CN113960236B - Method for determining skatole and dimethyl isoborneol in fish body based on rapid pretreatment technology - Google Patents
Method for determining skatole and dimethyl isoborneol in fish body based on rapid pretreatment technology Download PDFInfo
- Publication number
- CN113960236B CN113960236B CN202111180121.5A CN202111180121A CN113960236B CN 113960236 B CN113960236 B CN 113960236B CN 202111180121 A CN202111180121 A CN 202111180121A CN 113960236 B CN113960236 B CN 113960236B
- Authority
- CN
- China
- Prior art keywords
- fish
- sample
- extraction
- temperature
- gsm
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/025—Gas chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
- G01N2030/146—Preparation by elimination of some components using membranes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- 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
- G01N2030/884—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 organic compounds
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention belongs to the technical field of aquaculture and food monitoring, discloses a method for measuring skatole and dimethyl isoborneol in fish body based on a rapid pretreatment technology, and establishes a detection method for measuring GSM and 2MIB in fish tissue by a dispersive solid phase extraction-gas chromatography-mass spectrometry. In the method, a sample is extracted by acetonitrile solution, salting-out is assisted in extraction, after purification, under the gradient heating condition, target analytes are qualitatively analyzed by comparing retention time with ion pair information in a selected ion monitoring mode, and the internal standard method is used for quantification. The method has the advantages that: the method has the advantages of small required sample quantity and reagent quantity, high analysis speed, high sensitivity and good reproducibility, can meet the requirement of mass detection of GSM and 2MIB in fish tissue samples, and provides a rapid detection technical guarantee for evaluating the occurrence condition of the earthy substances in fish tissues and exploring the reduction and removal of typical earthy substances in fish bodies.
Description
Technical Field
The invention relates to a method for measuring earthy substances in fish bodies based on rapid pretreatment, belongs to the technical field of aquaculture and food monitoring, and mainly relates to freshwater fish, seawater fish and the like. The method is mainly based on a dispersion solid phase extraction technology, adopts an improved dispersion extraction and purification method to complete the effective extraction of the skatole and the dimethyl isoborneol in the sample, and then carries out GC/MS quantitative analysis, so that the rapid extraction and detection of the skatole and the dimethyl isoborneol in farmed fish and wild fish bodies can be realized, the analysis time can be effectively saved, and the analysis cost can be saved.
Background
The problem of the earthy smell in the aquatic products is long, the earthy smell substances in the aquatic products have profound effects on links such as cultivation, processing, sales and consumption of the aquatic products, and serious economic losses can be caused to related industries. The earthy smell substance is a generic term for various chemical substances, and has a complex composition, and among them, geosmin (GSM) and dimethyl isoborneol (2-MIB) are considered as two main substances responsible for earthy smell, and their related researches are also the greatest. Most of the production of GSM and 2-MIB is caused by environmental factors and also comprises related biochemical reactions occurring in aquatic animals, such as oxidation of fatty acid, enzymatic conversion of sulfur-containing and nitrogen-containing precursor substances, degradation of trimethylamine in fish under the action of microorganisms and enzymes, and the like.
Research shows that the contents of GSM and 2-MIB in water environment reach 9ng/L and 4ng/L respectively, so that the aquatic products can generate earthy smell. The content of either GSM or 2-MIB in fish exceeds 0.7 mug/kg, and the fish has a earthy taste. At present, a great deal of research is focused on the content analysis of GSM and 2-MIB in water, and standard inspection methods for odor substances of domestic drinking water, namely GSM and 2-MIB, GB/T32470-2016 are promulgated in 2016 in China, and relatively few research on effective analysis of GSM and 2-MIB in fish bodies is carried out, so that the methods developed at present mainly comprise a sensory analysis method and an instrumental analysis method. The sensory analysis method is considered to be the most direct and simple method, but the aim of quantitative detection cannot be achieved, and the experimental result cannot be repeated for a plurality of times, so that the method has great limitation; and the instrument analysis method can achieve accurate qualitative and quantitative analysis. At present, the Gas Chromatography (GC) or the gas chromatography-mass spectrometry (GC-MS) is often adopted to carry out instrument analysis on the GSM and the 2-MIB, but because the earthy substances in the fish body exist in trace amounts, a series of pretreatment such as enrichment, concentration and the like on the GSM and the 2-MIB in the sample are particularly important. The existing pretreatment methods for the fish tissue samples mainly comprise a microwave distillation extraction method (MAD-SE), a stirring rod adsorption method (SBSE), a solid-phase microextraction method (SPME), a headspace solid-phase microextraction method (HSPME), a purging trapping method (P & T) and the like, and the development of the pretreatment technologies directly influences the application and popularization of the detection of the earthy taste substances in the fish tissues.
