CN111982975A - Method for noninvasive evaluation of donkey whey protein anti-aging performance by using odor fingerprint spectrum - Google Patents
Method for noninvasive evaluation of donkey whey protein anti-aging performance by using odor fingerprint spectrum Download PDFInfo
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Abstract
The invention belongs to the technical field of functional food efficacy rapid evaluation, and discloses a method for noninvasive evaluation of donkey whey protein anti-aging performance by using an odor fingerprint. The method comprises the following steps: (1) respectively taking donkey whey protein to intervene mouse excrement in containers at different time periods, and sealing and standing the containers to obtain headspace gas of volatile odor substances; (2) contacting the electronic nose sensor array with a headspace gas to generate a sensor response signal and obtain odor fingerprint spectrums of mouse excrement interfered by donkey whey protein at different time; (3) extracting characteristic data from the odor fingerprint, qualitatively classifying the donkey whey protein intervention at different time and the feces of the control group mice, establishing the correlation between the odor fingerprint and the week age of the mice by utilizing multivariate linear regression analysis, and establishing a model for predicting the week age of the mice. The method utilizes the odor fingerprint spectrum to rapidly judge different stages of donkey whey protein intervention, and can non-invasively evaluate the donkey whey protein oxidation resistance.
Description
Technical Field
The invention relates to the technical field of rapid evaluation of functional food efficacy, relates to a method for evaluating food functionality based on excrement smell, and particularly relates to a method for noninvasive evaluation of donkey whey protein anti-aging performance by using smell fingerprint.
Background
The donkey milk is rich in protein and unsaturated fatty acid, has high linoleic acid content and low cholesterol content, and contains more vitamin C and trace elements. Donkey milk belongs to whey protein milk, and multiple researches show that various bioactive components of whey protein have antioxidant activity, for example, reaction products of beta-lactoglobulin and some saccharides have stronger activities of scavenging free radicals and resisting oxidation. In addition, lactalbumin, lactoferrin and the like in the whey protein are rich in cystine residues, can enter a cell membrane through a digestive tract, are reduced into raw material cysteine of GSH, and maintain the GSH level of cells and tissues, so that the oxidation resistance of an organism is enhanced, and the whey protein belongs to soluble protein and is easier to digest and absorb by a human body. At present, the research on the functional characteristics of whey protein is mainly carried out by establishing animal model experiments and human clinical experiments, but the research method has large dependence on experimental animals, so that the use amount of the experimental animals is on the rise trend year by year, the experimental animals need to be killed for acquiring physiological, biochemical and morphological indexes, and the research method runs counter to animal protection. Therefore, the method has important scientific significance for carrying out quick and noninvasive evaluation on experimental animals in the functional characteristic research of the whey protein.
The electronic nose utilizes the response of the gas sensor array to volatile odor substances to identify simple and complex odor information, and is widely applied to quality detection of food and agricultural products. Feces is one of the main ways of outputting final products of the whole metabolism of the body, and the change of metabolites of the feces can reflect the characteristics of the whole metabolism of the body and also reflect the external manifestations of dietary differences and nutrition regulation influences. However, at present, the research based on the flavor development electronic nose detection of volatile components in metabolites mainly comprises the evaluation of the functional components in foods, and the research of non-invasive evaluation of the functions in foods by using the odor information of the volatile odor substances in feces has a large blank.
Disclosure of Invention
The invention aims to overcome the defects of the background technology and provide a method for non-invasively evaluating the anti-aging performance of donkey whey protein by using an odor fingerprint spectrum. The method utilizes the odor fingerprint spectrum to rapidly judge different stages of donkey whey protein intervention, and can realize rapid judgment and prediction of the week age of donkey whey protein intervention mice.
In order to achieve the purpose of the invention, the method for non-invasively evaluating the anti-aging performance of donkey whey protein by using the odor fingerprint comprises the following steps:
(1) respectively taking donkey whey protein to intervene mouse excrement in containers at different time periods, sealing and standing to obtain headspace gas of volatile odor substances;
(2) contacting the electronic nose sensor array with a headspace gas to generate a sensor response signal, and obtaining odor fingerprint spectrums of mouse excrement interfered by donkey whey protein at different times;
(3) extracting characteristic data from the odor fingerprint, qualitatively classifying the feces of the control group mice at different time intervals after donkey whey protein intervention by using a mode identification method, establishing the correlation between the odor fingerprint and the week age of the mice by using multivariate linear regression analysis, and establishing a model for predicting the week age of the mice.
Further, in some embodiments of the present invention, 100-400 mg/(Kg · d) donkey whey protein is taken in the step (1).
