CN107179375A - A kind of method of volatile flavor substance in ultrasonic method auxiliary measuring brown rice - Google Patents
A kind of method of volatile flavor substance in ultrasonic method auxiliary measuring brown rice Download PDFInfo
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- CN107179375A CN107179375A CN201710547236.0A CN201710547236A CN107179375A CN 107179375 A CN107179375 A CN 107179375A CN 201710547236 A CN201710547236 A CN 201710547236A CN 107179375 A CN107179375 A CN 107179375A
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- 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
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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
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Abstract
The invention discloses a kind of method of volatile flavor substance in ultrasonic method auxiliary measuring brown rice, belong to volatile flavor substance analysis technical field.Methods described includes：(1) brown rice powder to be measured is taken, is mixed with inorganic salt solution, the obtained mixed liquor of ultrasound；(2) sorption extraction is carried out to the volatile flavor substance in mixed liquor using headspace solid-phase microextraction technology；(3) volatile flavor substance is identified using gaschromatographic mass spectrometric analysis technology.The present invention adds saturated sodium-chloride water solution into brown rice sample, improves the accumulation rate of flavor substance；Promote the developability of volatile flavor substance using ultrasonic technique, improve extraction efficiency；Extraction conditions is adjusted, the volatile materials in brown rice is effectively collected；Volatile compound to be measured is accurately have detected using gas-chromatography and mass spectral analysis, the confidence level of result is improved.
Description
Technical field
The present invention relates to volatile flavor substance analysis technical field, and in particular in a kind of ultrasonic method auxiliary measuring brown rice
The method of volatile flavor substance.
Background technology
For brown rice is with respect to polished rice, there is higher nutritive value, with antioxidant and anti-aging, improve metabolism, drop
A series of health-care efficacies such as blood pressure blood fat and strengthen immunity are rough with the lifting that people are pursued nutritive value in food
Rice is gradually by big well-established, therefore research of the recent domestic to brown rice gradually increases.
Its edible quality of the brown rice just harvested and processing quality are poor, after the storage of a period of time, and its quality then can
Make moderate progress.But brown rice quality can also change with storage time extension or storage condition, and occur corresponding change, volatility
The change of material is a reflection of its quality comparison.The volatile materials main source that brown rice is produced in storage has：
Some volatile materials that brown rice has in itself, such as esters, alcohols and a small amount of carbonyls；Free amino acid in brown rice
Automatic degraded；The metabolin that microorganism and stored product insect vital movement are produced；The oxidative degradation generation of lipid material in brown rice
The material such as aldehyde, ketone, alcohol.Wherein, the material that lipid material oxidative degradation is produced is the typical volatile matter in brown rice storage.
Therefore, volatile flavor component comprehensively in analysis brown rice, the change to brown rice quality is provided fundamental basis, and is the storage of brown rice
Effective foundation is provided, with far-reaching realistic meaning.
At present, the research for unpolished-rice nutrient composition is more, and the research for its volatile flavor component is less.Head space
SPME (Headspace-Solid Phase Micro-extraction, HS-SPME) contains sampling, extraction, dense
The processes such as contracting, sample introduction, compared with other volatile materials abstraction techniques, the method is selectively good, and required amount of samples is few, nothing
Organic solvent is needed, can be directly combined with GC/MS, powerful technical advantage is shown in flavour of food products analysis.
Zhang Yurong et al. (headspace solid-phase microextractions-Gc-mss wheat preservation such as Zhang Yurong, Gao Yanna, woods man be brave
During volatile ingredient change analytical chemistry, 38 (7), in July, 2010,953~957) use headspace solid-phase microextraction-gas
Volatile materials in phase chromatography-mass spectroscopy (HS-SPME-GC-MS) (strong) muscle wheat weak to different storage times is extracted, reflected
It is fixed with analysis, as a result show that the trend for first dropping and increasing afterwards is presented in 6 months in storage in volatile ingredient total content.
Influence the volatilization intensity that one of key factor of SPME bioaccumulation efficiency is flavor substance, some flavor substances
It is difficult to separate with sample substrate, therefore the sensitivity and the degree of accuracy of measurement result can be reduced, often using auxiliary in above method
The means such as heating and addition inorganic salts are helped to promote flavor substance to be volatilized from sample substrate.But volatility in cereal is detected at present
The method of material does not have with reference to above-mentioned supplementary means, it is therefore necessary to which flavor substances in brown rice can be significantly improved by probing into one kind
The volatile method of matter.
The content of the invention
It is an object of the invention to provide a kind of method for determining volatile flavor substance in brown rice, using ultrasonic wave added method
Promote flavor substance to be volatilized from brown rice matrix, significantly improve the sensitivity of volatile flavor substance detection.