Among pretreatment technologies related at home and abroad, a microwave distillation and solid-phase microextraction method is a pretreatment method mainly adopted at present. The microwave distillation has relatively simple operation steps, but has poor enrichment effect on organic matters with stronger volatility, such as earthy substances and the like. The microwave distillation-solid phase microextraction can fully extract volatile components in fish bodies, does not need an organic solvent or concentration, but has longer extraction time, more expensive extraction head and shorter service life, and is not suitable for mass detection and analysis. The microwave distillation-sweeping and trapping does not need an organic solvent, so that the time is saved, but the extraction solvent sodium chloride concentration is too high, the instrument is blocked, and the technology is not completely mature. The stirring rod is used for adsorption, an organic solvent is not needed, but the adsorption balance can be achieved only for a longer time, and according to the sample quantity, the short-term balance needs 1h, and the long-term balance can achieve the adsorption balance only for 3-4 h. Dynamic headspace extraction can perform continuous gas phase extraction, but the sample matrix can interfere with the analysis result, and the loss of sample components may occur in the adsorption and desorption processes, so that the reproducibility is poor. In conclusion, the pretreatment methods have large dependency on matched instruments, high detection time and detection cost, and low applicability to large-scale sampling analysis.
Also for the above reasons, there is an ongoing interest in actively developing rapid, inexpensive, efficient pretreatment techniques for GSM and 2-MIB in fish organizations to meet large-scale detection analysis. Among them, the liquid-liquid extraction and the dispersion solid phase extraction method are attracting attention again because of simple operation, rapidness and high repeatability. In recent process studies, researchers developed pretreatment techniques using n-hexane extraction. However, in the extraction of normal hexane, besides the extraction of the target substances GSM and 2-MIB, the lipophilicity of the normal hexane can cause the extraction of fat and lipophilic compounds in fish tissues, so that the purification treatment process of an SPE column is required to be added in the operation, the pretreatment extraction time is prolonged, the process is increased, and the detection cost is increased. The research technology is based on the dispersion solid phase extraction principle, acetonitrile and a water system are adopted, the rapid extraction of the target objects GSM and 2-MIB can be completed at normal temperature and normal pressure, and then the rapid purification is carried out for instrument analysis, so that the detection cost can be obviously reduced, the detection time is saved, the repeatability is high, and the method is suitable for large-batch rapid detection of the GSM and 2-MIB in the fish organization.
Fish is one of the main food sources for human consumption, and the earthy taste substances in the fish body always affect the whole process from cultivation (or fishing) to dining tables for a long time, and the objective evaluation of the earthy taste substances in the fish body and the development of the reduction technology are seriously dependent on the development of the related detection technology. In order to reduce the detection cost and save the detection time, it is very necessary to develop a rapid, simple and convenient pretreatment technology suitable for mass detection aiming at typical earthy substances GSM and 2-MIB.
Disclosure of Invention
The invention aims to provide a method for rapidly, simply and efficiently measuring GSM and 2-MIB in fish based on the prior art.
