CN113176227A - Method for rapidly predicting adulteration of dendrobium huoshanense in dendrobium hunan - Google Patents
Method for rapidly predicting adulteration of dendrobium huoshanense in dendrobium hunan Download PDFInfo
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- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
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- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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Abstract
The invention discloses a method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense, which comprises the following steps: (1) preparing a pseudo-dendrobium huoshanense powder sample; (2) collecting infrared spectrum; (3) preprocessing a spectrum; (4) screening spectral characteristic wavelengths; (5) and (5) predicting the adulteration amount. According to the invention, by adopting the same sample for multiple times of collection and spectrum pretreatment, the manual sampling error and the influence of the scanning times, resolution, temperature and optical path on the mid-infrared spectrogram can be effectively reduced; the characteristic wavelength variable is screened by three different wavelength selection methods, namely a moving window partial least square method, a Monte Carlo non-information variable elimination method and an interval random frog method, so that useless variables and interference information in the intermediate infrared spectrogram can be effectively compressed; meanwhile, a spectrum pretreatment method and a wavelength screening method which are most suitable for the prediction model are obtained, the accurate and reliable adulteration prediction model is established, and the rapid determination of the adulteration amount in the adulteration dendrobium huoshanense sample is realized.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine identification, in particular to a method for quickly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense.
Background
Dendrobium huoshanense (Dendrobium huoshanense Tang et Cheng) belongs to the genus Dendrobium of Orchidaceae, is mainly produced in Anhui Huoshan, Yuexi, Xishan, etc., and contains polysaccharide, alkaloid and abundant microelements. Has multiple efficacies of promoting the production of body fluid, benefiting the stomach, resisting tumors, resisting cataract and the like, and is one of important genuine medicinal materials in Anhui. Due to the low germination rate and slow growth of dendrobium huoshanense in a natural state and the long-term excessive mining, wild resources are endangered to be extinct, the market price is as high as 30-40 ten thousand yuan/kg, and the dendrobium huoshanense is valuable and marketable. At present, most of dendrobium huoshanense sold in the market is cultivated products of tissue culture dendrobium huoshanense through field domestication, the market price still reaches 10-15 ten thousand yuan/kg, and more of dendrobium huoshanense counterfeits in the market. The dendrobium hunan is the most common counterfeit of dendrobium huoshanense, and is often blended in the dendrobium huoshanense to be impersonated as the dendrobium huoshanense for sale. Because stem and its powdery processing product mix the false differentiation more difficult from the outward appearance hardly discerning in dendrobium huoshanense and Henan dendrobe, only hardly differentiatees whether mixing the false from the outward appearance form, both properties of a drug efficiency are very far away simultaneously, this kind mixes the quality that the pseudo-mode directly influences the medicinal material, seriously threatens the clinical medication safety of dendrobium huoshanense, has disturbed dendrobium huoshanense medicinal material market, and is very big to the long-term development harm of dendrobium huoshanense industry. Therefore, a detection method for adulterating dendrobium huoshanense to dendrobium heonanthum is urgently needed to be established.
The current common methods for identifying the authenticity of the traditional Chinese medicine comprise character identification, microscopic identification, chromatographic identification, molecular biological technology identification and the like. However, the trait and microscopic identification depend on the experience of the practitioner, and are highly subjective. Patent CN105779628A discloses an SNP marker for identifying dendrobium huoshanense and a molecular detection method thereof. Patent CN110951912A discloses a method for identifying dendrobium huoshanense based on DNA barcode; patent CN106282372A discloses a method for identifying dendrobium huoshanense and dendrobium huoshanense by comparison; patent CN106282373A discloses a method for comparing and identifying dendrobium huoshanense and dendrobium henryi. The technology adopts a molecular biology technology to identify dendrobium huoshanense and a relative species dendrobium huoshanense, but identification methods such as molecular and chromatographic methods are complex to operate, time and labor are wasted, and although the true and false of medicinal materials can be identified, the adulteration amount of a adulteration sample is difficult to detect.
