CN111982975B - Method for noninvasively evaluating anti-aging performance of donkey whey protein by using odor fingerprint spectrum - Google Patents
Method for noninvasively evaluating anti-aging performance of donkey whey protein by using odor fingerprint spectrum Download PDFInfo
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Abstract
本发明属于功能食品功效快速评价技术领域，公开了一种利用气味指纹图谱无创评价驴乳清蛋白抗衰老性能的方法。该方法包含以下步骤：(1)分别取驴乳清蛋白干预不同时间段小鼠粪便于容器中，密封静置获得挥发性气味物质的顶空气体；(2)将电子鼻传感器阵列与顶空气体接触，产生传感器响应信号，获得驴乳清蛋白干预不同时间小鼠粪便的气味指纹图谱；(3)从气味指纹图谱中提取特征数据，对驴乳清蛋白干预不同时间及对照组小鼠粪便进行定性分类，利用多元线性回归分析建立气味指纹图谱与小鼠周龄之间的相关性，并建立预测小鼠周龄的模型。这种方法利用气味指纹图谱对驴乳清蛋白干预不同阶段进行快速判别，可以无创评价驴乳清蛋白抗氧化性。
The invention belongs to the technical field of rapid evaluation of the efficacy of functional foods, and discloses a method for non-invasively evaluating the anti-aging performance of donkey whey protein by using odor fingerprints. The method comprises the following steps: (1) taking donkey whey protein intervention mouse feces for different periods of time, putting them in containers, sealing and standing still to obtain headspace gas of volatile odor substances; (2) combining the electronic nose sensor array with the headspace (3) extracting characteristic data from the odor fingerprints, and the donkey whey protein intervention time and control group mouse feces Qualitative classification was carried out, and the correlation between odor fingerprints and mouse week age was established by multiple linear regression analysis, and a model for predicting mouse week age was established. This method uses odor fingerprints to quickly identify different stages of donkey whey protein intervention, and can non-invasively evaluate the antioxidant activity of donkey whey protein.
Description
Technical Field
The invention relates to the technical field of rapid evaluation of efficacy of functional foods, relates to a method for evaluating food functionality based on fecal odor, and in particular relates to a method for noninvasively evaluating anti-aging performance of donkey whey protein by utilizing odor fingerprint.
Background
Donkey milk is rich in proteins and unsaturated fatty acids, 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 various researches show that various bioactive components of whey protein have antioxidant activity, such as 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 cell membranes through the alimentary canal, reduce into raw material cysteine of GSH, and maintain the GSH level of cells and tissues, thereby enhancing the oxidation resistance of organisms, and the whey protein belongs to soluble proteins and is easier to digest and absorb by human bodies. At present, the research on the functional characteristics of the whey protein is mainly carried out by establishing animal model experiments and human clinical experiments, but the research method has great dependence on experimental animals, so that the use amount of the experimental animals is in an ascending trend year by year, the experimental animals need to be sacrificed for obtaining physiological and biochemical and morphological indexes, and the method runs counter to animal protection sense, and in addition, the analysis process is complicated, and consumes a great amount of manpower, material resources and financial resources. Therefore, the method has important scientific significance for rapid and noninvasive evaluation of experimental animals in functional property research of whey protein.
The electronic nose is used for identifying simple and complex smell information by utilizing the response of the gas sensor array to volatile smell substances, and has been widely applied to quality detection of foods and agricultural products. Faeces are one of the main ways of outputting the final products of the whole metabolism of the body, and the change of the metabolites can reflect the characteristics of the whole metabolism of the body, and also the external manifestations of diet difference and nutrition regulation influence. However, the current research based on the detection of the volatile components in metabolites by the flavor electronic nose mainly comprises in-vivo efficacy evaluation of the functional components of foods, and the research of noninvasively evaluating the in-vivo functionality of foods by utilizing the odor information of the volatile odor substances of feces has a large blank.
Disclosure of Invention
The invention aims to overcome the defects of the background technology and provides a method for noninvasively evaluating the anti-aging performance of donkey whey protein by using odor fingerprint. According to the method, the odor fingerprint spectrum is utilized to rapidly judge different stages of the intervention of the donkey whey protein, so that the rapid judgment and prediction of the week age of the mice with the intervention of the donkey whey protein can be realized.
