CN112180004A

CN112180004A - Method for identifying high-volatility traditional Chinese medicinal materials on site by using surface acoustic wave gas chromatograph

Info

Publication number: CN112180004A
Application number: CN202010900164.5A
Authority: CN
Inventors: 赵军宁; 何世堂; 王剑波; 陆艳艳; 朱宏伟; 孙林; 刘久玲
Original assignee: Institute of Acoustics CAS; Sichuan Academy of Chinese Medicine Sciences SACMS
Current assignee: Institute of Acoustics CAS; Sichuan Academy of Chinese Medicine Sciences SACMS
Priority date: 2020-08-31
Filing date: 2020-08-31
Publication date: 2021-01-05
Also published as: WO2022041796A1

Abstract

The invention belongs to the technical field of quality control of traditional Chinese medicines, and particularly relates to a method for identifying high-volatility traditional Chinese medicinal materials on site by using a surface acoustic wave gas chromatograph. Aiming at the problem that a rapid and accurate traditional Chinese medicine field detection method is lacked in the prior art, the technical scheme of the invention is as follows: (1) judging the volatility of the sample under the pretreatment condition of normal-temperature saturation treatment; (2) pretreating the sample by a proper method; (3) obtaining a standard fingerprint spectrum; (4) detecting the sample to obtain a sample map of volatile components in the Chinese medicinal sample to be detected, and comparing the sample map with the standard fingerprint map to identify the authenticity of the Chinese medicinal sample to be detected. The method can effectively eliminate uncertainty caused by factors such as geographic environment, climate condition, weather condition and instrument state in field detection, and has the advantages of simple sample pretreatment, high analysis speed, field detection and the like.

Description

Method for identifying high-volatility traditional Chinese medicinal materials on site by using surface acoustic wave gas chromatograph

Technical Field

The invention belongs to the technical field of quality control of traditional Chinese medicines, and particularly relates to a method for identifying high-volatility traditional Chinese medicines on site by using a surface acoustic wave gas chromatograph.

Background

The traditional Chinese medicine can be used for preventing and treating various diseases, the quality of the traditional Chinese medicine is influenced by the production place, climate, environment, processing, transportation and storage, so that the quality of the traditional Chinese medicine from different sources is easy to be uneven, and particularly, for rare traditional Chinese medicines, some lawbreakers are mixed with more and more false for obtaining illegal benefits, so that the traditional Chinese medicines are hard to distinguish, the quality is unstable, and the safety and the effectiveness of the medicine are difficult to ensure. False and inferior products are misused, so that the drug effect is not good for the mild patients, the toxic and side effects are generated for the severe patients, and even the life of people is endangered, thus greatly damaging the quality safety of the traditional Chinese medicine and threatening the physical health of people. Meanwhile, the image of the traditional Chinese medicine is seriously damaged, and the healthy growth of the traditional Chinese medicine industry in China is hindered. Therefore, the active development of Chinese medicine true and false identification and quality evaluation and the development of various quick, reliable and easily popularized Chinese medicine true and false detection technologies and equipment are urgent needs in the modern development stage of Chinese medicine.

The traditional detection method of the traditional Chinese medicine at present comprises character identification, microscopic identification, physicochemical identification, gas chromatography, high performance liquid chromatography, near infrared spectroscopy and other instrument analysis methods, although the intuitive experience identification and the microscopic identification are simple and rapid, an identifier is required to have equivalent experience, microscope and other equipment, and the method is only suitable for the traditional Chinese medicine and the powder traditional Chinese medicine with obvious characteristics in appearance character, while other identification methods are complex in operation, pretreatment is required before testing, the method comprises crushing, extraction, separation, concentration, drying and the like, the testing time is long (30-60 min is required), the instrument and equipment requirements are high, the method needs to be carried out in a laboratory, the popularization is difficult, and the purpose of the rapid detection cannot be achieved. Due to the fact that the traditional Chinese medicines are various in variety and complex in source, the market detection sample size is large, the detection time can be effectively shortened through rapid field detection, and the labor cost is reduced. The existing field rapid detection method for traditional Chinese medicines comprises appearance identification, character identification, chemical identification, thin-layer identification and the like. But the sensitivity is poor, the anti-interference capability is weak, and false positive or false negative results are easy to occur. Therefore, the establishment of a rapid and accurate field detection method for traditional Chinese medicines has very important practical significance.

Disclosure of Invention

Aiming at the problem that a rapid and accurate traditional Chinese medicine field detection method is lacked in the prior art, the invention provides a method for identifying high-volatility traditional Chinese medicine on site by using a surface acoustic wave gas chromatograph, and aims to: provides a method for distinguishing the truth of traditional Chinese medicines on site (such as traditional Chinese medicine producing areas, traditional Chinese medicine markets and the like) without leaving laboratory conditions. According to the method, after the volatility of components in the traditional Chinese medicinal materials is simply judged, the traditional Chinese medicinal materials are subjected to simple pretreatment, retention index calibration and headspace sample injection are adopted, and a surface acoustic wave rapid gas chromatograph (GC-SAW) is combined, so that the characteristic components of the traditional Chinese medicinal materials are analyzed by utilizing the rapid analysis and high-sensitivity characteristics of the surface acoustic wave rapid gas chromatograph. The method has the advantages of simple sample pretreatment, high analysis speed, field detection and the like, and can be applied to field authenticity identification of traditional Chinese medicines.

