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Introduction shrimp
paste is a traditional fermented condiment in China, Malaysia, Singapore, Thailand and other Southeast Asian regions, which is made
by grinding low-value shrimp with salt and then grinding into a viscous fermentation.
Shrimp paste is rich in protein, calcium, iron, selenium, VA and astaxanthin and other nutrients, and moderate consumption is beneficial to health
.
Shrimp paste is a fermented food in which proteins are broken down into amino acids and fat is converted into fatty acids, resulting in a unique and delicious flavor
.
However, the production of shrimp paste in China usually adopts the way of natural fermentation, the whole process is very long, and it is easily affected by uncontrollable factors such as season, weather, temperature, etc.
, resulting in unstable
product quality, especially flavor quality.
At present, the research on the flavor of fermented foods mainly focuses on the finished product, and there are few reports
on the flavor changes during the whole process of fermentation of shrimp paste.
The flavor formation process of shrimp paste is quite complicated, in order to obtain stable flavor quality and provide the possibility for the regulation of flavor quality during shrimp paste fermentation, it is necessary to comprehensively analyze the flavor formation mechanism
of shrimp paste fermentation.
At present, a variety of detection technologies have been widely used in the study
of food volatile flavor.
Electronic nose (E-nose) is an aroma detection technology that can quickly distinguish samples quickly, sensitively, and non-destructively, but cannot accurately analyze volatile flavor components
in samples.
Solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) technology has the characteristics of high sensitivity for the determination of polymer substances, but it has limitations
in the detection of low molecular substances and trace substances.
In recent years, headspace-gas chromatography-ion mobility spectroscopy (HS-GC-IMS) has been widely used
in many fields such as food flavor analysis and quality testing.
HS-GC-IMS can improve the accuracy and sensitivity of flavor detection, and effectively solve the problem of slow GC-MS analysis speed and loss of
aroma substances caused by pretreatment.
However, the limitations of its database hinder its qualitative analysis
of volatile compounds.
The combination of SPME-GC-MS and E-nose is conducive to the comprehensive study of food flavor and is the main method
for detecting volatile flavor substances in food at present.
However, few studies have combined these three assays to analyze the flavor composition
of shrimp paste.
Professor Gao Ruichang and Master Li Ying, School of Food and Bioengineering, Jiangsu University, combined three detection methods: electronic nose, SPME-GC-MS and HS-GC-IMS, to comprehensively analyze the changes of volatile flavor compounds during the fermentation of
shrimp paste.
The method of calculating the relative aroma activity value (ROAV) was used to screen the key volatile flavor compounds in the fermentation process of shrimp paste, which provided a basis for the analysis of the formation path and mechanism of volatile flavor of traditional shrimp paste, and provided a theoretical basis
for the regulation of flavor quality of shrimp paste products.
Results and Discussion
used electronic nose analysis to analyze the overall flavor of shrimp paste samples at different fermentation stages and had almost no response to the four sensors W1C, W3C, W6S and W5C, indicating that aromatic compounds, ammonia and short-chain alkanes
were rarely produced during fermentation.
Although the signal intensity of the W5S, W2W, and W3S sensors to the sample is low, there are still differences between samples, indicating the production of small amounts of organosulfur compounds, nitrogen oxides, and long-chain alkanes
.
The shrimp paste sample has a high response value to W1S, W1W, and W2S, and these three sensors can significantly distinguish shrimp paste samples
at different stages of fermentation.
With the progress of fermentation, the content of short-chain alkanes such as methane, inorganic sulfides, alcohols, aldehydes, ketones and other substances generally showed an upward trend, and the response values of the three sensors remained basically unchanged
in the last two months of fermentation.
The PCA results show that the sample is roughly divided into three relatively separate regions
.
The samples in the two periods of m0 and m1 were relatively concentrated, m4 was relatively independent, the samples of m5 and m6 were similar, and the similarity of m7 and m8 was higher
.
Fig.
