CN111077262B - Method for identifying milk nutrition quality - Google Patents
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Abstract
The invention relates to a method for identifying the nutritional quality of milk, which comprises the following steps: step 1), extracting tryptophan and metabolites thereof from a milk sample to obtain an extract; step 2), performing chromatographic separation-mass spectrometry scanning on the extract in the step 1), performing quantitative analysis on tryptophan and metabolites thereof in the milk sample, and determining the quantitative limit and the recovery rate; step 3), analyzing the milk sample according to the content data of tryptophan and metabolites thereof in the milk sample in the step 2); the method provided by the invention has the advantages of good recovery rate, low quantitative limit and simple and easy operation, can quickly detect the content of tryptophan and metabolites thereof in the milk product, and can perform nutritional evaluation on the milk quality.
Description
Technical Field
The invention relates to the technical field of food inspection, in particular to a method for identifying the nutritional quality of milk.
Background
Amino acids are a class of important substances in the body, and are the basic units that constitute proteins; they are involved in the metabolism of fat, saccharides and the like in organisms, have important physiological functions for human beings and animals, and the lack of amino acids can interfere the normal metabolism of the human bodies. Among them, tryptophan is one of eight essential amino acids required by human body, cannot be produced in vivo, not only participates in the synthesis of protein in vivo, but also has certain effects on mental stability and depression relieving. In addition, tryptophan has three metabolic pathways in the organism, namely a serotonin metabolic pathway, a kynurenine pathway and an indole pyruvate pathway. Tryptophan can be further metabolized into serotonin (serotonin) on a serotonin metabolic pathway, and the substance can improve sleep, reduce impatient mood and bring joyful feeling and happiness to people; the kynurenic acid generated in the kynurenine pathway has a neuroprotective effect and can resist the neurotoxicity caused by the overexcitation of quinolinic acid, and the final product nicotinic acid in the pathway has the functions of promoting the absorption of iron in vivo and the generation of blood cells, maintaining the normal functions of skin, the secretion of digestive glands and the like. In addition, indole formaldehyde and indole acetic acid generated by indole pyruvic acid which is rarely concerned by people also play an important role in human health.
In recent years, the consumption demand of high-end dairy products in China is rising continuously, and consumers pay more attention to the types and contents of nutrient substances when buying the dairy products. Due to the low content of small-molecule nutrients such as tryptophan and metabolites thereof in milk, the industry neglects such nutrients in production. However, the functions of the substances are not small and varied, so that from the viewpoint of tryptophan and metabolites thereof, a method for detecting the content and the types of the nutritional ingredients in the dairy product is established, on one hand, a new idea and a new method can be provided for nutritional evaluation of the dairy product, and on the other hand, an idea and a suggestion can be provided for producing high-quality and various dairy products in the industry.
The high performance liquid chromatography-mass spectrometry combined technology plays an important role in food component safety detection, compared with the traditional fluorescence chromatography method for detecting tryptophan and serotonin, the technology has simple and convenient pretreatment method, can realize the comprehensive analysis of various metabolites of the tryptophan in the dairy product, and has low detection quantitative limit and accurate quantification. Can provide powerful technical support for the nutritional evaluation of the dairy.
Disclosure of Invention
Aiming at the defects and shortcomings in the prior art, the invention provides a method for identifying the nutritional quality of milk (by using a high performance liquid chromatography-mass spectrometry technology). The invention aims to solve the problem that the existing milk nutritional value evaluation system focuses on the evaluation of main nutritional substances such as protein, fatty acid and the like, and establishes a detection method for small molecular nutritional substances such as tryptophan and the like in milk by using a high performance liquid chromatography-mass spectrometry combined technology to realize the nutritional quality identification of different milks. Provides evaluation indexes of micromolecular nutrients in milk for the academic world and the industry.
