CN107144656B - Method for measuring 3-acetyl-2, 5-dimethyl thiophene in edible essence by GC-MS/MS (gas chromatography-Mass Spectrometry/Mass Spectrometry) - Google Patents

Method for measuring 3-acetyl-2, 5-dimethyl thiophene in edible essence by GC-MS/MS (gas chromatography-Mass Spectrometry/Mass Spectrometry) Download PDF

Info

Publication number
CN107144656B
CN107144656B CN201710390827.1A CN201710390827A CN107144656B CN 107144656 B CN107144656 B CN 107144656B CN 201710390827 A CN201710390827 A CN 201710390827A CN 107144656 B CN107144656 B CN 107144656B
Authority
CN
China
Prior art keywords
acetyl
dimethylthiophene
edible essence
extraction solvent
determining
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
Application number
CN201710390827.1A
Other languages
Chinese (zh)
Other versions
CN107144656A (en
Inventor
茅佩卿
罗金文
陈碧莲
王展华
施贝
岳超
王一涵
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
ZHEJIANG INSTITUTE FOR FOOD AND DRUG CONTROL
Original Assignee
ZHEJIANG INSTITUTE FOR FOOD AND DRUG CONTROL
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by ZHEJIANG INSTITUTE FOR FOOD AND DRUG CONTROL filed Critical ZHEJIANG INSTITUTE FOR FOOD AND DRUG CONTROL
Priority to CN201710390827.1A priority Critical patent/CN107144656B/en
Publication of CN107144656A publication Critical patent/CN107144656A/en
Application granted granted Critical
Publication of CN107144656B publication Critical patent/CN107144656B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

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
    • 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

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

A method for measuring 3-acetyl-2, 5-dimethyl thiophene in edible essence by GC-MS/MS belongs to the technical field of food additive detection. Which comprises the following steps: ultrasonically extracting an edible essence and spice sample by using an extraction solvent, cooling, fixing the volume by using the extraction solvent, uniformly mixing, standing, and filtering an upper solution by using a microporous filter membrane, wherein the extraction solvent is n-hexane or n-heptane; and (4) measuring by adopting GC-MS/MS, and quantifying by adopting an external standard method. The invention adopts GC-MS/MS to detect 3-acetyl-2, 5-dimethylthiophene in the edible essence for the first time; the matrix effect can be effectively eliminated by adopting an MRM mode, and the sensitivity and the accuracy are improved; the external standard method has good linearity and accurate quantification, and the detection limit is 0.1 mg/kg. The method has the advantages of simple pretreatment, reliable result and high sensitivity, and has certain practical significance for monitoring the use condition of the 3-acetyl-2, 5-dimethylthiophene in the edible essence and protecting the health of consumers.