The mid-infrared spectrum technology has the advantages of high analysis speed, low cost, simple sample pretreatment and the like, and is applied to analysis of various samples such as modern medicines, foods, agricultural products and the like. However, in the aspect of adulteration quantitative research, the mid-infrared spectrum mainly researches liquid samples such as grease, honey, milk and the like, solid samples mainly focus on quantitative analysis research of chemical additives, and adulteration quantitative detection of two plant solid samples is more complex and has fewer related researches. Although the intermediate infrared spectrum has the problems of short wavelength range, serious spectrum overlapping, high background noise, more irrelevant waveband interference information and the like in detection, the effective wavelength screening can improve the performance of the intermediate infrared spectrum for constructing a prediction model. Common wavelength screening methods are mainly applied to quantitative analysis of near infrared spectra, researches on adulteration quantitative analysis of solid plant samples by combining the intermediate infrared spectra with the wavelength screening methods are only reported, and in adulteration researches of traditional Chinese medicine dendrobium huoshanense, the researches on the quantitative analysis of the adulteration dendrobium huoshanense are not found yet.
Disclosure of Invention
Aiming at the problems, the invention provides a rapid, convenient, accurate and reliable method for measuring the adulteration amount of the adulteration dendrobium sample by combining the mid-infrared spectrum technology with the chemometrics method.
The invention solves the technical problems through the following technical means:
a method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense comprises the following steps:
(1) preparation of pseudo-dendrobium huoshanense powder sample
Picking up collected fresh dendrobium huoshanense stems and fresh dendrobium huoshanense stems, cleaning, removing impurities, cutting, freeze-drying, crushing and sieving, collecting dendrobium huoshanense stem powder and dendrobium huoshanense stem powder with the particle size of 60-100 meshes, mixing the dendrobium huoshanense stem powder and the dendrobium huoshanense stem powder according to different proportions according to a mass ratio to obtain 21 groups of adulterated samples, setting 10 parallel samples for each group of adulterated samples, and sealing and storing for later use, wherein the mass proportion range of the dendrobium huoshanense stem powder and the dendrobium huoshanense stem powder is 0-100%;
(2) infrared spectroscopy collection
Collecting the mid-infrared spectrogram of each adulteration sample by using 210 adulteration samples prepared in the step (1) through a Fourier transform attenuated total reflection mid-infrared spectrometer (Nicolet iS50ATR-FTIR infrared spectrometer), and collecting each sample for multiple times to obtain an original spectrum for calculating an average spectrogram;
(3) spectral preprocessing
Performing baseline correction and average spectrogram calculation on the intermediate infrared spectrogram of the adulterated sample to eliminate spectrogram difference caused by artificial sampling error, then moving the average spectrum by 15 data points, adopting a spectrum smoothing method to eliminate spectral noise caused by detection environment, and comparing R of a least square method (PLS) cross validation model2Comparing R of cross-validation model with R of root mean square error RMSECV value by 8 spectrum preprocessing methods2Screening the value and the root mean square error value RMSECV by a spectrum pretreatment method;
(4) screening of spectral characteristic wavelengths
Three different wavelength selection methods, namely a moving window partial least square method, a Monte Carlo non-information variable elimination method and an interval random frog method, are compared to obtain effective wavelengths for establishing a prediction model so as to improve the calculation workload and accuracy of the model;
(5) adulteration prediction
Predicting the adulteration amount of the adulteration sample by using a adulteration quantitative model; and establishing 105 adulteration dendrobium samples as a correction set by a partial least square method, establishing a adulteration quantitative model between the characteristic wavelength variable and the adulteration amount of the spectrogram, and verifying the adulteration amount of the adulteration samples of the 105 prediction sets by using the adulteration quantitative model.
The method for determining the adulteration amount of the adulteration dendrobium huoshanense sample is established by combining the mid-infrared spectrum technology, and the influence of manual sampling errors and scanning times, resolution, temperature and optical path on a mid-infrared spectrogram can be effectively reduced by adopting the method for collecting the same sample for multiple times and preprocessing the spectrum; the characteristic wavelength variable is screened by three different wavelength selection methods, namely a moving window partial least square method, a Monte Carlo non-information variable elimination method and an interval random frog method, so that useless variables and interference information in the intermediate infrared spectrogram can be effectively compressed; meanwhile, a spectrum pretreatment method and a wavelength screening method which are most suitable for the prediction model are obtained, the accurate and reliable adulteration prediction model is established, and the rapid determination of the adulteration amount in the adulteration dendrobium huoshanense sample is realized.
Preferably, in the 21 adulteration samples in the step (1), the mass ratio of the dendrobium huinan stem powder to the dendrobium huoshanense stem powder is respectively as follows: 0%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%.
Preferably, the total mass of each adulteration sample in the step (1) is controlled to be 500 mg.
Preferably, the sample in the step (1) is sealed and stored at-20 ℃ for later use, so as to ensure the stability of the dendrobium.
Preferably, the infrared spectrometer in step (2) is equipped with an AIR accessory.
Preferably, the parameters related to the spectrum acquisition of the infrared spectrometer in the step (2) are set as follows: the measurement range is 4000--1The number of scanning times is 32, and the resolution is 4.0cm-1And 7467 data points are collected, and the same sample is repeatedly collected for 6 times to obtain a mid-infrared original spectrogram.
Preferably, in the step (2), the baseline correction and the average spectrum calculation are performed on the raw spectra of the mid-infrared spectrum acquired 6 times from the same sample, so as to obtain the average spectrum, so as to eliminate the spectrum difference caused by the manual sampling error.
Preferably, in the step (3), the average spectrum is shifted by 15 data points, and a spectrum smoothing method is adopted to reduce the spectrum noise caused by the detection environment.
Preferably, the 8 spectrum preprocessing methods in the step (3) are a multivariate scattering correction method, a standard normal variation method, a first derivative method, a second derivative method, a multivariate scattering correction plus first derivative method, a multivariate scattering correction plus second derivative method, a standard normal variation plus first derivative method and a standard normal variation plus second derivative method.
Preferably, the adulteration quantitative model is used for predicting the adulteration amount of the adulteration dendrobium sample in the step (5), specifically, the adulteration quantitative model between characteristic variables of the spectrogram of the total sample number correction set adulteration dendrobium sample and the adulteration amount is established 1/2 by a partial least square method, and the adulteration quantitative model is used for verifying the adulteration amount of the adulteration dendrobium sample in the 1/2 total sample number prediction set.
The invention has the following beneficial effects: the method for determining the adulteration amount of the adulteration dendrobium huoshanense sample is established by combining the mid-infrared spectrum technology, and the influence of manual sampling errors and scanning times, resolution, temperature and optical path on a mid-infrared spectrogram can be effectively reduced by adopting the method for collecting the same sample for multiple times and preprocessing the spectrum; the characteristic wavelength variable is screened by three different wavelength selection methods, namely a moving window partial least square method, a Monte Carlo non-information variable elimination method and an interval random frog method, so that useless variables and interference information in the intermediate infrared spectrogram can be effectively compressed; meanwhile, a spectrum pretreatment method and a wavelength screening method which are most suitable for the prediction model are obtained, the accurate and reliable adulteration prediction model is established, and the rapid determination of the adulteration amount in the adulteration dendrobium huoshanense sample is realized.
Drawings
FIG. 1 is a flow chart of a method for rapidly predicting the content of Dendrobium huoshanense doped in Dendrobium hanense of the invention;
FIG. 2 is an original mid-infrared spectrum of several samples of pseudo-dendrobium huoshanense of example 3 of the present invention;
FIG. 3 is a mid-infrared spectrum pretreatment chart of several adulterated dendrobium huoshanense samples and the positions of the three wavelength selection methods in the embodiment 3 of the invention;
fig. 4 is an effect diagram of the adulteration amount detection model of the adulteration dendrobium huoshanense sample in embodiment 3 of the invention.
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 will be clearly and completely described below with reference to the embodiments of the present invention and the drawings in the specification, and it is obvious that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Test materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The specific techniques or conditions not specified in the examples can be performed according to the techniques or conditions described in the literature in the field or according to the product specification.
Example 1
A method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense comprises the following steps:
(1) preparation of pseudo-blended dendrobium huoshanense sample
Preparing a pseudo-dendrobium huoshanense sample: picking up the collected fresh dendrobium huoshanense stems and dendrobium huoshanense stems, cleaning with purified water to remove surface impurities, then, respectively cutting dendrobium huoshanense stems and dendrobium heonantha stems into small segments, freeze-drying the small segments in a low-temperature freeze dryer, crushing the small segments by using a crusher, sieving the crushed small segments by using a 60-mesh sieve to obtain dendrobium huoshanense stem powder and dendrobium heonantha stem powder with the particle size of 60 meshes, respectively mixing the dendrobium heonantha stem powder and the dendrobium heonantha stem powder according to the mass ratio of 0%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% and 100% to obtain 21 groups of adulteration samples, setting 10 parallel samples in each group of adulteration samples, and setting 210 parts of adulteration samples in total, and the quality of each adulteration sample is controlled to be 500mg, and finally 210 adulteration samples are sealed and stored at the temperature of minus 20 ℃ for standby;
(2) collection of original spectrum of pseudo-dendrobium huoshanense sample
And (3) collecting the mid-infrared spectrogram of the adulterated dendrobium huoshanense sample from a plurality of adulterated dendrobium samples obtained in the subsidy (1) by using a mid-infrared spectrometer (Nicolet iS50ATR-FTIR infrared spectrometer).
The procedure for spectrum acquisition was as follows: placing the sample mixed with the dendrobium huoshanense powder into a sample cell of a diamond single reflection AIR accessory equipped in an FTIR spectrometer, and setting the measurement range of the mid-infrared spectrometer to be 4000-400cm-1Setting the scanning times of the mid-infrared spectrometer to be 32 times and the resolution to be 4.0cm-17467 data points are collected, and the same sample is repeatedly collected for 6 times to obtain a mid-infrared original spectrogram;
(3) spectral preprocessing
Performing baseline correction and average spectrogram calculation on the original spectrogram of the intermediate infrared spectrum acquired 6 times by the same pseudo-dendrobium huoshanense sample to obtain an average spectrum, eliminating the spectrogram caused by artificial sampling error, moving the average spectrum by 15 points averagely, and eliminating the spectrum noise caused by the detection environment by adopting a smoothing method;
8 spectral pretreatment methods were applied: the method comprises the following steps of screening a pretreatment method by combining a multivariate scattering correction method, a standard normal variation method, a first derivative method, a second derivative method, a multivariate scattering correction plus first derivative method, a multivariate scattering correction plus second derivative method, a standard normal variation plus first derivative method, a standard normal variation plus second derivative method and a partial least squares algorithm (PLS); by comparing R of PLS cross validation models2Value and root mean square error value RMSECV, to obtain a higher R obtained by using a spectral preprocessing method of standard normal variation plus first derivative2Obtaining a mid-infrared spectrogram of the adulterated dendrobium huoshanense sample after the spectral pretreatment of standard normal change plus first-order derivative;
(4) screen for spectral characteristic variable
Three different wavelength selection methods, namely a moving window partial least square method, a Monte Carlo non-information variable elimination method and an interval random frog method, are compared; the three wavelength variable screening methods are combined to establish the effective wavelength of the PLS model so as to improve the calculation workload and accuracy of the model.
(5) Adulteration prediction
Predicting the adulteration amount of the adulteration dendrobium huoshanense sample by adopting a adulteration quantitative model according to the characteristic wavelength variable of the spectrogram of the adulteration dendrobium huoshanense sample; and establishing a adulteration quantitative model between the characteristic wavelength variable and the adulteration amount of the spectrogram of 105 correction set adulterated dendrobium huoshanense samples by a partial least square method, and verifying the adulteration amount of the 105 prediction set adulteration dendrobium huoshanense samples by using the adulteration quantitative model.
Example 2
Preparing a pseudo-dendrobium huoshanense sample: picking up the collected fresh dendrobium huoshanense stems and dendrobium huoshanense stems, cleaning with purified water to remove surface impurities, then, respectively cutting dendrobium huoshanense stems and dendrobium heonantha stems into small segments, freeze-drying the small segments in a low-temperature freeze dryer, crushing the small segments by using a crusher, sieving the small segments by using an 80-mesh sieve to obtain dendrobium huoshanense stem powder and dendrobium heonantha stem powder with the particle size of 80 meshes, respectively mixing the dendrobium heonantha stem powder and the dendrobium heonantha stem powder according to the mass ratio of 0%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% and 100% to obtain 21 groups of adulteration samples, setting 10 parallel samples in each group of adulteration samples, and setting 210 parts of adulteration samples in total, and the quality of each adulteration sample is controlled to be 500mg, and finally 210 adulteration samples are sealed and stored at the temperature of minus 20 ℃ for standby;
(2) collection of original spectrum of pseudo-dendrobium huoshanense sample
And (3) collecting the mid-infrared spectrogram of the adulterated dendrobium huoshanense sample from a plurality of adulterated dendrobium samples obtained in the subsidy (1) by using a mid-infrared spectrometer (Nicolet iS50ATR-FTIR infrared spectrometer).
The procedure for spectrum acquisition was as follows: placing a sample doped with pseudo dendrobium huoshanense powderSetting the measuring range of the mid-infrared spectrometer 4000- & 400cm in a sample cell of a diamond single reflection AIR accessory equipped with an FTIR spectrometer-1Setting the scanning times of the mid-infrared spectrometer to be 32 times and the resolution to be 4.0cm-17467 data points are collected, and the same sample is repeatedly collected for 6 times to obtain a mid-infrared original spectrogram;
(3) spectral preprocessing
Performing baseline correction and average spectrogram calculation on the original spectrogram of the intermediate infrared spectrum acquired 6 times by the same pseudo-dendrobium huoshanense sample to obtain an average spectrum, eliminating the spectrogram caused by artificial sampling error, moving the average spectrum by 15 points averagely, and eliminating the spectrum noise caused by the detection environment by adopting a smoothing method;
8 spectral pretreatment methods were applied: the method comprises the following steps of screening a pretreatment method by combining a multivariate scattering correction method, a standard normal variation method, a first derivative method, a second derivative method, a multivariate scattering correction plus first derivative method, a multivariate scattering correction plus second derivative method, a standard normal variation plus first derivative method, a standard normal variation plus second derivative method and a partial least squares algorithm (PLS); by comparing R of PLS cross validation models2Value and root mean square error value RMSECV, to obtain a higher R obtained by using a spectral preprocessing method of standard normal variation plus first derivative2Obtaining a mid-infrared spectrogram of the adulterated dendrobium huoshanense sample after the spectral pretreatment of standard normal change plus first-order derivative;
(4) screen for spectral characteristic variable
Three different wavelength selection methods, namely a moving window partial least square method, a Monte Carlo non-information variable elimination method and an interval random frog method, are compared; the three wavelength variable screening methods are combined to establish the effective wavelength of the PLS model so as to improve the calculation workload and accuracy of the model.
(5) Adulteration prediction
Predicting the adulteration amount of the adulteration dendrobium huoshanense sample by adopting a adulteration quantitative model according to the characteristic wavelength variable of the spectrogram of the adulteration dendrobium huoshanense sample; and establishing a adulteration quantitative model between the characteristic wavelength variable and the adulteration amount of the spectrogram of 105 correction set adulterated dendrobium huoshanense samples by a partial least square method, and verifying the adulteration amount of the 105 prediction set adulteration dendrobium huoshanense samples by using the adulteration quantitative model.
Example 3
A method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense comprises the following steps:
(1) preparation of pseudo-blended dendrobium huoshanense sample
Picking up the collected fresh dendrobium huoshanense stems and dendrobium huoshanense stems, cleaning with purified water to remove surface impurities, then, respectively cutting dendrobium huoshanense stems and dendrobium heonantha stems into small segments, freeze-drying the small segments in a low-temperature freeze dryer, crushing the small segments by using a crusher, and then sieving the small segments by using a 100-mesh sieve to obtain dendrobium huoshanense stem powder and dendrobium heonantha stem powder with the particle size of 100 meshes, respectively mixing the dendrobium heonantha stem powder and the dendrobium heonantha stem powder according to the mass ratio of 0%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% and 100% to obtain 21 groups of adulteration samples, setting 10 parallel samples in each group of adulteration samples, and setting 210 parts of adulteration samples in total, and the quality of each adulteration sample is controlled to be 500mg, and finally 210 adulteration samples are sealed and stored at the temperature of minus 20 ℃ for standby;
(2) collection of original spectrum of pseudo-dendrobium huoshanense sample
And (3) collecting a plurality of adulterated dendrobium samples obtained in the last step by using a mid-infrared spectrometer (Nicolet iS50ATR-FTIR infrared spectrometer) to collect mid-infrared spectrograms of the adulterated dendrobium huoshanense samples.
The procedure for spectrum acquisition was as follows: placing the sample mixed with the dendrobium huoshanense powder into a sample cell of a diamond single reflection AIR accessory equipped in an FTIR spectrometer, and setting the measurement range of the mid-infrared spectrometer to be 4000-400cm-1Setting the scanning times of the mid-infrared spectrometer to be 32 times and the resolution to be 4.0cm-17467 data points are collected, and the same sample is repeatedly collected for 6 times to obtain a mid-infrared original spectrogram as shown in figure 2;
(3) spectral preprocessing
Performing baseline correction and average spectrogram calculation on the original spectrogram of the intermediate infrared spectrum acquired 6 times by the same pseudo-dendrobium huoshanense sample to obtain an average spectrum, eliminating the spectrogram caused by artificial sampling error, moving the average spectrum by 15 points averagely, and eliminating the spectrum noise caused by the detection environment by adopting a smoothing method;
8 spectral pretreatment methods were applied: the method comprises the following steps of screening a pretreatment method by combining a multivariate scattering correction method, a standard normal variation method, a first derivative method, a second derivative method, a multivariate scattering correction plus first derivative method, a multivariate scattering correction plus second derivative method, a standard normal variation plus first derivative method, a standard normal variation plus second derivative method and a partial least squares algorithm (PLS);
by comparing R of PLS cross validation models2Values and root mean square error values RMSECV, results are shown in Table 1, and it can be seen that the spectral pretreatment method using standard normal plus first derivative yields a higher R2The value is 0.9487, the minimum RMSECV value is 9.58%, and a mid-infrared spectrogram of the dendrobium huoshanense-doped sample after the spectral pretreatment of standard normal change plus first derivative is obtained, as shown in FIG. 3;
table 1 shows the results of 8 spectral pretreatment methods
(4) Feature variable extraction
Screening of spectral characteristic variables: three different wavelength selection methods, namely a moving window partial least square method, a Monte Carlo non-information variable elimination method and an interval random frog method, are compared;
the effective wavelength of the PLS model is established by combining the three wavelength variable screening methods to improve the calculation workload and the accuracy of the model, the result is shown in Table 2, only 238 data points are used for comparatively using the interval random frog method to screen the characteristic wavelength, and higher R is obtained2Value 0.9721, minimum RMSECV value 7.37%, determined characteristic wavelength screening using the interval random frog method as shown in fig. 3;
table 2 shows the results of the 3 wavelength variable screening methods
(5) Adulteration prediction
Predicting the adulteration amount of the adulteration dendrobium huoshanense sample by adopting a adulteration quantitative model according to the characteristic wavelength variable of the spectrogram of the adulteration dendrobium huoshanense sample; establishing a adulteration quantitative model between characteristic wavelength variables and adulteration amounts of spectrograms of 105 correction set adulterated dendrobium huoshanense samples by a partial least square method, and verifying the adulteration amounts of the 105 prediction set adulterated dendrobium huoshanense samples by using the adulteration quantitative model, wherein the result is shown in fig. 4;
according to the results of fig. 4, the correlation coefficient of the correction set obtained by the method is 0.9951, the root mean square error is 2.98%, the correlation coefficient of the prediction set is 0.9556, and the root mean square error is 6.44%, and the constructed method can provide a reliable method for the determination of the adulteration amount of the dendrobium heshanense in the dendrobium huoshanense.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (10)
1. A method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense is characterized by comprising the following steps:
(1) preparation of pseudo-dendrobium huoshanense powder sample
Picking up collected fresh dendrobium huoshanense stems and fresh dendrobium huoshanense stems, cleaning, removing impurities, cutting, freeze-drying, crushing and sieving, collecting dendrobium huoshanense stem powder and dendrobium huoshanense stem powder with the particle size of 60-100 meshes, mixing the dendrobium huoshanense stem powder and the dendrobium huoshanense stem powder according to different proportions according to a mass ratio to obtain 21 groups of adulterated samples, setting 10 parallel samples for each group of adulterated samples, and sealing and storing for later use, wherein the mass proportion range of the dendrobium huoshanense stem powder and the dendrobium huoshanense stem powder is 0-100%;
(2) infrared spectroscopy collection
Collecting the mid-infrared spectrogram of each adulteration sample by using the Fourier transform attenuated total reflection mid-infrared spectrometer of 210 adulteration samples prepared in the step (1), wherein each sample is collected for multiple times to obtain an original spectrum for calculating an average spectrogram;
(3) spectral preprocessing
Performing baseline correction and average spectrogram calculation on the intermediate infrared spectrogram of the adulterated sample to eliminate spectrogram difference caused by artificial sampling error, then moving the average spectrum by 15 data points, adopting a spectrum smoothing method to eliminate spectral noise caused by detection environment, and performing cross validation on R of the model by comparing a least square method2Comparing R of cross-validation model with R of root mean square error RMSECV value by 8 spectrum preprocessing methods2Screening the value and the root mean square error value RMSECV by a spectrum pretreatment method;
(4) screening of spectral characteristic wavelengths
Three different wavelength selection methods, namely a moving window partial least square method, a Monte Carlo non-information variable elimination method and an interval random frog method, are compared to obtain effective wavelengths for establishing a prediction model so as to improve the calculation workload and accuracy of the model;
(5) adulteration prediction
Predicting the adulteration amount of the adulteration sample by using a adulteration quantitative model; and (3) establishing 105 adulterated dendrobium samples as a correction set by a partial least square method, and establishing adulteration quantification between characteristic wavelength variables and adulteration quantities of a spectrogram.
2. The method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense according to claim 1, which is characterized in that: in the 21 adulterated samples in the step (1), the mass ratio of the dendrobium huinan stem powder to the dendrobium huoshanense stem powder is respectively as follows: 0%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%.
3. The method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense according to claim 2, which is characterized in that: the total mass of each adulteration sample in the step (1) is controlled to be 500 mg.
4. The method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense according to claim 1, which is characterized in that: and (2) hermetically storing the sample in the step (1) at-20 ℃ for later use, so as to ensure the stability of the dendrobium medicinal material.
5. The method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense according to claim 1, which is characterized in that: and (3) the infrared spectrometer in the step (2) is provided with an AIR accessory.
6. The method of claim 5, wherein the method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense comprises the following steps: the related parameters of the infrared spectrometer spectrum acquisition in the step (2) are set as follows: the measurement range is 4000--1The number of scanning times is 32, and the resolution is 4.0cm-1And 7467 data points are collected, and the same sample is repeatedly collected for 6 times to obtain a mid-infrared original spectrogram.
7. The method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense according to claim 1, which is characterized in that: and (3) performing baseline correction and average spectrogram calculation on the original spectrogram of the mid-infrared spectrum acquired 6 times by the same sample in the step (2) to obtain an average spectrum of the original spectrogram, so as to eliminate spectrogram difference caused by artificial sampling errors.
8. The method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense according to claim 1, which is characterized in that: in the step (3), the average spectrum is shifted by 15 data points, and the spectrum smoothing method is adopted to reduce the spectrum noise caused by the detection environment.
9. The method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense according to claim 1, which is characterized in that: the 8 spectrum preprocessing methods in the step (3) are a multivariate scattering correction method, a standard normal variation method, a first derivative method, a second derivative method, a multivariate scattering correction plus first derivative method, a multivariate scattering correction plus second derivative method, a standard normal variation plus first derivative method and a standard normal variation plus second derivative method.
10. The method for rapidly predicting the adulteration of dendrobium huoshanense in dendrobium huoshanense according to claim 1, which is characterized in that: and (5) predicting the adulteration amount of the adulterated dendrobium sample by using a adulteration quantitative model, specifically, establishing 1/2 a adulteration quantitative model between a characteristic variable of a spectrogram of the adulterated dendrobium sample in the total sample number correction set and the adulteration amount by using a partial least square method, and verifying the adulteration amount of the adulterated dendrobium sample in the 1/2 total sample number prediction set by using the adulteration quantitative model.
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