CN105021743B - A kind of method of volatile flavor substance in acid-base method auxiliary measuring Sanguis Trionycis meat - Google Patents
A kind of method of volatile flavor substance in acid-base method auxiliary measuring Sanguis Trionycis meat Download PDFInfo
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Abstract
The invention discloses in a kind of acid-base method auxiliary measuring Sanguis Trionycis meat volatile flavor substance method, the method comprises the following steps：(1) take Sanguis Trionycis meat powder to be measured, mixed with aqueous hydrochloric acid solution or sodium hydrate aqueous solution, obtain mixed liquor；(2) sorption extraction is carried out to the volatile flavor substance in mixed liquor using headspace solid-phase microextraction technology；(3) using gaschromatographic mass spectrometric analysises technology, volatile flavor substance is identified.The present invention adds acid or alkaline aqueous solution before detection in Sanguis Trionycis meat sample, have adjusted the acid-base value of detected sample, promotes the developability of volatile flavor substance in Sanguis Trionycis meat, improves extraction efficiency, and then improves the sensitivity of detection method.
Description
Technical field
The present invention relates to acid-base method assisted Solid-state micro-extraction technique, belong to Trionyx sinensis Wiegmann volatile flavor analysis field, especially relate to
And in a kind of acid-base method auxiliary measuring Sanguis Trionycis meat volatile flavor substance method.
Background technology
The delicious meat of Trionyx sinensis Wiegmann, and there is antioxidation, improve immunity of organisms, promote wound healing, YIN nourishing nourishing, improve
Many effects such as serum protein content, promotion hemopoietic function, are the good merchantable brands of China's tradition dietetic therapy nourishing since ancient times.
In recent years, Trionyx sinensis Wiegmann intensive processing has started to walk and fast-developing, has developed small molecule Trionyx sinensis Wiegmann Gly-His-Lys, complete
A series of converted productss such as Trionyx sinensis Wiegmann powder, Trionyx sinensis Wiegmann quintessence oil, vacuum packaging instant type processed Trionyx sinensis Wiegmann.But, with living standard
Improve constantly the transformation with consumption idea, people it is also proposed higher requirement to Trionyx sinensis Wiegmann and Trionyx sinensis Wiegmann product special flavour.Trionyx sinensis Wiegmann and first
In fishery -ies product, the bad flavor such as fulsome fishy smell, terramycin slage is difficult to be pursued nutrition, health, delicious modern consumption by modern
Person is accepted.Therefore, analyze the volatile flavor component in Sanguis Trionycis meat comprehensively, theory is established for suppression or removing bad flavor
Basis, the product for exploitation a new generation provides effective foundation, has far-reaching realistic meaning.
Research currently for soft-shelled turtle nourishing composition is more, and the research for its volatile flavor component is less.Head space
Solid-phase microextraction (Headspace-Solid Phase Micro-extraction, HS-SPME) collection is sampled, extracts, concentrating, entering
The pre- one of the processes such as sample, shows powerful technical advantage in flavour of food products analysis.With other volatile material abstraction techniques
Compare, the method selectivity is good, without organic solvent, required amount of samples is few, directly can be combined with GC/MS, wave in aquatic products
Have in the property sent out flavor substance research and be widely used.
Xie Wancui et al. (Xie Wancui, Yang Xihong, Zhang Chaohua etc. headspace solid-phase microextraction-Gas Chromatography-Mass Spectrometry
The volatile ingredient of arctic shrimp prawn head. analytical chemistry, 39 (12), in December, 2011,1852～1857) adopt the micro- extraction of head space solid phase
Take-volatile flavor component separately and in Preliminary Identification arctic shrimp prawn head for the gas chromatography-mass spectrum (HS-SPME-GC-MS), right
62 kinds of volatile compounds in prawn head have carried out qualitative and semi-quantitative analyses.
However, the volatilization intensity of flavor substance is one of key factor of impact solid-phase microextraction bioaccumulation efficiency, Mou Xiefeng
Taste material is difficult to separate with sample substrate, thus reducing sensitivity and the accuracy of measurement result, in above method often
Flavor substance is promoted to volatilize from sample substrate using the means such as auxiliary heating and interpolation inorganic salt.
Zhao Qingxi et al. (Zhao Qingxi, Xue Changhu, Xu Jie etc. microwave distillation-solid-phase microextraction-gas chromatography-mass spectrum-smell
Feel the volatile ingredient in the detector combination analysis Aristichthys nobiliss flesh of fish. chromatograph, 25 (2), in March, 2007,267～261) adopt microwave
Auxiliary heating promotes the separation of volatile ingredient and enrichment in Aristichthys nobiliss, the 53 kinds of compounds detecting altogether in the Aristichthys nobiliss flesh of fish.
