/L (25 DEG C), be soluble in Ergol and diethyl phthalate, water insoluble, glycerine and propylene glycol, have dry sweet Moschus sample animal fragrance, imposing manner is more weak but lasting is long.Musk xylene, as high persistence, high bioconcentration material, is classified as high attention rate material (SubstancesofVeryHighConcern, SVHC) by EU chemicals management administration (ECHA).European Union's " health care and the environmental science council " is carcinogenic, mutagenesis or cause genotoxicity (CMR) expert panel, in the 29th meeting, musk xylene is classified as the 3rd class material.International daily use chemicals fragrance association (IFRA) plan to advise that musk xylene does not use as daily use chemicals fragrance component in IFRA standard the 44th revised draft.International daily use chemicals spices research institute (RIFM) also advise that musk xylene must not be used for lip product or dental care product.China's cosmetics health specification and the middle regulation musk xylene of European Union's regulatory affairs (CouncilDirec2tive76/768/EEC) are regulated substance in components of cosmetics (must not be used for dental care product), and the limitation in essence, floral water and other products is respectively 1.0%, 0.4% and 0.03%.
Sesamol (molecular structural formula is as follows), also known as 3,4-methylenedioxyphenol, chemical formula is C
7h
6o
3molecular weight is 138.12, No. CAS is 533-31-3, and fusing point is 62-65 DEG C, and boiling point is 113-116 DEG C, slightly water-soluble, present white to Light yellow crystals powder or particle, be soluble in ethanol, ether, the synthetic technology of sesamol mainly contains three: the first is extracted from sesame oil, it two is entirely synthesize from pepper amine, and it three is semi-synthetic routes from heliotropin.The tumour data that cause of sesamol show: rat oral TDLo is 874mg/kg/2Y-C, the RTECS standard of knurl-carcinogenic, intestines and stomach-tumour; Its mouse oral TDLo is 1092mg/kg/2Y-C, the RTECS standard of knurl-carcinogenic, intestines and stomach-tumour; Rat oral TD is 1092mg/kg/2Y-C, the RTECS standard of knurl-carcinogenic, intestines and stomach-tumour; Its mouse oral TD is 1612mg/kg/2Y-C, the RTECS standard of knurl-carcinogenic, intestines and stomach-tumour.2006, Japanese MHLW announced just to comprise sesamol in 42 kinds of food additives of forbidding.
Flavoring essence is chemical combination material indispensable in daily life, production in light industry, its security is the very important aspect of food overall security, and the source that foodsafety controls, be raw materials for production at all, for this reason, must the chemical combination material paid close attention in flavoring essence be monitored.Literature survey shows, the method for existing detection imidazoles, musk xylene and sesamol mainly contains gas chromatography and liquid chromatography.In view of imidazoles, musk xylene and sesamol are likely used in flavoring essence, therefore a kind of method simultaneously detecting imidazoles, musk xylene and sesamol is fast and accurately explored, the residual quantity of this series material in flavoring essence is control effectively, and then ensure the safety in utilization of edible additive, just seem very urgent and very necessary.
Summary of the invention
The present invention is intended to overcome existing technological deficiency, a kind of method adopting gas chromatography tandem mass spectrometry instrument to measure imidazoles, musk xylene and sesamol residual quantity in flavoring essence is provided, the method energy accurate quantification analyzes the residual quantity of trace imidazoles, musk xylene and sesamol in flavoring essence, measurement result is accurate, highly sensitive, and matrix interference is few.