In the invention, tryptophan is one of eight essential amino acids required by human bodies, cannot be generated in vivo, not only participates in the synthesis of protein in organisms, but also has certain effects on mental stability and depression relief. In addition, tryptophan has three metabolic pathways in organisms, various produced bioactive substances have important biological functions for human bodies, such as serum quality improvement, sleep reduction, dysphoria reduction, and nicotinic acid has the functions of promoting iron absorption and blood cell generation in the bodies, maintaining normal functions of the skin, secretion of digestive glands and the like. The milk also contains trace bioactive substances besides tryptophan, so that the invention establishes a detection method of the nutrient components in the dairy product from the perspective of tryptophan and metabolites thereof, so as to provide a new thought and method for dairy product nutrition evaluation and provide a thought and suggestion for producing high-quality and multi-type dairy products in the dairy industry.
One of the purposes of the invention is to provide a method for identifying the nutritional quality of milk, which comprises the following steps:
the method comprises the following steps:
step 1), extracting tryptophan and metabolites thereof from a milk sample to obtain an extract;
step 2), performing chromatographic separation-mass spectrometry scanning on the extract in the step 1), performing quantitative analysis on tryptophan and metabolites thereof in the milk sample, and determining the quantitative limit and the recovery rate;
and 3) analyzing the milk sample according to the content data of the tryptophan and the metabolites thereof in the milk sample in the step 2).
According to some preferred embodiments of the present invention, the tryptophan and metabolites thereof are selected from one or more of melatonin, 5-hydroxytryptophan, serotonin, indole-3-ethylamine, tryptamine, 3-hydroxyanthranilic acid, nicotinic acid, xanthurenic acid, tryptophan, kynurenine, kynurenic acid, indolyldehyde and indolylacetic acid, preferably 5-hydroxytryptophan, indole-3-ethylamine, tryptamine, 3-hydroxyanthranilic acid, nicotinic acid, xanthurenic acid, kynurenine, kynurenic acid, indolyldehyde and indolacetic acid.
According to some preferred embodiments of the present invention, in step 1), the milk sample comprises pasteurized milk, ultra-high temperature sterilized milk, reconstituted milk sample, and yogurt sample; preferably, the reconstituted milk sample is whole milk powder and is prepared according to the protein content of 3.0g/100 mL; and/or performing subsequent treatment after the yoghurt sample is diluted by 1: 1.
According to some preferred embodiments of the invention, in step 1), acetonitrile formate is added to the milk sample, vortexed; adding an extraction salt bag, performing vortex and centrifugation, transferring the upper layer solution, adding a purification reagent, performing vortex and centrifugation, taking the upper layer solution, and drying by using nitrogen; adding the complex solution to carry out water bath ultrasound, carrying out vortex mixing, dissolving again, centrifuging, taking the upper layer solution, and passing through a membrane for the next detection; preferably, 5-10 parts of milk sample are taken, and the volume of each part of milk sample is 5-10 ml; the volume ratio of the milk sample, the acetonitrile formate and the complex solution is (5-10): (5-15): 0.5-3), and preferably 5:10: 1.
According to some preferred embodiments of the present invention, in step 1), the acetonitrile formate is a mixed solution of formic acid and acetonitrile, and the volume ratio of formic acid to acetonitrile is 3-8: 97-92, preferably 5: 95; and/or the double solution is 10% acetonitrile in water; and/or centrifuging for 8-10 min at 8000-10000 rpm; and/or the water bath ultrasound condition is that the water bath ultrasound is carried out for 8-10 min at the temperature of 20-30 ℃.
According to some preferred embodiments of the present invention, the extraction salt package is selected from one or more of sodium sulfate, sodium chloride and magnesium sulfate, and the extraction salt package is selected from one or more of sodium sulfate, sodium chloride and magnesium sulfate, preferably 2 to 8g of sodium sulfate and 0.5 to 3g of sodium chloride, and more preferably 4g of sodium sulfate and 1g of sodium chloride.
According to some preferred embodiments of the present invention, the purifying agent is selected from one or more of sodium sulfate, magnesium sulfate, C18 and PSA, preferably 200-1500 mg sodium sulfate and 50-300 mg C18; more preferably 900mg sodium sulfate and 150mg C18.