1 Volatile components in shrimp paste samples at different fermentation stages based on electronic nose data SPME-GC-MS Analysis SPME-GC-MS is used to analyze volatile flavor compounds
in shrimp paste samples at different fermentation stages.
A total of 75 volatile flavor compounds were identified in the samples at different stages of fermentation, including 21 alcohols, 11 ketones, 8 esters, 1 acid, 4 hydrocarbons, 2 sulfides, 3 amines, 9 pyrazines, 4 aldehydes and 12 other compounds
.
From the stacking plot (Figure 2A), it can be intuitively seen that the types of volatile compounds in different shrimp paste samples change significantly as fermentation progresses
.
In general, the composition of volatile compounds in the early stage of fermentation was relatively similar, and the flavor components of the samples changed significantly in the fourth month of fermentation, and the volatile components in the late fermentation stage were small
.
This is consistent with PCA results for
electronic nose data.
Alcohols and amines were high
in volatile compounds in all samples.
Aldehydes and pyrazine
begin to be produced in the fifth month of fermentation.
Changes in specific volatile compounds can be seen through heat maps (Figure 2B).
The relative content of volatile compounds does not accurately reflect their true contribution
to the overall aroma profile.
Therefore, the contribution
of volatile compounds to the overall aroma profile was identified by calculating the ROAV value.
The higher the ROAV value, the greater
the compound's contribution to the overall flavor.
The substance of ROAV≥1 is the key flavor compound of the sample, and the substance of 0.
1≤ROAV<1 is the compound that changes the overall flavor of the sample.
During the fermentation of shrimp paste, SPME-GC-MS identified four volatiles of 0≤ROAV<1, including 1-pentanol, 1-pentan-3-ol, (2Z)-2-octen-1-ol and benzaldehyde, which contribute to the formation<b136> of overall flavor 。 Thirteen volatile components of ROAV≥1 were identified, of which 9 were key volatile components in the middle and late stages of fermentation, namely 1-octen-3-ol, isobutanol, 1-nonanol, isoamyl alcohol, ethyl acetate, dimethylthiomethane, dimethyl sulfide, trimethylamine, 3-ethyl-2,5-methylpyrazine, 2,3,5-trimethylpyrazine, 2-methyl-3-isopropylpyrazine, isovaleraldehyde and 2-methylbutyraldehyde
。 Fig.
2 Volatile components in samples of shrimp paste at different fermentation stages based on GC-MS data HS-GC-IMS was used to analyze the volatile organic components (VOCs) of shrimp paste at different fermentation stages (Fig.
3A).
It can be seen that with the progress of fermentation, the volatile components in shrimp paste are gradually enriched with the extension of fermentation time, and the types and contents have changed significantly
.
According to the identification results of HS-GC-IMS, THE VOCs in shrimp paste at different fermentation stages mainly include alcohols, ketones, esters, aldehydes and heteroatoms (pyrazines and sulfide ethers).
VOCs, which have a greater influence on the flavor of shrimp paste, are mainly produced in the middle and late stages
of fermentation.
The overall trend of volatile component changes is consistent
with the SPME-GC-MS results.
In Figure 3, 3-methylbutyraldehyde, 2-methylbutyraldehyde, butanone and other substances in the red frame increased significantly when the shrimp paste fermented to the M4 stage, and then gradually decreased
.
Substances such as 2,5-dimethylpyrazine, ethyl isovalerate, ethyl acetate, propionaldehyde, etc.
in the green box increased significantly in the M5 phase and then gradually decreased
.
Ethyl hexanoate, 1-pentanol, heptaldehyde, isobutyraldehyde, 1-butanol and other substances in the yellow frame showed an overall downward trend with the fermentation of shrimp paste, among which the content of 2-pentanone M4 stage was the highest, and then decreased
.
PCA results indicate high similarity between m0 and m1 samples, m5 and m6 samples, and m7 and m8
.
However, M4 is quite
different from samples from other periods.