Specifically, present invention employs following technical scheme:
Measure a method for imidazoles in flavoring essence, musk xylene and sesamol residual quantity, it is characterized in that, said method comprising the steps of:
(1) preparation of inner mark solution: take phenylethyl propionate as internal standard compound, use ethanol is solvent, prepares inner mark solution;
(2) preparation of standard working solution: with the standard items of imidazoles, musk xylene and sesamol for object, use ethanol is solvent, be prepared into Standard Stock solutions through stepwise dilution, then add a certain amount of inner mark solution respectively, be prepared into standard working solution;
(3) preparation of sample solution: take a certain amount of flavoring essence sample in centrifuge tube, add a certain amount of water, more accurately move into the inner mark solution of certain volume, be uniformly dispersed with eddy mixer; Then add the ethanol of certain volume again, ultrasonic extraction, add a certain amount of anhydrous sodium sulfate, shake up rear leaving standstill, centrifugal in hydro-extractor afterwards, get supernatant liquor and cross organic phase filter membrane, obtain sample solution;
(4) gas chromatography tandem mass spectrometry instrument is analyzed: carry out detection with gas chromatography tandem mass spectrometry instrument to standard working solution and sample solution and analyze;
(5) drafting of standard working curve and the calculating of sample result.
Wherein, the preparation of step (1) inner mark solution comprises:
(1a) mark storing solution in: take 0.2g phenylethyl propionate, be accurate to 0.0001g, dissolve with ethanol and be settled in 100mL volumetric flask, shaking up;
(1b) inner mark solution: accurately pipette mark storing solution in 0.1mL, be settled in 100mL volumetric flask with ethanol, shake up.
In addition, the preparation of step (2) standard working solution comprises:
(2a) primary standard storing solution: take each 0.1g of imidazoles, musk xylene and sesamol respectively, be accurate to 0.0001g, dissolve with ethanol and be settled in 100mL volumetric flask, shaking up;
(2b) secondary standard storing solution: accurately pipette 1mL primary standard storing solution, in the volumetric flask of 100mL, dilutes with ethanol and is settled to scale;
(2c) standard working solution: accurately pipette in the volumetric flask of secondary standard storing solution 20 μ L, 50 μ L, 100 μ L, 200 μ L, 500 μ L and 1000 μ L to 10mL respectively, accurately add 100 μ L inner mark solutions more respectively, dilute with ethanol and be settled to scale, obtaining series standard working solution.
Further, the preparation of step (3) sample solution comprises: take 0.5g sample, be accurate to 0.0001g, be placed in 50mL centrifuge tube, add 1mL water, accurate immigration 100 μ L inner mark solution, be uniformly dispersed with eddy mixer, add 8mL ethanol again, be placed in ultrasonic extraction 20min in ultrasonic generator, add 0.5g anhydrous sodium sulfate, shake up rear standing 5min, afterwards in hydro-extractor under 4000r/min rotating speed centrifugal 5min, get supernatant liquor and cross 0.45 μm of organic phase filter membrane, obtain sample feeding solution.
Preferably, the instrumental conditions that step (4) GC-MS/MS analyzes is: adopt specification to be the HP-INNOWAX capillary chromatographic column of 30m × 0.25mm × 0.25 μm; Carrier gas is helium, and constant current speed is 1.0mL/min; Input mode: sample size is 1 μ L, pulse Splitless injecting samples, and sample introduction pulse is 25psi, and the time is 1min; Injector temperature is 250 DEG C; Transmission line temperature is 260 DEG C; Heating schedule is initial temperature is 100 DEG C, is increased to 240 DEG C with the speed of 3 DEG C/min, keeps 5min; Mass spectrophotometry condition is: ionization mode is EI source, positive ion mode; Ion source temperature: 230 DEG C; Quadrupole rod temperature: be 150 DEG C; Collision gas: nitrogen, flow velocity 1.5mL/min, carrier gas helium flow velocity is 2.25mL/min; Multiple-reaction monitoring MRM pattern, detail parameters is listed in the table below:
The MRM parameter situation of object
In above method, step (5) Specification Curve of Increasing and result are calculated as follows: with the ratio of object in standard working solution and interior target concentration for horizontal ordinate, with the ratio of object in chromatogram and interior target peak area for ordinate, carry out linear regression analysis, obtain standard working curve, by object and interior target chromatographic peak area ratio in the sample solution that records under the same terms, substitute into standard working curve, try to achieve the content of object in flavoring essence sample according to following formula:
In formula:
W---imidazoles, musk xylene and sesamol residual quantity in flavoring essence, unit is mg/kg;
The peak area of A---imidazoles, musk xylene and sesamol;
As---the peak area of internal standard compound phenylethyl propionate;
The intercept of b---standard working curve;
Ms---add the quality of internal standard compound phenylethyl propionate, unit is μ g;
The slope of a---standard working curve;
M---the quality of flavoring essence sample, unit is for propping up.