A method for identifying traditional Chinese medicinal materials on site by using a surface acoustic wave gas chromatograph comprises the following steps:

(1) and (3) determining the volatility of the sample: taking a traditional Chinese medicine sample to be detected, and carrying out normal-temperature saturation treatment to obtain a judgment sample to be detected; detecting a to-be-detected judgment sample by using a surface acoustic wave gas chromatograph to obtain a spectrum of volatile components in the to-be-detected judgment sample; judging the traditional Chinese medicine sample to be detected as a high-volatility sample, a medium-volatility sample or a low-volatility sample according to the response value of the maximum peak of the response values;

(2) sample pretreatment: preprocessing a traditional Chinese medicine sample to be detected and a standard product corresponding to the traditional Chinese medicine sample to be detected according to a judgment result to obtain a sample to be detected and a standard sample, wherein the preprocessing condition of the high-volatility sample is normal-temperature saturation processing; the pretreatment condition of the medium volatile sample is high-temperature saturation treatment; the pretreatment condition of the low-volatility sample is solid-phase extraction;

(3) obtaining a standard fingerprint spectrum: detecting the standard sample by using a surface acoustic wave gas chromatograph to obtain a standard fingerprint spectrum of volatile components in the standard sample;

(4) and (3) detection of the sample: and (4) detecting the sample to be detected by using a surface acoustic wave gas chromatograph to obtain a sample map of volatile components in the traditional Chinese medicine sample to be detected, and comparing the sample map with the standard fingerprint map obtained in the step (3) to identify the authenticity of the traditional Chinese medicine to be detected.

Preferably, before the step (1), a surface acoustic wave gas chromatograph is also used for detecting the normal alkane mixed standard substance, and recording the peak time of each normal alkane standard substance so as to calculate the retention index of each volatile component in the process of obtaining the map in the step (1), the step (3) or the step (4);

the retention index is calculated as follows:

wherein RI is retention index of volatile component, t_xN and n +1 are respectively the number of carbon atoms of the standard substance of two normal alkanes with spectral peaks adjacent to the spectral peak of the volatile component, and t is the retention time of the volatile component_nAnd t_n+1Are respectively a spectrumRetention time of two normal paraffin standard peaks adjacent to the volatile component spectrum peak.

Preferably, in step (1), the criterion for determining the volatility of the sample is as follows: the response value of the maximum peak of the response values in the atlas is greater than or equal to 2000Hz, and the sample is high volatility; the response value of the maximum peak of the response values in the atlas is less than 2000Hz, and the medium volatility sample is obtained when the response value is more than or equal to 100 Hz; the response value of the peak with the maximum response value in the map is less than 100Hz, and the sample with low volatility is obtained.

Preferably, in the step (1) or the step (2), the normal temperature saturation treatment specifically comprises the steps of putting 20-1000mg of the traditional Chinese medicine sample into a 10-40ml headspace bottle, and saturating for 1-60min at normal temperature; in the step (2), the high-temperature saturation treatment specifically comprises the steps of putting 20-1000mg of the traditional Chinese medicine sample into a 10-40ml headspace bottle, and saturating at 30-100 ℃ for 1-60 min; in the step (2), the solid phase extraction process comprises the steps of putting 20-1000mg of the traditional Chinese medicine sample into a 10-40ml headspace bottle, and extracting for 5s-5min by using a solid phase extraction needle.

Preferably, in step (1), step (3) or step (4), the testing process of the surface acoustic wave gas chromatograph is as follows: in a sample introduction state, connecting a sampling port with a sample to be detected, starting a sampling pump, and sucking the sample to be detected into a pre-concentration pipe; and in the detection state, switching the six-way valve to enable the carrier gas to flow through the pre-concentration pipe and carry the sample to be detected in the pre-concentration pipe to enter the chromatographic column and the detector in sequence to finish detection.

Preferably, the surface acoustic wave gas chromatography column is selected from DB-5, SPB-5, Rtx-5, BP-5, OV-5, 007-2(MPS-5), SE-52, SE-54, XTI-5, PTE-5, ZB-5, AT-5, MDN-5 or ZB-5, and the length of the column is 1 to 10 meters.

Preferably, the detection conditions of the surface acoustic wave gas chromatograph are as follows: the initial temperature of the chromatographic column is 40-50 ℃, and in the detection process, the temperature of the chromatographic column is increased to 200 ℃ according to a temperature increasing program of 0-20 ℃/s; the flow rate of the chromatographic column is 1-7mL/min, and the carrier gas is nitrogen or helium.

Preferably, the working time of the sampling pump is 5-60s in the sample injection state of the surface acoustic wave gas chromatograph.

Preferably, the detection conditions of the surface acoustic wave gas chromatograph are as follows: the temperature of the detector is 25-60 ℃.

Preferably, in the step (4), the sample spectrum is compared with the standard fingerprint spectrum obtained in the step (3) by using a similarity evaluation or principal component analysis method so as to identify the authenticity of the traditional Chinese medicine to be detected.

Preferably, aiming at the high-volatility sample, the method for identifying the high-volatility Chinese medicinal material on site by using the surface acoustic wave gas chromatograph comprises the following steps of:

(1) and (3) determining the volatility of the sample: taking a traditional Chinese medicine sample to be detected, and carrying out normal-temperature saturation treatment to obtain a judgment sample to be detected; detecting a to-be-detected judgment sample by using a surface acoustic wave gas chromatograph to obtain a spectrum of volatile components in the to-be-detected judgment sample; judging whether the sample to be detected is a high-volatility sample or not according to the response value of the maximum peak of the response values; the standard for determining the volatility of the sample is as follows: the response value of the maximum peak of the response values in the atlas is greater than or equal to 2000Hz, and the sample is high volatility;

(2) sample pretreatment: after determining that the sample to be detected is a high-volatility sample, pre-treating the traditional Chinese medicine sample to be detected and a standard sample corresponding to the traditional Chinese medicine sample to be detected to obtain the sample to be detected and the standard sample, wherein the pre-treating condition is normal-temperature saturation treatment;

(4) and (3) detection of the sample: and (4) detecting the sample to be detected by using a surface acoustic wave gas chromatograph to obtain a sample map of volatile components in the traditional Chinese medicine sample to be detected, and comparing the sample map with the standard fingerprint map obtained in the step (3) to identify the authenticity of the traditional Chinese medicine to be detected. The traditional Chinese medicine sample, the traditional Chinese medicine sample to be detected and the standard substance are processed into the traditional Chinese medicine with proper size in the modes of pre-cutting, crushing, shearing or trimming and the like. The to-be-detected judgment sample, the to-be-detected sample and the standard sample refer to gaseous or solid phase extraction samples which can be directly used for the surface acoustic wave gas chromatograph test after the traditional Chinese medicinal materials are treated by the pretreatment method provided by the invention. The term "normal temperature" as used herein means a condition where no additional heating or cooling is performed at the air temperature of the inspection site.