In order to achieve the aim of the invention, the method for noninvasively evaluating the anti-aging performance of the donkey whey protein by using the odor fingerprint spectrum comprises the following steps:
(1) Respectively taking donkey whey protein to interfere with mouse excrement in different time periods in a container, sealing and standing to obtain headspace gas of volatile odor substances;
(2) The electronic nose sensor array is contacted with a headspace gas to generate a sensor response signal, so as to obtain odor fingerprint spectra of mouse faeces at different times when donkey whey protein intervenes;
(3) Characteristic data are extracted from odor fingerprint patterns, qualitative classification is carried out on donkey whey protein intervention at different times and mouse faeces of a control group by using a pattern recognition method, correlation between the odor fingerprint patterns and the week age of a mouse is established by using multiple linear regression analysis, and a model for predicting the week age of the mouse is established.
Further, in some embodiments of the present invention, 100 to 400 mg/(kg.d) donkey whey protein is taken in the step (1).
Further, in some embodiments of the invention, the mouse feces in step (1) are 1 to 3 grains.
Further, in some embodiments of the present invention, the sealing and standing time in the step (1) is 5 to 10 minutes.
Further, in some embodiments of the invention, the volume of the headspace gas in step (1) is 150 to 500mL.
Further, in some embodiments of the present invention, the carrier gas flow rate is 200-400 mL/min when the electronic nose sensor array is contacted with the headspace gas in step (2).
Further, in some embodiments of the present invention, the pattern recognition method in the step (3) is typically discriminant analysis and multiple linear regression analysis.
Compared with the prior art, the method provided by the invention can be used for noninvasively evaluating the oxidation resistance of donkey whey protein, fills up the research blank of odor fingerprint analysis in the aspect of food functionality evaluation, widens the method for evaluating animal experiment effect, and avoids killing experimental animals. The method does not need a pretreatment step, is simple to operate, has high detection efficiency and sensitivity, can realize rapid judgment and prediction of the donkey whey protein intervening mice for the week age, and is suitable for being used as a real-time and rapid method for evaluating the food functionality.
Drawings
FIG. 1 is a graph of the donkey whey protein intervention at various times in the mouse fecal gas-gustatory radar;
FIG. 2 is a graph showing a typical discriminant analysis of the odor of feces of mice 7 weeks after a group of donkey whey proteins of the present invention and a control group, wherein the low concentration is 100 mg/(kg.d) per donkey whey protein, the medium concentration is 200 mg/(kg.d) per donkey whey protein, and the high concentration is 400 mg/(kg.d) per donkey whey protein;
FIG. 3 is a graph showing two-dimensional score of the classical discriminant analysis of the odor of the feces of mice at different times during the intervention of donkey whey protein according to the present invention.
Detailed Description
The present invention will be described in further detail with reference to the drawings and examples, in order to make the objects, technical solutions and advantages of the present invention more apparent. 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 intended to be illustrative of the invention and not restrictive.
The terms "comprising," "including," "having," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, step, 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, step, method, article, or apparatus.
When an equivalent, concentration, or other value or parameter is expressed as a range, preferred range, or a range bounded by a list of 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 ranges of "1 to 5" are disclosed, the described ranges should be construed to include ranges of "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a numerical range is described herein, unless otherwise indicated, the range is intended to include its endpoints and all integers and fractions within the range.
Furthermore, the descriptions of the terms "one embodiment," "some embodiments," "examples," "particular examples," or "some examples," etc., described below mean 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 present invention. In this specification, schematic representations of the above terms are not necessarily for the same embodiment or example. The technical features of the respective embodiments of the present invention may be combined with each other as long as they do not collide with each other.