Further, in some embodiments of the present invention, the stool of the mouse in the step (1) is 1-3 pieces.
Further, in some embodiments of the present invention, the time for the sealing and standing in the step (1) is 5-10 min.
Further, in some embodiments of the present invention, the volume of the headspace gas in the step (1) is 150-500 mL.
Further, in some embodiments of the present invention, the carrier gas flow rate when the electronic nose sensor array is in contact with the head space gas in step (2) is 200-400 mL/min.
Further, in some embodiments of the present invention, the pattern recognition method in the step (3) is canonical discriminant analysis and multiple linear regression analysis.
Compared with the prior art, the method provided by the invention can be used for non-invasive evaluation of the oxidation resistance of donkey whey protein, fills up the blank of research on the aspect of food functionality evaluation of odor fingerprint analysis, widens the method for evaluating animal experiment effect, and avoids the death of experimental animals. The method disclosed by the invention does not need a pretreatment step, is simple to operate, has high detection efficiency and sensitivity, can realize rapid judgment and prediction of donkey whey protein intervention mouse week age, and is suitable for being used as a real-time and rapid method for evaluating food functionality.
Drawings
FIG. 1 is a radar chart of odor perception of mouse feces at different times of donkey whey protein intervention;
FIG. 2 is a typical discriminant analysis of the odor of mouse feces after 7 weeks in the donkey whey protein groups with different concentrations and the control group, wherein the low concentration of the gavage drug is 100 mg/(kg. d) per mouse, the medium concentration of the gavage drug is 200 mg/(kg. d) per mouse, and the high concentration of the gavage drug is 400 mg/(kg. d) per mouse;
FIG. 3 is a two-dimensional score chart of discriminant analysis of odor of mouse feces at different times in intervention of donkey whey protein of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. 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. It is to be understood that the following description is only illustrative of the present invention and is not intended to limit the present invention.
As used herein, the terms "comprises," "comprising," "includes," "including," "has," "having," "contains," "containing," or any other variation thereof, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.
When an amount, concentration, or other value or parameter is expressed as a range, preferred range, or as a range of values, including upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when a range of "1 to 5" is disclosed, the described range should be interpreted to include the ranges "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a range of values is described herein, unless otherwise stated, the range is intended to include the endpoints thereof and all integers and fractions within the range.
Furthermore, the description below of the terms "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., 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 are not necessarily intended to refer to the same embodiment or example. Further, the technical features of the embodiments of the present invention may be combined with each other as long as they do not conflict with each other.
Example 1
A method for non-invasively evaluating the anti-aging performance of donkey whey protein by using an odor fingerprint spectrum comprises the following steps:
(1) respectively taking 100-400 mg/(Kg. d) donkey whey protein to intervene 1-3 mouse excrement in 150-500 mL beakers at different time periods, sealing and standing for 5-10 min to obtain headspace gas of volatile odor substances;
(2) under the condition that the flow rate of carrier gas is 200-400 mL/min, the electronic nose sensor array is in contact with the headspace gas of a sample, a sensor response signal is generated, and odor fingerprint spectrums of mouse excrement at different time of donkey whey protein intervention are obtained;
(3) extracting characteristic data from the odor fingerprint, qualitatively classifying the feces of the control group mice at different time intervals after donkey whey protein intervention by using a mode identification method, establishing the correlation between the odor fingerprint and the week age of the mice by using multivariate linear regression analysis, and establishing a model for predicting the week age of the mice.
Example 2
A method for processing donkey whey protein interference mouse feces and a method for processing and modeling odor fingerprint data. An electronic nose based on an array of metal odor sensors was used, the sensor array consisting of 10 sensors, each sensor having the name and performance given in table 1.
TABLE 1 odor information and corresponding sensors and sensitive substances
The function of the sensors is to convert donkey whey protein into measurable electric signals to interfere the action of different odor substances in the mouse excrement on the surface of the mouse excrement.
And (3) intervening the mouse by using 100-400 mg/(Kg. d) donkey whey protein, collecting feces intervening at different time periods ( weeks 0, 1, 3, 5 and 7), taking 1 particle of a donkey whey protein interference mouse feces sample, sealing and standing in a 150mL beaker for 10 min. 40 parallel samples are prepared for donkey whey protein interference mouse fecal samples in each time period in a modeling set and a verification set, the detection time of an electronic nose is set to be 60s, the sampling interval is set to be 80s, and the response value of the steady state 59s of the sensor is selected for analysis.