Description

Method for measuring 3-acetyl-2, 5-dimethyl thiophene in edible essence by GC-MS/MS (gas chromatography-Mass Spectrometry/Mass Spectrometry)
Technical Field
The invention belongs to the technical field of food additive detection, and particularly relates to a method for determining 3-acetyl-2, 5-dimethylthiophene in edible essence by GC-MS/MS.
Background
Synthetic flavor 3-acetyl-2, 5-dimethylthiophene has a roasted, nut-like odor and was used as a food flavor in 2002 by safety evaluation of the committee on food additive joint experts, which was established under the world health organization and the food and agriculture organization of the united nations. At present, China allows the spice to be used as a synthetic spice for food, and the spice is included in the national standard GB 2760-2014. On 6.4.2013, the European food safety administration publishes the opinion on the reevaluation of the substance, and the substance is considered to show mutagenicity in vivo and in vitro tests, so that the substance has a safety problem when used as a food spice. On 14.6.2013, the use of this substance as a food flavor was prohibited by the eu committee under regulation (ec) No 545/2013, and was deleted from the list of food flavors in food flavor code appendix i. Aiming at the situation, the Chinese Association for the industries of essence, spice and cosmetics rapidly makes a response, and issues an autonomous document (No. 2013 of the harmonization co-ordination character [2013] 62) in 8 and 14 months in 2013, so that spice manufacturers are required not to produce 3-acetyl-2, 5-dimethylthiophene used as a food spice, and the food spice manufacturers are required not to use the substance by adjusting the formula.
At present, the detection of 3-acetyl-2, 5-dimethylthiophene is still blank in the national standard, few quantitative detection researches for the substance are carried out in China, and only one patent report is measured by a gas chromatography-mass spectrometer, and the publication number is as follows: CN1046557775A, which discloses Selective Ion Monitoring (SIM) mode detection, internal standard method for quantification. The main defects of the method are as follows: because the edible essence has complex components and generally contains compounds with similar structures, the components cannot be completely separated in the detection process and are easy to interfere with each other, and the accuracy and the sensitivity are remained to be examined by adopting the monopole mass spectrometry for detection.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to design and provide a method for determining 3-acetyl-2, 5-dimethylthiophene in edible essence by GC-MS/MS. Compared with a single-stage mass spectrum SIM mode, the invention can be more accurately qualitative by adopting a multi-stage mass spectrum MRM mode, and meanwhile, because the parent ions are extracted from the first-stage fragments with serious interference and are further fragmented to obtain the daughter ions for quantification, the sensitivity is higher than that of the single-stage mass spectrum, and the invention is more suitable for detecting trace and trace substances.
The GC-MS/MS method for determining 3-acetyl-2, 5-dimethylthiophene in the edible essence is characterized by comprising the following steps of:
1) ultrasonically extracting an edible essence and spice sample by using an extraction solvent, cooling, fixing the volume by using the extraction solvent, uniformly mixing, standing, and filtering an upper solution by using a microporous filter membrane, wherein the extraction solvent is n-hexane or n-heptane;
2) taking the filtrate obtained in the step 1), determining by adopting GC-MS/MS, and quantifying by adopting an external standard method.
The GC-MS/MS method for determining 3-acetyl-2, 5-dimethylthiophene in the edible essence is characterized in that the ultrasonic extraction time in the step 1) is at least 15 min.
The GC-MS/MS method for determining 3-acetyl-2, 5-dimethylthiophene in the edible essence is characterized in that the microporous filter membrane in the step 1) is an organic filter membrane, and the pore diameter is 0.22 mu m or 0.45 mu m.
The GC-MS/MS method for determining 3-acetyl-2, 5-dimethyl thiophene in the edible essence is characterized in that the GC-MS/MS chromatographic conditions in the step 2) are as follows:
a chromatographic column: HP-INNOWax or equivalent;
sample inlet temperature: 240 ℃;
temperature programming conditions: keeping at 50 deg.C for 1min, increasing the temperature to 200 deg.C at 20 deg.C/min, keeping for 3min, increasing the temperature to 240 deg.C at 25 deg.C/min, and keeping for 7 min;
the split ratio is 10: 1;
the column flow rate was 1.0 mL/min;
the sample injection amount is 1.0 mu L;
the GC-MS/MS mass spectrum conditions are as follows:
an ion source: an electron bombardment source;
electron ionization energy: 70 eV;
ion source temperature: 230 ℃;
quadrupole temperature: 150 ℃;
collision gas (N)2) Flow rate: 1.5 mL/min;
quench gas (He) flow rate: 2.25 mL/min;
solvent retardation: 8 min;
the scanning mode is as follows: MRM;
parent, daughter and collision energies are as in table 1:
table 1 parent ion, daughter ion and collision energy table (.' is a quantitative ion)
Figure 137028DEST_PATH_IMAGE001