The technical scheme of the invention is as follows:
a method for determining the content of skatole and dimethyl isoborneol in fish based on a rapid pretreatment technology comprises the following steps:
(1) Liquid-solid extraction: acetonitrile is added into the homogenized fish tissue sample for extraction, and vortex oscillation is carried out for 30-60 s;
(2) Salting out and back extraction: adding sodium chloride into the sample obtained in the step (1), adding 1-2 g of sodium chloride into every 2g of fish tissue sample, carrying out vortex oscillation for 30-60 s, and centrifuging for 3-5 min at 8000-12000 r/min;
(3) Dehydrating and purifying: adding anhydrous sodium sulfate into the solution to be purified obtained in the step (2), adding 4-6 g of anhydrous sodium sulfate into every 2g of fish tissue sample, carrying out vortex oscillation for 30-60 s, centrifuging for 3-5 min at 8000-12000 r/min, and transferring supernatant;
(4) Adsorption purification: adding an adsorption purifying agent into the supernatant obtained in the step (3), carrying out vortex oscillation for 30-60 s, centrifuging for 3-5 min at 8000-12000 r/min, and transferring the supernatant; the adsorption purifying agent is PSA and activated carbon, and 0.1g of the adsorption purifying agent is added into each 2g of fish tissue sample;
(5) And (3) membrane filtration and purification: filtering the supernatant obtained in the step (4) by a filtering membrane, purifying and loading the filtered supernatant into a machine;
(6) Preparing a standard solution: first, 10mg of 2-MIB was weighed and fixed to 10mL with methanol to prepare a standard stock solution of 1mg/mL, and 10mg of GSM was weighed and fixed to 10mL with methanol to prepare a standard stock solution of 1 mg/mL. And (3) respectively weighing a proper amount of GSM and 2-MIB, and preparing a mixed standard intermediate solution with the concentration of 1mg/L by using methanol. Sucking a proper amount of mixed standard intermediate solution, and diluting the mixed standard intermediate solution into standard series working solutions containing GSM and 2-MIB of 1ng/mL, 2ng/mL, 4ng/mL, 10ng/mL, 20ng/mL and 40ng/mL respectively by using methanol, wherein the content of an internal standard 2-isobutyl-3-methoxy pyrazine is 4ng/mL;
(7) Qualitative and quantitative analysis: and (3) feeding the liquid to be analyzed into a gas chromatograph-mass spectrometer, monitoring by adopting a SIM scanning mode, and quantifying by an internal standard method.
a) Qualitative analysis: determining a sample by using a GC-MS instrument and establishing a standard working curve, wherein if the retention time of a chromatographic peak detected in the sample is consistent with the retention time of a chromatographic peak of a corresponding standard substance, the allowable deviation is less than +/-2.5%; comparing the relative abundance of each component qualitative ion in the sample with the relative abundance of the corresponding qualitative ion in the mixed standard working solution with the concentration close to the relative abundance, and judging that the corresponding object to be detected exists in the sample if the deviation is not more than the range specified in the table 1.
TABLE 1 maximum allowable deviation of relative ion abundance in qualitative validation
| Relative ion abundance | ＞50％ | > 20% to 50% | > 10% to 20% | ≦10％ |
| Maximum allowable deviation | ±20％ | ±25％ | ±30％ | ±50％ |
b) Quantitative analysis: determining mixed standard series working solution by using a GC-MS method to obtain chromatographic peak areas of corresponding standard solutions, and drawing a standard curve by taking the concentration of the mixed standard working solution as an abscissa and the peak areas of the chromatographic peaks as an ordinate; and then the sample to be measured is measured under the same condition, the chromatographic peak area of the sample to be measured is obtained, and the concentration of each component in the liquid to be measured is obtained according to the standard curve. The response value of each component to be measured in the sample should be within the linear response range of the standard curve.
(7) And (3) calculating results:
the calculation formula is as follows:
in the formula (1):
x-the content of the component to be measured in micrograms per kilogram (μg/kg) in the sample;
c-the concentration of each component to be measured in micrograms per liter (μg/L) in the measurement solution read from the standard curve;
m-mass of sample weighing in grams (g);
v-the volume of the sample solution is determined in milliliters (mL).
The standard stock solution can be stored for 3 months at the temperature of minus 18 ℃ in a dark place.
The mixed standard series working solution is prepared temporarily.
The gas chromatography-mass spectrometry test conditions of the invention are as follows:
the chromatographic conditions of the GC-MS are as follows:
a) Chromatographic column: DB-35MS quartz capillary column, 30m x 0.32mm (i.d), film thickness 0.25 μm, or equivalent.
b) Chromatographic column temperature: starting at 100deg.C, heating to 220deg.C at 12deg.C/min, and heating to 280deg.C at 30deg.C/min (holding for 5 min).
c) Sample inlet temperature: 230 ℃.
d) Chromatographic-mass spectrometry interface temperature: 280 ℃.
e) Carrier gas: helium with purity of 99.999% or more and 1.5mL/min.
f) Sample injection amount: 1 mul.
g) Sample injection mode: sample introduction is not carried out in a split way, and the valve is opened after 0.75 min.
The mass spectrum conditions of the GC-MS are as follows:
k) Ionization mode: EI.
l) ionization energy: 70eV.
m) detection mode: ion monitoring mode (SIM) is selected.
n) solvent delay: 3.0min.