To achieve the above object, the present invention is adopted the following technical scheme that：
The method of volatile flavor substance, comprises the following steps in a kind of ultrasonic method auxiliary measuring brown rice：
(1) brown rice powder to be measured is taken, is mixed with inorganic salt solution, the obtained mixed liquor of ultrasound；
(2) sorption extraction is carried out to the volatile flavor substance in mixed liquor using headspace solid-phase microextraction technology；
(3) volatile flavor substance is identified using gas chromatography-mass spectrometry analysis technology.
In step (1), the inorganic salt solution is saturated sodium-chloride water solution.Salinity is to head space gas chromatography
Response influence is very big, sodium chloride is added in the solution, it is possible to decrease the solubility of volatile materials, makes test substance be easier to be extracted
Take.
The liquid ratio of the saturated sodium-chloride water solution and brown rice powder is 1.6~2.8mL:1g.Preferably, liquid ratio
For 2.4mL:1g.
The present invention makes the flavor substance in brown rice fully volatilize using ultrasonic technique, improves extraction efficiency.Described ultrasound
Condition is：Frequency 40000Hz, the time is 5~60min.
Preferably, the ultrasonic time is 30min.Research is found, with the increase of ultrasonic time, volatile materials
Extracting and enriching effect present first increase after reduction trend, in supersonic frequency 40000Hz, chromatogram total peak when the time is 30min
Area substantially rises.
In step (2), headspace solid-phase microextraction is using 50/30 μm of DVB/PDMS/CAR extracting head.
During headspace solid-phase microextraction, extraction temperature is 65~75 DEG C, and extraction time is 30~60min, incubator concussion speed
Spend for 250rpm.Preferably, extraction temperature is 65 DEG C, extraction time is 60min.
In step (3), gas-chromatography uses chromatographic column for HP-5MS, 30m, 0.25mm, i.d.0.25 μm, and carrier gas is high-purity
Helium (99.999%) is spent, flow velocity is 0.8mL/min, and injection port is using not shunting mode sample introduction pattern.Complete the extraction of extraction
Head carries out the parsing of adsorbent in gas chromatographic sample introduction mouthful, and resolution temperature is 260 DEG C, and the parsing time is 4min.
During gas chromatographic analysis, column oven heating degree is：60 DEG C of initial temperature is kept after 4min, is heated up with 5 DEG C/min
240 DEG C are warming up to 180 DEG C of holding 5min, then with 20 DEG C/min, 5min is kept.
Mass spectral analysis condition is：Using EI sources, positive ion detection, ionizing energy is 70eV, and 260 DEG C of ion source temperature is passed
Defeated 280 DEG C of line temperature, scan pattern is full scan, 20~550m/z of scanning range.
The beneficial effect that the present invention possesses：
The present invention adds saturated sodium-chloride water solution into brown rice sample, improves the accumulation rate of flavor substance；Utilize ultrasound
Technology promotes the developability of volatile flavor substance, improves extraction efficiency；Extraction conditions is adjusted, waving in brown rice is effectively collected
Volatile material；Volatile compound to be measured is accurately have detected using gas-chromatography and mass spectral analysis, the confidence level of result is improved.
Brief description of the drawings
Fig. 1 is the comparative result of SPME bioaccumulation efficiency under different ultrasonic times.
Fig. 2 is total for what is detected afterwards to volatile flavor substance progress SPME (not aided in using ultrasonic method) in brown rice
Ion flow graph.
Fig. 3 is to carry out ultrasonic method auxiliary (ultrasonic time is 15 minutes) SPME to volatile flavor substance in brown rice
The total ion current figure detected afterwards.
Fig. 4 is to carry out ultrasonic method auxiliary (ultrasonic time is 30 minutes) SPME to volatile flavor substance in brown rice
The total ion current figure detected afterwards.
Fig. 5 is to carry out ultrasonic method auxiliary (ultrasonic time is 45 minutes) SPME to volatile flavor substance in brown rice
The total ion current figure detected afterwards.
Fig. 6 be in brown rice volatile flavor substance carry out ultrasonic method auxiliary (ultrasonic time be 60 minutes) detect always from
Subflow figure.
Fig. 7 is the peak of main flavor compound in brown rice under the conditions of different SPMEs measured by different ultrasonic times
Area principal component analysis result (PCA), wherein ZC1.1 refers to ultrasonic time 0 minute, the testing result under extraction conditions 1.
Embodiment
With reference to specific embodiment, the present invention is further described, and its specific embodiment is interpreted as being only for example
Illustrate, and it is non-limiting, it is impossible to limit protection scope of the present invention with following illustrations.