It may be due to the significant increase in ambient temperature during the M4 period that drastically changes in the composition and activity of the bacterial community, eventually leading to the formation
of a large number of volatile metabolites.
These results are consistent
with those of electronic nose and SPME-GC-MS.
The method of calculating ROAV values is also used to screen key volatile compounds
in VOCs identified by HS-GC-IMS.
A TOTAL OF 3 VOCs of 0≤ROAV<1 AND 8 VOCs<b151> of ROAV≥1 WERE IDENTIFIED BY HS-GC-IMS.
The high sensitivity of HS-GC-IMS technology compensates for the lack
of SME-GC-MS detection of trace substances.
In addition to ethyl acetate, dimethyl sulfide, 2-methylbutyraldehyde, and isovaleraldehyde identified by SME-GC-MS, nonanaldehyde, ethyl caproate, ethyl isovalerate and propionaldehyde have also been identified as key volatile flavor compounds
.
Fig.
3 Volatile components in samples of shrimp paste at different fermentation stages based on GC-IMS data Conclusion
A total of 17 key volatile aroma components
were identified in traditional shrimp paste at different fermentation stages.
The combination of electronic nose, SPME-GC-MS and HS-GC-IMS technologies can compensate for eachSelf-limitation, more fully reflect the change
of volatile components.
The high efficiency and sensitivity of HS-GC-IMS offers the possibility
to use it as a visualization tool for monitoring flavor changes.
In the process of research, it was found that the production of most flavor substances is related
to the degradation and metabolism of flavor precursors such as amino acids and fatty acids.
Since the fermentation process is closely related to the biochemical metabolic pathways of various microorganisms and enzymes, the correlation between key flavor compounds, enzymes and microorganisms will be further explored as the focus of the next research, which is of great significance
for flavor quality control and industrialization of shrimp paste production.
Reference:
paste is a traditional fermented condiment in China, Malaysia, Singapore, Thailand and other Southeast Asian regions, which is made
by grinding low-value shrimp with salt and then grinding into a viscous fermentation.
Shrimp paste is rich in protein, calcium, iron, selenium, VA and astaxanthin and other nutrients, and moderate consumption is beneficial to health
.
Shrimp paste is a fermented food in which proteins are broken down into amino acids and fat is converted into fatty acids, resulting in a unique and delicious flavor
.
However, the production of shrimp paste in China usually adopts the way of natural fermentation, the whole process is very long, and it is easily affected by uncontrollable factors such as season, weather, temperature, etc.
, resulting in unstable
product quality, especially flavor quality.
At present, the research on the flavor of fermented foods mainly focuses on the finished product, and there are few reports
on the flavor changes during the whole process of fermentation of shrimp paste.
The flavor formation process of shrimp paste is quite complicated, in order to obtain stable flavor quality and provide the possibility for the regulation of flavor quality during shrimp paste fermentation, it is necessary to comprehensively analyze the flavor formation mechanism
of shrimp paste fermentation.
At present, a variety of detection technologies have been widely used in the study
of food volatile flavor.
Electronic nose (E-nose) is an aroma detection technology that can quickly distinguish samples quickly, sensitively, and non-destructively, but cannot accurately analyze volatile flavor components
in samples.
Solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) technology has the characteristics of high sensitivity for the determination of polymer substances, but it has limitations
in the detection of low molecular substances and trace substances.
In recent years, headspace-gas chromatography-ion mobility spectroscopy (HS-GC-IMS) has been widely used
in many fields such as food flavor analysis and quality testing.
HS-GC-IMS can improve the accuracy and sensitivity of flavor detection, and effectively solve the problem of slow GC-MS analysis speed and loss of
aroma substances caused by pretreatment.
However, the limitations of its database hinder its qualitative analysis
of volatile compounds.
The combination of SPME-GC-MS and E-nose is conducive to the comprehensive study of food flavor and is the main method
for detecting volatile flavor substances in food at present.
However, few studies have combined these three assays to analyze the flavor composition
of shrimp paste.
