CN114705773A - Method for measuring aroma substances in apples by using headspace solid-phase microextraction technology - Google Patents
Method for measuring aroma substances in apples by using headspace solid-phase microextraction technology Download PDFInfo
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- CN114705773A CN114705773A CN202210311650.2A CN202210311650A CN114705773A CN 114705773 A CN114705773 A CN 114705773A CN 202210311650 A CN202210311650 A CN 202210311650A CN 114705773 A CN114705773 A CN 114705773A
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- 238000000034 method Methods 0.000 title claims abstract description 34
- 239000000126 substance Substances 0.000 title claims abstract description 29
- 235000021016 apples Nutrition 0.000 title claims abstract description 28
- 238000005516 engineering process Methods 0.000 title claims abstract description 12
- 238000001319 headspace solid-phase micro-extraction Methods 0.000 title claims abstract description 12
- 244000141359 Malus pumila Species 0.000 title 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 52
- 241000220225 Malus Species 0.000 claims abstract description 33
- 239000011780 sodium chloride Substances 0.000 claims abstract description 26
- 238000002470 solid-phase micro-extraction Methods 0.000 claims abstract description 19
- 238000000605 extraction Methods 0.000 claims abstract description 17
- JHIVVAPYMSGYDF-UHFFFAOYSA-N cyclohexanone Chemical compound O=C1CCCCC1 JHIVVAPYMSGYDF-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 claims abstract description 11
- 238000002347 injection Methods 0.000 claims abstract description 9
- 239000007924 injection Substances 0.000 claims abstract description 9
- 239000002131 composite material Substances 0.000 claims abstract description 5
- 238000005520 cutting process Methods 0.000 claims abstract description 5
- 239000004205 dimethyl polysiloxane Substances 0.000 claims abstract description 5
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims abstract description 5
- 238000002156 mixing Methods 0.000 claims abstract description 5
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 claims abstract description 5
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims abstract description 5
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims abstract description 5
- 238000005303 weighing Methods 0.000 claims abstract description 4
- 241000220324 Pyrus Species 0.000 claims description 10
- 235000021017 pears Nutrition 0.000 claims description 4
- 239000012159 carrier gas Substances 0.000 claims description 3
- 239000001307 helium Substances 0.000 claims description 3
- 229910052734 helium Inorganic materials 0.000 claims description 3
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 claims description 3
- 150000002500 ions Chemical class 0.000 claims description 3
- 238000001819 mass spectrum Methods 0.000 claims description 3
- 239000004810 polytetrafluoroethylene Substances 0.000 claims description 3
- 229920001343 polytetrafluoroethylene Polymers 0.000 claims description 3
- 125000006850 spacer group Chemical group 0.000 claims description 3
- 239000003205 fragrance Substances 0.000 claims 1
- 238000002546 full scan Methods 0.000 claims 1
- 235000013399 edible fruits Nutrition 0.000 abstract description 10
- 238000000265 homogenisation Methods 0.000 description 8
- 235000014443 Pyrus communis Nutrition 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 230000008569 process Effects 0.000 description 5
- 235000013305 food Nutrition 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000019634 flavors Nutrition 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 238000012764 semi-quantitative analysis Methods 0.000 description 3
- 102000030523 Catechol oxidase Human genes 0.000 description 2
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Images
Classifications
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- 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
-
- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
-
- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8679—Target compound analysis, i.e. whereby a limited number of peaks is analysed
-
- 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
- G01N2030/065—Preparation using different phases to separate parts of sample
Abstract
The invention belongs to the technical field of extraction, and provides a method for measuring aroma substances in apples by using a headspace solid-phase microextraction technology, which comprises the following steps: 1, removing cores of apples, cutting the apples into blocks of about 1-2cm, mixing the blocks with sodium chloride (NaCl, 1:1, m/m) and homogenizing; step 2, weighing the homogenate sample to an SPME sample injection bottle; step 3, adding cyclohexanone internal standard, and screwing down the cap of the SPME sample injection bottle; step 4, SPME method: incubating a DVB/CAR/PDMS composite material extraction head at 80 ℃ for 15min, then extracting for 15min, feeding the extraction head into a gas chromatography-mass spectrometry (GC-MS) sample inlet, and desorbing at 200 ℃ for 1 min. By using the method, the browning of the fruit pulp is inhibited, the change of the components of the aroma substances is avoided, and the volatilization of the aroma substances can be promoted; the method is suitable for measuring the aroma substances in a broad range by optimizing the extraction conditions.
Description
Technical Field
The invention belongs to the technical field of extraction, and particularly relates to a method for measuring aroma substances in apples by using a headspace solid-phase microextraction technology.