Example 1
A method for noninvasively evaluating the anti-aging performance of donkey whey protein by using odor fingerprint, which comprises the following steps:
(1) Taking 100-400 mg/(Kg.d) donkey whey protein to intervene 1-3 grains of mouse faeces in different time periods in a 150-500 mL beaker respectively, sealing and standing for 5-10 min to obtain headspace gas of volatile odor substances;
(2) Contacting the electronic nose sensor array with the sample headspace gas under the condition that the carrier gas flow rate is 200-400 mL/min, generating a sensor response signal, and obtaining odor fingerprint spectra of the donkey whey protein interfering with the mouse faeces at different times;
(3) Characteristic data are extracted from odor fingerprint patterns, qualitative classification is carried out on donkey whey protein intervention at different times and mouse faeces of a control group by using a pattern recognition method, correlation between the odor fingerprint patterns and the week age of a mouse is established by using multiple linear regression analysis, and a model for predicting the week age of the mouse is established.
Example 2
A donkey whey protein interference mouse faeces processing method and an odor fingerprint spectrum data processing and modeling method. An electronic nose based on a metal sensor odor sensor array, the sensor array of which consists of 10 sensors, the names and properties of each sensor are shown in table 1, was used.
TABLE 1 smell information and corresponding sensor and sensitive substance
The function of these sensors is to convert donkey whey proteins interfering with the action of different odour substances in mouse faeces on their surface into a measurable electrical signal.
The donkey whey protein is used for interfering mice by 100-400 mg/(Kg.d), faeces interfering in different time periods ( weeks 0, 1, 3, 5 and 7) are collected, 1 particle of the donkey whey protein interfering mouse faeces sample is taken and placed in a 150mL beaker in a sealing way for 10min. The modeling set and the verification set prepare 40 parallel samples for each time period donkey whey protein interference mouse fecal sample, the detection time of the electronic nose is set to be 60s, the sampling interval is 80s, and the 59 th response value of the sensor steady state is selected for analysis.
As shown in fig. 1, the odor fingerprint information of the donkey whey protein interference mouse feces in the sensors S1, S2, S3, S4, S5, S8, S9 and S10 in different time periods is less different; there is a large difference in the odor fingerprint information at sensors S6 and S7.
FIG. 2 is a graph showing a typical discriminant analysis of the smell of mouse faeces 7 weeks after intervention with donkey whey protein and control. The smell of the feces of different intervening mice can be basically identified by utilizing the smell of the electronic nose of the feces, and a basis is provided for evaluating the in-vivo functionality of foods based on smell information.
FIG. 3 is a two-dimensional score chart of a typical discriminant analysis of the odor of the feces of mice at different times during 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 fig. 3, the mouse fecal samples of donkey whey protein intervention at weeks 0, 1, 3, 5 and 7 are regularly distributed, namely the longer the intervention time is, the smaller the 1 st principal component score is, and the donkey whey protein intervention time can be well distinguished by using classical discriminant analysis.
Example 3
Based on classical discriminant analysis, a multiple linear regression analysis is further used to establish a correlation between smell information and the age of the mice. The odor information of the feces of the mice at 5 intervention times ( weeks 0, 1, 3, 5, 7) was used as a modeling set, and 12.5% of the data was used as a prediction set. Regression is carried out by taking the smell information of the electronic nose as a parameter of multiple linear regression analysis, and a model for predicting the week age of the mice is established.
The method comprises the steps of obtaining a mice week-age prediction model by adopting multiple linear regression analysis:
mice week age = -35.986S 1 +1.234S 2 -21.064S 3 -1.664S 4 +0.131S 5 -1.879S 6 +1.384S 7 +4.309S 8 -7.928S 9 -7.657S 10 +71.717
In the above formula, S1 to S10 are the odors such as aromatic components, alkanes, and organic sulfides in the odor fingerprint information.
Determining coefficient R of prediction model 2 = 0.8901, indicating that the predictive model established by multiple linear regression analysis is valid.
The prediction results of the prediction model established by the multiple linear regression analysis on the modeling set sample and the prediction set sample are shown in the 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 results can show that the relation between smell fingerprint information and the week age of the mice can be established, and the invention is proved to be feasible for the donkey whey protein to intervene in the week age prediction of the mice.
Table 2 prediction results of multiple linear regression analysis model on modeled and predicted set samples
It will be readily appreciated by those skilled in the art that the foregoing description is merely a preferred embodiment of the invention and is not intended to limit the invention, but any modifications, equivalents, improvements or alternatives falling within the spirit and principles of the invention are intended to be included within the scope of the invention.
Claims (7)
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