As shown in fig. 1, odor fingerprint information of mouse feces interfered by donkey whey protein at sensors S1, S2, S3, S4, S5, S8, S9 and S10 in different time periods is less different; there is a large difference in the smell fingerprint information at the sensors S6 and S7.
FIG. 2 is a classic discriminant analysis of mouse stool odor 7 weeks after intervention with donkey whey protein and controls. The odor of the excrement of different intervening mice can be basically identified by using the electronic nose odor of the excrement and by discriminant analysis, and a foundation is provided for the functional evaluation of food in vivo based on odor information.
FIG. 3 is a two-dimensional score chart of discriminant analysis of odor of mouse feces at different time after donkey whey protein intervention. The contribution rates of the first two main components are 73.39% and 17.41%, respectively, and the total contribution rate reaches 90.80%. As can be seen from the attached figure 3, the mouse excrement samples of 0, 1, 3, 5 and 7 weeks after intervention of donkey whey protein are regularly distributed, namely the score of the 1 st principal component is smaller as the intervention time is longer, and the intervention time of donkey whey protein can be well distinguished by discriminant analysis by utilizing classics.
Example 3
On the basis of classical discriminant analysis, multivariate linear regression analysis is further adopted to establish the correlation between the smell information and the mouse week age. Odor information of 5 intervention time ( weeks 0, 1, 3, 5, 7) mouse feces was used as a modeling set, and 12.5% of the data was used as a prediction set. And (3) performing regression by using the odor information of the electronic nose as a parameter of the multiple linear regression analysis, and establishing a model for predicting the week age of the mouse.
Obtaining a mouse week age prediction model by adopting multivariate linear regression analysis:
mice week-old ═ 35.986S1+1.234S2-21.064S3-1.664S4+0.131S5-1.879S6 +1.384S7+4.309S8-7.928S9-7.657S10+71.717
In the above formula, S1-S10 represent the odor of aromatic components, alkanes, organic sulfides, etc. in the odor fingerprint information.
Coefficient of determination R of prediction model20.8901, the predictive model established by multiple linear regression analysis is shown to be valid.
The prediction results of the prediction model established by the multiple linear regression analysis on the modeling set samples and the prediction set samples are shown in table 2, the error range of the prediction results is allowed to fluctuate within +/-1 (the animal experiment difference is large), and the prediction accuracy is 100%. The model prediction result shows that the relationship between the smell fingerprint information and the mouse week age can be established, which shows that the method is feasible for the donkey whey protein to intervene in the mouse week age prediction.
TABLE 2 prediction results of multiple linear regression analysis model on modeling set samples and prediction set samples
It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (7)
1. A method for non-invasively evaluating the anti-aging performance of donkey whey protein by using an odor fingerprint spectrum is characterized by comprising the following steps:
(1) respectively taking donkey whey protein to intervene mouse excrement in containers at different time periods, sealing and standing to obtain headspace gas of volatile odor substances;
(2) contacting the electronic nose sensor array with a headspace gas to generate a sensor response signal and obtain odor fingerprint spectrums of mouse excrement interfered by donkey whey protein at different time;
(3) extracting characteristic data from the odor fingerprint, qualitatively classifying the feces of the control group mice at different time intervals after donkey whey protein intervention by using a mode identification method, establishing the correlation between the odor fingerprint and the week age of the mice by using multivariate linear regression analysis, and establishing a model for predicting the week age of the mice.
2. The method for non-invasively evaluating the anti-aging performance of donkey whey protein by using the odor fingerprint as claimed in claim 1, wherein 100-400 mg/(Kg-d) donkey whey protein is taken in the step (1).
3. The method for non-invasively evaluating the anti-aging performance of donkey whey protein by using the odor fingerprint as claimed in claim 1, wherein 1-3 mouse stools are obtained in the step (1).
4. The method for non-invasively evaluating the anti-aging performance of donkey whey protein by using the odor fingerprint spectrum according to claim 1, wherein the time for sealing and standing in the step (1) is 5-10 min.
5. The method for non-invasively evaluating the anti-aging performance of donkey whey protein by using the odor fingerprint spectrum according to claim 1, wherein the volume of headspace gas in the step (1) is 150-500 mL.
6. The method for non-invasively evaluating the anti-aging performance of donkey whey protein by using the odor fingerprint spectrum according to claim 1, wherein the flow rate of the carrier gas is 200-400 mL/min when the electronic nose sensor array is in contact with the headspace gas in the step (2).
7. The method for non-invasively evaluating the anti-aging performance of donkey whey protein by using the odor fingerprint as claimed in claim 1, wherein the mode identification method in the step (3) is canonical discriminant analysis and multiple linear regression analysis.
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