Compared with the traditional laboratory detection, the difficulty of field detection is that due to the influences of factors such as geographic environment, weather conditions, current weather conditions and the like, the uncertainty of the most appropriate pretreatment condition and detection condition for the specific traditional Chinese medicinal materials is enhanced, the state change of instruments and equipment is large, and the detection accuracy is further influenced. Based on the difficulties, the volatility of the sample is judged before testing, and a proper medicinal material pretreatment method is determined under the conditions of local geography, climate and weather according to certain standards, so that the content of each volatile component in the sample to be tested is ensured. Furthermore, through comparison with a standard sample, the fluctuation of the state of the instrument can be overcome, and the detection accuracy is improved. Through the calibration of the normal paraffin mixed standard sample, the peak-out time of the volatile components can be converted into a retention index, so that each spectrum peak is more visual and standardized, and a standard fingerprint spectrum can be obtained conveniently, and the qualitative judgment on the spot is facilitated.

Therefore, the method provided by the embodiment of the invention at least has the following advantages:

1. the most effective pretreatment condition and detection condition can be selected, and the detection accuracy is ensured.

2. The pretreatment process of the sample is simple, the analysis speed of the used portable surface acoustic wave gas chromatograph is high, one sample is completely tested for only 2-8min, and the method is suitable for field rapid detection and screening work;

3. the method has the advantages of high testing speed, simple and convenient operation, no need of organic reagents, environmental friendliness and no secondary pollution;

4. the running cost of the instrument is very low, the consumption products are very few, and the instrument is suitable for large-batch field test work;

5. for medium-volatility samples, the high-temperature saturation method greatly improves the detection sensitivity of the portable surface acoustic wave gas chromatograph, and can complete detection on traditional Chinese medicines with few volatile components. For low-volatility samples, the detection sensitivity of the portable surface acoustic wave gas chromatograph is greatly improved by adopting solid phase microextraction, and the detection can be completed for traditional Chinese medicines with few volatile components.

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 a schematic structural diagram of a SAW gas chromatograph in a sample injection state;

FIG. 2 shows a schematic structural diagram of a SAW gas chromatograph in a detection state;

FIG. 3 shows the spectrum of the detection result of musk in the traditional Chinese medicine in example 1;

figure 4 shows the chromatogram of musk standard 1 in example 1 together with the retention index and the response value of each peak;

FIG. 5 shows the detection result spectrum of the traditional Chinese medicine turmeric in example 2;

FIG. 6 shows the chromatogram of Sichuan Jiantang turmeric of example 2 and the retention index and response value of each peak;

FIG. 7 shows the spectrum of the detection result of Cordyceps sinensis in example 3;

FIG. 8 shows the chromatogram of Cordyceps sinensis, Anhui, in example 3, and the retention index and response value of each peak;

FIG. 9 shows the detection result spectrum of the Chinese medicinal Notoginseng of example 4;

FIG. 10 shows the chromatogram of Notoginseng flower 8-3 of example 4 and the retention index and response value of each peak

FIG. 11 shows a spectrum of the detection result of bezoar bovis as a Chinese medicine in example 5;

FIG. 12 shows a chromatogram of 5-1 bezoar bovis in example 5 and retention indices and response values of the respective peaks;

FIG. 13 shows a spectrum of the detection results of the traditional Chinese medicine fingered citron in example 6;

FIG. 14 shows the chromatogram of 10-2 finger citron in example 6, and the retention index and response value of each peak.

Detailed Description

The surface acoustic wave gas chromatograph adopted by the embodiment of the invention is a portable surface acoustic wave gas chromatograph. As shown in fig. 1 and 2, the surface acoustic wave gas chromatograph includes a sampling port 1, a sampling pump 2, a preconcentration pipe 3, a six-way valve 4, a gas carrying cylinder 5, a chromatographic column 6, a detector 7, and a data processing device 8.

The surface acoustic wave gas chromatograph can be in a sample injection state or a detection state, and can be switched between the two states.

As shown in fig. 1, when the surface acoustic wave gas chromatograph is in the sample injection state, the first gas port 41 of the six-way valve 4 is connected to the first gas port 31 of the pre-concentration tube 3, the second gas port 42 of the six-way valve 4 is connected to the carrier gas cylinder 5, the third gas port 43 of the six-way valve 4 is connected to the gas inlet of the chromatographic column 6, the fourth gas port 44 of the six-way valve 4 is connected to the second gas port 32 of the pre-concentration tube 3, the fifth gas port 45 of the six-way valve 4 is connected to the sampling pump 2, and the sixth gas port 46 of the six-way valve 4 is connected to.

The sample enters the pre-concentration pipe 3 through the sampling port 1 by the air pressure difference formed by the sampling pump 2 to finish sampling.

After the sample injection is completed, the six-way valve 4 is rotated to switch the surface acoustic wave gas chromatograph to the sample injection state, as shown in fig. 2. In the sample injection state, the first gas port 41 of the six-way valve 4 is connected with the second gas port 32 of the pre-concentration tube 3, the second gas port 42 of the six-way valve 4 is connected with the sampling pump 2, the third gas port 43 of the six-way valve 4 is connected with the sampling port 1, the fourth gas port 44 of the six-way valve 4 is connected with the first gas port 31 of the pre-concentration tube 3, the fifth gas port 45 of the six-way valve 4 is connected with the gas carrying bottle 5, and the sixth gas port 46 of the six-way valve 4 is connected with the gas inlet of the chromatographic column 6.

At this time, a gas path is formed from the carrier gas cylinder 5 through the preconcentration tube 3, the column 6, and the detector 7 in this order. During the detection process, the pre-concentration tube 3 is warmed to vaporize the sample therein. The gasified sample in the pre-concentration pipe 3 can enter the chromatographic column 6 along with the carrier gas to complete gas phase separation, and then enter the detector 7 to complete detection of each gas phase component after sample separation. The data detected by the detector 7 can be sent to the data processing device 8, and the data processing device 8 processes and presents the data so that the detection personnel can know the detection result.