The GC-MS/MS method for determining 3-acetyl-2, 5-dimethylthiophene in the edible essence is characterized in that in the step 2), the external standard method comprises the steps of dissolving 3-acetyl-2, 5-dimethylthiophene by using an extraction solvent, diluting the solution to a required concentration by using the extraction solvent, measuring on a computer, and drawing a standard curve by using a peak area or a peak height.
The GC-MS/MS method for determining 3-acetyl-2, 5-dimethylthiophene in the edible essence is characterized in that the specification of the chromatographic column is 30m multiplied by 0.25mm multiplied by 0.25 mu m or equivalent.
The invention adopts GC-MS/MS to detect 3-acetyl-2, 5-dimethylthiophene in the edible essence for the first time; the matrix effect can be effectively eliminated by adopting an MRM mode, and the sensitivity and the accuracy are improved; the external standard method has good linearity and accurate quantification, and the detection limit is 0.1mg/kg, as shown in FIG. 3. The method has the advantages of simple pretreatment, reliable result and high sensitivity, and has certain practical significance for monitoring the use condition of the 3-acetyl-2, 5-dimethylthiophene in the edible essence and protecting the health of consumers.
Drawings
FIG. 1 is a typical profile of a standard;
FIG. 2 is a typical spectrum of 3-acetyl-2, 5-dimethylthiophene in a spiked recovery test sample;
FIG. 3 is a detection limit (plus scalar 0.1 mg/kg) profile.
Detailed Description
The technical solution of the present invention is further described below with reference to the accompanying drawings, but not limited thereto, and modifications or equivalent substitutions may be made to the technical solution of the present invention without departing from the spirit and scope of the technical solution of the present invention.
Example (b): the sample in the embodiment of the invention is powdered meat flavor
First, preparation of Standard stock solution
Accurately weighing a 3-acetyl-2, 5-dimethylthiophene standard substance, dissolving the standard substance by using normal hexane or normal heptane, fixing the volume and preparing standard stock solution with the concentration of 1 mg/mL.
Preparation of Linear Standard solution
100.0 mu L of 1mg/mL standard stock solution is diluted by n-hexane or n-heptane to 10mL, and 10 mu g/mL standard use solution is obtained.
Taking 10. mu.L, 20.0. mu.L, 40.0. mu.L, 80.0. mu.L, 100.0. mu.L, 150.0. mu.L, 200.0. mu.L and 0.25mL of standard use solution, respectively diluting with n-hexane and fixing the volume to 10mL to obtain linear standard solutions with the concentrations of 10ng/mL, 20ng/mL, 40ng/mL, 80ng/mL, 100ng/mL, 150 ng/mL, 200 ng/mL and 250 ng/mL for GC-MS/MS measurement.
Third, gas chromatography/mass spectrometry conditions
The chromatographic conditions are that Agilent HP-INNOWax chromatographic column is adopted for separation; the temperature of a sample inlet is 240 ℃; the programmed temperature rise condition is that the temperature is kept for 1min at 50 ℃, is increased to 200 ℃ at 20 ℃/min, is kept for 3min, is increased to 240 ℃ at 25 ℃/min, and is kept for 7 min; the split ratio is 10: 1; the column flow rate was 1.0 mL/min; the amount of sample was 1.0. mu.L.
Mass spectrum conditions: the ion source is an electron bombardment source; electron ionization energy is 70 eV; the ion source temperature is 230 ℃; the temperature of the four-level bar is 150 ℃; collision gas (N)2) The flow rate is 1.5 mL/min; the flow rate of the quenching gas (He) was 2.25 mL/min; the solvent delay time is 8 min; the scanning mode is MRM; the parent ions, the daughter ions, and the collision energy are shown in the following table.
Figure 874039DEST_PATH_IMAGE001
Fourth, sample determination
Accurately weighing 0.1g of sample into a 10mL volumetric flask, adding about 7mL of n-hexane, carrying out ultrasonic extraction for 15min, standing and cooling, then carrying out constant volume with the n-hexane, uniformly mixing, standing for several minutes, filtering the upper extracting solution with a 0.22 mu m organic phase filter membrane, taking the subsequent filtrate, and carrying out on-machine determination. And simultaneously, making a reagent blank.
The test shows that the reagent blank has no interference, and the sample is not detected.
Fifth, adding the standard to reclaim the test sample to determine
Accurately weighing 0.1g of the sample in a 10mL volumetric flask, adding the sample according to the adding amount listed in the table 2, then treating the sample, and measuring the subsequent filtrate on a machine. The recovery rate was examined. Each spiked level was measured in parallel at 3 parts.
TABLE 2 SANHUITARGETING TEST SCALAR AND AVERAGE RECOVERY METER
Figure 272791DEST_PATH_IMAGE002
As can be seen from Table 2, the recovery rate of the sample is between 94.9 and 100.3, which indicates that the recovery rate of the method is better.
FIG. 1 is a typical profile of a standard; FIG. 2 is a typical spectrum of 3-acetyl-2, 5-dimethylthiophene in a spiked recovery test sample. It can be obtained from the figure that the target object has symmetrical peak pattern and good response.
Sixth, method precision test
The above-mentioned spiked (medium level) samples were prepared in 6 portions in parallel and the results are shown in Table 3.
Table 3 method precision test results table
Figure 578001DEST_PATH_IMAGE003
As can be seen from Table 3, the method is highly reproducible.