But, the high temperature that above-mentioned supplementary meanss produce has affected on the accuracy of testing result, and for promotion local flavor
The effect that material volatilizees from sample substrate still has much room for improvement.Therefore, it is necessary to probe into one kind can significantly improve in Sanguis Trionycis meat
The volatile method of flavor substance, to solve the above problems.
Content of the invention
The invention provides in a kind of acid-base method auxiliary measuring Sanguis Trionycis meat volatile flavor substance method, the method can
Significantly improve the volatility of flavor substance in Sanguis Trionycis meat, and then improve the sensitivity of volatile flavor substance detection.
In a kind of acid-base method auxiliary measuring Sanguis Trionycis meat, the method for volatile flavor substance, comprises the following steps：
(1) take Sanguis Trionycis meat powder to be measured, mixed with aqueous hydrochloric acid solution or sodium hydrate aqueous solution, obtain mixed liquor；
(2) sorption extraction is carried out to the volatile flavor substance in mixed liquor using headspace solid-phase microextraction technology；
(3) using gas chromatography-mass spectrometry analysis technology, volatile flavor substance is identified.
Adjust the acid-base value of detected sample, promote the volatilization of flavor substance by making the mechanism such as protein denaturation, carry
High extraction efficiency, and then improve the sensitivity of detection method.
Preferably, the pH of described aqueous hydrochloric acid solution is 2～4；The pH of described sodium hydrate aqueous solution is 9～11.
It is further preferred that the pH of described aqueous hydrochloric acid solution is 2；The pH of described sodium hydrate aqueous solution is 9.
Preferably, described aqueous hydrochloric acid solution or sodium hydrate aqueous solution are 1 with the mass ratio of Sanguis Trionycis meat powder:1.
In step (2), headspace solid-phase microextraction adopts 50/30 μm of DVB/PDMS/CAR extracting head.
Preferably, before headspace solid-phase microextraction, mixed liquor is placed in preheating 30min at 50 DEG C.
Preferably, during headspace solid-phase microextraction, extraction temperature is 50 DEG C, extraction time is 30min.
In step (3), during gas chromatographic analysiss, heated up using following procedure：After 40 DEG C of initial temperature keeps 2min, with 5
DEG C/min is warming up to 180 DEG C, then is warming up to 280 DEG C with 10 DEG C/min, keeps 10min.
In step (3), mass spectral analyses condition is：Using EI source, positive ion detection, ionizing energy is 70eV, ion source temperature
230 DEG C of degree, 280 DEG C of transmission line temperature, scan pattern is full scan, sweep limitss 33～450m/z.
Compared with prior art, the invention has the advantages that：
The present invention adds acid or alkaline aqueous solution before detection in Sanguis Trionycis meat sample, have adjusted detected sample
Acid-base value, promote the developability of volatile flavor substance in Sanguis Trionycis meat, improve extraction efficiency, and then improve detection
The sensitivity of method.
Brief description
Fig. 1 is the comparative result of solid-phase microextraction bioaccumulation efficiency under different Acid-Base Systems.
Fig. 2 be volatile flavor substance in Sanguis Trionycis meat is carried out acid system auxiliary (pH=2) solid-phase microextraction detection always from
Subflow figure.
Fig. 3 be volatile flavor substance in Sanguis Trionycis meat is carried out acid system auxiliary (pH=4) solid-phase microextraction detection always from
Subflow figure.
Fig. 4 be volatile flavor substance in Sanguis Trionycis meat is carried out subtraction auxiliary (pH=9) solid-phase microextraction detection always from
Subflow figure.
Fig. 5 be volatile flavor substance in Sanguis Trionycis meat is carried out subtraction auxiliary (pH=11) solid-phase microextraction detection always from
Subflow figure.
Fig. 6 is that volatile flavor substance in Sanguis Trionycis meat is carried out with the total ion current that common solid-phase microextraction (pH=7) detects
Figure.
Fig. 7 is the peak of main flavor compound in the Sanguis Trionycis meat measured by different pH systems under the conditions of different solid-phase microextraction
Area principal component analysiss result (PCA).
Specific embodiment
With reference to specific embodiments, the present invention is further described, and its specific embodiments is interpreted as being only for example
Illustrate, and indefiniteness is it is impossible to limit protection scope of the present invention with following illustrations.
Embodiment 1
In a kind of acid-base method auxiliary measuring Sanguis Trionycis meat, the method for volatile flavor substance, comprises the following steps：
(1) take Sanguis Trionycis meat sample liquid nitrogen to be measured to freeze completely, be broken into powder immediately after, then use aqueous hydrochloric acid solution
Or sodium hydrate aqueous solution and Sanguis Trionycis meat powder in mass ratio 1:1 mixing, obtains mixed liquor.