The method has the advantages that after the sample is subjected to solvent extraction by acetonitrile, salting-out back extraction is assisted, and after the extracting solution is adsorbed and purified by the purifying agent, gas analysis is performed under the temperature programming condition. The average recovery rate of the compound is 87.2% -115.7% and the Relative Standard Deviation (RSD) is 0.6% -6.2%. When the sample is 2g and the constant volume is 5mL, the detection limit of GSM is 0.37 mug/kg, and the detection limit of 2-MIB is 0.51 mug/kg; the quantitative limit of GSM was 1.23. Mu.g/kg and the quantitative limit of 2-MIB was 1.69. Mu.g/kg.
The invention has the advantages of simple pretreatment, less consumption of samples and solvents, less matrix interference, high analysis speed, high sensitivity, good reproducibility and the like, can be suitable for the rapid detection of a large number of samples, and meets the requirements of daily detection work.
Drawings
FIG. 1 (a) is a mass spectrum (qualitative and quantitative ion selective) of GSM according to an embodiment of the present invention;
FIG. 1 (b) is a mass spectrum (qualitative and quantitative ion selection) of MIB according to example 2 of the present invention;
FIG. 2 is a Selected Ion Monitoring (SIM) chromatogram of GSM and 2-MIB in a fish organization according to an embodiment of the invention.
Detailed Description
In order to make the technical solution of the present invention better understood by those skilled in the art, the technical solution of the present invention is further explained below with reference to the accompanying drawings and the following examples, but is not to be construed as limiting the scope of the present invention.
Example 1
(1) Liquid-solid dispersion extraction: weighing 2.0g of sample, placing the sample into a 20mL polypropylene centrifuge tube, adding 2mL of water, carrying out vortex oscillation for 30s, dispersing the tissue sample, adding 5mL of acetonitrile extraction solution, carrying out vortex oscillation for 30s, adding 1g of sodium chloride for back extraction, carrying out vortex oscillation for 30s, adding 4g of anhydrous sodium sulfate, centrifuging for 5min at 8000r/min, and transferring supernatant into the 20mL polypropylene centrifuge tube for purification.
(2) Liquid-solid adsorption purification: to the supernatant, 0.1g of PSA powder and 0.1g of C18 powder were added, and after vortexing for 30s and centrifugation at 8000r/min for 1min, the supernatant was taken and passed through a microporous filter membrane for analysis.
(3) Preparing a standard solution: first, 0.01g (accurate to 0.0001 g) of GSM and 2-MIB were weighed, and each was fixed to 10mL with methanol to prepare a 1mg/mL stock solution of standard substance. And (3) respectively weighing a proper amount of GSM and 2-MIB, and preparing a mixed standard intermediate solution with the concentration of 1mg/L by using methanol. Absorbing proper amount of mixed standard intermediate solution, diluting the mixed standard intermediate solution with methanol to obtain standard series working solutions containing GSM and 2-MIB of 1ng/mL, 2ng/mL, 4ng/mL, 10ng/mL, 20ng/mL and 40ng/mL respectively, wherein the content of internal standard 2-isobutyl-3-methoxy pyrazine is 4ng/mL
(4) Qualitative and quantitative analysis: and (3) feeding the liquid to be analyzed into a gas chromatograph-mass spectrometer (GC-MS), and adopting a Selective Ion Monitoring (SIM) mode for monitoring and quantifying by an internal standard method. Specific test conditions for qualitative and quantitative analysis are as follows:
gas chromatography conditions: DB-35MS quartz capillary column chromatography (30 m×0.32mm, film thickness 0.25 μm, or equivalent) is selected, and the temperature programming condition of the column is: starting at 100deg.C, heating to 220deg.C at 12deg.C/min, and heating to 280deg.C at 30deg.C/min (holding for 5 min). Sample inlet temperature: 230 ℃. Chromatographic-mass spectrometry interface temperature: 280 ℃. Carrier gas: helium with purity of 99.999% or more and 1.5mL/min. Sample injection mode: sample introduction is not carried out in a split way, and the valve is opened after 0.75 min. Sample injection amount: 1 mul. Mass spectrometry parameter conditions: ionization mode is Electrospray Ionization (EI), ionization energy: 70eV. The detection mode is a selective ion monitoring mode (SIM). Solvent delay: 3.0min.