Claims (5)

  1. The method for determining 3-acetyl-2, 5-dimethyl thiophene in edible essence by GC-MS/MS is characterized by comprising the following steps:
    1) ultrasonically extracting an edible essence and spice sample by using an extraction solvent, cooling, fixing the volume by using the extraction solvent, uniformly mixing, standing, and filtering an upper solution by using a microporous filter membrane, wherein the extraction solvent is n-hexane or n-heptane;
    2) taking the filtrate obtained in the step 1), determining by adopting GC-MS/MS, and quantifying by adopting an external standard method;
    the GC-MS/MS mass spectrum conditions are as follows:
    an ion source: an electron bombardment source;
    electron ionization energy: 70 eV;
    ion source temperature: 230 ℃;
    quadrupole temperature: 150 ℃;
    collision gas N2Flow rate: 1.5 mL/min;
    flow rate of quench gas He: 2.25 mL/min;
    solvent retardation: 8 min;
    the scanning mode is as follows: MRM;
    parent, daughter and collision energies are as in table 1:
    table 1 parent ion, daughter ion and collision energy table, as quantitative ions
    Figure 519222DEST_PATH_IMAGE001
    ；
    The chromatographic conditions for GC-MS/MS were:
    a chromatographic column: HP-INNOWax;
    sample inlet temperature: 240 ℃;
    temperature programming conditions: keeping at 50 deg.C for 1min, increasing the temperature to 200 deg.C at 20 deg.C/min, keeping for 3min, increasing the temperature to 240 deg.C at 25 deg.C/min, and keeping for 7 min;
    the split ratio is 10: 1;
    the column flow rate was 1.0 mL/min;
    the amount of sample was 1.0. mu.L.
  2. 2. The method for determining 3-acetyl-2, 5-dimethylthiophene in a flavor according to claim 1, wherein the ultrasonic extraction time in step 1) is at least 15 min.
  3. 3. The method for determining 3-acetyl-2, 5-dimethylthiophene in a flavor according to claim 1, wherein the microfiltration membrane in step 1) is an organic filtration membrane with a pore size of 0.22 μm or 0.45 μm.
  4. 4. The method for determining 3-acetyl-2, 5-dimethylthiophene in a flavor according to claim 1 by GC-MS/MS, wherein the external standard method in the step 2) comprises the steps of dissolving 3-acetyl-2, 5-dimethylthiophene by using an extraction solvent, diluting the solution to a required concentration by using the extraction solvent, and performing on-machine determination to draw a standard curve by using a peak area or a peak height.
  5. 5. The method for determining 3-acetyl-2, 5-dimethylthiophene in a flavor according to claim 1, wherein the chromatographic column has a size of 30m x 0.25mm x 0.25 μm.
CN201710390827.1A 2017-05-27 2017-05-27 Method for measuring 3-acetyl-2, 5-dimethyl thiophene in edible essence by GC-MS/MS (gas chromatography-Mass Spectrometry/Mass Spectrometry) Active CN107144656B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710390827.1A CN107144656B (en) 2017-05-27 2017-05-27 Method for measuring 3-acetyl-2, 5-dimethyl thiophene in edible essence by GC-MS/MS (gas chromatography-Mass Spectrometry/Mass Spectrometry)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710390827.1A CN107144656B (en) 2017-05-27 2017-05-27 Method for measuring 3-acetyl-2, 5-dimethyl thiophene in edible essence by GC-MS/MS (gas chromatography-Mass Spectrometry/Mass Spectrometry)

Publications (2)

Publication Number Publication Date
CN107144656A CN107144656A (en) 2017-09-08
CN107144656B true CN107144656B (en) 2020-03-31

Family

ID=59779334

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710390827.1A Active CN107144656B (en) 2017-05-27 2017-05-27 Method for measuring 3-acetyl-2, 5-dimethyl thiophene in edible essence by GC-MS/MS (gas chromatography-Mass Spectrometry/Mass Spectrometry)

Country Status (1)

Country Link
CN (1) CN107144656B (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110082441B (en) * 2019-04-12 2022-03-01 中国检验检疫科学研究院 Method and kit for detecting forbidden essences
CN110568103A (en) * 2019-09-20 2019-12-13 浙江省食品药品检验研究院 Deuterium-substituted marker and application thereof in detection of edible essence 3-acetyl-2, 5-dimethylthiophene