Beneficial effect: detection method of the present invention is optimized confirmation to sample treatment and chromatograph mass spectrum analysis condition.Compared with prior art, the present invention has following excellent results:
(1) the inventive method creative use gas chromatography tandem mass spectrometry instrument measures imidazoles, musk xylene and other residual quantity of sesamol trace level in flavoring essence, solve such material residual quantity level low, and the impact of the factor such as the Analysis interference of complex matrices on object be serious.
(2) the present invention is in sample pretreatment process, the feature that, complexity extensive for flavoring essence sample source differs, use the dicyandiamide solution of few dosage water+multiple dose ethanol to dissolve, then be combined eddy mixer and mixed, object is extracted.Method is novel, and reliably easy, strong innovation, compensate for the deficiency of current methods.
(3) the inventive method utilizes inner mark method ration, can accurate constant volume, and can reduce the error brought by pre-treating method reappearance and instrument precision problem.
Accompanying drawing explanation
Fig. 1 is the process flow diagram of assay method of the present invention;
Fig. 2 is the Ion-pair chromalography figure of imidazoles, musk xylene and sesamol and interior mark phenylethyl propionate in standard working solution;
Fig. 3 is the Ion-pair chromalography figure of imidazoles, musk xylene and sesamol and interior mark phenylethyl propionate in flavoring essence (mark-on) sample.
Embodiment
The present invention discloses a kind of method of imidazoles, musk xylene and sesamol residual quantity in quantitative analysis flavoring essence, comprises the following steps:
(1) preparation of inner mark solution: take phenylethyl propionate as internal standard compound, use ethanol is solvent, prepares inner mark solution;
(2) preparation of standard working solution: with the standard items of the materials such as imidazoles, musk xylene and sesamol for object, use ethanol is solvent, be prepared into Standard Stock solutions through stepwise dilution, then add a certain amount of inner mark solution respectively, be prepared into standard working solution;
(3) preparation of sample solution: take a certain amount of flavoring essence sample in centrifuge tube, add a certain amount of water, more accurately move into the inner mark solution of certain volume, be uniformly dispersed with eddy mixer; Then add the ethanol of certain volume again, ultrasonic extraction, add a certain amount of anhydrous sodium sulfate, shake up rear leaving standstill.Centrifugal in hydro-extractor afterwards, get supernatant liquor and cross organic phase filter membrane, obtain sample solution;
(4) gas chromatography tandem mass spectrometry instrument is analyzed: carry out detection with gas chromatography tandem mass spectrometry instrument (GC-MS/MS) to standard working solution and sample solution and analyze;
(5) drafting of standard working curve and the calculating of sample result.
The preparation of described inner mark solution, specifically comprises the following steps: mark storing solution in (1): take 0.2g (being accurate to 0.0001g) phenylethyl propionate, dissolve and be settled in 100mL volumetric flask, shake up with ethanol.(2) inner mark solution: accurately pipette mark storing solution in 0.1mL, be settled in 100mL volumetric flask with ethanol, shake up.
The preparation of described standard working solution specifically comprises the following steps: (1) primary standard storing solution: take each 0.1g of imidazoles, musk xylene and sesamol respectively, be accurate to 0.0001g, dissolve with ethanol and be settled in 100mL volumetric flask, shaking up.(2) secondary standard storing solution: accurately pipette 1mL primary standard storing solution, in the volumetric flask of 100mL, dilutes with ethanol and is settled to scale.(3) standard working solution: accurately pipette in the volumetric flask of secondary standard storing solution 20 μ L, 50 μ L, 100 μ L, 200 μ L, 500 μ L and 1000 μ L to 10mL respectively, accurately add 100 μ L inner mark solutions more respectively, dilute with ethanol and be settled to scale, obtaining series standard working solution.