In the present embodiment, 4 different disposal are set, are to take the aqueous hydrochloric acid solution that pH is 2 or 4 or pH to be 9 or 11 respectively
Sodium hydrate aqueous solution and Sanguis Trionycis meat powder in mass ratio 1:1 mixing, obtains mixed liquor, and remaining treatment conditions is identical.
Above-mentioned aqueous hydrochloric acid solution or sodium hydrate aqueous solution before use in 4 DEG C of Temperature drop in refrigerators to 4 DEG C
(2) weigh rapidly the mixed liquor 5g after mixing as in 20mL ml headspace bottle, add 1g NaCl, with PTFE/ indigo plant
The hollow magnetic metal cover seal of color silica gel dottle pin, by multi-functional full-automatic sample pre-treatments platform MPS XT (German Gerstel
Company) carry out full-automatic headspace solid-phase microextraction operation.
In headspace solid-phase microextraction operation, the ml headspace bottle equipped with mixed liquor first preheats 30min at 50 DEG C；Then, adopt
1cm50/30 μm of DVB/PDMS/CAR extracting head, under conditions of temperature is 50 DEG C, extracts 30min；In extraction process, hatching
Device concussion speed is 250rpm.
(3) gas chromatography-mass spectrometry analysis：After completing extraction, extracting head is in gas chromatogram (Agilent GC 7890) sample introduction
Mouth is using 250 DEG C of parsing 5min of Splitless injecting samples pattern, right with mass spectrograph (Agilent 5977A) after completing desorption and separating
Each volatile ingredient is identified.
The adopted chromatographic column of gas chromatogram is 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：After 40 DEG C of initial temperature keeps 2min, be warming up to 180 DEG C with 5 DEG C/min, then with 10 DEG C/
Min is warming up to 280 DEG C, keeps 10min.
Mass Spectrometry Conditions：Using EI source, positive ion detection, ionizing energy is 70eV, 230 DEG C of ion source temperature, transmission line temperature
280 DEG C of degree, scan pattern is full scan, sweep limitss 33～450m/z, solvent-free delay.
(4) data analysiss and identification：The retention time of chromatogram and integral area are automatically performed by work station, using NIST
11 spectrum storehouses carry out qualitative analyses to each volatile ingredient.The relative amount of each volatile ingredient is obtained by area normalization method,
Result takes the meansigma methodss of different disposal.
Result such as table 1 and Fig. 1～6 show.
The composition of volatile ingredient and relative amount (n=3) in table 1. Sanguis Trionycis meat.
As seen from Figure 1, with the increase of the acidity in Sanguis Trionycis meat solid-phase microextraction system or basicity, volatility wind
The extracting and enriching effect of taste material increases, and measured by said method, the abundance of result substantially rises, so that the sensitivity of method
Improve.
Referring to Fig. 2～6, acid system assisted Solid-state micro-extraction and volatile material composition measured by subtraction assisted Solid-state micro-extraction
Basically identical with content results, identify compound in selected Sanguis Trionycis meat volatile component and (take matching score >=85 for 44
Confirm as this compound, maximum 100), identifiable volatile ingredient and its retention time are listed in table 1.Retention time and peak
Area is calculated with Agilent MassHunter work station, and the method for relative amount area normalization calculates.
Embodiment 2
Flavor substance in acid-base method auxiliary measuring Sanguis Trionycis meat under different condition, comprises the following steps：
(1) take Sanguis Trionycis meat sample liquid nitrogen to be measured to freeze completely, be broken into powder immediately after, then use aqueous hydrochloric acid solution
Or sodium hydrate aqueous solution and Sanguis Trionycis meat powder in mass ratio 1:1 mixing, obtains mixed liquor.
The different pH process of setting 5 in the present embodiment, be respectively the aqueous hydrochloric acid solution that takes pH to be 2 or 4, pH be 9 or 11
Sodium hydrate aqueous solution and deionized water (pH is 7), with Sanguis Trionycis meat powder in mass ratio 1:1 mixing, obtains mixed liquor.
Above-mentioned aqueous hydrochloric acid solution or sodium hydrate aqueous solution before use in 4 DEG C of Temperature drop in refrigerators to 4 DEG C.
(2) weigh rapidly the mixed liquor 5g after mixing as in 20mL ml headspace bottle, add 1g NaCl, with PTFE/ indigo plant
The hollow magnetic metal cover seal of color silica gel dottle pin, by multi-functional full-automatic sample pre-treatments platform MPS XT (German Gerstel
Company) carry out full-automatic headspace solid-phase microextraction operation.
3 different solid-phase microextraction conditions are set in this example, as follows respectively：
Condition 1：50 DEG C of preheating temperature, equilibration time 30min, 50 DEG C of extraction temperature, extraction time 60min.
