Gas chromatography-mass spectrometry detection method for moschus wanshanensis in essence

Abstract

The invention provides a gas chromatography-mass spectrometry detection method for moschus wanshanensis in essence, which specifically comprises the following steps: adding an organic solvent for extraction, carrying out vortex treatment, and then carrying out shaking treatment to obtain a liquid A; adding ultrapure water, magnetic particles and carbon nanotubes into the liquid A, and carrying out centrifugation, vortex oscillation and solid-liquid separation; adding n-butane into an organic layer obtained by separation, carrying out ultrasonic treatment, separation and elution, and collecting an eluent to obtain a solution B; carrying out water bath concentration on the liquid B under the protection of nitrogen, and obtaining a liquid C after constant volume treatment and a microfiltration membrane treatment; and then analyzing and detecting under a specific gas chromatography-mass spectrometry condition. The detection method disclosed by the invention is simple and easy to implement, good in separation effect, high in sensitivity and good in detection effect.

Description

Technical Field

[0001] The invention belongs to the field of detection technology, and specifically relates to a gas chromatography-mass spectrometry detection method for Wanshan Musk in essence. Background Art

[0002] As an important additive, flavors are used in different products to give them fragrance and functionality, thereby increasing the added value of the products. Flavors can be divided into natural flavors and synthetic flavors according to their sources, which are extracted from natural substances and synthesized artificially and obtained in industrial production. Flavors can be divided into three categories according to their uses: daily flavors, edible flavors, and flavors for other purposes; flavors can be divided into six categories according to their fragrance: floral, non-floral, fruity, wine, tobacco, and food; and they can be divided into two categories according to their form: liquid flavors and powdered flavors. Liquid flavors can be divided into three types: water-soluble flavors, oil-soluble flavors, and emulsified flavors.

[0003] Wanshan Musk, scientific name: 1,1,4,4-tetramethyl-6-ethyl-7-acetyl-1,2,3,4-tetrahydronaphthalene, also known as Musk-36A. It is white or flaky crystals with an elegant musk aroma, similar to macrocyclic musk. It can be mixed with almost all spices, soluble in diethyl phthalate (62%), soluble in ethanol (8%), and insoluble in water. With ethylbenzene and 2,5-dichloro-2,5-dimethylhexane as raw materials and ferric chloride as catalyst, 1,1,4,4-tetramethyl-6-ethyl-tetrahydronaphthalene is obtained through reaction, and then acylation reaction is carried out under the action of aluminum chloride catalyst to obtain Wanshan Musk.

[0004] Patent CN103869039B provides a method for simultaneously detecting seven prohibited compounds in flavors and fragrances. The method uses a CH2Cl2 extract containing an internal standard for static filtration pretreatment, and uses gas chromatography-mass spectrometry to analyze and detect musk components, but the separation and detection effects are poor; Patent CN105467053B provides a method for rapid detection of artificial musk residues in daily chemical products. The method uses a mixture of dichloromethane and ethyl acetate as an extract and a microwave pretreatment method, and uses a purification column to remove interfering substances such as grease, wax, and pigments. The residual content of 11 artificial musks is detected at the same time, but the pretreatment time is long, and the separation efficiency and detection effect need to be improved; Patent CN113552254A provides a method for detecting polycyclic musks in water bodies. By improving the MPS2 multi-functional automatic sampler, the electromagnetic stirring device is combined with the automatic sampler to realize the simultaneous magnetic stirring and dynamic extraction processes, thereby improving the mass transfer rate, extraction efficiency, and analysis speed, but no pretreatment is performed, and the detection sensitivity and effect are average.

[0005] Therefore, there are few detection methods for the single component of Wanshan Musk in flavors in the existing technology, and the traditional detection of musk has problems such as complicated pre-treatment process and poor treatment effect. It is urgent to explore a method that can quickly and accurately detect the Wanshan Musk component in flavors. Summary of the invention

[0006] The object of the present invention is to provide a method for detecting the content of musk dew in essence, by pre-treating the sample with a mixture of ether / n-hexane / petroleum ether and performing gas chromatography-mass spectrometry analysis under preferred gas chromatography-mass spectrometry conditions, thereby achieving rapid, simple and accurate detection of musk dew in essence.