According to some preferred embodiments of the invention, in step 2), the conditions of the chromatographic separation are: the adopted mobile phase A is an aqueous solution containing 0.1 percent of formic acid, and the mobile phase B is a methanol solution containing 0.1 percent of formic acid; and/or, the chromatographic separation-mass spectrometry scan comprises a primary scan and a secondary ion fragment scan.
According to some preferred embodiments of the present invention, step 2) is performed by gradient elution, preferably, the elution gradient is:
according to some preferred embodiments of the invention, in step 2), the conditions of mass spectrometry are: multiple reaction detection (MRM) positive ion mode, air curtain gas (CUR)35psi, jet collision gas (CAD)10psi, ionization voltage (IS)5500V, Temperature (TEM)500 ℃; nebulizer (GS1)50 psi; supplemental heater (GS2)50 psi.
According to some preferred embodiments of the present invention, in step 3), according to the content data of each metabolite in the milk sample, performing variance analysis and partial least squares discriminant analysis on the milk sample to determine compounds with different contents in different milk samples.
The invention has the beneficial effects that: the method provided by the invention has the advantages of good recovery rate, low quantitative limit and simple and easy operation, can quickly detect the content of tryptophan and metabolites thereof in the milk product, provides a method for performing nutritional evaluation on the milk quality, provides a thought for producing high-quality milk and milk products, and provides a reference for producing related food health products.
Drawings
FIG. 1 is a bar graph showing the content of each characterization factor in pasteurized milk, UHT milk, reconstituted milk and yogurt.
Fig. 2 is a partial least squares discriminant analysis two-dimensional (a) and three-dimensional (b) plot of pasteurized milk, UHT milk, reconstituted milk, and yogurt.
FIG. 3 is a cross-validation result graph of partial least squares discriminant analysis; in the figure, R2 and Q2 are respectively a judgment coefficient and a cross validation coefficient.
FIG. 4 is a sample hierarchical clustering result graph based on the discriminant model.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are a part of the embodiments of the present invention, but not all of the embodiments. The technical solution of the present invention is not limited to the following specific embodiments, and includes any combination of the specific embodiments, and all other embodiments obtained by a person of ordinary skill in the art without creative efforts based on the embodiments of the present invention belong to the protection scope of the present invention.
In the present invention, the specific techniques or conditions not specified in the examples are performed according to the techniques or conditions described in the literature in the art or according to the product specification. The instruments and the like are conventional products which are purchased by normal distributors and are not indicated by manufacturers. The chemical raw materials used in the invention can be conveniently purchased in domestic chemical product markets, the used C18 and PSA are common purification reagents in the field, the purification reagents are purchased from Agilent company, and the used compound solution is a 10% acetonitrile aqueous solution.
According to some preferred embodiments of the present invention, the specific steps are as follows:
the specific method for pretreatment in the step 1) comprises the following steps:
taking 5-10 parts of milk sample, adding 10 parts of acetonitrile formate into 5 parts of sample by volume, and carrying out vortex; then adding 4g of sodium sulfate and 1g of sodium chloride, and carrying out vortex and centrifugation; transferring the upper layer solution; adding 900mg of sodium sulfate and 150mgC18, and performing vortex and centrifugation; taking the upper solution and drying by using nitrogen; adding 1 volume part of the complex solution, performing water bath ultrasound, performing vortex mixing to dissolve the sample again, centrifuging, and taking the upper layer solution to pass through a membrane for the next detection.
Wherein, the reconstituted milk sample is prepared by using whole milk powder according to the protein content of 3.0g/100 mL; and diluting the yoghourt sample by 1:1 and then carrying out subsequent treatment.
Wherein, the formic acid acetonitrile contains 5 percent of formic acid; the composite solution is 0.1% formic acid acetonitrile.
Wherein the centrifugation condition is 8000-10000 rpm for 8-10 min.