When the chromatographic column is selected, three different chromatographic column fillers, namely an HP-5 capillary column chromatographic column (30 m multiplied by 0.25mm, film thickness of 0.25 mu m), a DB-35MS quartz capillary column chromatographic column (30 m multiplied by 0.32mm, film thickness of 0.25 mu m) and a DB-35MS quartz capillary column chromatographic column (30 m multiplied by 0.53mm, film thickness of 3.0 mu m), are respectively selected. The result shows that the GSM and 2-MIB have good chromatographic retention behaviors on three chromatographic columns, the peak shape, response, separation degree and commodity price of each compound to be tested are comprehensively considered, and finally, a DB-35MS quartz capillary column chromatographic column (30 m multiplied by 0.32mm, film thickness of 0.25 μm) is selected.
In the invention, during the selection of the extraction solvent, the related compounds are easily dissolved in organic reagents such as methanol, acetonitrile, n-hexane and the like, so that the methanol, the acetonitrile, the n-hexane and the ethyl acetate which are commonly used in a laboratory are firstly selected as the extraction solvent in the process of sample extraction. Experimental results show that the extraction efficiency of methanol, acetonitrile n-hexane and ethyl acetate on GSM and 2-MIB can reach more than 80%, in order to save the use amount of an organic solvent and consider that acetonitrile has obvious precipitation effect on free proteins containing samples, and in order to achieve a better purification effect, the invention determines acetonitrile as an extraction solvent.
The invention respectively examines 100mg PSA/100mg C during the selection of sample purification 18 100mg PSA/100mg neutral alumina, 100mg C 18 100mg neutral alumina, effect of 3 combinations on sample purification. PSA is a commonly used dispersed solid phase extraction scavenger that can remove fatty acids, organic acids, polar substances, etc. from tissues by weak anion exchange or polar action. C (C) 18 Can effectively adsorb compounds such as fat and fatty acid. Neutral alumina is a strong polar adsorbent, the surface of which is neutral, and is easy to retain electron-rich compounds such as heterocycles, organic amine and the like. By applying to different C 18 Examination of neutral alumina and PSA combination, 100mg PSA/100mg C 18 When the combination is a purifying agent, the recovery rate of GSM and 2-MIB is 80-120 percent. Thus, 100mg of PSA/100mg of C is ultimately employed 18 Sample purification treatment was performed in combination.
In the invention, the influence of anhydrous sodium sulfate and anhydrous magnesium sulfate is examined when the dehydrating agent is selected, the influence of the anhydrous sodium sulfate and the anhydrous magnesium sulfate on the recovery rate of GSM and 2-MIB is not obvious, and the anhydrous sodium sulfate is adopted as the dehydrating agent in view of strong water absorption capacity and weak adsorption effect of the anhydrous sodium sulfate.
In the embodiment of the invention, GSM is purchased from Shanghai microphone Biochemical technology Co., ltd, and the purity is more than or equal to 97%;2-MIB was purchased from Chem Service company, U.S.A., and the purity was not less than 99%.
Results and analysis of the five
a) Regression analysis is carried out under the selected chromatographic and mass spectrum conditions according to standard solutions with the determined series of concentrations, the concentration of the target object is taken as an abscissa (x), the peak area of a chromatographic peak is taken as an ordinate (y), the results are shown in Table 2, GSM and 2-MIB have good linearity within the range of 0.5-500 mug/L, and the correlation coefficient r is more than 0.997. According to the definition of the detection limit by the International Union of Pure and Applied Chemistry (IUPAC), serial mixed standard solutions are diluted and tested, the signal to noise ratio (S/N) is calculated, S/n=3 as the detection Limit (LOD) and S/n=10 as the quantification Limit (LOQ). A mixed standard solution was quantitatively added to a blank sample containing no target substance, and the mixture was treated and tested, and the LOD of GSM and 2-MIB was classified into 0.37. Mu.g/kg and 0.51. Mu.g/kg, and the LOQ of the method was 1.23. Mu.g/kg and 1.69. Mu.g/kg.
Table 2 linear equations, correlation coefficients, linear range (n=6), detection limit and quantitative limit of GSM and 2-MIB
| Analyte(s) | Linear equation | r | Linear range/. Mu.g/L | LOD(μg/kg) | LOQ(μg/kg) |
| GSM | y＝79.037x-78.481 | 0.999 | 0.5-500 | 0.37 | 1.23 |
| 2-MIB | y＝80.022x+19.192 | 0.999 | 0.5-500 | 0.51 | 1.69 |
b) The fish tissue samples without GSM and 2-MIB were selected, additive recovery and precision tests were performed at three concentration levels, 6 replicates for each level, and measurements were made according to the procedure specified for the method, and the average recovery for the method was examined, as shown in table 3. The result shows that the average recovery rate of GSM and 2-MIB is 87.2% -115.7% and the Relative Standard Deviation (RSD) is 0.6% -6.2%, which all meet the trace analysis requirement of the sample.