[0007] In order to achieve the above object, the present invention provides the following technical solutions: A gas chromatography-mass spectrometry detection method for Wanshan Musk in essence comprises the following steps: S1: Sample extraction: adding an organic solvent to the essence for extraction, vortexing and then shaking to obtain liquid A; S2: extraction separation: ultrapure water, magnetic particles and carbon nanotubes are added to the liquid A in step S1 for centrifugation, vortex oscillation, and solid-liquid separation; n-butane is then added to the separated organic layer, ultrasonically treated, separated, eluted, and the eluate is collected to obtain liquid B; S3: Concentration and filtration: Liquid B obtained in step S2 is concentrated in a water bath under nitrogen protection, and liquid C is obtained after constant volume and microporous filtration; S4: Gas chromatography-mass spectrometry analysis: Liquid C obtained in step S3 is analyzed and detected under gas chromatography-mass spectrometry conditions.

[0008] Preferably, step S1 is as follows: by mass g, 10-20 g of the flavor sample is added to 50-100 g of a mixture of ether / n-hexane / petroleum ether, wherein the volume ratio of ether / n-hexane / petroleum ether in the mixture is 1:1.1-1.3:1.5-2; extraction is performed at a temperature of 80-100° C. and a speed of 250-450 r / min, vortexed for 2-6 min, and then shaken for 10-20 min to obtain liquid A.

[0009] Preferably, by mass g, 12-16 g of the flavor sample is added to 65-85 g of ether / n-hexane / petroleum ether mixture, wherein the volume ratio of the three in the ether / n-hexane / petroleum ether mixture is 1:1.2:1.6; extraction is performed at a temperature of 90-95°C and a speed of 300-400 r / min, vortexed for 3-4 min, and then shaken for 14-16 min to obtain liquid A.

[0010] Preferably, by mass g, 5-10 g of ultrapure water, 1-3 g of magnetic particles and 0.5-1 g of carbon nanotubes are added to the liquid A in step S1 for centrifugation, vortex oscillation, and solid-liquid separation; by mass g, 15-30 g of n-butane is added to the 10-20 g organic layer obtained by separation, ultrasonically treated, separated, eluted, and the eluate is collected to obtain liquid B.

[0011] Preferably, by mass g, 7-9 g of ultrapure water, 1-3 g of magnetic particles and 0.5-1 g of carbon nanotubes are added to the liquid A in step S1, and the solution is centrifuged at a rate of 6000-8000 rps for 5-10 min, vortexed for 2-5 min, and shaken for 3-6 min for solid-liquid separation.

[0012] Preferably, by mass, 16-22 g of n-butane is added to 15-20 g of the separated organic layer, and ultrasonic treatment is performed at a temperature of 40-60° C. for 10-20 min. After separation and elution 2-4 times, the eluate is collected to obtain liquid B.

[0013] Preferably, in step S3, under nitrogen protection, the solution is concentrated in a water bath at a temperature of 35-55° C. to 0.2-0.5 mL, the residue is dissolved with ethanol and the volume is fixed to 1.0 mL, and the solution is filtered through a 0.25 μm microporous filter membrane to obtain liquid C.

[0014] Preferably, the gas chromatography conditions in step S4 are: using an HP-5MS type 60m×0.25mm×0.25μm fused quartz capillary column; injection volume 0.5-0.8μL; carrier gas: He; flow rate: 1-3mL / min; injection port temperature 260-320℃; programmed temperature conditions: 80-100℃, hold for 2-4min, increase to 200-250℃ at 20-30℃ / min, then increase to 260-320℃ at a rate of 5-8℃ / min, and hold for 10-15min.

[0015] Preferably, the injection volume is 0.7 μL; carrier gas: He; flow rate: 1.8 mL / min; injection port temperature is 285°C; programmed temperature conditions: 90°C, hold for 3 min, increase to 250°C at 25°C / min, then increase to 285°C at a rate of 5°C / min, and hold for 12 min.

[0016] Preferably, the mass spectrometry conditions in step S4 are: electron impact ion source (EI), electron current of 70ev; ion source temperature of 250-280°C, quadrupole temperature of 150-170°C, transmission line temperature of 260-280°C; filament current: 35μA; multiple reaction monitoring (MRM) mode is selected; internal standard method is used for quantification; characteristic ion fragment mass-to-charge ratios during identification are: m / z 243, 258, 186.