Embodiment 1
The method of volatile flavor substance, comprises the following steps in a kind of ultrasonic method auxiliary measuring brown rice：
(1) take brown rice sample to be measured to be broken into powder with Cyclone mill, matter is pressed with the saturated common salt aqueous solution and brown rice powder
Amount compares 2.4:1(v：M) mix in 20mL ml headspace bottles, mixed liquor is obtained, with the hollow magnetic with PTFE/ blue silica gel dottle pins
Metal cover is sealed, then mixed liquor is placed in the environment that supersonic frequency is 40000Hz, adjusts ultrasonic time.
In the present embodiment set 5 different disposals, respectively be set ultrasonic time be 0 minute, 15 minutes, 30 minutes, 45
Minute and 60 minutes, remaining treatment conditions is identical.
(2) full-automatic head space is carried out by multi-functional full-automatic sample pre-treatments platform MPS XT (German Gerstel companies)
SPME is operated.In headspace solid-phase microextraction operation, using 50/30 μm of DVB/PDMS/CAR extracting head of 1cm, in temperature
Under conditions of 75 DEG C, 30min is extracted；In extraction process, incubator concussion speed is 250rpm.
(3) gas chromatography-mass spectrometry analysis：Extracting head is in gas-chromatography (Agilent GC 7890) sample introduction after completion extraction
Mouth uses mass spectrograph (Agilent 5977A) right using 260 DEG C of parsing 4min of Splitless injecting samples pattern after completing desorption and separation
Each volatile ingredient is identified.
Gas-chromatography uses chromatographic column for HP-5MS, 30m, 0.25mm, i.d.0.25 μm, and high-purity helium is carrier gas, stream
Speed is 0.8mL/min；
Column oven heating schedule：60 DEG C of initial temperature is kept after 4min, and 180 DEG C of holding 5min are warming up to 5 DEG C/min, then
240 DEG C are warming up to 20 DEG C/min, 5min is kept.
Mass Spectrometry Conditions：Using EI sources, positive ion detection, ionizing energy is 70eV, 260 DEG C of ion source temperature, transmission line temperature
280 DEG C of degree, scan pattern is full scan, 20~550m/z of scanning range.
(4) data analysis and identification：The retention time and integral area of chromatogram are automatically performed by work station, utilize NIST
11 spectrum storehouses carry out qualitative analysis to each volatile ingredient.The relative amount of each volatile ingredient is obtained by area normalization method,
As a result the average value of different disposal is taken.
As a result as table 1 and Fig. 1~6 show.
The composition of volatile ingredient and relative amount (n=3) in the brown rice of table 1.
* in many places, retention time is detected ,-it is not detect
As seen from Figure 1, with the increase of ultrasonic time, the extracting and enriching effect of volatile materials is presented after first increasing
The trend of reduction, the measured total peak area when ultrasonic time is 30 minutes of the above method substantially rises, so that the spirit of method
Sensitivity is improved.
Referring to Fig. 2~6, determine, reflected in brown rice volatile component through ultrasonic method assisted Solid-state micro-extraction and gas chromatography mass spectrometry method
Compound is made for 36 (take matching score >=80 confirms as the compound, maximum 100), identifiable volatile ingredient
And its retention time is listed in table 1.Retention time and peak area are calculated with Agilent MassHunter work stations, relative amount face
The normalized method of product is calculated.
Embodiment 2
Volatile materials in ultrasonic method auxiliary measuring brown rice, comprises the following steps under different condition：
(1) take brown rice sample to be measured to be broken into powder with Cyclone mill, matter is pressed with the saturated common salt aqueous solution and brown rice powder
Amount compares 2.4:1(v：M) mix in 20mL ml headspace bottles, mixed liquor is obtained, with the hollow magnetic with PTFE/ blue silica gel dottle pins
Metal cover is sealed, then mixed liquor is placed in the environment that supersonic frequency is 40000Hz, adjusts ultrasonic time.
In the present embodiment set 5 different disposals, respectively be set ultrasonic time be 0 minute, 15 minutes, 30 minutes, 45
Minute and 60 minutes, remaining treatment conditions is identical.
(2) full-automatic head space is carried out by multi-functional full-automatic sample pre-treatments platform MPS XT (German Gerstel companies)
SPME is operated.
3 different SPME conditions are set in this example, it is as follows respectively：
Condition 1：65 DEG C of extraction temperature, extraction time 30min.
Condition 2：65 DEG C of extraction temperature, extraction time 60min.
Condition 3：75 DEG C of extraction temperature, extraction time 30min.
In headspace solid-phase microextraction operation, using 50/30 μm of DVB/PDMS/CAR extracting head of 1cm, incubator concussion speed
Spend for 250rpm.
(3) gas chromatography-mass spectrometry analysis：Extracting head is in gas-chromatography (Agilent GC 7890) sample introduction after completion extraction
Mouth uses mass spectrograph (Agilent 5977A) right using 260 DEG C of parsing 4min of Splitless injecting samples pattern after completing desorption and separation
Each volatile ingredient is identified.