The preparation of described sample solution specifically comprises following process: take 0.5g (being accurate to 0.0001g) in 50mL centrifuge tube, add 1mL water, accurate immigration 100 μ L inner mark solution, be uniformly dispersed with eddy mixer, add 8mL ethanol again, be placed in ultrasonic extraction 20min in ultrasonic generator, add 0.5g anhydrous sodium sulfate, shake up rear standing 5min.Afterwards in hydro-extractor under 4000r/min rotating speed centrifugal 5min, get supernatant liquor and cross 0.45 μm of organic phase filter membrane, obtain sample feeding solution.
Described GC-MS/MS analyzes, and its instrumental conditions is: adopt HP-INNOWAX (30m × 0.25mm × 0.25 μm) capillary chromatographic column; Carrier gas is helium, and constant current speed is 1.0mL/min; Input mode: sample size is 1 μ L, pulse Splitless injecting samples, and sample introduction pulse is 25psi, and the time is 1min; Injector temperature is 250 DEG C; Transmission line temperature is 260 DEG C; Heating schedule is initial temperature is 100 DEG C, is increased to 240 DEG C with the speed of 3 DEG C/min, keeps 5min.Its mass spectrophotometry condition is: ionization mode is EI source, positive ion mode; Ion source temperature: 230 DEG C; Quadrupole rod temperature: be 150 DEG C; Collision gas: nitrogen, flow velocity 1.5mL/min, carrier gas (helium) flow velocity is 2.25mL/min; Multiple-reaction monitoring (MRM) pattern, detail parameters is listed in the table below.
The MRM parameter situation of object
Described Specification Curve of Increasing and result are calculated as follows: with the ratio of object in standard working solution and interior target concentration for horizontal ordinate, with the ratio of object in chromatogram and interior target peak area for ordinate, carry out linear regression analysis, obtain standard working curve.By object and interior target chromatographic peak area ratio in the sample solution that records under the same terms, substitute into standard working curve, try to achieve the content of object in flavoring essence sample according to following formula:
In formula:
W---imidazoles, musk xylene and sesamol residual quantity in flavoring essence, unit is milligrams per kilogram (mg/kg);
The peak area of A---imidazoles, musk xylene and sesamol;
As---the peak area of internal standard compound phenylethyl propionate;
The intercept of b---standard working curve;
Ms---add the quality of internal standard compound phenylethyl propionate, unit is microgram (μ g);
The slope of a---standard working curve;
M---the quality of flavoring essence sample, unit is for propping up.
The invention will be further described in conjunction with specific embodiments more below.
Embodiment 1
The present embodiment is to the detection method of trace imidazoles, musk xylene and sesamol residual quantity in flavoring essence following (process flow diagram of described detection method is as shown in Figure 1):
(1) preparation of inner mark solution
1. mark storing solution in: accurately take 0.2060g phenylethyl propionate, in the volumetric flask of 100mL, dissolve with ethanol and be settled to scale.2. inner mark solution: accurately pipette mark storing solution in 0.1mL, in the volumetric flask of 100mL, dilute with ethanol and be settled to scale.Wherein the concentration of phenylethyl propionate is 2.06 μ g/mL.
(2) preparation of standard working solution
1. primary standard storing solution: primary standard storing solution: take imidazoles, musk xylene and sesamol each 0.1000g, 0.1020g and 0.1019g respectively, dissolve with ethanol and be settled in 100mL volumetric flask, shaking up.2. secondary standard storing solution: accurately pipette 1mL primary standard storing solution, in the volumetric flask of 100mL, dilutes with ethanol and is settled to scale.3. standard working solution: accurately pipette in the volumetric flask of secondary standard storing solution 20 μ L, 50 μ L, 100 μ L, 200 μ L, 500 μ L and 1000 μ L to 10mL respectively, accurately add 100 μ L inner mark solutions more respectively, dilute with ethanol and be settled to scale, obtaining series standard working solution.Wherein the concentration range of imidazoles is 0.02 ~ 1.00 μ g/mL, the concentration range of musk xylene be the concentration range of 0.0204 ~ 1.02 μ g/mL and sesamol is 0.0202 ~ 1.019 μ g/mL, and wherein, the concentration of mark phenylethyl propionate is 0.0206 μ g/mL.