[0017] Compared with the prior art, the advantages and beneficial effects of the present invention are: 1. In the sample pretreatment stage, ether / n-hexane / petroleum ether is used as the organic extract, and ultrapure water, magnetic particles and carbon nanotubes are added for centrifugation. After vortex oscillation, ultrasound and other processes, the extraction and separation rate is accelerated and the separation effect is good. At the same time, the unique structure, physical and chemical properties of multi-walled carbon nanotubes are used to strongly adsorb and purify impurities.

[0018] 2. In the gas chromatography and mass spectrometry detection stage, by optimizing the gas chromatography and mass spectrometry conditions, the content of Wanshan Musk in the essence can be detected more efficiently and accurately, with high sensitivity and good detection effect. DETAILED DESCRIPTION

[0019] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0020] Example 1 This embodiment provides a gas chromatography-mass spectrometry detection method for Wanshan Musk in a fragrance, comprising the following steps: S1: Sample extraction: 10 g of flavor sample was added into 50 g of a mixture of ether / n-hexane / petroleum ether, wherein the volume ratio of ether / n-hexane / petroleum ether in the mixture was 1:1.1:1.5; the mixture was extracted at a temperature of 80°C and a speed of 250 r / min, vortexed for 2 min, and then shaken for 10 min to obtain liquid A; S2: Extraction separation: add 5 g ultrapure water, 1 g magnetic particles and 0.5 g carbon nanotubes to the liquid A in step S1 and centrifuge at a rate of 6000 rps for 5 min, vortex for 2 min, oscillate for 6 min, and separate the solid and liquid; then add 15 g n-butane to the 10 g organic layer obtained by separation, ultrasonically treat at a temperature of 60° C. for 10 min, separate and elute twice, and collect the eluate to obtain liquid B; S3: Concentration and filtration: Under nitrogen protection, concentrate in a water bath at 35°C to 0.2 mL, dissolve the residue with ethanol and make up to 1.0 mL, and filter through a 0.25 μm microporous membrane to obtain liquid C; S4: Gas chromatography-mass spectrometry (GC-MS) analysis: HP-5MS 60m×0.25mm×0.25μm fused quartz capillary column was used; injection volume was 0.5μL; carrier gas: He; flow rate: 1mL / min; injection port temperature was 260℃; programmed temperature conditions: 80℃, maintained for 2min, increased to 200℃ at 20℃ / min, then increased to 260℃ at a rate of 5℃ / min, and maintained for 10min.

[0021] Example 2 This embodiment provides a gas chromatography-mass spectrometry detection method for Wanshan Musk in a fragrance, comprising the following steps: S1: Sample extraction: 20 g of flavor sample was added to 100 g of a mixture of ether / n-hexane / petroleum ether, wherein the volume ratio of ether / n-hexane / petroleum ether in the mixture was 1:1.3:1.5; extraction was performed at a temperature of 100°C and a speed of 450 r / min, vortexed for 6 min, and then shaken for 20 min to obtain liquid A; S2: Extraction separation: add 10 g ultrapure water, 3 g magnetic particles and 1 g carbon nanotubes to the liquid A in step S1 and centrifuge at a rate of 8000 rps for 10 min, vortex for 2 min, oscillate for 6 min, and separate the solid and liquid; then add 30 g n-butane to the 20 g organic layer obtained by separation, ultrasonically treat at a temperature of 40°C for 20 min, separate and elute 4 times, and collect the eluate to obtain liquid B; S3: Concentration and filtration: Under nitrogen protection, concentrate the solution in a water bath at 55°C to 0.5 mL, dissolve the residue in ethanol and adjust the volume to 1.0 mL, and filter through a 0.25 μm microporous filter to obtain liquid C; S4: Gas chromatography-mass spectrometry (GC-MS) analysis: HP-5MS 60m×0.25mm×0.25μm fused quartz capillary column was used; injection volume was 0.8μL; carrier gas: He; flow rate: 3mL / min; injection port temperature was 320℃; programmed temperature conditions: 100℃, maintained for 4min, increased to 250℃ at 30℃ / min, and then increased to 320℃ at a rate of 8℃ / min, and maintained for 15min.