Wherein the condition of the water bath ultrasound is that the water bath ultrasound is carried out for 8-10 min at the temperature of 20-30 ℃.
The chromatographic separation conditions in the step 2) are as follows:
the mobile phase A used was an aqueous solution containing 0.1% formic acid, and the mobile phase B was a methanol solution containing 0.1% formic acid.
More preferably, the elution gradient is:
further, step 3) comprises:
i.quantitative limit of tryptophan and its metabolites in milk
The limit of detection (LOD) and limit of quantitation (LOQ) of each species were determined from the signal-to-noise ratio (S/N) of the compound peaks. (Table 1)
TABLE 1 detection and quantitation limits (unit: ng/ml) for tryptophan and its metabolites in different milks
Recovery rate of method for detecting tryptophan and its metabolites in milk
TABLE 2 recovery rates of the detection methods for tryptophan and its metabolites in different milks
Further, step 4) comprises:
the significance of the content difference of tryptophan and metabolites thereof in the four milks is analyzed by a one-way ANOVA method (Table 3), and the characterization factors of the four milks are found out. According to the content original data of each characterization factor, partial least squares discriminant analysis is carried out on the four kinds of milk, and the result shows that the variance cumulative contribution rate of the first three main components to the sample reaches 78.6%, and pasteurized milk, UHT milk, reconstituted milk and yoghourt can be completely distinguished.
TABLE 3 analysis of tryptophan and its metabolites content and significance in four milks
In the table, the contents are expressed as "mean ± standard deviation", and the letters a, b, c, d indicate that the significance of each value differs within a 95% confidence interval based on the data from the analysis of variance and the duncan test.
Example 1
1) Pretreatment of samples
5 brands of pasteurized milk, 10 brands of UHT milk, 10 brands of milk powder and 10 brands of yoghourt are purchased from respective supermarkets of Beijing, wherein the reconstituted milk sample is prepared from the purchased milk powder according to the protein content of 3.0g/100mL, and the yoghourt sample is prepared by the following steps of 1:1, diluting and then carrying out subsequent treatment; respectively taking 5ml of each milk sample, adding 10ml of acetonitrile solution containing 5% formic acid (the volume ratio of formic acid to acetonitrile is 5:95), and vortexing; then adding 4g of sodium sulfate and 1g of sodium chloride, and centrifuging for 10min at 10000rpm by vortex; transferring 7ml of the upper layer solution, adding 900mg of sodium sulfate and 150mgC18, and centrifuging for 5min at 10000rpm by vortex; taking 6ml of upper layer solution and drying by using nitrogen; adding 1ml of the re-solution, performing water bath ultrasound for 5min, performing vortex mixing to re-dissolve the sample, centrifuging to obtain the upper layer solution, passing the upper layer solution through a membrane for the next detection, and repeating the experiment for 3 times for each sample. FIG. 1 is a bar graph showing the content of partial tryptophan metabolites in pasteurized milk, UHT milk, reconstituted milk and yogurt.
2) Qualitative and quantitative analysis of various characterization factors in different kinds of milk:
detecting by using a high performance liquid chromatography-mass spectrometry combined technology:
the mass spectrometer used for the experiment was UPLC-QTRAP 4500(AB SCIEX), the chromatographic column used was ZORBAX Eclipse plus C18(150 mm. times.3.0 mm, particle size 1.8 μm) in Agilent, and the column oven temperature was 30 ℃.
And (3) pretreating the sample, and detecting the sample by using high performance liquid chromatography-mass spectrometry. The mobile phase A and the mobile phase B are respectively an aqueous solution containing 0.1% formic acid and a methanol solution containing 0.1% formic acid, and the elution gradient is as follows:
the conditions of mass spectrum are: multiple reaction detection (MRM) positive ion mode, air curtain gas (CUR)35psi, jet collision gas (CAD)10psi, ionization voltage (IS)5500V, Temperature (TEM)500 ℃; nebulizer (GS1)50 psi; supplemental heater (GS2)50 psi.