(3) preparation of sample solution
With the flavoring essence of a market pin pin for analytic target, take 0.5132g sample in 50mL centrifuge tube, add 1mL water, accurate immigration 100 μ L inner mark solution, be uniformly dispersed with eddy mixer, then add 8mL ethanol, be placed in ultrasonic extraction 20min in ultrasonic generator, add 0.5g anhydrous sodium sulfate, shake up rear standing 5min.Afterwards in hydro-extractor under 4000r/min rotating speed centrifugal 5min, get supernatant liquor and cross 0.45 μm of organic phase filter membrane, obtain sample feeding solution.
(4) gas chromatography tandem mass spectrometry analysis
Standard working solution and the sample solution to be measured of getting 6 variable concentrations respectively carry out gas chromatography tandem mass spectrometry analysis (in described standard working solution, typical mark-on sample solution, the chromatogram of object and internal standard compound as shown in Figure 2 to 3).
Its instrumental conditions is: adopt HP-INNOWAX (30m × 0.25mm × 0.25 μm) capillary chromatographic column; Carrier gas is helium, and constant current speed is 1.0mL/min; Input mode: sample size is 1 μ L, pulse Splitless injecting samples, and sample introduction pulse is 25psi, and the time is 1min; Injector temperature is 250 DEG C; Transmission line temperature is 260 DEG C; Heating schedule is initial temperature is 100 DEG C, is increased to 240 DEG C with the speed of 3 DEG C/min, keeps 5min.Its mass spectrophotometry condition is: ionization mode is EI source, positive ion mode; Ion source temperature: 230 DEG C; Quadrupole rod temperature: be 150 DEG C; Collision gas: nitrogen, flow velocity 1.5mL/min, carrier gas (helium) flow velocity is 2.25mL/min; Multiple-reaction monitoring (MRM) pattern, detail parameters is listed in the table below.
The MRM parameter situation of object
(5) Specification Curve of Increasing and result calculate
First, with the ratio of object in standard working solution and interior target concentration for horizontal ordinate, with the ratio of object in chromatogram and interior target peak area for ordinate, carry out linear regression analysis, obtain standard working curve.Get the standard working solution of least concentration, do 9 Parallel testing analyses, calculate its standard deviation, with the concentration that the standard deviation of 3 times is corresponding, the detection limit of the method that draws that converts.The data such as the regression equation corresponding with standard working curve, related coefficient, detection limit see the following form.
The working curve of analytical approach and detection limit
By object and interior target chromatographic peak area ratio in the sample solution that records under the same terms, substitute into standard working curve, try to achieve the content of object in flavoring essence sample according to following formula.
In formula:
W---imidazoles, musk xylene and sesamol residual quantity in flavoring essence, unit is milligrams per kilogram (mg/kg);
The peak area of A---imidazoles, musk xylene and sesamol;
As---the peak area of internal standard compound phenylethyl propionate;
The intercept of b---standard working curve;
Ms---add the quality of internal standard compound phenylethyl propionate, unit is microgram (μ g);
The slope of a---standard working curve;
M---the quality of flavoring essence sample, unit is for propping up.
In the present embodiment, in flavoring essence sample main flume, 8 kinds of main Pyrazine fragrance matter testing results see the following form:
| Material title | Content (mg/kg) |
| Imidazoles | ND |
| Musk xylene | ND |
| Sesamol | ND |
Remarks: ND, for not detect, represents that result is lower than detection limit.