[0022] Example 3 This embodiment provides a gas chromatography-mass spectrometry detection method for Wanshan Musk in a fragrance, comprising the following steps: S1: Sample extraction: 12 g of flavor sample was added into 80 g of a mixture of ether / n-hexane / petroleum ether, wherein the volume ratio of ether / n-hexane / petroleum ether in the mixture was 1:1.2:1.7; the mixture was extracted at 90°C and 300 r / min, vortexed for 4 min, and then shaken for 15 min to obtain liquid A; S2: Extraction separation: add 7 g ultrapure water, 2 g magnetic particles and 0.5 g carbon nanotubes to the liquid A in step S1, centrifuge at a rate of 7000 rps for 8 min, vortex for 3 min, oscillate for 5 min, and separate the solid and liquid; then add 28 g n-butane to the 15 g organic layer obtained by separation, ultrasonically treat at a temperature of 50° C. for 15 min, separate and elute 3 times, and collect the eluate to obtain liquid B; S3: Concentration and filtration: Under nitrogen protection, concentrate in a water bath at 40°C to 0.4 mL, dissolve the residue with ethanol and make the volume to 1.0 mL, and filter through a 0.25 μm microporous membrane to obtain liquid C; S4: Gas chromatography-mass spectrometry (GC-MS) analysis: HP-5MS 60m×0.25mm×0.25μm fused quartz capillary column was used; injection volume was 0.6μL; carrier gas: He; flow rate: 1.5mL / min; injection port temperature was 280℃; programmed temperature conditions: 85℃, maintained for 2min, increased to 250℃ at 25℃ / min, and then increased to 280℃ at a rate of 7℃ / min, and maintained for 12min.

[0023] Example 4 This embodiment provides a gas chromatography-mass spectrometry detection method for Wanshan Musk in a fragrance, comprising the following steps: S1: Sample extraction: 18 g of flavor sample was added into 60 g of ether / n-hexane / petroleum ether mixture, wherein the volume ratio of ether / n-hexane / petroleum ether in the mixture was 1:1.15:1.6; extraction was performed at 95°C and 400 r / min, vortexed for 5 min, and then shaken for 18 min to obtain liquid A; S2: Extraction separation: add 8 g ultrapure water, 3 g magnetic particles and 1 g carbon nanotubes to the liquid A in step S1, centrifuge at a rate of 7500 rps for 7 min, vortex for 4 min, oscillate for 5 min, and separate the solid and liquid; then add 25 g n-butane to the 16 g organic layer obtained by separation, ultrasonically treat at a temperature of 55°C for 15 min, separate and elute 3 times, and collect the eluate to obtain liquid B; S3: Concentration and filtration: Under nitrogen protection, concentrate in a water bath at 40°C to 0.4 mL, dissolve the residue with ethanol and make the volume to 1.0 mL, and filter through a 0.25 μm microporous membrane to obtain liquid C; S4: Gas chromatography-mass spectrometry (GC-MS) analysis: HP-5MS 60m×0.25mm×0.25μm fused quartz capillary column was used; injection volume was 0.7μL; carrier gas: He; flow rate: 2mL / min; injection port temperature was 300℃; programmed temperature conditions: 85℃, maintained for 3min, increased to 250℃ at 25℃ / min, and then increased to 300℃ at a rate of 5℃ / min, and maintained for 10-15min.

