Method for efficiently and rapidly analyzing characteristic volatile components of eucommia leaves and fermentation product thereof
阅读说明：本技术 一种高效快速分析杜仲叶及其发酵产物特征挥发性成分的方法 (Method for efficiently and rapidly analyzing characteristic volatile components of eucommia leaves and fermentation product thereof ) 是由 王志宏 彭密军 杨秋玲 张命龙 于 2019-11-05 设计创作，主要内容包括：本发明公开了一种高效快速分析杜仲叶及其发酵产物特征挥发性成分的方法。通过将杜仲叶及其发酵产物不同样品进行气相-离子迁移谱的分析检测,根据迁移速率、保留时间和相对离子峰强度进行三维定性分析,绘制相应的谱图,建立指纹图谱,能够直观比较杜仲叶以及发酵产物不同样品之间的差异,同时根据不同样品的特征信息,选取谱图中特征区域,定性识别特征挥发性成分,进一步利用数据预处理方法和化学计量学手段,实现样品的快速分类。本方法无需对样品进行复杂的前处理过程,操作简单,特征区域完全可视化,在准确高效识别杜仲叶及其发酵产物中的特征挥发性成分等方面具有广阔的应用前景,对于杜仲资源的高值化利用与可持续开发具有重要的意义。(The invention discloses a method for efficiently and quickly analyzing characteristic volatile components of eucommia ulmoides leaves and fermentation products of the eucommia ulmoides leaves. The method comprises the steps of carrying out analysis and detection on the eucommia ulmoides leaves and different samples of fermentation products of the eucommia ulmoides leaves by using a gas phase-ion mobility spectrometry, carrying out three-dimensional qualitative analysis according to migration rate, retention time and relative ion peak intensity, drawing corresponding spectrograms, establishing fingerprint spectrograms, visually comparing differences between the eucommia ulmoides leaves and different samples of the fermentation products, selecting characteristic areas in the spectrograms according to characteristic information of different samples, qualitatively identifying characteristic volatile components, and further realizing rapid classification of the samples by using a data preprocessing method and a chemometric means. The method does not need to carry out a complex pretreatment process on the sample, is simple to operate, has complete visualization of characteristic regions, has wide application prospects in the aspects of accurately and efficiently identifying characteristic volatile components in the eucommia leaves and fermentation products thereof and the like, and has important significance for high-valued utilization and sustainable development of eucommia resources.)
1. A method for efficiently and rapidly analyzing characteristic volatile components of folium cortex eucommiae and fermentation products thereof is characterized by comprising the following steps:
(1) respectively preparing samples to be detected of eucommia ulmoides leaves and fermentation products of the eucommia ulmoides leaves;
(2) performing headspace-gas chromatography-ion mobility spectrometry on a sample to be detected, collecting data of migration rate, retention time and relative ion peak intensity, and establishing a fingerprint according to the migration rate, the retention time and the relative ion peak intensity;
(3) extracting characteristic areas in fingerprint spectra of different samples by a comparison method, and performing qualitative analysis on characteristic volatile components of the different samples by adopting gas phase ion mobility chromatograph database software;
(4) and establishing a classification model by adopting a chemometrics method, and performing orthogonal transformation and dimension reduction on relative intensity data of volatile component ion peak signals corresponding to all characteristic regions of different samples to realize rapid classification of different samples.
2. The method for efficiently and rapidly analyzing the characteristic volatile components of the eucommia ulmoides leaves and the fermentation products thereof according to claim 1, wherein the sample to be tested of the eucommia ulmoides leaves is a sample obtained by cutting fresh eucommia ulmoides leaves into a proper size or a powder sample obtained by drying, crushing and sieving fresh eucommia ulmoides leaves; the sample to be detected of the eucommia ulmoides leaf fermentation product is a fermentation product obtained by fermenting eucommia ulmoides leaf powder and edible fungi or microbial strains together, or a powder sample obtained by drying, crushing and sieving the fermentation product.
3. The method for efficiently and rapidly analyzing characteristic volatile components of folium cortex eucommiae and fermentation products thereof according to claim 1, wherein the conditions for performing headspace-gas chromatography-ion mobility spectrometry on the sample to be tested in the step (2) are as follows:
the headspace sample injection amount is 1.0-5.0 g, the headspace heating temperature is 70-95 ℃, and the equilibration time is 10-30 min;
gas ion chromatograph G.A.S. Co
4. The method for efficiently and rapidly analyzing characteristic volatile components of folium cortex eucommiae and fermentation products thereof as claimed in claim 1, wherein the characteristic regions in the fingerprint spectra of different samples are extracted by a comparison method in step (3), specifically, the spectrum characteristic regions with obvious corresponding color change or large relative intensity difference of ion peak signals in the volatile component spectra of different samples are analyzed according to the color difference change or the relative intensity of ion peak signals of the characteristic volatile components of different samples in the fingerprint spectra.
5. The method for efficiently and rapidly analyzing characteristic volatile components of eucommia ulmoides leaves and fermentation products thereof according to claim 1, wherein in the step (3), qualitative analysis is performed on the characteristic volatile components of different samples by using gas phase ion mobility chromatograph database software, and specifically, qualitative analysis is performed on the characteristic volatile compounds of different samples by performing two-dimensional qualitative analysis according to a NIST 2014 gas phase retention index database built in GC x IMS Library Search software and an IMS mobility time database of G.A.S.
6. The method for efficiently and rapidly analyzing characteristic volatile components of eucommia ulmoides leaves and fermentation products thereof according to claim 1, wherein the step (4) of establishing the classification model by using a chemometric method is to establish the classification model by using a principal component analysis method, the migration rate, retention time and relative ion peak intensity data of a fingerprint spectrogram need to be preprocessed before establishing the classification model, the data are introduced into LAV software, data filtering is carried out by using a standard deviation option, then normalization is carried out, median normalization is selected by normalization, logarithmic data conversion is adopted, and finally, the classification model is established by using the principal component analysis method after full-distance scaling treatment is adopted.
Technical Field
The invention belongs to the field of rapid analysis and detection, and particularly relates to a method for efficiently and rapidly analyzing characteristic volatile components of folium cortex eucommiae and fermentation products of the folium cortex eucommiae.
Background
Eucommia ulmoides (Eucommia ulmoides Oliv.) is a traditional nourishing medicinal material in China. The eucommia ulmoides leaves contain a plurality of natural active ingredients, including iridoid, lignanoid, flavonoid and other compounds, and have good effects on regulating blood pressure, reducing blood sugar, reducing blood fat, enhancing immunity, resisting oxidation, inhibiting bacteria, resisting aging and the like. The eucommia ulmoides leaves are very rich in resources in China, and are collected in Chinese pharmacopoeia from 2005 in 4 months in 2018, and the national Weijian committee plans to manage the eucommia ulmoides leaves as materials of food and traditional Chinese medicinal materials, so the eucommia ulmoides leaves have wide application prospects in the industries of food, medicines and cosmetics. At present, the chemical composition, the biological activity and the like of the eucommia ulmoides leaves are always the hot points of attention of people. However, there are few reports on volatile components of eucommia ulmoides leaves, including the composition of volatile components in products after biological fermentation. The composition of the volatile components often influences the characteristics of corresponding products such as sense, flavor and the like, and can be used as an important reference index for product classification. Therefore, the composition and relative content of characteristic volatile components in the eucommia ulmoides leaves and fermentation products thereof have important research values.
The ion mobility spectrometry technology is a rapid separation detection technology. The working principle is that under the atmospheric pressure condition, volatile or semi-volatile substances move in an external electric field after being ionized, and have different moving speeds according to the differences of different ions in the aspects of mass, charge number, collision cross section, molecular space structure and the like, so that the time required by the ions to reach an ion detector is different, and the effective separation of different volatile components is finally realized. The ion mobility spectrometry has the advantages of high analysis speed, high sensitivity, simple operation, portability, high-throughput analysis and the like, and is widely applied to the field rapid detection of drugs and dangerous substances. In recent years, with the continuous development of the technology, the gas-phase ion mobility spectrometry technology has been applied to the aspects of rapid screening of environmental pollutants, nondestructive inspection of freshness of meat products, identification of adulteration of edible oil and the like.
At present, few reports are made on the research of volatile components in the eucommia ulmoides leaves and fermentation products thereof by utilizing gas phase ion mobility spectrometry. Therefore, a method suitable for analyzing volatile components in folium cortex eucommiae and fermentation products thereof can be established by optimizing analysis and detection conditions, so that a characteristic component fingerprint is rapidly established, the difference of the composition and relative content of the volatile components among different samples is analyzed, and meanwhile, the characteristic compounds can be qualitatively analyzed according to a related database; in addition, the rapid classification of different eucommia leaf samples can be realized by combining a chemometrics method. The method has important reference value for high-value utilization and comprehensive development of eucommia ulmoides leaf resources.
Disclosure of Invention
The invention aims to provide a method for efficiently and quickly analyzing characteristic volatile components of eucommia ulmoides leaves and fermentation products of the eucommia ulmoides leaves.
The technical scheme adopted by the invention is as follows:
a method for efficiently and rapidly analyzing characteristic volatile components of folium cortex eucommiae and fermentation products thereof comprises the following steps:
(1) respectively preparing samples to be detected of eucommia ulmoides leaves and fermentation products of the eucommia ulmoides leaves;
(2) performing headspace-gas chromatography-ion mobility spectrometry on a sample to be detected, collecting data of migration rate, retention time and relative ion peak intensity, and establishing a fingerprint according to the migration rate, the retention time and the relative ion peak intensity;
(3) extracting characteristic areas in fingerprint spectra of different samples by a comparison method, and performing qualitative analysis on characteristic volatile components of the different samples by adopting gas phase ion mobility chromatograph database software;
(4) and establishing a classification model by adopting a chemometrics method, and performing orthogonal transformation and dimension reduction on relative intensity data of volatile component ion peak signals corresponding to all characteristic regions of different samples to realize rapid classification of different samples.
Preferably, the sample to be detected of the eucommia ulmoides leaves is a sample obtained by cutting fresh eucommia ulmoides leaves into pieces with proper sizes, or a powder sample obtained by drying, crushing and sieving fresh eucommia ulmoides leaves; the sample to be detected of the eucommia ulmoides leaf fermentation product is a fermentation product obtained by fermenting eucommia ulmoides leaf powder and edible fungi (such as lucid ganoderma, grifola frondosa and hericium erinaceus) or microbial strains together, or a powder sample obtained by drying, crushing and sieving the fermentation product.
Preferably, the conditions for performing headspace-gas chromatography-ion mobility spectrometry on the sample to be detected in the step (2) are as follows:
the headspace sample injection amount is 1.0-5.0 g, the headspace sample injection bottle volume is 20-40mL, the headspace heating temperature is 70-95 ℃, and the equilibration time is 10-30 min;
gas ion chromatograph G.A.S. Co
Preferably, in the step (3), the characteristic regions in the fingerprint spectra of different samples are extracted by a comparison method, and specifically, the spectrum characteristic regions with obvious corresponding color change or large relative intensity difference of ion peak signals in the spectrum of the volatile components in different samples are analyzed according to the change of the color difference of the characteristic volatile components of different samples in the fingerprint spectra or the relative intensity of the ion peak signals.
Preferably, in the step (3), qualitative analysis is performed on characteristic volatile components of different samples by using gas ion mobility chromatograph database software, specifically, two-dimensional qualitative analysis is performed on characteristic volatile compounds of different samples according to a NIST 2014 gas retention index database built in GC × IMS Library Search software and an IMS mobility time database of g.a.s.
Preferably, the step (4) of establishing the classification model by using a chemometrics method is to establish the classification model by using a principal component analysis method, wherein the migration rate, retention time and relative ion peak intensity data of a fingerprint spectrogram need to be preprocessed before establishing the classification model, the data is imported into LAV software, data filtering is performed by using a standard deviation option, then normalization is performed, median normalization is selected for normalization, logarithm data conversion is performed, and finally, after full-distance scaling processing is performed, the principal component analysis method is used for establishing the classification model.
The invention has the beneficial effects that: the invention utilizes gas phase ion mobility spectrometry technology to detect and analyze volatile components in eucommia leaves and fermentation products thereof. The result shows that the method can rapidly analyze the volatile components in the eucommia ulmoides leaves and the fermentation products thereof by the gas phase ion mobility spectrometry technology, can realize the accurate determination of the characteristic volatile components by the GC separation and the IMS secondary separation, and is an effective method for accurately, efficiently and easily detecting the volatile components in the eucommia ulmoides leaves and the fermentation products thereof. The method provided by the invention has the advantages of high accuracy, rapidness, high efficiency and easiness in operation, can realize high-throughput analysis on the sample, and greatly improves the efficiency of sample identification and analysis.
The innovations of the invention comprise: (1) the gas-phase ion mobility spectrometry technology is applied to the detection of volatile components in the eucommia leaves and fermentation products thereof for the first time, and a new method is provided for quickly identifying the characteristic volatile components in the eucommia leaves; (2) the fingerprint of volatile components of the eucommia ulmoides leaves and fermentation products thereof is quickly established for the first time through a gas phase ion mobility spectrometry technology, and different eucommia ulmoides leaf samples can be quickly classified through a chemometrics model, so that the method has an important reference value for high-value utilization of eucommia ulmoides resources.
The advantages of the invention include: (1) the sample does not need any pretreatment process, so that the interference of a solvent on a detection result is avoided; (2) the sample is heated and volatilized before analysis, the heating temperature is 70-95 ℃, and the time is 10-30min, so that volatile components in the sample can be ensured to volatilize as far as possible, and the problem that the structure of the volatile components is changed due to excessive pretreatment conditions to influence the analysis result can be avoided; (3) GC and IMS technologies are combined, three-dimensional qualitative analysis is carried out according to retention indexes, migration rates and ion peak relative intensities, then a corresponding spectrogram is drawn, analysis results are efficient and accurate, and the analysis results can be visually presented; (4) and the rapid classification of the samples can be realized according to a chemometrics method by combining the fingerprint spectrum.
Drawings
FIG. 1 is a comparison graph of the differences of characteristic volatile components of eucommia ulmoides leaves and different fermentation products thereof.
FIG. 2 is a peak diagram of characteristic volatile component signals of folium Eucommiae.
FIG. 3 is a diagram showing the results of principal component analysis of different samples.
Detailed Description
The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
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