In the detection process, the experimental conditions of the surface acoustic wave gas chromatograph are as follows:

a DB-5 chromatographic column is adopted; the temperature rising procedure of the chromatographic column is as follows: maintaining the initial temperature at 40-50 deg.C, and raising the temperature to 200 deg.C at 0-20 deg.C/s;

the flow rate of the DB-5 chromatographic column is 1-7 ml/min;

the length of the DB-5 chromatographic column is 1-10 meters;

the temperature of the sample inlet is 120-200 ℃;

the temperature of the six-way valve 4 is 120-165 ℃;

the temperature of the detector is 25-60 ℃;

the pumping time of the sampling pump 2 is 5-60 s;

the gas carrying bottle 5 is filled with nitrogen or helium.

As a preferable mode, the experimental conditions of the surface acoustic wave gas chromatograph used in examples 1 to 6 are as follows:

a DB-5 chromatographic column is adopted; the temperature rising procedure of the chromatographic column is as follows: initial temperature 40 deg.C, 10 deg.C/s to 200 deg.C;

the flow rate of the DB-5 chromatographic column is 4 ml/min;

the size of the DB-5 chromatographic column is 1m multiplied by 0.25mm multiplied by 0.25 mu m;

the temperature of a sample inlet is 200 ℃;

the temperature of the six-way valve 4 is 160 ℃;

the temperature of the detector is 40 ℃;

the pumping time of the sampling pump 2 is 10 s;

the gas-carrying bottle 5 is filled with nitrogen.

The technical solution of the present invention is further illustrated by the following specific examples.

Example 1

In this example, the musk is detected, which includes the following steps:

(1) and (4) retention index calibration: in the embodiment, the digital fingerprint is prepared by using the retention index, so that errors caused by operation factors and experimental condition differences can be effectively avoided. Taking 20mg of 13 normal paraffin mixed standard products (C6-C18), adding into a 40ml headspace bottle, saturating for 5min at normal temperature, connecting the headspace bottle with a portable surface acoustic wave gas chromatograph, so that a gas sample can be sucked into the chromatograph through a pump, and separating on the chromatographic column. And detecting, namely obtaining a detection signal on a surface acoustic wave detector, and recording the peak-appearing retention time of each normal paraffin standard. The peak time of the normal alkane standard product is used for calculating the retention index of the volatile component of the sample to be detected by adopting a retention index RI calculation formula of Kovats in the subsequent step. Among them, the RI calculation formula of Kovats is shown in formula (1).

Wherein RI is retention index of volatile component, t_xN and n +1 are respectively the number of carbon atoms of the standard substance of two normal alkanes with spectral peaks adjacent to the spectral peak of the volatile component, and t is the retention time of the volatile component_nAnd t_n+1Respectively is the retention time of two normal paraffin standard products with peaks adjacent to the volatile component peaks.

(2) And (3) determining the volatility of the sample: in order to ensure that signals with strong enough intensity are collected, different pretreatment modes are needed for traditional Chinese medicines with different volatility. When the detection personnel need to detect the traditional Chinese medicine with unknown volatility, the following scheme can be adopted to determine the appropriate pretreatment method of the traditional Chinese medicine.

Firstly, a normal-temperature saturation treatment method is used for pretreating a standard substance of a traditional Chinese medicine sample to be detected. The normal temperature saturation treatment method specifically comprises the steps of putting 20mg of a sample into a 40ml headspace bottle, and obtaining a sample to be detected after saturation for 5min at normal temperature (the detection field temperature can be regarded as 25 ℃ approximately in the embodiment). And detecting the sample to be detected by using a surface acoustic wave gas chromatography instrument, and observing a detection signal.

In this embodiment, the response value of the peak with the largest response value is greater than 2000Hz, and it can be determined that the traditional Chinese medicine sample is a high-volatility traditional Chinese medicine sample, and the suitable pretreatment method is normal-temperature saturation treatment.

(3) Detecting the background of the headspace bottle: in this embodiment, the process of detecting the blank background of the headspace bottle using the portable surface acoustic wave gas chromatograph is as follows: a40 ml headspace bottle is connected with a portable surface acoustic wave gas chromatograph, air in the headspace bottle is sucked into the chromatograph through a pump, separated on a chromatographic column, and a detection signal is obtained on a surface acoustic wave detector. The obtained blank background has no signal interference at the characteristic peak of the traditional Chinese medicine.

(4) Standard analytical experiments: in the embodiment, musk purchased from different manufacturers is used as standard substances and respectively marked as a musk standard substance 1, a musk standard substance 2, a musk standard substance 3, a musk standard substance 4 and a musk standard substance 5, 20mg of each of the five standard substances is taken and added into a 40ml headspace bottle, after the five standard substances are saturated at normal temperature for 5min, the headspace bottle is connected with a portable surface acoustic wave gas chromatograph, gas to be detected is sucked into the chromatograph through a pump, the gas to be detected is separated on the chromatographic column, and a detection signal is obtained on the surface acoustic wave detector. After obtaining the detection signal, the data processing device may calculate the retention index of the active ingredient in the musk standard product according to the retention time of the peak of the normal paraffin standard product calibrated in step (1) and the retention time of the active ingredient in the musk standard product. The data processing means may also calculate the response (peak area) of the active ingredient in the musk standard. According to the sample peak retention index and response obtained in the embodiment, a traditional Chinese medicine musk fingerprint spectrum database is established. Wherein the response, i.e. the peak area, may reflect the content of the corresponding active ingredient.

(5) Analysis experiment of traditional Chinese medicine sample to be tested: in the embodiment, an unknown musk sample is used as a traditional Chinese medicine sample to be detected, 20mg of the traditional Chinese medicine sample to be detected is taken and added into a 40ml headspace bottle, the headspace bottle is connected with a portable surface acoustic wave gas chromatograph after the traditional Chinese medicine sample is saturated at normal temperature for 5min, gas to be detected is sucked into the chromatograph through a pump, is separated on the chromatographic column, and a detection signal is obtained on a surface acoustic wave detector. After obtaining the detection signal, the data processing device may calculate the retention index of the active ingredient in the detected musk according to the retention time of the peak of the normal alkane standard calibrated in step (1) and the retention time of the active ingredient in the detected musk. The data processing means may also calculate the response (peak area) of the active ingredient in the musk standard. And (4) comparing the obtained sample peak retention index with the traditional Chinese medicine musk fingerprint spectrum database established in the step (4), and identifying the authenticity by observing the similarity of the positions and relative sizes of the spectrum peaks of the main components.