Embodiment 2
The detection method of the present embodiment to precision of the present invention and recovery of standard addition is as follows:
With essence sample used in embodiment 1 for analytic target, appropriate standard solution is added to it and obtains test specimen.Then, carry out withinday precision experiment respectively, withinday precision experiment is same sample replicate determination 6 times (same batch processed) under identical conditions, and calculate the relative standard deviation (RSD) of 6 parallel determinations respectively, measurement result sees the following form.Result display in table, the RSD<10% of the in a few days repeatability of this experimental technique, for the quantitative test of trace materials, shows that method has good precision.
With replica test test specimen used for analytic target, process according to above-mentioned sample-pretreating method, add the standard specimen of the suitable content of its burst size, carry out the test of matrix mark-on, the results are shown in following table.As can be seen from the table, the recovery of standard addition of sample is between 84.50% ~ 118.88%, and illustration method has good accuracy.
| Material title | RSD(％) |
| Imidazoles | 89.65 |
| Musk xylene | 94.37 |
| Sesamol | 104.24 |
Standard solution used by the present embodiment is only described for one of them concentration, the typical curve that standard solution prepared by other concentration value obtains through the analysis of gas chromatography tandem mass spectrometry instrument and regression equation same as the previously described embodiments, do not enumerating at this.Illustrated embodiment just method for a better understanding of the present invention, do not have any restriction, namely the method for said method or equivalent above-mentioned situation is all included in the protection domain of technical scheme of the present invention.
By reference to the accompanying drawings embodiments of the present invention are described in detail above, but the invention is not restricted to above-mentioned embodiment, in the ken that art those of ordinary skill possesses, can also make a variety of changes under the prerequisite not departing from present inventive concept.
Claims (6)
1. measure a method for imidazoles in flavoring essence, musk xylene and sesamol residual quantity, it is characterized in that, said method comprising the steps of:
(1) preparation of inner mark solution: take phenylethyl propionate as internal standard compound, use ethanol is solvent, prepares inner mark solution;
(2) preparation of standard working solution: with the standard items of imidazoles, musk xylene and sesamol for object, use ethanol is solvent, be prepared into Standard Stock solutions through stepwise dilution, then add a certain amount of inner mark solution respectively, be prepared into standard working solution;
(3) preparation of sample solution: take a certain amount of flavoring essence sample in centrifuge tube, add a certain amount of water, more accurately move into the inner mark solution of certain volume, be uniformly dispersed with eddy mixer; Then add the ethanol of certain volume again, ultrasonic extraction, add a certain amount of anhydrous sodium sulfate, shake up rear leaving standstill, centrifugal in hydro-extractor afterwards, get supernatant liquor and cross organic phase filter membrane, obtain sample solution;
(4) gas chromatography tandem mass spectrometry instrument is analyzed: carry out detection with gas chromatography tandem mass spectrometry instrument to standard working solution and sample solution and analyze;
(5) drafting of standard working curve and the calculating of sample result.
2. the method for imidazoles, musk xylene and sesamol residual quantity in mensuration flavoring essence according to claim 1, it is characterized in that, the preparation of step (1) inner mark solution comprises:
(1a) mark storing solution in: take 0.2g phenylethyl propionate, be accurate to 0.0001g, dissolve with ethanol and be settled in 100mL volumetric flask, shaking up;
(1b) inner mark solution: accurately pipette mark storing solution in 0.1mL, be settled in 100mL volumetric flask with ethanol, shake up.
3. the method for imidazoles, musk xylene and sesamol residual quantity in mensuration flavoring essence according to claim 1, it is characterized in that, the preparation of step (2) standard working solution comprises:
(2a) primary standard storing solution: take each 0.1g of imidazoles, musk xylene and sesamol respectively, be accurate to 0.0001g, dissolve with ethanol and be settled in 100mL volumetric flask, shaking up;
(2b) secondary standard storing solution: accurately pipette 1mL primary standard storing solution, in the volumetric flask of 100mL, dilutes with ethanol and is settled to scale;
(2c) standard working solution: accurately pipette in the volumetric flask of secondary standard storing solution 20 μ L, 50 μ L, 100 μ L, 200 μ L, 500 μ L and 1000 μ L to 10mL respectively, accurately add 100 μ L inner mark solutions more respectively, dilute with ethanol and be settled to scale, obtaining series standard working solution.