[0024] Example 5 This embodiment provides a gas chromatography-mass spectrometry detection method for Wanshan Musk in a fragrance, comprising the following steps: S1: Sample extraction: 15 g of flavor sample was added into 70 g of a mixture of ether / n-hexane / petroleum ether, wherein the volume ratio of ether / n-hexane / petroleum ether in the mixture was 1:1.2:1.6; the mixture was extracted at a temperature of 90°C and a speed of 400 r / min, vortexed for 4 min, and then shaken for 15 min to obtain liquid A; S2: Extraction and separation: add 8 g of ultrapure water, 1.5 g of magnetic particles and 0.8 g of carbon nanotubes to the liquid A in step S1, centrifuge at a rate of 7000 rps for 8 min, vortex for 4 min, oscillate for 5 min, and separate the solid and liquid; then add 20 g of n-butane to the 15 g organic layer obtained by separation, ultrasonically treat at a temperature of 55° C. for 15 min, separate and elute 3 times, and collect the eluate to obtain liquid B; S3: Concentration and filtration: Under nitrogen protection, concentrate in a water bath at 50°C to 0.4 mL, dissolve the residue with ethanol and make up to 1.0 mL, and filter through a 0.25 μm microporous filter to obtain liquid C; S4: Gas chromatography-mass spectrometry (GC-MS) analysis: HP-5MS 60m×0.25mm×0.25μm fused quartz capillary column was used; injection volume was 0.65μL; carrier gas: He; flow rate: 2mL / min; injection port temperature was 285°C; programmed temperature conditions: 100°C, maintained for 2min, increased to 200°C at 25°C / min, then increased to 285°C at a rate of 5°C / min, and maintained for 15min.

[0025] Comparative Example 1 The difference between this comparative example and Example 1 is that in the pre-treatment process of step S1, only ether is used as the organic extract.

[0026] Comparative Example 2 The difference between this comparative example and Example 1 is that no magnetic particles and carbon nanotubes are added in step S2.

[0027] Performance Testing The internal standard method was used for quantitative analysis, that is, the chromatographic peak area of ​​each Musk dew was used for regression analysis on its corresponding concentration to obtain a standard curve with a correlation coefficient greater than or equal to 0.999. The extracted samples were measured, and the chromatographic peak area of ​​the detected Musk dew was measured and substituted into the standard curve to obtain the residual amount of Musk dew in the sample.

[0028] In the present invention, the preparation method of the standard working solution is as follows: accurately weigh 10 mg of Wanshan Musk in a 100 mL volumetric flask, accurate to 0.1 mg, dissolve and dilute with methanol to prepare a standard stock solution with a concentration of 100 μg / mL; transfer 1000 μL of the standard stock solution to a 10 mL volumetric flask, dilute with methanol, and the concentration of this standard solution is 10.0 μg / mL; transfer 100, 200, 500, 1000, 2000, 5000 μL of the standard solution to a 10 mL volumetric flask, and add 100 μL of the internal standard working solution, dilute with methanol to make up to the volume, and prepare standard working solutions of different concentrations. The concentrations in the series of standard working solutions are: 100 ng / mL, 200 ng / mL, 500 ng / mL, 1000 ng / mL, 2000 ng / mL and 5000 ng / mL.

[0029] Table 1 Performance test results

[0030] From the above performance test results, it can be seen that the linear coefficients of Examples 1-5 are all above 0.999, the detection limits are also below 1.2, the relative standard deviation is small, the recovery rate is high, and the detection effect and effectiveness are obvious. In particular, the comprehensive performance of Example 3 is the most outstanding.

[0031] However, the comparative example does not adopt the necessary technical solution, resulting in its performance being significantly worse than that of the embodiment in the corresponding performance test. The above experimental results further prove the importance of the technical solution defined in the present invention to its technical effect.

[0032] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A gas chromatography-mass spectrometry detection method for Wanshan Musk in essence, It is characterized in that The following steps are included: S1: Sample extraction: add organic solvent to the essence for extraction, vortex treatment and then shake treatment to obtain liquid A; S2: extraction separation: ultrapure water, magnetic particles and carbon nanotubes are added to the liquid A in step S1 for centrifugation, vortex oscillation, and solid-liquid separation; n-butane is then added to the separated organic layer, ultrasonically treated, separated, eluted, and the eluate is collected to obtain liquid B; S3: Concentration and filtration: Liquid B obtained in step S2 is concentrated in a water bath under nitrogen protection, and liquid C is obtained after constant volume and microporous filtration; S4: Gas chromatography-mass spectrometry analysis: Liquid C obtained in step S3 is analyzed and detected under gas chromatography-mass spectrometry conditions.