Wherein, fig. 3 shows the chromatogram of each musk detected in step (5). Figure 4 shows the chromatogram of musk standard 1 and the retention index and response value of each peak. The comparison of the maps in the figure can obviously identify that the traditional Chinese medicine sample to be detected is the genuine musk. The experimental results of the embodiment show that the method can accurately detect and identify whether the traditional Chinese medicine sample to be detected is a genuine product or a counterfeit product.

Example 2

The embodiment of the method for detecting the turmeric comprises the following steps:

Wherein RI is retention index of volatile component, t_xFor the retention time of the volatile components, n and n +1 are eachThe number of carbon atoms, t, of the standard substance of two normal alkanes with spectral peaks adjacent to the volatile component spectral peak_nAnd t_n+1Respectively is the retention time of two normal paraffin standard products with peaks adjacent to the volatile component peaks.

Firstly, a normal-temperature saturation treatment method is used for pretreating a standard substance of a traditional Chinese medicine sample to be detected. The normal temperature saturation treatment method specifically comprises the steps of putting 450mg of a sample into a 40ml headspace bottle, and obtaining a sample to be detected after saturation for 5min at normal temperature (the detection site temperature can be regarded as 25 ℃ approximately in the embodiment). And detecting the sample to be detected by using a surface acoustic wave gas chromatography instrument, and observing a detection signal.

(4) Standard analytical experiments: respectively taking 450mg of standard substance Guangxi turmeric, standard substance Anhui turmeric, standard substance Sichuan Ji Antang turmeric and standard substance Sichuan Tongrentang turmeric, adding the standard substance Sichuan Ji Antang turmeric and the standard substance Sichuan Tongrentang turmeric into a 40ml headspace bottle, after the mixture is saturated at normal temperature for 5min, connecting the headspace bottle with a portable surface acoustic wave gas chromatograph, sucking gas to be detected into the chromatograph through a pump, separating the gas on the chromatographic column, and obtaining a detection signal on a surface acoustic wave detector. After obtaining the detection signal, the data processing device may calculate the retention index of the active ingredient in the turmeric standard according to the peak retention time of the normal paraffin standard calibrated in step (1) and the retention time of the active ingredient in the turmeric standard. The data processing device may also calculate the response (peak area) of the active ingredient in the turmeric standard. According to the sample peak retention index and response obtained in the embodiment, a fingerprint spectrum database of the traditional Chinese medicine turmeric is established. Wherein the response, i.e. the peak area, may reflect the content of the corresponding active ingredient.

(5) Analysis experiment of traditional Chinese medicine sample to be tested: in the embodiment, an unknown sample is used as a traditional Chinese medicine sample to be detected, 450mg of the traditional Chinese medicine sample to be detected is taken and added into a 40ml headspace bottle, the headspace bottle is connected with a portable surface acoustic wave gas chromatograph after the traditional Chinese medicine sample is saturated at normal temperature for 5min, gas to be detected is sucked into the chromatograph through a pump, is separated on the chromatographic column, and a detection signal is obtained on a surface acoustic wave detector. After obtaining the detection signal, the data processing device may calculate the retention index of the active ingredient in the detected turmeric based on the peak retention time of the normal paraffin standard calibrated in step (1) and the retention time of the active ingredient in the detected turmeric. The data processing device may also calculate the response (peak area) of the active ingredient in the turmeric standard. And (4) comparing the obtained sample peak retention index with the traditional Chinese medicine turmeric fingerprint spectrum database established in the step (4), and identifying the authenticity by observing the similarity of the positions and relative sizes of the spectrum peaks of the main components.

Wherein, fig. 5 shows the chromatogram of each turmeric detected in step (5). FIG. 6 shows the chromatogram of standard Sichuan Jiantang turmeric and the retention index and response value of each peak. The comparison of the maps in the figure can obviously identify that the traditional Chinese medicine sample to be detected is genuine turmeric. The experimental results of the embodiment show that the method can accurately detect and identify whether the traditional Chinese medicine sample to be detected is a genuine product or a counterfeit product.

Example 3

In this embodiment, the detection of cordyceps sinensis comprises the following steps:

(1) and (4) retention index calibration: in the embodiment, the digital fingerprint is prepared by using the retention index, so that errors caused by operation factors and experimental condition differences can be effectively avoided. Taking 20mg of 13 normal paraffin mixed standard products (C6-C18), putting the standard products into a 40ml headspace bottle, saturating the bottle in a constant temperature device at 60 ℃ for 5min, connecting the headspace bottle with a portable surface acoustic wave gas chromatograph, sucking air in the headspace bottle into the chromatograph through a pump, and separating the air on the chromatographic column. And detecting, namely obtaining a detection signal on a surface acoustic wave detector, and thus obtaining and recording the retention time of each normal alkane standard product. The peak time of the normal alkane standard product is used for calculating the retention index of the volatile component of the sample to be detected by adopting a retention index RI calculation formula of Kovats in the subsequent step.

Among them, the RI calculation formula of Kovats is shown in formula (1).

(2) The volatility of the sample was determined in the same manner as in example 1. In this embodiment, the response value of the peak with the largest response value is between 100Hz and 2000Hz, and it can be determined that the traditional Chinese medicine sample is a medium volatile traditional Chinese medicine sample, and the suitable pretreatment method is high temperature saturation treatment.

(3) Detecting a high-temperature background of the headspace bottle: in this embodiment, the process of detecting a blank traditional Chinese medicine sample by using a portable surface acoustic wave gas chromatograph is as follows: taking a 40ml headspace bottle, saturating in a constant temperature device at 60 ℃ for 5min, connecting with a portable surface acoustic wave gas chromatograph, sucking air in the headspace bottle into the chromatograph through a pump, separating on a chromatographic column, and obtaining a detection signal on a surface acoustic wave detector. The obtained blank background has no signal interference at the characteristic peak of the traditional Chinese medicine.