4. imidazoles in mensuration flavoring essence according to claim 1, the method of musk xylene and sesamol residual quantity, it is characterized in that, the preparation of step (3) sample solution comprises: take 0.5g sample, be accurate to 0.0001g, be placed in 50mL centrifuge tube, add 1mL water, accurate immigration 100 μ L inner mark solution, be uniformly dispersed with eddy mixer, add 8mL ethanol again, be placed in ultrasonic extraction 20min in ultrasonic generator, add 0.5g anhydrous sodium sulfate, shake up rear standing 5min, afterwards in hydro-extractor under 4000r/min rotating speed centrifugal 5min, get supernatant liquor and cross 0.45 μm of organic phase filter membrane, obtain sample feeding solution.
5. the method for imidazoles, musk xylene and sesamol residual quantity in mensuration flavoring essence according to claim 1, it is characterized in that, the instrumental conditions that step (4) GC-MS/MS analyzes is: adopt specification to be the HP-INNOWAX capillary chromatographic column of 30m × 0.25mm × 0.25 μm; Carrier gas is helium, and constant current speed is 1.0mL/min; Input mode: sample size is 1 μ L, pulse Splitless injecting samples, and sample introduction pulse is 25psi, and the time is 1min; Injector temperature is 250 DEG C; Transmission line temperature is 260 DEG C; Heating schedule is initial temperature is 100 DEG C, is increased to 240 DEG C with the speed of 3 DEG C/min, keeps 5min; Mass spectrophotometry condition is: ionization mode is EI source, positive ion mode; Ion source temperature: 230 DEG C; Quadrupole rod temperature: be 150 DEG C; Collision gas: nitrogen, flow velocity 1.5mL/min, carrier gas helium flow velocity is 2.25mL/min; Multiple-reaction monitoring MRM pattern, detail parameters is listed in the table below:
The MRM parameter situation of object
6. imidazoles in mensuration flavoring essence according to claim 1, the method of musk xylene and sesamol residual quantity, it is characterized in that, step (5) Specification Curve of Increasing and result are calculated as follows: with the ratio of object in standard working solution and interior target concentration for horizontal ordinate, with the ratio of object in chromatogram and interior target peak area for ordinate, carry out linear regression analysis, obtain standard working curve, by object and interior target chromatographic peak area ratio in the sample solution that records under the same terms, substitute into standard working curve, the content of object in flavoring essence sample is tried to achieve according to following formula:
In formula:
W---imidazoles, musk xylene and sesamol residual quantity in flavoring essence, unit is mg/kg;
The peak area of A---imidazoles, musk xylene and sesamol;
As---the peak area of internal standard compound phenylethyl propionate;
The intercept of b---standard working curve;
Ms---add the quality of internal standard compound phenylethyl propionate, unit is μ g;
The slope of a---standard working curve;
M---the quality of flavoring essence sample, unit is for propping up.
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| CN111220761A (en) * | 2020-03-11 | 2020-06-02 | 宁夏倬昱新材料科技有限公司 | High performance liquid chromatography analysis method for determining imidazole content in alkaline reaction solution |
| CN115343392A (en) * | 2022-08-23 | 2022-11-15 | 福建省纤维检验中心 | Method for measuring 1-vinyl imidazole in textile by gas chromatography-mass spectrometry |
| WO2023209006A1 (en) * | 2022-04-27 | 2023-11-02 | Givaudan Sa | Organic compounds |
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| CN107064399A (en) * | 2016-12-26 | 2017-08-18 | 上海烟草集团有限责任公司 | A kind of assay method of musk xylene in tobacco additive and sesamol |
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| CN115343392B (en) * | 2022-08-23 | 2024-10-01 | 福建省纤维检验中心 | Method for determining 1-vinyl imidazole in textile by gas chromatography-mass spectrometry |
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