2. The gas chromatography-mass spectrometry detection method of Wanshan Musk in a flavor according to claim 1, It is characterized in that Specifically, step S1 includes: taking 10-20 g of the essence sample by mass, adding 50-100 g of ether / n-hexane / petroleum ether mixture, wherein the volume ratio of ether / n-hexane / petroleum ether in the mixture is 1:1.1-1.3:1.5-2; extracting at a temperature of 80-100° C. and a speed of 250-450 r / min, vortexing for 2-6 min, and shaking for 10-20 min to obtain liquid A.

3. The gas chromatography-mass spectrometry detection method of Wanshan Musk in a flavor according to claim 2, It is characterized in that 12-16 g of flavor sample was added into 65-85 g of ether / n-hexane / petroleum ether mixture, wherein the volume ratio of ether / n-hexane / petroleum ether mixture was 1:1.2:1.6; the mixture was extracted at 90-95°C and 300-400 r / min, vortexed for 3-4 min, and then shaken for 14-16 min to obtain liquid A.

4. The method for detecting the Wanshan Musk in a flavor by gas chromatography-mass spectrometry according to claim 1, It is characterized in that In terms of mass g, 5-10 g of ultrapure water, 1-3 g of magnetic particles and 0.5-1 g of carbon nanotubes are added to the liquid A in step S1, and the mixture is centrifuged, vortexed and subjected to solid-liquid separation; 15-30 g of n-butane is added to the 10-20 g organic layer obtained by separation, and the mixture is ultrasonically treated, separated and eluted, and the eluate is collected to obtain liquid B.

5. The method for detecting the Wanshan Musk in the essence by gas chromatography-mass spectrometry according to claim 4, It is characterized in that In terms of mass g, 7-9 g of ultrapure water, 1-3 g of magnetic particles and 0.5-1 g of carbon nanotubes were added to the liquid A in step S1, and the mixture was centrifuged at a rate of 6000-8000 rps for 5-10 min, vortexed for 2-5 min, and shaken for 3-6 min for solid-liquid separation.

6. The method for detecting the Wanshan Musk in the essence by gas chromatography-mass spectrometry according to claim 4, It is characterized in that In terms of mass, 16-22 g of n-butane was added to 15-20 g of the separated organic layer, and the mixture was ultrasonically treated at a temperature of 40-60° C. for 10-20 min. After separation and elution for 2-4 times, the eluate was collected to obtain liquid B.

7. The method for detecting the Wanshan Musk in a flavor by gas chromatography-mass spectrometry according to claim 1, It is characterized in that In step S3, under nitrogen protection, the liquid B obtained in step S2 is concentrated to 0.2-0.5 mL in a water bath at a temperature of 35-55° C., the residue is dissolved with ethanol and the volume is fixed to 1.0 mL, and filtered through a 0.25 μm microporous filter membrane to obtain liquid C.

8. The method for detecting the Wanshan Musk in a flavor by gas chromatography-mass spectrometry according to claim 1, It is characterized in that The gas chromatography conditions in step S4 are: using a HP-5MS type 60m×0.25mm×0.25μm fused quartz capillary column; injection volume 0.5-0.8μL; carrier gas: He; Flow rate: 1-3mL / min; injection port temperature 260-320℃; programmed temperature conditions: 80-100℃, hold for 2-4min, increase to 200-250℃ at 20-30℃ / min, then increase to 260-320℃ at a rate of 5-8℃ / min, and hold for 10-15min.

9. The method for detecting the Wanshan Musk in the essence by gas chromatography-mass spectrometry according to claim 8, It is characterized in that Injection volume: 0.7 μL; carrier gas: He; Flow rate: 1.8 mL / min; injection port temperature 285°C; programmed temperature conditions: 90°C, hold for 3 min, increase to 250°C at 25°C / min, then increase to 285°C at a rate of 5°C / min, and hold for 12 min.

10. The method for detecting the Wanshan Musk in a flavor by gas chromatography-mass spectrometry according to claim 1, It is characterized in that The mass spectrometry conditions in step S4 are: electron impact ion source (EI), electron current of 70 ev; ion source temperature of 250-280°C, quadrupole temperature of 150-170°C, transfer line temperature of 260-280°C; filament current: 35 μA; multiple reaction monitoring (MRM) mode is selected; internal standard method is used for quantification; characteristic ion fragment mass-to-charge ratios during identification are: m / z 243, 258, 186.

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