TABLE 3 recovery of GSM and 2-MIB in different fish tissue samples and precision (n=6)
Claims (2)
1. A method for determining the content of skatole and dimethyl isoborneol in fish based on a rapid pretreatment technology is characterized by comprising the following steps:
(1) Liquid-solid extraction: acetonitrile is added into the homogenized fish tissue sample for extraction, and vortex oscillation is carried out for 30-60 s;
(2) Salting out and back extraction: adding sodium chloride into the sample obtained in the step (1), adding 1-2 g of sodium chloride into every 2g of fish tissue sample, and centrifuging for 3-5 min under vortex oscillation 30~60 s,8000~12000 r/min;
(3) Dehydrating and purifying: adding anhydrous sodium sulfate into the solution to be purified obtained in the step (2), adding 4-6 g of anhydrous sodium sulfate into each 2g of fish tissue sample, centrifuging for 3-5 min by vortex oscillation 30~60 s,8000~12000 r/min, and transferring supernatant;
(4) Adsorption purification: adding an adsorption purifying agent into the supernatant obtained in the step (3), carrying out vortex oscillation for 30-60 s, centrifuging for 3-5 min at 8000-12000 r/min, and transferring the supernatant; the adsorption purifying agent is PSA and activated carbon, and 0.1g of the adsorption purifying agent is added into each 2g of fish tissue sample;
(5) And (3) membrane filtration and purification: filtering the supernatant obtained in the step (4) by a filtering membrane, purifying and loading the filtered supernatant into a machine;
(6) Standard series solution preparation: preparing standard series on-machine working solutions containing the skatole and the dimethyl isoborneol with the concentration of 0.5 ng/mL, 1ng/mL, 2ng/mL, 5ng/mL, 10ng/mL and 20ng/mL respectively by using a methanol solution;
(7) Qualitative and quantitative analysis: the liquid to be analyzed enters a gas chromatograph-mass spectrometer, is monitored by adopting a SIM scanning mode, and is quantified by an internal standard method;
specific test conditions for qualitative and quantitative analysis are as follows:
gas chromatography conditions: DB-35MS quartz capillary column chromatography (30 m' -0.32 mm, film thickness 0.25mm or equivalent) is selected, and the temperature programming condition of the column is that: 100. starting at the temperature of 12 ℃/min to 220 ℃, and then 30 ℃/min to 280 ℃ (keeping for 5 min); sample inlet temperature: 230. the temperature is lower than the temperature; chromatographic-mass spectrometry interface temperature: 280. the temperature is lower than the temperature; carrier gas: helium with purity of 99.999% or more and 1.5. 1.5 mL/min; sample injection mode: sampling without diversion, and opening a valve after 0.75 min; sample injection amount: 1mL; mass spectrometry parameter conditions: ionization mode is Electrospray Ionization (EI), ionization energy: 70 eV; the detection mode is a selected ion monitoring mode (SIM); solvent delay: 3.0 And (5) min.