Gas-chromatography uses chromatographic column for HP-5MS, 30m, 0.25mm, i.d.0.25 μm, and high-purity helium is carrier gas, stream
Speed is 0.8mL/min；
Column oven heating schedule：60 DEG C of initial temperature is kept after 4min, and 180 DEG C of holding 5min are warming up to 5 DEG C/min, then
240 DEG C are warming up to 20 DEG C/min, 5min is kept.
Mass Spectrometry Conditions：Using EI sources, positive ion detection, ionizing energy is 70eV, 260 DEG C of ion source temperature, transmission line temperature
280 DEG C of degree, scan pattern is full scan, 20~550m/z of scanning range.
(4) data analysis and identification：The retention time and integral area of chromatogram are automatically performed by work station, utilize NIST
11 spectrum storehouses carry out qualitative analysis to each volatile ingredient, as a result as shown in table 2.
To volatility in different ultrasound condition assisted Solid-state micro-extraction is determined under the conditions of different SPMEs brown rice
Material result carries out principal component analysis (PCA), as a result as Fig. 7 shows.
By Fig. 7 to find out, the contribution rate 79.3% of first principal component (PC-1), lower 5 ultrasound conditions of 3 groups of extraction conditions
Result measured by assisted Solid-state micro-extraction is sequentially distributed with axle.The sample of different ultrasound conditions auxiliary is surveyed under different extraction conditions
Determine result to be clearly separated, wherein 65 DEG C of extraction temperature, 60 minutes highest scorings of extraction time, do not use ultrasonic method to aid in
The measurement result score of (ZC1.1, ZC1.2, ZC1.3) is relatively low.As can be seen here under different extraction conditions, using ultrasonic method
Auxiliary can significantly improve the concentration effect of volatile materials and detection abundance in brown rice.
The composition of volatile ingredient and relative amount (n=3) in the brown rice of table 2.
Claims (10)
1. a kind of method of volatile flavor substance in ultrasonic method auxiliary measuring brown rice, it is characterised in that comprise the following steps：
(1) brown rice powder to be measured is taken, is mixed with inorganic salt solution, the obtained mixed liquor of ultrasound；
(2) sorption extraction is carried out to the volatile flavor substance in mixed liquor using headspace solid-phase microextraction technology；
(3) volatile flavor substance is identified using gas chromatography-mass spectrometry analysis technology.
2. the method as described in claim 1, it is characterised in that the inorganic salt solution is saturated sodium-chloride water solution.
3. method as claimed in claim 2, it is characterised in that the liquid ratio of the saturated sodium-chloride water solution and brown rice powder
For 1.6~2.8mL:1g.
4. the method as described in claim 1, it is characterised in that in step (1), described ultrasound condition is：Frequency
40000Hz, the time is 5~60min.
5. method as claimed in claim 4, it is characterised in that the ultrasonic time is 30min.
6. the method as described in claim 1, it is characterised in that in step (2), headspace solid-phase microextraction uses 50/30 μ
MDVB/PDMS/CAR extracting heads.
7. the method as described in claim 1, it is characterised in that during headspace solid-phase microextraction, extraction temperature is 65~75 DEG C, extraction
It is 30~60min to take the time, and incubator concussion speed is 250rpm.
8. the method as described in claim 1, it is characterised in that in step (3), the resolution temperature of volatile flavor substance is
260 DEG C, the parsing time is 4min.
9. the method as described in claim 1, it is characterised in that in step (3), during gas chromatographic analysis, flow rate of carrier gas is
0.8mL/min, heating degree is：60 DEG C of initial temperature is kept after 4min, and 180 DEG C of holding 5min are warming up to 5 DEG C/min, then with
20 DEG C/min is warming up to 240 DEG C, keeps 5min.
10. the method as described in claim 1, it is characterised in that in step (3), mass spectral analysis condition is：Using EI sources, just
Ion detection, ionizing energy is 70eV, 260 DEG C of ion source temperature, 280 DEG C of transmission line temperature, and scan pattern is full scan, scanning
20~550m/z of scope.
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| CN110596255A (en) * | 2019-04-03 | 2019-12-20 | 北京工商大学 | Method for detecting volatile components of tomatoes and products thereof |
| CN115508486A (en) * | 2022-10-19 | 2022-12-23 | 天津春发生物科技集团有限公司 | Method for determining flavor substances in chicken soup based on SAFE-GC-MS technology and application thereof |
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| CN115508486A (en) * | 2022-10-19 | 2022-12-23 | 天津春发生物科技集团有限公司 | Method for determining flavor substances in chicken soup based on SAFE-GC-MS technology and application thereof |
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