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104655753A (en) * 2015-02-10 2015-05-27 广西中烟工业有限责任公司 Method for determining 3-acetyl-2, 5-thioxene in food additive
CN104655775A (en) * 2015-02-10 2015-05-27 广西中烟工业有限责任公司 Method for measuring 3-acetyl-2, 5-dimethyl thiophene in edible flavor and fragrance

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104655753A (en) * 2015-02-10 2015-05-27 广西中烟工业有限责任公司 Method for determining 3-acetyl-2, 5-thioxene in food additive
CN104655775A (en) * 2015-02-10 2015-05-27 广西中烟工业有限责任公司 Method for measuring 3-acetyl-2, 5-dimethyl thiophene in edible flavor and fragrance

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Green Synthesis, Characterization, Photophysical and Electrochemical Properties of Bis-chalcones;Abdullah M. Asiri等;《Int. J. Electrochem. Sci.》;20141231;第9卷;全文 *
Synthesis and Anti-Bacterial Activities of a Bis-Chalcone Derived from Thiophene and Its Bis-Cyclized Products;Abdullah M. Asiri等;《Molecules》;20111231;第16卷;全文 *

Also Published As

Publication number Publication date
CN107144656A (en) 2017-09-08

Similar Documents

Publication Publication Date Title
CN107764917B (en) Method for determining key volatile components in cigarette blasting beads
CN103983725B (en) The rapid assay methods of cumarin and safrole in a kind of essence and flavoring agent
CN107907600B (en) Method for simultaneously measuring aflatoxin and flavoring agent in vegetable oil based on liquid-liquid extraction-liquid chromatography-tandem mass spectrometry
CN110133156A (en) The analysis method of super more target flavor components in a kind of high throughput assay tobacco juice for electronic smoke
CN105403630A (en) Method for detecting concentration of nine compound coloring agents in auxiliary materials for reconstituted tobacco
Xiang et al. Multivariate statistical analysis of tobacco of different origin, grade and variety according to polyphenols and organic acids
CN107144656B (en) Method for measuring 3-acetyl-2, 5-dimethyl thiophene in edible essence by GC-MS/MS (gas chromatography-Mass Spectrometry/Mass Spectrometry)
Zhang et al. Screening multimycotoxins in food-grade gums by stable isotope dilution and liquid chromatography/tandem mass spectrometry
CN113325103A (en) Analysis method for simultaneously measuring gelsemium, gelsemine and gelsemine in hair
CN108956812B (en) Method for detecting water-soluble organic chloride pesticide residue in soil
CN111855866A (en) Method for rapidly determining formaldehyde in tobacco additive
CN116953112A (en) Analysis method for simultaneously determining sugar, organic acid, amino acid and Maillard reaction product in tobacco flavor
CN108982703B (en) Liquid chromatography-mass spectrometry detection method for polyphenol substances
CN110596229A (en) Detection method for element impurities in hemp extract and hemp oil product
CN111024872B (en) Method for rapidly detecting 3-methylthio propanol in sesame-flavor liquor
CN113866305A (en) Method for rapidly and accurately analyzing theanine in fresh tea leaves based on liquid chromatography-mass spectrometry technology
CN104713976A (en) Method for determining nitrite in food by employing headspace-gas chromatography/mass spectrography
Tešević et al. Development and validation of an LC-MS/MS method with a multiple reactions monitoring mode for the quantification of vanillin and syringaldehyde in plum brandies
CN114235988A (en) High performance liquid chromatography for determining content of maleic hydrazide
CN114236026A (en) Non-targeting rapid screening method for pesticide residues in fruits and vegetables
Ebeler Analysis of grapes and wines: An overview of new approaches and analytical tools
CN114113350A (en) Method for distinguishing aged and fragrant yellow years
CN111208227A (en) Method for simultaneously determining content of 8 carbonyl compounds in cigarette blasting beads
CN114814009B (en) Analysis method simultaneously applicable to phenolic compounds in novel tobacco products and metabolites of phenolic compounds in urine
CN103983731B (en) A kind of method measuring ortho-aminobenzoic acid cinnamic ester in essence and flavoring agent

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