Quantitative analysis of tryptophan and its metabolites in different milks:
due to different matrix effects of tryptophan and metabolites thereof in milk, the tryptophan and the metabolites thereof in different milk are quantitatively analyzed by utilizing a matrix standard curve.
3) one-way ANOVA analysis and partial least square method discriminant analysis
And (3) performing one-way ANOVA analysis and partial least squares discriminant analysis processing based on the content data of tryptophan and metabolites thereof in different milks, and extracting a partial least squares discriminant analysis chart shown in figure 2. FIG. 3 is a graph of the cross-validation results of the partial least squares discriminant analysis showing that the metabolites subjected to the partial least squares discriminant analysis were not overfitted; fig. 4 is a sample hierarchical clustering result chart based on a discriminant model, and the result shows that metabolites with significant differences are obviously distinguished from four kinds of milk. The first three main components from the figure can completely distinguish pasteurized milk, UHT milk, reconstituted milk and yoghurt. This indicates that ten factors (5-hydroxytryptophan, indole-3-ethylamine, tryptamine, 3-hydroxyanthranilic acid, nicotinic acid, xanthurenic acid, kynurenine, kynurenic acid, indole carbaldehyde, indole acetic acid) differ significantly in the classification of the four milks. Based on the content data of the nine characterization factors in different milks, 7-fold cross validation and replacement inspection are carried out on the partial least square discrimination model, and the accuracy rate reaches 100%.
Example 2
This example was carried out in the same manner as example 1 except that the extraction solvent used was pure acetonitrile solvent. Through inspection, the recovery rates of the melatonin, the serotonin, the indole-3-ethylamine, the tryptamine, the indole formaldehyde, the indole acetic acid and the 3-hydroxy anthranilic acid are 60 to 120 percent. The recovery rate of tryptophan, 5-hydroxytryptophan, nicotinic acid, xanthurenic acid, kynurenine and kynurenine is only 2-30%.
Example 3
This example was carried out in the same manner as example 1 except that the extraction solvent used was an acetonitrile solvent containing 2% formic acid. Through inspection, the recovery rate of tryptophan, nicotinic acid, xanthurenic acid, kynurenine, melatonin, serotonin, indole-3-ethylamine, tryptamine, indole formaldehyde, indole acetic acid and 3-hydroxy anthranilic acid is 64-121%; while the recovery rate of the 5-hydroxytryptophan and the kynurenic acid is only 3 to 40 percent.
Comparative example 1
The method adopted by the comparative example specifically comprises the following steps: taking 5ml of each milk sample, precipitating protein with an aqueous solution containing 1% trichloroacetic acid (TCA), centrifuging, adjusting pH to 7, extracting tryptophan and metabolites thereof from the solution by using a C18 or HLB solid phase extraction column, eluting with 6ml of methanol, receiving the eluate, and drying with nitrogen; adding 1ml of the compound solution, carrying out water bath ultrasound for 5min, carrying out vortex mixing, re-dissolving the sample, centrifuging, taking the upper layer solution, and passing the upper layer solution through a membrane for LC-MS detection. The recovery rate of the method is only 5-50%.
By comparing the comparative example 1 with the example 1, it can be found that the recovery rate of tryptophan and the metabolites thereof is higher and the quantitative accuracy is higher in the example 1.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. A method for identifying the quality of milk products is characterized by comprising the following steps:
step 1), extracting tryptophan metabolites from a milk sample to obtain an extract; the tryptophan metabolites are 5-hydroxytryptophan, indole-3-ethylamine, tryptamine, 3-hydroxy anthranilic acid, nicotinic acid, xanthurenic acid, kynurenine, kynurenic acid, indole formaldehyde and indole acetic acid;
step 2), performing chromatographic separation-mass spectrometry scanning on the extract in the step 1), performing quantitative analysis on tryptophan metabolites in the milk sample, and determining a quantitative limit and a recovery rate;
step 3), analyzing the milk sample according to the content data of the tryptophan metabolites in the milk sample in the step 2);
in the step 1), the milk samples comprise pasteurized milk, ultrahigh-temperature sterilized milk, reconstituted milk samples and yogurt samples;
in the step 1), adding acetonitrile formate into a milk sample, wherein the acetonitrile formate is a mixed solution of formic acid and acetonitrile, and the volume ratio of the formic acid to the acetonitrile is 3-8: 97-92;
in the step 1), adding acetonitrile formate into a milk sample, and vortexing; adding an extraction salt bag, performing vortex and centrifugation, transferring the upper layer solution, adding a purification reagent, performing vortex and centrifugation, taking the upper layer solution, and drying by using nitrogen; adding the complex solution to carry out water bath ultrasound, carrying out vortex mixing, dissolving again, centrifuging, taking the upper layer solution, and passing through a membrane for the next detection; the extraction salt bag is sodium sulfate and sodium chloride; the purifying reagent is sodium sulfate and C18;
in the step 2), the chromatographic separation conditions are as follows: the adopted mobile phase A is aqueous solution containing 0.1 percent of formic acid, the mobile phase B is methanol solution containing 0.1 percent of formic acid, and gradient elution is adopted;
in step 2), the elution gradient is:
in the step 2), the conditions of mass spectrum are as follows: multiple reaction detection (MRM) positive ion mode, air curtain gas (CUR)35psi, jet collision gas (CAD)10psi, ionization voltage (IS)5500V, Temperature (TEM)500 ℃; nebulizer (GS1)50 psi; supplemental heater (GS2)50 psi.
2. The method of claim 1, wherein the reconstituted milk sample is whole milk powder formulated to have a protein content of 3.0g/100 mL; and/or performing subsequent treatment after the yoghurt sample is diluted by 1: 1.
3. The method according to claim 1, characterized in that 5-10 portions of milk sample are taken, each portion of milk sample having a volume of 5-10 ml; the volume ratio of the milk sample, the acetonitrile formate and the complex solution is (5-10): 5-15): 0.5-3.
4. The method according to claim 3, wherein the volume ratio of the milk sample, the acetonitrile formate and the complex solution is 5:10: 1.
5. The method of claim 1, wherein the extraction salt package is 2-8 g sodium sulfate and 0.5-3 g sodium chloride.
6. The method of claim 5, wherein the purifying agent is 200-1500 mg sodium sulfate and 50-300 mg C18.
7. The method of claim 6, wherein the extraction salt package is 4g sodium sulfate and 1g sodium chloride; the detergent reagent was 900mg sodium sulfate and 150mg C18.
8. The method according to claim 5, wherein in the step 1), the acetonitrile formate is a mixed solution of formic acid and acetonitrile, and the volume ratio of the formic acid to the acetonitrile is 5: 95; and/or the double solution is 10% acetonitrile in water; and/or centrifuging for 8-10 min at 8000-10000 rpm; and/or the water bath ultrasound condition is that the water bath ultrasound is carried out for 8-10 min at the temperature of 20-30 ℃.
9. The method of claim 1, wherein in step 2), the chromatographic separation-mass spectrometry scan comprises a primary scan and a secondary ion fragmentation scan.
10. The method according to any one of claims 1 to 9, wherein in step 3), the content data of each metabolite in the milk sample is subjected to analysis of variance and partial least squares discriminant analysis, so as to determine the compounds with different contents in different milk samples.
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| CN111912915A (en) * | 2020-05-26 | 2020-11-10 | 中国农业大学 | Sample rapid pretreatment method for determining melatonin in milk, serum and products thereof |
| CN112986431B (en) * | 2021-02-18 | 2022-09-16 | 中国农业科学院农业质量标准与检测技术研究所 | Method for identifying organic milk and conventional milk based on metabonomics |
| CN113960185A (en) * | 2021-02-18 | 2022-01-21 | 四川省人民医院 | Method for detecting tryptophan and metabolite thereof |
| CN114137097A (en) * | 2021-09-16 | 2022-03-04 | 中国农业大学 | Method for detecting melatonin in milk by liquid chromatography-tandem mass spectrometry and performance evaluation thereof |
| CN114137105B (en) * | 2021-11-15 | 2024-01-19 | 北京奶牛中心 | Method for analyzing melatonin content in sample by liquid chromatography-tandem mass spectrometry |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102440989A (en) * | 2010-10-13 | 2012-05-09 | 四川科伦药物研究有限公司 | Compound amino acid injecta, and preparation method and detection method thereof |
| CN103149317A (en) * | 2012-12-21 | 2013-06-12 | 江南大学 | Method for promptly detecting biogenic amine in food |
| CN105606750A (en) * | 2016-01-06 | 2016-05-25 | 大连海事大学 | Aquatic product producing area tracing method based on fatty acid carbon stable isotopes |
| CN109187826A (en) * | 2018-08-06 | 2019-01-11 | 安徽中医药大学 | A kind of detection method of the outer component of candida albicans biofilm extracellular matrix protein matter |
| CN109738551A (en) * | 2019-02-27 | 2019-05-10 | 北京市营养源研究所 | A kind of method of imidaclothiz content in detection livestock meat |
| CN110018266A (en) * | 2019-02-15 | 2019-07-16 | 广州市妇女儿童医疗中心 | A kind of method of 48 kinds of amino acid of fast quantitative analysis |
| CN110470781A (en) * | 2019-09-05 | 2019-11-19 | 中国农业科学院农业质量标准与检测技术研究所 | Identify the method for reconstituted milk and ultra-high-temperature sterilized milk |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120238030A1 (en) * | 2011-02-07 | 2012-09-20 | Laboratory Corporation Of America Holdings | Methods and Systems for Multiplex Analysis of Biomolecules by Liquir Chromatography-Mass Spectrometry |
| CN109324132A (en) * | 2018-10-31 | 2019-02-12 | 杭州佰辰医学检验所有限公司 | Kit and its application of high throughput detection tryptophan and its metabolite |
-
2019
- 2019-12-30 CN CN201911397797.2A patent/CN111077262B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102440989A (en) * | 2010-10-13 | 2012-05-09 | 四川科伦药物研究有限公司 | Compound amino acid injecta, and preparation method and detection method thereof |
| CN103149317A (en) * | 2012-12-21 | 2013-06-12 | 江南大学 | Method for promptly detecting biogenic amine in food |
| CN105606750A (en) * | 2016-01-06 | 2016-05-25 | 大连海事大学 | Aquatic product producing area tracing method based on fatty acid carbon stable isotopes |
| CN109187826A (en) * | 2018-08-06 | 2019-01-11 | 安徽中医药大学 | A kind of detection method of the outer component of candida albicans biofilm extracellular matrix protein matter |
| CN110018266A (en) * | 2019-02-15 | 2019-07-16 | 广州市妇女儿童医疗中心 | A kind of method of 48 kinds of amino acid of fast quantitative analysis |
| CN109738551A (en) * | 2019-02-27 | 2019-05-10 | 北京市营养源研究所 | A kind of method of imidaclothiz content in detection livestock meat |
| CN110470781A (en) * | 2019-09-05 | 2019-11-19 | 中国农业科学院农业质量标准与检测技术研究所 | Identify the method for reconstituted milk and ultra-high-temperature sterilized milk |
Non-Patent Citations (3)
| Title |
|---|
| Evolution of tryptophan and its foremost metabolites’ concentrations in milk and fermented dairy products;Antonella Bertazzo 等;《PharmaNutrition》;20161231;第1-8页 * |
| QuEChERS-HPLC-MS/MS 法检测液态乳中9种β2-受体激动剂残留量;李婧妍 等;《中国乳品工业》;20161231;第44卷(第11期);第49-52页 * |
| 高效液相色谱-串联质谱法测定人血浆中色氨酸及其代谢物浓度;刘仪滨 等;《实用药物与临床》;20190731;第22卷(第7期);第742-749页 * |
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