(4) Standard analytical experiments: taking 20mg of aweto, cordyceps sinensis and cordyceps sinensis, adding the aweto, cordyceps sinensis and cordyceps sinensis standard into a 40ml headspace bottle, saturating for 5min in a constant temperature device at 60 ℃, connecting the headspace bottle with a portable surface acoustic wave gas chromatograph, sucking gas to be detected into the chromatograph through a pump, separating on the chromatographic column, and obtaining a detection signal on a surface acoustic wave detector. After the detection signal is obtained, the data processing device can calculate the retention index of the active ingredient in the cordyceps sinensis standard according to the retention time of the peak of the normal paraffin standard substance calibrated in the step (1) and the retention time of the active ingredient in the cordyceps sinensis standard substance. The data processing device can also calculate the response (peak area) of the active ingredients in the cordyceps sinensis standard. According to the sample peak retention index and response obtained in the embodiment, a Chinese medicine cordyceps sinensis fingerprint spectrum database is established. Wherein the response, i.e. the peak area, may reflect the content of the corresponding active ingredient.

(5) Analysis experiment of traditional Chinese medicine sample to be tested: in the embodiment, the method takes the false cordyceps sinensis (counterfeit product) as a traditional Chinese medicine sample to be detected, 20mg of the false cordyceps sinensis (counterfeit product) is added into a 40ml headspace bottle, the headspace bottle is connected with a portable surface acoustic wave gas chromatograph after the headspace bottle is saturated in a constant temperature device at 60 ℃ for 5min, gas to be detected is sucked into the chromatograph through a pump, the gas to be detected is separated on the chromatographic column, and a detection signal is obtained on a surface acoustic wave detector. After obtaining the detection signal, the data processing device may calculate the retention index of the active ingredient in the detected cordyceps sinensis according to the retention time of the peak of the normal paraffin standard substance calibrated in step (1) and the retention time of the active ingredient in the detected cordyceps sinensis. The data processing device can also calculate the response (peak area) of the active ingredients in the cordyceps sinensis standard. And (4) comparing the obtained sample peak retention index with the Chinese caterpillar fungus fingerprint spectrum database established in the step (4), and identifying the authenticity by observing the similarity of the positions and relative sizes of the spectrum peaks of the main components.

Wherein, FIG. 7 shows the chromatogram of each Cordyceps sinensis detected in step (5). FIG. 8 shows the chromatogram of Cordyceps Anhui and the retention index and response value of each peak. The Chinese medicinal sample to be detected can be obviously identified as the false cordyceps sinensis by comparing the maps in the figure. The experimental results of the embodiment show that the method can accurately detect and identify whether the traditional Chinese medicine sample to be detected is a genuine product or a counterfeit product.

In addition, in this embodiment, the pretreatment of the volatile sample by the high-temperature saturation method greatly improves the sensitivity of the surface acoustic wave gas chromatography detection compared with the pretreatment of the sample by normal-temperature saturation, and the detection time of each traditional Chinese medicine sample is only 7 min.

Example 4

The method for detecting the notoginseng flowers comprises the following steps:

Among them, the RI calculation formula of Kovats is shown in formula (1).

(4) Standard analytical experiments: the method comprises the steps of purchasing notoginseng flowers from three different manufacturers, wherein the notoginseng flowers are respectively numbered as 8-1 notoginseng flower, 8-2 notoginseng flower and 8-3 notoginseng flower, taking 20mg of bezoar as a standard substance, adding the bezoar into a 40ml headspace bottle, saturating for 5min in a constant temperature device at 60 ℃, connecting the headspace bottle with a portable surface acoustic wave gas chromatograph, sucking gas to be detected into the chromatograph through a pump, separating on a chromatographic column, and obtaining detection signals on a surface acoustic wave detector. After obtaining the detection signal, the data processing device can calculate the retention index of the active ingredient in the pseudo-ginseng flower standard according to the retention time of the peak of the normal paraffin standard substance calibrated in the step (1) and the retention time of the active ingredient in the pseudo-ginseng flower standard substance. The data processing means may also calculate the response (peak area) of the active ingredient in the notoginseng flower standard. According to the sample peak retention index and response obtained in the embodiment, a fingerprint spectrum database of the traditional Chinese medicine notoginseng flower is established. Wherein the response, i.e. the peak area, may reflect the content of the corresponding active ingredient.

(5) Analysis experiment of traditional Chinese medicine sample to be tested: in the embodiment, an unknown true and false sanchi flower is taken as a traditional Chinese medicine sample to be detected, 20mg of the traditional Chinese medicine sample to be detected is taken and added into a 40ml headspace bottle, the headspace bottle is connected with a portable surface acoustic wave gas chromatograph after the traditional Chinese medicine sample is saturated in a constant temperature device at 60 ℃ for 5min, gas to be detected is sucked into the chromatograph through a pump, is separated on the chromatographic column, and a detection signal is obtained on a surface acoustic wave detector. After obtaining the detection signal, the data processing device may calculate the retention index of the active ingredient in the detected notoginseng flower according to the retention time of the normal paraffin standard substance peak calibrated in step (1) and the retention time of the active ingredient in the detected notoginseng flower. The data processing means may also calculate the response (peak area) of the active ingredient in the notoginseng flower standard. And (4) comparing the obtained sample peak retention index with the fingerprint database of the traditional Chinese medicine notoginseng flower established in the step (4), and identifying the authenticity by observing the similarity of the positions and relative sizes of the spectral peaks of the main components.

Wherein, fig. 9 shows the chromatogram of each notoginseng flower detected in step (5). Fig. 10 shows the chromatogram of notoginseng flowers in anhui and the retention index and response value of each peak. The Chinese medicine sample to be detected can be obviously identified as the genuine pseudo-ginseng flower by comparing the maps in the figure. The experimental results of the embodiment show that the method can accurately detect and identify whether the traditional Chinese medicine sample to be detected is a genuine product or a counterfeit product.

In addition, in the embodiment, compared with the pretreatment of the sample by normal-temperature saturation, the pretreatment of the volatile sample by the high-temperature saturation method has greatly improved sensitivity of the surface acoustic wave gas chromatography detection, and the detection time of each traditional Chinese medicine sample is only 7 min.