Professor Gao Ruichang and Master Li Ying, School of Food and Bioengineering, Jiangsu University, combined three detection methods: electronic nose, SPME-GC-MS and HS-GC-IMS, to comprehensively analyze the changes of volatile flavor compounds during the fermentation of
shrimp paste.
The method of calculating the relative aroma activity value (ROAV) was used to screen the key volatile flavor compounds in the fermentation process of shrimp paste, which provided a basis for the analysis of the formation path and mechanism of volatile flavor of traditional shrimp paste, and provided a theoretical basis
for the regulation of flavor quality of shrimp paste products.
Results and Discussion
used electronic nose analysis to analyze the overall flavor of shrimp paste samples at different fermentation stages and had almost no response to the four sensors W1C, W3C, W6S and W5C, indicating that aromatic compounds, ammonia and short-chain alkanes
were rarely produced during fermentation.
Although the signal intensity of the W5S, W2W, and W3S sensors to the sample is low, there are still differences between samples, indicating the production of small amounts of organosulfur compounds, nitrogen oxides, and long-chain alkanes
.
The shrimp paste sample has a high response value to W1S, W1W, and W2S, and these three sensors can significantly distinguish shrimp paste samples
at different stages of fermentation.
With the progress of fermentation, the content of short-chain alkanes such as methane, inorganic sulfides, alcohols, aldehydes, ketones and other substances generally showed an upward trend, and the response values of the three sensors remained basically unchanged
in the last two months of fermentation.
The PCA results show that the sample is roughly divided into three relatively separate regions
.
The samples in the two periods of m0 and m1 were relatively concentrated, m4 was relatively independent, the samples of m5 and m6 were similar, and the similarity of m7 and m8 was higher
.
Fig.
1 Volatile components in shrimp paste samples at different fermentation stages based on electronic nose data SPME-GC-MS Analysis SPME-GC-MS is used to analyze volatile flavor compounds
in shrimp paste samples at different fermentation stages.
A total of 75 volatile flavor compounds were identified in the samples at different stages of fermentation, including 21 alcohols, 11 ketones, 8 esters, 1 acid, 4 hydrocarbons, 2 sulfides, 3 amines, 9 pyrazines, 4 aldehydes and 12 other compounds
.
From the stacking plot (Figure 2A), it can be intuitively seen that the types of volatile compounds in different shrimp paste samples change significantly as fermentation progresses
.
In general, the composition of volatile compounds in the early stage of fermentation was relatively similar, and the flavor components of the samples changed significantly in the fourth month of fermentation, and the volatile components in the late fermentation stage were small
.
This is consistent with PCA results for
electronic nose data.
Alcohols and amines were high
in volatile compounds in all samples.
Aldehydes and pyrazine
begin to be produced in the fifth month of fermentation.
Changes in specific volatile compounds can be seen through heat maps (Figure 2B).
The relative content of volatile compounds does not accurately reflect their true contribution
to the overall aroma profile.
Therefore, the contribution
of volatile compounds to the overall aroma profile was identified by calculating the ROAV value.
The higher the ROAV value, the greater
the compound's contribution to the overall flavor.
The substance of ROAV≥1 is the key flavor compound of the sample, and the substance of 0.
1≤ROAV<1 is the compound that changes the overall flavor of the sample.
During the fermentation of shrimp paste, SPME-GC-MS identified four volatiles of 0≤ROAV<1, including 1-pentanol, 1-pentan-3-ol, (2Z)-2-octen-1-ol and benzaldehyde, which contribute to the formation<b136> of overall flavor 。 Thirteen volatile components of ROAV≥1 were identified, of which 9 were key volatile components in the middle and late stages of fermentation, namely 1-octen-3-ol, isobutanol, 1-nonanol, isoamyl alcohol, ethyl acetate, dimethylthiomethane, dimethyl sulfide, trimethylamine, 3-ethyl-2,5-methylpyrazine, 2,3,5-trimethylpyrazine, 2-methyl-3-isopropylpyrazine, isovaleraldehyde and 2-methylbutyraldehyde
。 Fig.