Condition 2：60 DEG C of preheating temperature, equilibration time 30min, 60 DEG C of extraction temperature, extraction time 60min.
Condition 3：60 DEG C of preheating temperature, equilibration time 30min, 60 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, couveuse concussion speed
Spend for 250rpm.
(3) gas chromatography-mass spectrometry analysis：After completing extraction, extracting head is in gas chromatogram (Agilent GC 7890) sample introduction
Mouth is using 250 DEG C of parsing 5min of Splitless injecting samples pattern, right with mass spectrograph (Agilent 5977A) after completing desorption and separating
Each volatile ingredient is identified.
The adopted chromatographic column of gas chromatogram is 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：After 40 DEG C of initial temperature keeps 2min, be warming up to 180 DEG C with 5 DEG C/min, then with 10 DEG C/
Min is warming up to 280 DEG C, keeps 10min.
Mass Spectrometry Conditions：Using EI source, positive ion detection, ionizing energy is 70eV, 230 DEG C of ion source temperature, transmission line temperature
280 DEG C of degree, scan pattern is full scan, sweep limitss 33～450m/z, solvent-free delay.
(4) data analysiss and identification：The retention time of chromatogram and integral area are automatically performed by work station, using NIST
11 spectrum storehouses carry out qualitative analyses to each volatile ingredient.
Each odor threshold and the average relative being measured in conjunction with each volatile ingredient in the Sanguis Trionycis meat measured by example 1 contain
Amount, calculates the local flavor contribution margin of each compound according to below equation (1), takes abnormal smells from the patient contribution margin main for Sanguis Trionycis meat more than 1
Flavor substance, thus obtaining main flavor substance in Trionyx sinensis Wiegmann to include 9 kinds of compounds such as aldehyde C-9, heptadienal, refers to table 2.
Abnormal smells from the patient contribution margin=relative amount/threshold odour number (ug/kg) (1)
With this 9 kinds of flavor substances as target compound, to pH system assisted Solid-states different under the conditions of different solid-phase microextraction
The result that 9 kinds of Trionyx sinensis Wiegmanns flesh of fish flavor substances of micro-extraction are measured carries out principal component analysiss (PCA), and result such as Fig. 7 shows.
Main flavor and its odor characteristics in table 2 Sanguis Trionycis meat
By Fig. 7 to find out, the contribution rate 80.9% of first principal component (PC-1), 3 groups of extraction conditionss lower 5 pH value auxiliary
Measured by solid-phase microextraction, result is sequentially distributed with axle.Under the conditions of each, the sample determination result of different pH value auxiliary is substantially divided
Open, score on first principal component for wherein pH2 and pH11 is close and apparently higher than other pH value, do not adopt acid-base method to assist
(pH=7) measurement result score is minimum.As can be seen here at different conditions, first can be significantly improved using acid-base method auxiliary
The concentration effect of flavor substance and detection abundance in the flesh of fish.
Claims (1)
1. in a kind of acid-base method auxiliary measuring Sanguis Trionycis meat the method for volatile flavor substance it is characterised in that comprising the following steps：
(1) take Sanguis Trionycis meat powder to be measured, the aqueous hydrochloric acid solution being 2 with pH or the sodium hydrate aqueous solution that pH is 9 to be mixed, obtain
To mixed liquor；Described aqueous hydrochloric acid solution or sodium hydrate aqueous solution are 1 with the mass ratio of Sanguis Trionycis meat powder:1；
(2) sorption extraction is carried out to the volatile flavor substance in mixed liquor using headspace solid-phase microextraction technology；
Headspace solid-phase microextraction adopts 50/30 μm of DVB/PDMS/CAR extracting head；Before headspace solid-phase microextraction, mixed liquor is put
30min is preheated at 50 DEG C；During headspace solid-phase microextraction, extraction temperature is 50 DEG C, and extraction time is 30min；
(3) using gas chromatography-mass spectrometry analysis technology, volatile flavor substance is identified；
During gas chromatographic analysiss, heated up using following procedure：After 40 DEG C of initial temperature keeps 2min, it is warming up to 180 with 5 DEG C/min
DEG C, then it is warming up to 280 DEG C with 10 DEG C/min, keep 10min；
Mass spectral analyses condition is：Using EI source, positive ion detection, ionizing energy is 70eV, 230 DEG C of ion source temperature, transmission line
280 DEG C of temperature, scan pattern is full scan, sweep limitss 33～450m/z.
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| CN111323505A (en) * | 2020-03-18 | 2020-06-23 | 中国农业科学院兰州畜牧与兽药研究所 | Method for detecting sulfur-containing compounds in volatile flavor substances of fermented meat products |
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