Background
The aroma is a key factor determining the flavor of the food, and directly influences the acceptance of consumers and the product competitiveness. The content of aroma substances in the agricultural products is low, the extraction is difficult, the agricultural products are easily influenced by weather, diseases, pests, weeds and the like, and the cost of the agricultural products can reach more than 10 times of that of the chemical synthetic aroma. The quality and sales profit of agricultural products are greatly different due to the influence of varieties, production areas and the like. Agricultural products are often found to be too good and fake, so that consumers can resist the mind and the economy is damaged; the food quality and safety are difficult to ensure, and great threat is formed to the health of consumers. As part of metabonomics, the research of aroma substances has great contribution in the fields of food quality identification and product traceability. The analysis of the components of the aromatic substances in the agricultural products can promote the improvement of the flavor quality of the agricultural products, and can also be used for food traceability research, so that the information of the aromatic substances in the agricultural products is collected, and the establishment of a database is the basis of the research. The apple and the pear are easy to brown, and the sample pretreatment processes such as cutting, homogenizing and the like are particularly serious, so that the types and the contents of aroma substances are changed. However, the influence of browning on the components of the aroma substances in the pretreatment process is of little concern in the existing research, the accurate determination of the aroma substances is not facilitated, and the healthy development of the apple and pear industry in China is hindered.
Disclosure of Invention
The invention aims to solve the problems recorded in the background technology and provides a method for measuring aroma substances in apples by utilizing a headspace solid-phase microextraction technology.
In order to achieve the purpose, the invention adopts the following technical scheme: a method for measuring aroma substances in apples by using a headspace solid phase microextraction technology comprises the following steps:
1, removing cores of apples, cutting the apples into blocks of about 1-2cm, and mixing the cut blocks with NaCl for homogenate;
step 2, weighing the homogenate sample to an SPME sample injection bottle;
step 3, adding cyclohexanone internal standard, and screwing down the cap of the SPME sample injection bottle;
step 4, using the SPME method: and (3) feeding the DVB/CAR/PDMS composite material extraction head into a GC-MS sample inlet, and desorbing at 200 ℃ for 1 min.
In a preferred embodiment of the invention, the cut apples are mixed with NaCl in a ratio of 1:1 m/m.
In a preferred embodiment of the invention, the SPME extraction head is aged at 250 ℃ for 10 min; SPME sample bottles were incubated at 80 ℃ for 15min before extraction for 15 min.
In a preferred embodiment of the present invention, in step 4, the parameters of the GC-MS are set: the initial temperature of the heating program is kept for 1min at 50 ℃, the temperature is increased to 180 ℃ at 2 ℃/min and kept for 1min, then the temperature is increased to 230 ℃ at 10 ℃/min and kept for 10min, the carrier gas is helium (with the purity of 99.999 percent), and the flow rate is 1.0 mL/min; the mass spectrum adopts a full-scanning mode; ion source energy was 70eV and temperature was 230 ℃, and NIST 17s database was used for aroma matching.
In a preferred embodiment of the invention, the cap of the SPME sampling vial is provided with a PTFE spacer.
In a preferred embodiment of the invention, the method is used for measuring the aroma substances of pears.
The principle and the beneficial effects of the invention are as follows: 1. the method mixes the cut apples or pears with NaCl (1:1, m/m) for homogenate, thereby not only inhibiting browning, but also promoting the volatilization of aroma substances;
2. the method adopts the DVB/CAR/PDMS composite material extraction head, optimizes the pretreatment conditions, and is suitable for measuring aroma substances in a broad range.
3. The linear relation of cyclohexanone as an internal standard is good, and R is within the range of 0.02-200.0 mg/kg FW (FW, fresh weight)2Above 0.99, semi-quantitative analysis of the aroma can be performed.
In conclusion, the method mixes the cut fruits with NaCl (1:1, m/m), and then homogenates, so as to inhibit browning, avoid changes of aroma components, promote volatilization of aroma substances and improve extraction efficiency; the method has good linear relation by using cyclohexanone as an internal standard, and can perform semi-quantitative analysis on the aroma substances.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
FIG. 1 is a photograph comparing browning phenomena of apples of the present invention before and after homogenization with NaCl.
FIG. 2 is a GC-MS chromatogram comparison of the aroma of apples with NaCl before and after homogenization.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.
The application provides a method for measuring aroma substances in apples by using a headspace solid-phase microextraction technology, which comprises the following steps:
1, removing cores of apples, cutting the apples into blocks of about 1-2cm, mixing the cut apples with NaCl (1:1, m/m) and homogenizing;
step 2, weighing 10g of homogenate sample to an SPME sample injection bottle;
step 3, adding cyclohexanone internal standard, screwing the SPME sample injection bottle cap (with PTFE spacer)
Step 4, using the SPME method: the method comprises the following steps of (1) aging a DVB/CAR/PDMS composite material extraction head at 250 ℃ for 10min before sample injection by using an AOC 6000(Shimadazu, Japan) automatic sample injector; SPME was incubated at 80 ℃ for 15min before extraction for 15min (shaker speed 300 rpm). And finally, the extraction head enters a GC sample inlet, and desorption is carried out for 1min at 200 ℃. Adjusting the parameters of GC-MS: GC-MS was Shimadazu GC-2010 gas phase coupled QP 2010 mass spectrometer (Shimadazu, Japan) and the column was HP-INNOWAX (60m 0.25mm 0.25 μm, Agilent Technologies). Temperature-raising program initial temperature 50 deg.C for 1min, temperature-raising at 2 deg.C/min to 180 deg.C for 1min, then temperature-raising at 10 deg.C/min to 230 deg.C (maintaining for 10 min). The carrier gas was helium (99.999% purity) at a flow rate of 1.0 mL/min; the mass spectrum is in a Full scan mode (Full scan mode, m/z 50-500); ion source (EI) energy was 70eV temperature was 230 ℃ and NIST 17s database was used for aroma matching (screening compounds with a degree of match of 90% and above).