2 Volatile components in samples of shrimp paste at different fermentation stages based on GC-MS data HS-GC-IMS was used to analyze the volatile organic components (VOCs) of shrimp paste at different fermentation stages (Fig.
3A).
It can be seen that with the progress of fermentation, the volatile components in shrimp paste are gradually enriched with the extension of fermentation time, and the types and contents have changed significantly
.
According to the identification results of HS-GC-IMS, THE VOCs in shrimp paste at different fermentation stages mainly include alcohols, ketones, esters, aldehydes and heteroatoms (pyrazines and sulfide ethers).
VOCs, which have a greater influence on the flavor of shrimp paste, are mainly produced in the middle and late stages
of fermentation.
The overall trend of volatile component changes is consistent
with the SPME-GC-MS results.
In Figure 3, 3-methylbutyraldehyde, 2-methylbutyraldehyde, butanone and other substances in the red frame increased significantly when the shrimp paste fermented to the M4 stage, and then gradually decreased
.
Substances such as 2,5-dimethylpyrazine, ethyl isovalerate, ethyl acetate, propionaldehyde, etc.
in the green box increased significantly in the M5 phase and then gradually decreased
.
Ethyl hexanoate, 1-pentanol, heptaldehyde, isobutyraldehyde, 1-butanol and other substances in the yellow frame showed an overall downward trend with the fermentation of shrimp paste, among which the content of 2-pentanone M4 stage was the highest, and then decreased
.
PCA results indicate high similarity between m0 and m1 samples, m5 and m6 samples, and m7 and m8
.
However, M4 is quite
different from samples from other periods.
It may be due to the significant increase in ambient temperature during the M4 period that drastically changes in the composition and activity of the bacterial community, eventually leading to the formation
of a large number of volatile metabolites.
These results are consistent
with those of electronic nose and SPME-GC-MS.
The method of calculating ROAV values is also used to screen key volatile compounds
in VOCs identified by HS-GC-IMS.
A TOTAL OF 3 VOCs of 0≤ROAV<1 AND 8 VOCs<b151> of ROAV≥1 WERE IDENTIFIED BY HS-GC-IMS.
The high sensitivity of HS-GC-IMS technology compensates for the lack
of SME-GC-MS detection of trace substances.
In addition to ethyl acetate, dimethyl sulfide, 2-methylbutyraldehyde, and isovaleraldehyde identified by SME-GC-MS, nonanaldehyde, ethyl caproate, ethyl isovalerate and propionaldehyde have also been identified as key volatile flavor compounds
.
Fig.
3 Volatile components in samples of shrimp paste at different fermentation stages based on GC-IMS data Conclusion
A total of 17 key volatile aroma components
were identified in traditional shrimp paste at different fermentation stages.
The combination of electronic nose, SPME-GC-MS and HS-GC-IMS technologies can compensate for eachSelf-limitation, more fully reflect the change
of volatile components.
The high efficiency and sensitivity of HS-GC-IMS offers the possibility
to use it as a visualization tool for monitoring flavor changes.
In the process of research, it was found that the production of most flavor substances is related
to the degradation and metabolism of flavor precursors such as amino acids and fatty acids.
Since the fermentation process is closely related to the biochemical metabolic pathways of various microorganisms and enzymes, the correlation between key flavor compounds, enzymes and microorganisms will be further explored as the focus of the next research, which is of great significance
for flavor quality control and industrialization of shrimp paste production.
Li Ying, female, Master of Engineering, School of Food and Biological Engineering, Jiangsu University, mainly researching food flavor chemistry and fermented aquatic flavor quality control, participating in one of the sub-projects of the National Key R&D Program of "Blue Granary Science and Technology Innovation" "New Technology and Product Development of High-quality Bioprocessing of Aquatic Products"; He presided over a project of
Jiangsu Province Graduate Practice Innovation Program.