2. The method of claim 1, wherein the membrane-passing purification material in step (5) is a 0.22mm filter membrane.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111180121.5A CN113960236B (en) | 2021-10-11 | 2021-10-11 | Method for determining skatole and dimethyl isoborneol in fish body based on rapid pretreatment technology |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111180121.5A CN113960236B (en) | 2021-10-11 | 2021-10-11 | Method for determining skatole and dimethyl isoborneol in fish body based on rapid pretreatment technology |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113960236A CN113960236A (en) | 2022-01-21 |
| CN113960236B true CN113960236B (en) | 2023-07-25 |
Family
ID=79463410
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111180121.5A Active CN113960236B (en) | 2021-10-11 | 2021-10-11 | Method for determining skatole and dimethyl isoborneol in fish body based on rapid pretreatment technology |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113960236B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117434186A (en) * | 2023-12-22 | 2024-01-23 | 中国市政工程华北设计研究总院有限公司 | Synchronous identification and quantification method for complex odor gas composition of sewage-carrying rainwater |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0538561U (en) * | 1991-10-28 | 1993-05-25 | 株式会社島津製作所 | Gas chromatograph detector |
| CN105784872A (en) * | 2016-05-05 | 2016-07-20 | 劲牌有限公司 | Gas chromatography/ mass spectrometry method for detecting dioctyl terephthalate in Luzhou-flavor Daqu base liquor |
Family Cites Families (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6902675B2 (en) * | 2001-10-19 | 2005-06-07 | Mississippi State University | Method to control off-flavor in water and aquaculture products |
| EP1664728A1 (en) * | 2003-09-07 | 2006-06-07 | Andrea Büttner | Detection of analytes in a defined area of the body |
| CN101625339A (en) * | 2008-07-07 | 2010-01-13 | 吴光红 | Method for detecting residue of seven synthetic antibacterial agents in aquatic products |
| CN102628844A (en) * | 2012-04-09 | 2012-08-08 | 湖南省食品药品检验研究院 | Content determining method for trichlorfon in dried fish |
| CN105021717A (en) * | 2014-04-29 | 2015-11-04 | 中国水产科学研究院 | Method for measuring residual quantity of ethyl 3-aminobenzoate in aquatic product |
| RU2578974C1 (en) * | 2015-01-23 | 2016-03-27 | Федеральное государственное бюджетное учреждение "Всероссийский государственный Центр качества и стандартизации лекарственных средств для животных и кормов" (ФГБУ "ВГНКИ") | Method of detecting residual quantities of triphenylmethane dyes in fish muscular tissue |
| CN104597189B (en) * | 2015-01-29 | 2016-01-06 | 山东出入境检验检疫局检验检疫技术中心 | A kind of assay method of Pyrifluquinazon residual quantity |
| CN105181839A (en) * | 2015-09-06 | 2015-12-23 | 中国农业科学院兰州畜牧与兽药研究所 | Method for detecting residual quantity of ivermectin in sheep muscle tissues by using liquid chromatograph/mass spectrometer with doramectin as internal standard substance |
| CN105259288B (en) * | 2015-10-30 | 2017-06-16 | 四川新希望畜牧科技有限公司 | It is a kind of at the same detect poultry tissues in multi-medicament residual quantity method |
| CN105352782B (en) * | 2015-11-23 | 2018-07-31 | 中国科学院南京地理与湖泊研究所 | The measurement pre-treating method of thioether class odor causing substance in a kind of deposit |
| CN107328889A (en) * | 2017-06-29 | 2017-11-07 | 安徽瑞思威尔科技有限公司 | The detection method of ground depth and 2 methyl isoborneols in Daqu |
| CN107576741B (en) * | 2017-09-14 | 2020-07-17 | 浙江省海洋水产研究所 | Method for detecting organophosphorus pesticide multi-component residues in aquatic product by solid phase extraction-gas chromatography tandem mass spectrometry |
| CN108195982A (en) * | 2017-12-19 | 2018-06-22 | 浙江省海洋水产研究所 | The detection method of persistence organic pollutant in a kind of aquatic products |
| CN109358129A (en) * | 2018-12-06 | 2019-02-19 | 安徽国科检测科技有限公司 | A kind of method that quinolones quickly detects in the flesh of fish |
| CN110108821B (en) * | 2019-04-22 | 2020-10-16 | 广西壮族自治区疾病预防控制中心 | Dispersed solid phase extraction material and preparation method and application thereof |
| CN110174470B (en) * | 2019-05-10 | 2021-10-12 | 宁波检验检疫科学技术研究院 | High-flux detection method for marine biotoxin in aquatic product |
| CN112014485A (en) * | 2019-05-30 | 2020-12-01 | 苏州见远检测技术有限公司 | Method for measuring geosmin and 2-methylisoborneol in sediment |
| CN110672774B (en) * | 2019-11-11 | 2022-07-26 | 临沂大学 | Method for conveniently and rapidly detecting smelly substances including geosmin and dimethyl isoborneol in water source |
| CN110940766A (en) * | 2019-11-28 | 2020-03-31 | 四川威尔检测技术股份有限公司 | Method for determining earthy substances in fish |