Example 5

In this embodiment, the method for detecting bezoar includes the following steps:

(1) and (4) retention index calibration: in the embodiment, the digital fingerprint is prepared by using the retention index, so that errors caused by operation factors and experimental condition differences can be effectively avoided. Taking 20mg of 13 normal paraffin mixed standard products (C6-C18), placing the standard products into a 40ml headspace sample bottle, extracting for 20s by using a solid phase micro-extraction needle (in the embodiment, fused quartz optical fiber is selected), connecting the solid phase micro-extraction needle with an adopted port of a portable surface acoustic wave gas chromatograph, sucking gas to be detected in the solid phase micro-extraction needle into the chromatograph through a pump, and separating on the chromatographic column. And detecting, namely obtaining a detection signal on a surface acoustic wave detector, and thus obtaining and recording the peak time of each normal alkane standard product. The peak retention time of the normal paraffin standard is used for calculating the retention index of the volatile component of the sample to be detected by adopting a retention index RI calculation formula of Kovats in the subsequent step. Among them, the RI calculation formula of Kovats is shown in formula (1).

(2) The volatility of the sample was determined in the same manner as in example 1. In this embodiment, the response value of the peak with the largest response value is less than 100Hz, and it can be determined that the traditional Chinese medicine sample is a low-volatility traditional Chinese medicine sample, and the suitable pretreatment method is solid phase extraction.

(3) Detecting a solid phase extraction background of the headspace bottle: in this embodiment, the process of detecting a blank traditional Chinese medicine sample by using a portable surface acoustic wave gas chromatograph is as follows: taking a 40ml headspace bottle, extracting for 20s by using a solid phase microextraction needle, connecting the headspace bottle with a portable surface acoustic wave gas chromatograph by using the solid phase microextraction needle, sucking the headspace bottle into the chromatograph by a pump, separating on a chromatographic column, and obtaining a detection signal on a surface acoustic wave detector. The obtained blank background has no signal interference at the characteristic peak of the traditional Chinese medicine.

(4) Standard analytical experiments: the method comprises the steps of purchasing bezoar from five different manufacturers, wherein the bezoar is respectively numbered as 5-1 bezoar, 5-2 bezoar, 5-3 bezoar, 5-4 bezoar and 5-5 bezoar, taking 20mg of the five bezoar as standard substances, adding the five bezoar into a 40ml headspace bottle, extracting for 20s by using a solid phase micro extraction needle, connecting the solid phase micro extraction needle with a portable surface acoustic wave gas chromatograph, sucking gas to be detected into the chromatograph through a pump, separating on the chromatographic column, and obtaining detection signals on a surface acoustic wave detector. After obtaining the detection signal, the data processing device may calculate the retention index of the active ingredient in the bezoar standard according to the retention time of the peak of the normal paraffin standard calibrated in step (1) and the retention time of the active ingredient in the bezoar standard. The data processing means may also calculate the response (peak area) of the active ingredient in the bezoar bovis standard. According to the sample peak retention index and response obtained in the embodiment, a traditional Chinese medicine bezoar fingerprint spectrum database is established. Wherein the response, i.e. the peak area, may reflect the content of the corresponding active ingredient.

(5) Analysis experiment of traditional Chinese medicine sample to be tested: in this example, a counterfeit bezoar (calculus bovis factitius) was used as a sample of the Chinese medicine to be tested. Adding 20mg of pseudo bezoar (calculus bovis pseudonarum) into a 40ml headspace bottle, extracting for 20s by using a solid phase micro-extraction device, connecting with a portable surface acoustic wave gas chromatograph, sucking gas to be detected into the chromatograph by a pump, separating on a chromatographic column, and obtaining a detection signal on a surface acoustic wave detector. After obtaining the detection signal, the data processing device may calculate the retention index of the active ingredient in the detected bezoar according to the retention time of the peak of the normal paraffin standard substance calibrated in step (1) and the retention time of the active ingredient in the detected bezoar. The data processing means may also calculate the response (peak area) of the active ingredient in the bezoar bovis standard. And (4) comparing the obtained sample peak retention index with the traditional Chinese medicine bezoar fingerprint database established in the step (4), and identifying the authenticity by observing the similarity of the positions and relative sizes of the spectral peaks of the main components.

FIG. 11 shows the chromatogram of each bezoar detected in step (5). FIG. 12 shows the chromatogram of 5-1 bezoar bovis and the retention index and response value of each peak. The Chinese medicine sample to be detected can be obviously identified as the false bezoar by comparing the maps in the figure. The experimental results of the embodiment show that the method can accurately detect and identify whether the traditional Chinese medicine sample to be detected is a genuine product or a counterfeit product.

In addition, in the embodiment, compared with the pretreatment of the sample by normal temperature or high temperature saturation, the pretreatment of the low-volatility sample by adopting the solid phase micro-extraction device has greatly improved sensitivity of the surface acoustic wave gas chromatography detection, and the detection time of each traditional Chinese medicine sample is only 2 min.

Example 6

The embodiment detects the fingered citron, and comprises the following steps:

(4) Standard analytical experiments: the method comprises the steps of purchasing fingered citrons from three different manufacturers, wherein the fingered citrons are respectively numbered as 10-1 fingered citron, 10-2 fingered citron and 10-3 fingered citron, taking 20mg of the three fingered citrons as standard products, adding the standard products into a 40ml headspace bottle, extracting for 20s by using a solid phase micro-extraction needle, connecting the solid phase micro-extraction needle with a portable surface acoustic wave gas chromatograph, sucking gas to be detected into the chromatograph through a pump, separating on a chromatographic column, and obtaining detection signals on a surface acoustic wave detector. After the detection signal is obtained, the data processing device can calculate the retention index of the active ingredients in the fingered citron standard according to the retention time of the normal paraffin standard substance peak and the retention time of the active ingredients in the fingered citron standard substance calibrated in the step (1). The data processing device can also calculate the response (peak area) of the active ingredients in the fingered citron standard. According to the sample peak retention index and response obtained in the embodiment, a traditional Chinese medicine fingered citron fingerprint spectrum database is established. Wherein the response, i.e. the peak area, may reflect the content of the corresponding active ingredient.