NaCl can promote aroma to volatilize from a sample matrix, the common usage method is adding after homogenizing apples, however, apple pulp browning is serious in the homogenizing process, and a rotten fruit smell is generated, which indicates that fruit aroma substance components are changed, and the influence of browning on aroma components in the pretreatment process is of fresh concern in the existing research.
In this example, as shown in fig. 1a, the sample rapidly underwent severe browning after homogenization, mainly due to the production of a large amount of brown material by enzymatic reactions such as polyphenol oxidase (PPO). The literature indicates that the protein configuration is changed and the enzyme activity is reduced in the presence of a large amount of salt. NaCl in an amount of 1:1(m/m) of the sample mass was added before homogenization, and almost no browning occurred in the color of the pulp (FIG. 1b), and the fruity flavor was stronger. FIG. 2 is a GC-MS chromatogram comparison of apple aroma after and after adding NaCl before and after homogenizing apple (note: in FIG. 2, a, 5g of fruit pulp without NaCl, b, 5g of fruit pulp + 5g of NaCl, c, 10g of NaCl in the same amount as the sample is added before homogenizing), and the measurement conditions are the same except that the time for adding NaCl is different. As can be seen from FIG. 2b in comparison with FIG. 2a, the amount and response of aroma compounds detected by GC-MS are significantly increased after NaCl is added, indicating that NaCl can promote the volatilization of aroma, and therefore the conventional usage is to add NaCl after homogenization; compared with the prior art, the NaCl is added before homogenization, the quantity and the response value of aroma substances are obviously further improved, and the NaCl is added before the homogenization to inhibit browning and preserve the original aroma of the fruits by combining the graph 1, so that the diced apples and the NaCl are mixed and then homogenized. Furthermore, pear is also an easy browning fruit. Through research, the pear is cut into blocks and then mixed with NaCl with the sample mass of 1:1(m/m), the browning of the pear fruit in the homogenization process can be successfully inhibited, and the influence of the NaCl on the quantity and response value of aroma compounds in the pear fruit detected by GC-MS is similar to that of the apple.
The internal standards for aroma determination used in current studies are inconsistent and few do linear analyses. In this example, cyclohexanone was used as an internal standard, and R was in the range of 0.02-200.0 mg/kg FW (FW, fresh weight)2And if the concentration is more than 0.99, the linear relation is good, the errors caused by pretreatment and instrument states can be effectively corrected, and the method is suitable for semi-quantitative analysis of aroma substances in apples and pears.
In the description herein, reference to the description of the terms "preferred embodiment," "one embodiment," "some embodiments," "an example," "a specific example" or "some examples" or the like means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
While embodiments of the invention have been shown and described, it will be understood by those of ordinary skill in the art that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.
Claims (6)
1. A method for measuring aroma substances in apples by using a headspace solid-phase microextraction technology is characterized by comprising the following steps:
1, removing cores of apples, cutting the apples into blocks of about 1-2cm, and mixing the cut blocks with NaCl for homogenate;
step 2, weighing the homogenate sample to an SPME sample injection bottle;
step 3, adding cyclohexanone internal standard, and screwing down the cap of the SPME sample injection bottle;
step 4, using the SPME method: and (3) desorbing the DVB/CAR/PDMS composite material extraction head for 1min at 200 ℃ in a GC-MS sample inlet.
2. The method for measuring aroma substances in apples by using the headspace solid-phase microextraction technology as claimed in claim 1, wherein the mixing ratio of the cut pieces to NaCl is 1:1 m/m.
3. The method for measuring aroma in apples by headspace solid phase microextraction technique according to claim 2, wherein the SPME extraction head is aged at 250 ℃ for 10 min; SPME sample vials were incubated at 80 ℃ for 15min, followed by extraction for 15 min.
4. The method for measuring aroma substances in apples by using the headspace solid phase microextraction technology as claimed in claim 3, wherein in step 4, the parameters of GC-MS are set as follows: the initial temperature of the temperature raising program is kept for 1min at 50 ℃, the temperature is raised to 180 ℃ at 2 ℃/min and kept for 1min, then the temperature is raised to 230 ℃ at 10 ℃/min and kept for 10min, the carrier gas is helium, and the flow rate is 1.0 mL/min; the mass spectrum adopts a full-scan mode; ion source energy was 70eV, temperature was 230 ℃ and fragrance matching used NIST 17s database.
5. The method for measuring aroma in apples by headspace solid phase microextraction technique according to claim 4, wherein the cap of the SPME sample bottle is provided with a PTFE spacer.
6. A method as claimed in any one of claims 1 to 4, for determining the aroma of pears.
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