| CN111289707A (en) * | 2020-03-11 | 2020-06-16 | 中国海洋大学 | Method for characterizing characteristic taste of steamed fish sample and application |
| CN113176356B (en) * | 2021-04-25 | 2022-11-04 | 中国水产科学研究院淡水渔业研究中心 | Pretreatment method for detecting soil odor substances in aquaculture water |
-
2021
- 2021-10-11 CN CN202111180121.5A patent/CN113960236B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0538561U (en) * | 1991-10-28 | 1993-05-25 | 株式会社島津製作所 | Gas chromatograph detector |
| CN105784872A (en) * | 2016-05-05 | 2016-07-20 | 劲牌有限公司 | Gas chromatography/ mass spectrometry method for detecting dioctyl terephthalate in Luzhou-flavor Daqu base liquor |
Non-Patent Citations (1)
| Title |
|---|
| 气相色谱-质谱法测定鱼油脂肪酸;王建,林秋萍;中国饲料(第17期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113960236A (en) | 2022-01-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | A review of pretreatment and analytical methods of biogenic amines in food and biological samples since 2010 | |
| Mohammed et al. | A critical overview on the chemistry, clean-up and recent advances in analysis of biogenic amines in foodstuffs | |
| Erim | Recent analytical approaches to the analysis of biogenic amines in food samples | |
| Braun et al. | Trace analysis of technical nonylphenol, bisphenol A and 17α-ethinylestradiol in wastewater using solid-phase microextraction and gas chromatography–mass spectrometry | |
| Rivero et al. | Quantitative determination of formaldehyde in cosmetics using a combined solid-phase microextraction–isotope dilution mass spectrometry method | |
| Zeng et al. | Application of QuEChERS-based purification coupled with isotope dilution gas chromatography-mass spectrometry method for the determination of N-nitrosamines in soy sauce | |
| Petronilho et al. | A critical review on extraction techniques and gas chromatography based determination of grapevine derived sesquiterpenes | |
| Cai et al. | Rapid determination of trans-resveratrol in red wine by solid-phase microextraction with on-fiber derivatization and multidimensional gas chromatography–mass spectrometry | |
| Yo | Analysis of volatile fatty acids in wastewater collected from a pig farm by a solid phase microextraction method | |
| Zhang et al. | Comprehensive profiling of mercapturic acid metabolites from dietary acrylamide as short-term exposure biomarkers for evaluation of toxicokinetics in rats and daily internal exposure in humans using isotope dilution ultra-high performance liquid chromatography tandem mass spectrometry | |
| CN113960236B (en) | Method for determining skatole and dimethyl isoborneol in fish body based on rapid pretreatment technology | |
| Bian et al. | Progress in the pretreatment and analysis of N-nitrosamines: an update since 2010 | |
| Siříšťová et al. | Changes in Quality Parameters of Vodka Filtered through Activated Charcoal. | |
| CN106018624A (en) | HPLC detection method for nitrosamines in food | |
| CN109696499A (en) | A kind of nitrosamine Sensitive Determination method in the water based on high resolution mass spec | |
| CN109781883A (en) | A method of based on QuEChERS- dispersive liquid-liquid microextraction trace detection 5 hydroxymethyl furfural | |
| Meyers et al. | Analysis of gamma-hydroxybutyric acid (GHB) in spiked water and beverage samples using solid phase microextraction (SPME) on fiber derivatization/gas chromatography-mass spectrometry (GC/MS) | |
| Zachariadis et al. | Effect of sample matrix on sensitivity of mercury and methylmercury quantitation in human urine, saliva, and serum using GC‐MS | |
| Kapsimali et al. | Comparison of tetraethylborate and tetraphenylborate for selenite determination in human urine by gas chromatography mass spectrometry, after headspace solid phase microextraction | |
| Zanella et al. | Advanced sample preparation techniques for pesticide residues determination by HRMS analysis | |
| Li et al. | Determination of methylmalonic acid and glutaric acid in urine by aqueous‐phase derivatization followed by headspace solid‐phase microextraction and gas chromatography‐mass spectrometry | |
| Olmo et al. | Development of methodologies for different degrees of resolution of linear alkylbenzene sulfonates in groundwater and wastewater by liquid chromatography using sodium dodecyl sulphate | |
| CN113866336A (en) | Integrated detection method for safrole and dihydrosafrole in ginger and ginger powder | |
| Rotich et al. | Optimization of high-performance liquid chromatography and solid-phase extraction for determination of organophosphorus pesticide residues in environmental samples | |
| CN114414713B (en) | Method for extracting and detecting main PAEs in tea based on response surface method optimization |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |