CN111220562A - Method for identifying different species of swertia - Google Patents
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- 241001530209 Swertia Species 0.000 title claims abstract description 59
- 238000000034 method Methods 0.000 title claims abstract description 26
- 241000894007 species Species 0.000 title description 12
- 238000002329 infrared spectrum Methods 0.000 claims abstract description 40
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 claims abstract description 32
- 238000001228 spectrum Methods 0.000 claims abstract description 29
- 238000005102 attenuated total reflection Methods 0.000 claims abstract description 22
- 238000002360 preparation method Methods 0.000 claims abstract description 5
- 238000002156 mixing Methods 0.000 claims abstract description 4
- 238000010521 absorption reaction Methods 0.000 claims description 60
- 238000012360 testing method Methods 0.000 claims description 40
- 241000977602 Swertia mussotii Species 0.000 claims description 20
- 241000974657 Swertia pseudochinensis Species 0.000 claims description 15
- 238000001035 drying Methods 0.000 claims description 5
- 241000967294 Swertia japonica Species 0.000 claims description 4
- 238000001559 infrared map Methods 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 2
- 239000000463 material Substances 0.000 abstract description 7
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- 238000010298 pulverizing process Methods 0.000 abstract description 2
- 238000007873 sieving Methods 0.000 abstract description 2
- 230000009102 absorption Effects 0.000 description 47
- 238000002835 absorbance Methods 0.000 description 12
- 238000004458 analytical method Methods 0.000 description 11
- 238000002834 transmittance Methods 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 5
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- 108090000056 Complement factor B Proteins 0.000 description 4
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- 238000001514 detection method Methods 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 235000008216 herbs Nutrition 0.000 description 2
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- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 1
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 241000501704 Swertia franchetiana Species 0.000 description 1
- 241000501703 Swertia tetraptera Species 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- 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
- G01N21/3563—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing solids; Preparation of samples therefor
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- 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
- G01N21/359—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light using near infrared light
Abstract
The invention provides an infrared spectrum identification method for different swertia. It comprises the following steps: a. sample preparation: pulverizing the sample, sieving, and oven drying at constant temperature; b. collecting an infrared spectrum: mixing the sample obtained in the step a with potassium bromide uniformly according to a proportion, tabletting, setting parameters of an infrared spectrometer, and scanning to obtain a middle infrared spectrum; c. collecting an ATR map: taking the sample in the step a, setting parameters of an infrared spectrometer, and scanning to obtain an ATR (attenuated total reflectance) spectrum; d. collecting a near-infrared spectrum: taking the sample in the step a, setting parameters of an infrared spectrometer, and scanning to obtain a near-infrared spectrum; e. and d, analyzing the maps obtained in the steps b to d. The method for identifying the false Chinese swertia by the infrared spectrum of the different kinds of false Chinese swertia has the advantages of simple operation, good repeatability and stability and high accuracy, and can identify false Chinese swertia and original medicinal materials of the false Chinese swertia of different kinds by infrared spectrum scanning, thereby quickly identifying the false Chinese swertia.
Description
Technical Field
The invention belongs to the technical field of medicinal material detection, and particularly relates to an infrared spectrum identification method for different kinds of swertia.
Background
Swertia (Swertia L.) plant, annual herb, thin root, brown-yellow. The stem is upright, round, hollow, and branched above the middle. Basal leaves wither during the flowering phase; the cauline leaves have no or short stem, the leaves are oval to be in the shape of an oval needle, the tip is long and tapered, the base is blunt, the back is obviously protruded, and the uppermost part is in the shape of a leaf bud. The large-scale conical compound umbrella inflorescence is loose. Is mainly distributed in northern Sichuan, Tibet, southwest Qinghai and Yunnan, is a folk single herbal medicine and is mainly used for treating dyspepsia, cholecystitis, various hepatitis and other diseases. As a wild plant with the treasure, the research on the medicinal components of the swertia pseudochinensis is deepened along with the rapid development of national medicines, and the demand is increased day by day.
The medicinal ingredients of different species of swertia davidi are different, and the contents of the medicinal ingredients are also obviously different. At present, the identification of the swertia pseudochinensis is limited to appearance and thin-layer chromatography, such as Liqi (2017), and the like (quality standard research of Tibetan medicine swertia pseudochinensis [ J ] Chinese medicine and clinic), the identification of the swertia pseudochinensis is realized by a microscopic identification and thin-layer identification method, the method for applying the method is multiple, the operation is complex, the method is influenced by knowledge reserve of technical personnel, and the aim of rapid and accurate analysis cannot be achieved.
Disclosure of Invention
In order to solve the problems, the invention provides an infrared spectrum identification method of different swertia davidi herbs, which is characterized by comprising the following steps: it comprises the following steps:
a. sample preparation: taking a test sample, crushing, screening by a 100-mesh screen and drying;
b. collecting an infrared spectrum: mixing the sample obtained in the step a with potassium bromide uniformly according to a proportion, tabletting, setting parameters of an infrared spectrometer, and scanning to obtain a middle infrared spectrum;
c. collecting an ATR map: taking the sample in the step a, setting parameters of an infrared spectrometer, and scanning to obtain an ATR (attenuated total reflectance) spectrum;
d. collecting a near-infrared spectrum: taking the sample in the step a, setting parameters of an infrared spectrometer, and scanning to obtain a near-infrared spectrum;
e. and d, analyzing the maps obtained in the steps b to d.
Further, the drying temperature in the step a is 45 ℃ and the time is 24 hours.
Further, the ratio of the sample to KBr in the step b is 1: 100.
Further, the parameters in steps b and c are: the wave number is 4000-400 cm-1Scanning times 8 times, resolution 6cm-1(ii) a And/or, the parameters in the step d are as follows: the wave number range is 10000-4000 cm-1Scanning times of 64 times and resolution of 6cm-1。
Furthermore, in the map in the step e, the middle infrared map shows that 10 common absorption peaks of swertia japonica are 3407cm respectively-1、2924cm-1、2854cm-1、1735cm-1、1621cm-1、1511cm-1、1418cm-1、1376cm-1、1268cm-1、1068cm-1。
Further, the middle infrared spectrum shows that the concentration of swertia pseudochinensis is 1210cm-1Special absorption is performed; the content of swertia pseudochinensis Franch is 1650cm-1Special absorption is performed; 1461cm of India swertia-1Special absorption is performed; herba Swertiae Bimaculatae at 589cm-1There is no absorption.
Further, in the map in the step e, the ATR map shows that the swertia herb has 9 common absorption peaks which are 3298cm respectively-1、2920cm-1、1611cm-1、1414cm-1、1368cm-1、1236cm-1、1016cm-1、830cm-1、518cm-1。
Further, the ATR map is shown as an India swertia in 1153cm-1And 559cm-1Has special absorption at 2852cm-1There is no absorption; herba Swertiae Bimaculatae at 1263cm-1Special absorption is performed; swertia mussotii Franch at 1732cm-1And 1509cm-1There is no absorption; only swertia pseudochinensis Franch and swertia davidii Franch are at 930cm-1There is absorption.
Furthermore, in the map in the step e, the near infrared map shows that the swertia japonica Makino has 6 common absorption peaks which are 6858cm respectively-1、5783cm-1、5176cm-1、4723cm-1、4325cm-1、4256cm-1。
Furthermore, the near-infrared spectrum shows that the concentration of swertia mussotii, swertia davidi and swertia davidi is 8338cm-1The absorption is at 8338cm-1There is no absorption.
The method for identifying the false Chinese swertia by the infrared spectrum of the different kinds of false Chinese swertia has the advantages of simple operation, good repeatability and stability and high accuracy, and can identify false Chinese swertia and original medicinal materials of the false Chinese swertia of different kinds by infrared spectrum scanning, thereby quickly identifying the false Chinese swertia.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1 shows infrared one-dimensional spectrogram of herba Swertiae Bimaculatae of different species
FIG. 2 ATR spectrogram of different species of swertia
FIG. 3 is a near-infrared spectrum of different species of swertia
FIG. 4 is an average spectrum of infrared orthogonal test spectrum
FIG. 5 comparison of theoretical optimal combination and actual optimal combination profiles
FIG. 6 is a graph of an average near-infrared spectrum of an orthogonal test
FIG. 72 No. 5 and No. 15 Norris first derivative spectra
FIG. 8 comparison of near-infrared theoretical optimal combination with actual optimal combination maps
FIG. 9 clustering chart of different species of swertia
Detailed Description
Materials and instruments
Swertia mussotii Franch is collected from Dudoku county of Qinghai province, Swertia davidi (Swertia franchetiana H.Smith) is collected from Yushu county of Qinghai province, Swertia davidi (Swertia tetraptera Maxim.) is collected from Xiaojin county of Sichuan province, Swertia davidi (Swertia punica Hemsl.) is collected from Xiaojin county of Sichuan province, and Swertia indica (Swertia chirality Karsten) is purchased from market.
Fourier transform infrared spectrum analyzer (model: IS 50, Thermo Nicolet Co., Ltd.), DTGS detector, pulverizer (Tester Co., Tianjin Co., Ltd.), oven (Shanghai-Hengchu scientific instruments Co., Ltd.), electronic balance (ME104,0.0001g), agate mortar, tablet die (diameter: 13mm, PIKE Co., USA), and press,
Example 1 identification of different species of swertia
a. Sample preparation: respectively pulverizing the medicinal materials, sieving with 100 mesh sieve, and oven drying at 45 deg.C for 24 hr;
b. collecting an infrared spectrum: and (b) taking the sample in the step a, uniformly mixing the sample with potassium bromide in a mortar according to the mass ratio of 1:100, filling the mixture into a mold, and tabletting the mixture on a tabletting machine. Setting spectrum acquisition conditions: the wave number is 4000-400 cm-1Scanning times 8 times, resolution 6cm-1Detecting the mid-infrared spectrum;
c. collecting ATR atlas: taking the sample in the step a, and setting spectrum acquisition conditions: the wave number is 4000-400 cm-1Scanning times 8 times, resolution 6cm-1Detecting an ATR spectrum;
d. collecting a near-infrared spectrum: taking the sample in the step a, and setting spectrum acquisition conditions: the wave number range is 10000-4000 cm-1Scanning times of 64 times and resolution of 6cm-1And detecting the near infrared spectrum.
The spectrum of the mid-infrared spectrum obtained by the detection in the step b is shown in a figure 1, and the mid-infrared absorption data is shown in a table 1.
As can be seen from fig. 1 and table 1: the common peaks of the mid-infrared spectrograms of the 5 medicinal materials are 10: 3407cm-1、2924cm-1、2854cm-1、1735cm-1、1621cm-1、1511cm-1、1418cm-1、1376cm-1、1268cm-1、1068cm-1. The difference between the intermediate infrared spectrograms is mainly concentrated at 1700-500cm-1And (3) removing the solvent. Wherein the peak of swertia pseudochinensis Franch is 1210cm-1(ii) a The peculiar peak of the swertia pseudochinensis Franch is 1650cm-1(ii) a 1461cm of India swertia-1Has special absorption at 533cm-1There is no absorption; herba Swertiae Bimaculatae at 589cm-1There is no absorption; the number of absorption peaks of swertia mussotii is the least, and the absorbance of swertia mussotii is the highest among 5 species, and the absorbance of swertia mussotii is the lowest.
The spectrum of the ATR obtained by the detection in the step c is shown in a figure 2, and ATR absorption data is shown in a table 2.
As can be seen from fig. 2 and table 2: the total peaks of ATR spectrogram of 5 medicinal materials are 9: 3298cm-1、2920cm-1、1611cm-1、1414cm-1、1368cm-1、1236cm-1、1016cm-1、830cm-1、518cm-1. The difference between the ATR spectra is mainly concentrated in 1600-560cm-1. ATR is 2854cm less than mid-infrared common peak-1、1735cm-1、1511cm-1. But is increased by 830cm-1、518cm-1The other common peaks are not very different. Wherein the India swertia Makino is 1153cm-1And 559cm-1Has special absorption at 2852cm-1There is no absorption; herba Swertiae Bimaculatae at 1263cm-1Special absorption is performed; swertia mussotii Franch at 1732cm-1And 1509cm-1There is no absorption; only swertia pseudochinensis Franch and swertia davidii Franch are at 930cm-1There is absorption. The absorption of the 5 Chinese swertia mussotii species is highest, and the absorption of the swertia pseudochinensis franch is lowest.
The near infrared spectrum detected in step d is shown in FIG. 3, and the near infrared absorption data is shown in Table 3.
As can be seen from FIG. 3 and Table 3, the total peaks of the near infrared spectrograms of 5 herbs are 6: 6858cm-1、5783cm-1、5176cm-1、4723cm-1、4325cm-1、4256cm-1. The absorption information between near infrared spectrograms is less, and various absorptions are similar, wherein swertia mussotii Franch, swertia davidii Franch, and swertia davidii Franch are in 8338cm-1The absorption is at 8338cm-1There is no absorption.
In conclusion, the middle infrared spectrum shows that the concentration of swertia pseudochinensis in 1210cm-1Characteristic absorption is formed; the content of swertia pseudochinensis Franch is 1650cm-1Characteristic absorption is formed; 1461cm of India swertia-1Characteristic absorption is formed; herba Swertiae Bimaculatae at 589cm-1There is no absorption. The ATR map is shown as India swertia in 1153cm-1And 559cm-1Has characteristic absorption at 2852cm-1There is no absorption; herba Swertiae Bimaculatae at 1263cm-1Characteristic absorption is formed; swertia mussotii Franch at 1732cm-1And 1509cm-1There is no absorption. The near infrared spectrum shows that the concentration of swertia mussotii, swertia davidi var.franch and swertia davidi var.tetraphylla is 8338cm-1The absorption is at 8338cm-1There is no absorption.
TABLE 1 absorption data of infrared one-dimensional spectrogram of different kinds of swertia
TABLE 2 ATR absorption data of different species of swertia
TABLE 3 near-infrared absorption data of different species of swertia
The following test examples specifically illustrate the advantageous effects of the present invention:
experimental example 1 optimization of collection conditions of infrared spectra of swertia
1. Quadrature test
Considering factors that may affect infrared spectral acquisition (table 4), a 3-factor 4-level orthogonal test table (table 5) was designed for 16 sets of tests. Randomly selecting a swertia mussotii sample to carry out full-waveband spectrum scanning, carrying out spectrum collection of 16 groups of tests according to conditions, calculating an average spectrum (n is 3, shown in figure 4) of each test, and selecting an actual optimal combination according to the maximum absorbance and the maximum transmittance.
TABLE 4 infrared orthogonal test factor horizon
TABLE 5 orthogonal test table for mid-infrared conditions
When selecting a sample spectrogram, the absorbance value and the transmittance of the sample need to be considered, namely the absorbance value of the sample is in the range of 0.8-1.2 and about 1.0 is the best, and the transmittance needs to be more than 75 percent. The results show that the results of test No. 5 are more appropriateThe absorbance value of the sample was 0.898 and the sample transmittance was 77.87%, so the actual optimum combination was No. 5, namely A2B1C2(sample to KBr ratio 1:100, scan times 8 times, resolution 4cm-1)。
2. Range analysis
Performing range analysis on the test results (table 6), wherein for the maximum absorbance value, the sequence of the influence factors from large to small is A > B > C, namely the ratio of the sample to KBr > the scanning times > resolution; for the maximum transmittance, the influence factors are sequentially A > C > B from large to small, namely the ratio of the sample to KBr > resolution > scanning times.
For maximum absorbance values, factor A gives the best results at level 2, factor B gives the best results at level 1, and factor C gives the best results at level 3, so the theoretically best combination is A2B1C3(i.e., sample to KBr ratio of 1:100, scan times of 8, resolution of 6cm-1) (ii) a For transmittance, factor A gives the best results at 4 levels, factor B gives the best results at 1 level, and factor C gives the best results at 4 levels, so the theoretical optimal combination is A4B1C4(i.e., sample to KBr ratio of 1:200, scan times of 8 times, resolution of 8cm-1)。
For factor A, the absorbance within the range of 0.8-1.2 is best with a result of about 1.0, so A is selected2(ii) a For factor B, both optimal combinations are B1Thus selecting B1(ii) a For factor C, the higher the resolution, the more apparent the pattern noise, so C is chosen3Therefore, the optimal theoretical combination is A2B1C3(i.e., sample to KBr ratio of 1:100, scan times of 8, resolution of 6cm-1)。
TABLE 6 results of pole difference analysis of mid-infrared orthogonal test
3. Comparison of theoretical optimal combination with actual optimal combination
FIG. 5 showsCompared with the actual optimal combined spectrogram, the comparison result of the theoretical optimal combined spectrogram is smoother and less in spectrogram burr. The analysis and comparison of the two results of 3 times of experiments show that the correlation coefficients of the actual optimal combination of the results of 3 times of experiments are respectively as follows: 1.0000, 0.9858, 0.9854, RSD value 0.832%; the correlation coefficients of the theoretical optimal combination of the 3 test results are respectively as follows: 1.0000, 0.9949, 0.9948, RSD value of 0.297%, indicating poor reproducibility of the actual optimum combination, so the theoretical optimum combination condition a was selected during the test2B1C3(i.e., sample to KBr ratio of 1:100, scan times of 8, resolution of 6cm-1)。
4. Analysis of variance
From the results of the anova (table 7), the ratio of the sample to KBr significantly affected the results, but the number of scans and the resolution did not significantly affect the results. And the influence of various factors on the result can be known according to the F value, and for the absorbance value: ratio > scan number > resolution, for transmittance: sample preparation: KBr ratio > resolution > scan number, which is consistent with the results of the range analysis.
Table 7 analysis results of variance in mid-infrared orthogonal test
Note: the influence of the expression factors on the results reaches a very significant level
5. Methodology validation
The methodological verification test is carried out by taking the aforementioned sample of swertia mussotii Franch as a test sample and testing under the condition of determining the collection condition of a pre-test spectrum, namely the ratio of the sample to KBr is 1:100, the scanning times are 8 times, and the resolution is 6cm-1。
5.1 repeatability
The same sample is continuously pressed for 6 times, and the correlation coefficients of 6 times are respectively 1.0000, 0.9979, 0.9977, 0.9972, 0.9967 and 0.9948 by taking one time as a standard, and the RSD value is 0.170%, which indicates that the test has better repeatability.
5.2 precision
The same tablet of the same sample is continuously measured for 6 times, and by taking one time as a standard, the correlation coefficients of the measured 6 times are respectively 1.0000, 0.9986, 0.9976, 0.9974, 0.9973 and 0.9960, and the RSD value is 0.135%, which indicates that the precision of the test is better.
5.3 stability
The same sample is pressed to obtain the same tablet, the tablet is placed in a dryer for storage, the tablet is measured once every 1 hour, the continuous measurement is carried out for 6 times, the correlation coefficients of the 6 times are respectively 1.0000, 0.9973, 0.9968, 0.9955, 0.9953, 0.9946 and the RSD value is 0.195% by taking one time as a standard, and the test has better stability.
Experimental example 2 optimization of near infrared spectrum collection conditions of swertia mussotii
1.1, orthogonal test
Considering factors that may affect the acquisition of near infrared spectra (table 8), a 3-factor 4-level orthogonal test table (table 9) was designed for 16 sets of experiments. Taking a swertia mussotii sample to carry out full-waveband spectrum scanning, carrying out spectrum collection of 16 groups of experiments according to conditions, calculating an average spectrum (n is 3, see figure 6) of each experiment, and selecting an actual optimal combination according to a peak value (the peak value reflects a signal-to-noise ratio to a certain extent).
TABLE 8 near-infrared orthogonal test factor horizon
As can be seen from table 9, the peak value was 5 # with the highest peak value, and was next 2 # and 15 # with the correlation coefficients of the spectra acquired three times for No. 2 being 1.0000, 0.9415, 0.9386(RSD of 3.61%), the correlation coefficients of the spectra acquired three times for No. 5 being 1.0000, 0.8413, 0.8319(RSD of 10.60%), and the correlation coefficients of the spectra acquired three times for No. 15 being 1.0000, 0.9960, 0.9959(RSD of 0.23%). Meanwhile, the near infrared spectrum and the near infrared Norris first-order derivative spectrum (figure 7) are combined, and the spectrum No. 15 is smoother than the spectrums No. 2 and No. 5, so that the spectrum No. 15 is selected as an actual optimal combination.
TABLE 9 near-infrared orthogonal test Table
1.2, range analysis
The results of the above tests were analyzed for range differences (Table 10), and the magnitude of the influence of each factor on the absorbance was: size of particle>Resolution ratio>The number of scans. The best results are obtained at level 1 for factor A, 1 for factor B and 2 for factor C, so the best theoretical combination is A1B1C2(i.e. 8 scans at 2cm resolution-1Particle size 100 mesh).
TABLE 10 results of range analysis in near-infrared orthogonal test
1.3 comparison of the actual optimal combination with the theoretical optimal combination
The correlation coefficients of the actual optimal combined cubic spectrograms are 1.0000, 0.9966 and 0.9959, the RSD value is 0.219%, the correlation coefficients of the theoretical optimal combined cubic spectrogram are 1.0000, 0.7349 and 0.7076, the RSD value is 16.151%, the theoretical and actual near infrared spectrum and the Norris first-order derivative spectrum (figure 8) are combined, the actual optimal combination is smoother than the theoretical optimal combination spectrogram, so the experimental acquisition conditions are acquired according to the actual optimal combination, namely the scanning times are 64 times, and the resolution is 6cm-1The granularity is 100 meshes.
1.4 analysis of variance
The analysis of variance in the near-infrared orthogonal test is shown in table 11, and it is known that the influence of the granularity on the result reaches an extremely significant level, and the influence of the resolution on the result reaches a significant level; according to the magnitude of the F value, the influence of the three factors on the result is as follows: granularity > resolution > scan times, consistent with range analysis results.
TABLE 11 analysis of variance in near-infrared orthogonal test
Note: indicates that the influence of the factors on the results reaches a very significant level
1.5 methodology validation
The methodology verification test is carried out by taking swertia mussotii Franch as a sample under the spectrum collection condition determined by a pre-test, namely: the scanning times are 64 times, and the resolution is 6cm-1The granularity is 100 meshes.
1.5.1 repeatability test
The same sample is continuously tested for 6 times by different samples, one time is taken as a standard, the correlation coefficients of the 6 times are respectively 1.0000, 0.9961, 0.9960 and 0.9958, and the RSD value is 0.163 percent, which indicates that the test has better repeatability.
1.5.2 precision test
The same sample of the same sample is continuously measured for 6 times, wherein one time is taken as a standard, the correlation coefficients of the 6 times are respectively 1.0000, 0.9959, 0.9959, 0.9959, 0.9956 and 0.9955, and the RSD value is 0.174%, which shows that the precision of the test is better.
1.5.3 stability test
The same sample of the same sample is continuously measured for 6 times, the sample is placed in a dryer for storage once after the measurement is finished, the sample is measured once every 1h, the total number of the measurement is 6, and the correlation coefficients of the 6 times are respectively 1.0000, 0.9979, 0.9974, 0.9973 and the RSD value is 0.106 percent by taking one of the times as a standard, which indicates that the test has better stability.
Experimental example 3 Cluster analysis of different species of Swertiae
Clustering analysis is carried out on 5 kinds of swertia davidi in the ATR spectral absorbance data (table 2) of example 1, the result is shown in figure 9, according to the result, when the preserved information is 75%, the 5 kinds of swertia davidi can be divided into 3 groups, wherein the first group is swertia davidi, swertia davidi with stem and red swertia; the second group is swertia mussotii; the third group is India swertia. According to the knowledge of 'Chinese plant record-62 book', swertia mussotii, swertia davidi and swertia davidi are large floral multi-branch systems, and swertia davidi is four different floral groups, so that the clustering method can effectively display various close relations.
To sum up: the infrared, ATR and near-infrared absorption conditions of the swertia are different, so that the types of 5 swertia samples can be judged by combining the absorption data of the three spectrograms.
Claims (10)
1. An infrared spectrum identification method for different types of swertia davidi is characterized by comprising the following steps: it comprises the following steps:
a. sample preparation: taking a test sample, crushing, screening by a 100-mesh screen and drying;
b. collecting an infrared spectrum: mixing the sample obtained in the step a with potassium bromide uniformly, tabletting, and scanning to obtain a middle infrared spectrum;
c. collecting an ATR map: taking the sample in the step a, and scanning to obtain an ATR (attenuated total reflectance) spectrum;
d. collecting a near-infrared spectrum: taking the sample in the step a, and scanning to obtain a near-infrared spectrum;
e. and d, analyzing the maps obtained in the steps b to d.
2. The authentication method according to claim 1, wherein: and the drying temperature in the step a is 45 ℃, and the drying time is 24 hours.
3. The authentication method according to claim 1, wherein: step b the ratio of the sample to KBr was 1: 100.
4. The authentication method according to claim 1, wherein: the acquisition parameters in the steps b and c are as follows: the wave number is 4000-400 cm-1Scanning times 8 times, resolution 6cm-1(ii) a And/or, the parameters in the step d are as follows: the wave number range is 10000-4000 cm-1Scanning times of 64 times and resolution of 6cm-1。
5. The authentication method according to claim 1, wherein: step e in the map, inThe infrared spectrum shows that the total absorption peaks of herba Swertiae Bimaculatae are 10, and are 3407cm respectively-1、2924cm-1、2854cm-1、1735cm-1、1621cm-1、1511cm-1、1418cm-1、1376cm-1、1268cm-1、1068cm-1。
6. The authentication method according to claim 5, wherein: the middle infrared spectrum shows that the concentration of swertia pseudochinensis is 1210cm-1Characteristic absorption is formed; the content of swertia pseudochinensis Franch is 1650cm-1Characteristic absorption is formed; 1461cm of India swertia-1Characteristic absorption is formed; herba Swertiae Bimaculatae at 589cm-1There is no absorption.
7. The authentication method according to claim 1, wherein: in the map in the step e, the ATR map shows that the swertia chinensis has 9 common absorption peaks which are 3298cm respectively-1、2920cm-1、1611cm-1、1414cm-1、1368cm-1、1236cm-1、1016cm-1、830cm-1、518cm-1。
8. The authentication method according to claim 7, wherein: the ATR map shows that the India swertia herb is 1153cm-1And 559cm-1Has characteristic absorption at 2852cm-1There is no absorption; herba Swertiae Bimaculatae at 1263cm-1Characteristic absorption is formed; swertia mussotii Franch at 1732cm-1And 1509cm-1There is no absorption.
9. The authentication method according to claim 1, wherein: in the map of the step e, the near infrared map shows that the swertia japonica Makino has 6 common absorption peaks which are 6858cm respectively-1、5783cm-1、5176cm-1、4723cm-1、4325cm-1、4256cm-1。
10. The method of claim 9An authentication method characterized by: the near infrared spectrum shows that the concentration of swertia mussotii, swertia davidi var.franch and swertia davidi var.tetraphylla is 8338cm-1The absorption is at 8338cm-1There is no absorption.
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