Gao Ruichang, male, is a professor and doctoral supervisor
at the School of Food and Biological Engineering, Jiangsu University.
National Modern Agricultural Industry Technology System Post Scientist, former Director of the Food Science Discipline Project of the National Natural Science Foundation of China, High-level Talent of "Six Talent Peaks" in Jiangsu Province, Leader of Food Bioengineering and Intelligent Equipment of Jiangsu University, Director of Chinese Bioengineering Society, Member of Youth Working Committee of China Food Science and Technology Society, Member of Jiangsu Yangtze River Rare Fish Standardization Committee, Editorial Board Member of Future Food Science, Youth Editorial Board Member of Food Science, Editorial Board Member
of Meat Research 。 He has been engaged in food protein processing and comprehensive utilization for a long time, mainly focusing on the research on the relationship between the structure and processing function of food protein; The direction of aquatic food components and nutritional efficacy mainly focuses on the research on the development of nutritional components and efficacy relationship of aquatic raw materials, especially the efficacy of active substances such as protein polypeptides.
The research on food flavor chemistry mainly carries out the flavor formation mechanism and control technology
of traditional aquatic products.
He has presided over more than 20 projects such as the National Natural Science Foundation of China and the National Modern Agricultural Industry Technology System Special Project
.
As the first author or corresponding author in Trends in Food Science & Technology, Critical Reviews in Food Science and Nutrition, Food Hydrocolloids, Journal of Agriculture and Food Chemistry, Food Chemistry, Food & Function, Food Research International, Food Science, Chinese Journal of Food Science and other authoritative journals at home and abroad have published more than 100 papers, including more than 60 papers included in SCI and EI, 2 ESI highly cited papers, chief editor of 1 general planning textbook for colleges and universities in the "Twelfth Five-Year Plan", and participated in the compilation of 5 general textbooks for other colleges and universities; 11 national invention patents were authorized, 22 national invention patents were applied for, and 3 PCT patents were applied; He has won 7 provincial, ministerial and municipal awards, such as Jiangsu Science and Technology Progress Award, China Food Science and Technology Society Science and Technology Progress Award, China Light Industry Federation Science and Technology Progress Award
, and Ministry of Education Higher Education Scientific Research Outstanding Achievement Award.
Ying Lia, Li Yuana, Huijie Liua, Hongying Liub, Yue Zhoua, Miaonan Lia, Ruichang Gaoa,*
a School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China
b Ocean College, Hebei Agriculture University, Qinhuangdao 066000, China
*Corresponding author.
Traditionally fermented shrimp paste has a long fermentation period and is susceptible of external factors, which leads to unstable quality and limits its development and application.
Therefore, the purpose of this study is to analyze the flavor changes in the shrimp paste fermentation process and screen out the key volatile aroma components in the shrimp paste to control the flavor quality of the shrimp paste products.
The overall odor profile was detected by the electronic nose.
A total of 106 volatile flavor compounds in the shrimp paste samples at different fermentation stages were identified by solid-phase microextraction-gas chromatography-mass spectrometry ( SPME-GC-MS) and headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS).
The main aroma components alcohols, aldehydes, pyrazines and other substances in the fermentation process showed an overall upward trend.
A total of 17 key volatile aroma components in shrimp paste at different fermentation stages were identified by the relative aroma activity value (ROAV) method.
The combination of electronic nose, SPME-GC-MS and HS-GC-IMS could comprehensively reflect the changes of volatile components in shrimp paste at different fermentation stages, which helps to further understand the mechanism of shrimp paste flavor formation and provides a basis for the regulation of the flavor quality of shrimp paste products.
Reference:
LI Y, YUAN L, LIU H J, et al.
Analysis of the changes of volatile flavor compounds in a traditional Chinese shrimp paste during fermentation based on electronic nose, SPME-GC-MS and HS-GC-IMS[J].
Food Science and Human Wellness, 2023, 12(1): 173-182.
DOI:10.
1016/j.
fshw.
2022.
07.
035.