(5) Analysis experiment of traditional Chinese medicine sample to be tested: in this embodiment, a counterfeit fingered citron (pseudo fingered citron) is used as a traditional Chinese medicine sample to be tested. Taking 20mg of a fake product fingered citron (fake fingered citron), adding the fake product fingered citron (fake fingered citron) into a 40ml headspace bottle, extracting for 20s by using a solid phase micro-extraction device, connecting the fake product fingered citron with a portable surface acoustic wave gas chromatograph, sucking gas to be detected into the chromatograph through a pump, separating the gas on a chromatographic column, and obtaining a detection signal on a surface acoustic wave detector. After the detection signal is obtained, the data processing device can calculate the retention index of the active ingredients in the detected fingered citron according to the retention time of the normal paraffin standard substance peak and the retention time of the active ingredients in the detected fingered citron, which are calibrated in the step (1). The data processing device can also calculate the response (peak area) of the active ingredients in the fingered citron standard. And (4) comparing the obtained sample peak retention index with the traditional Chinese medicine fingered citron fingerprint spectrum database established in the step (4), and identifying the authenticity by observing the similarity of the positions and relative sizes of the spectrum peaks of the main components.

Wherein, fig. 13 shows the chromatogram of each finger citron detected in step (5). FIG. 14 shows the chromatogram of 5-1 finger citron and retention index and response value of each peak. The Chinese medicine sample to be detected can be obviously identified as the false fingered citron by comparing the maps in the figure. The experimental results of the embodiment show that the method can accurately detect and identify whether the traditional Chinese medicine sample to be detected is a genuine product or a counterfeit product.

In conclusion, the method for identifying the traditional Chinese medicine by using the surface acoustic wave gas chromatograph provided by the invention. Compared with the prior art, the method can effectively eliminate uncertainty caused by factors such as geographic environment, weather conditions and instrument states in field detection, and has the advantages of simple sample pretreatment, high analysis speed, field detection and the like.

Claims

1. A method for identifying high-volatility traditional Chinese medicinal materials on site by using a surface acoustic wave gas chromatograph is characterized by comprising the following steps of:

(1) and (3) determining the volatility of the sample: taking a standard substance of a traditional Chinese medicine sample to be detected, and carrying out normal-temperature saturation treatment to obtain a judgment sample to be detected; detecting a to-be-detected judgment sample by using a surface acoustic wave gas chromatograph to obtain a spectrum of volatile components in the to-be-detected judgment sample; judging whether the sample to be detected is a high-volatility sample or not according to the response value of the maximum peak of the response value, wherein the standard of sample volatility judgment is as follows: the response value of the maximum peak of the response values in the atlas is greater than or equal to 2000Hz, and the sample is high volatility;

(2) sample pretreatment: after determining that the sample to be detected is a high-volatility sample according to the step (1), pre-treating the traditional Chinese medicine sample to be detected and a standard sample corresponding to the traditional Chinese medicine sample to be detected to obtain the sample to be detected and the standard sample, wherein the pre-treating condition is normal-temperature saturation treatment;

2. The method for identifying the high-volatility traditional Chinese medicinal material in situ by using the surface acoustic wave gas chromatograph as claimed in claim 1, wherein the method comprises the following steps: the normal temperature saturation treatment in the step (1) or the step (2) comprises the specific process that 20-1000mg of traditional Chinese medicine samples are placed into a 10-40ml headspace bottle and saturated for 1-60min at normal temperature.

3. The method for identifying the high-volatility traditional Chinese medicinal material in situ by using the surface acoustic wave gas chromatograph as claimed in claim 1, wherein the method comprises the following steps: in the step (1), the step (3) or the step (4), the testing process of the surface acoustic wave gas chromatograph is as follows: in a sample introduction state, connecting a sampling port with a sample to be detected, starting a sampling pump, and sucking the sample to be detected into a pre-concentration pipe; and in the detection state, switching the six-way valve to enable the carrier gas to flow through the pre-concentration pipe and carry the sample to be detected in the pre-concentration pipe to enter the chromatographic column and the detector in sequence to finish detection.

4. The method for identifying the high-volatility traditional Chinese medicinal material in situ by using the surface acoustic wave gas chromatograph as claimed in claim 1 or 3, wherein the surface acoustic wave gas chromatographic column is selected from DB-5, SPB-5, Rtx-5, BP-5, OV-5, 007-2(MPS-5), SE-52, SE-54, XTI-5, PTE-5, ZB-5, AT-5, MDN-5 or ZB-5, and the length of the chromatographic column is 1-10 meters.

5. The method for identifying the high-volatility traditional Chinese medicinal material in situ by using the surface acoustic wave gas chromatograph as claimed in claim 4, wherein the method comprises the following steps: the detection conditions of the surface acoustic wave gas chromatography are as follows: the initial temperature of the chromatographic column is 40-50 ℃, and the temperature of the chromatographic column is increased to 200 ℃ according to a temperature increasing program of 0-20 ℃/s in the detection process; the flow rate of the chromatographic column is 1-7mL/min, and the carrier gas is nitrogen or helium.

6. The method for identifying the high-volatility traditional Chinese medicinal material in situ by using the surface acoustic wave gas chromatograph as claimed in claim 1 or 3, wherein the method comprises the following steps: and under the sample introduction state of the surface acoustic wave gas chromatograph, the working time of the sampling pump is 5-60 s.

7. The method for identifying the high-volatility traditional Chinese medicinal material in situ by using the surface acoustic wave gas chromatograph as claimed in claim 1 or 3, wherein the method comprises the following steps: the detection conditions of the surface acoustic wave gas chromatograph are as follows: the temperature of the detector is 25-60 ℃.

8. The method for identifying the high-volatility traditional Chinese medicinal material in situ by using the surface acoustic wave gas chromatograph as claimed in claim 1, wherein the method comprises the following steps: in the step (4), a similarity evaluation or principal component analysis method is used for comparing the sample spectrum with the standard fingerprint spectrum obtained in the step (3) so as to identify the authenticity of the traditional Chinese medicine to be detected.