As the growers of Chinese herbal medicines lack the knowledge about the rational use of pesticides, and the pesticide market is filled with many fake and shoddy products, growers use long-term high-toxicity and high-residue pesticides. Many of the above reasons have caused many highly toxic and high-residue pesticides in Chinese herbal medicines. In the residue, which seriously restricts the Chinese herbal medicines to the international herbal market. However, it is encouraging to note that the 5 State Administration of Drug Administration promulgated on March 18, 2002 and implemented on June 1 5 Quality Management Practices for Producing Chinese Medicinal Plants (Trial)6 and the Ministry of Foreign Trade and Economic Cooperation in July 2001. The implementation of the 5 green industrial standards for the import and export of medicinal plants and preparations on the 1st will surely make China's medicinal materials production lawful and will certainly promote the modernization and internationalization of Chinese medicine.
1 Problems with pesticide residues in medicinal herbs The quality of Chinese herbal medicines directly affects the quality of Chinese herbal medicines. In addition to the active ingredients of Chinese herbs, the quality standards of Chinese herbal medicines also include the pollution of chemical pesticides and heavy metals. The issue of pesticide residues in medicinal herbs is increasingly attracting the attention of countries in the world. Since 1970, the issue of pesticide residues in medicinal herbs has been studied internationally. In 1980, the World Health Organization listed the pesticide residue determination as a test item, which has become a worldwide research hotspot in recent years. Egyptian scholars analyzed organochlorine and organophosphorus pesticides in 303 samples of 20 species of plants. Since 1986, researchers in our country have gradually carried out research on pesticide residue testing, and have gradually stipulated the limit standards for pesticide residues in various Chinese medicines. Li Qingmin et al measured the content of 666 pesticides in ginseng. In addition to determining the content of 666 ginseng, the content of DDT was also determined. The residues of pesticides (BHC, DDT) in five medicinal materials such as safflower were analyzed. Han Guiru et al [analyzed residues of organochlorine pesticides and organophosphorus pesticides in Chinese medicinal materials such as honeysuckle, gardenia, Daqingye, Zhimu and Codonopsis pilosula; established gas chromatograms for determination of organochlorine pesticides in American ginseng, astragalus and licorice law. The contents of various organochlorine pesticides in Chinese medicinal materials such as licorice and astragalus were analyzed by capillary gas chromatography. Tian Jinjiao et al. analyzed the residues of organochlorine pesticides in proprietary Chinese medicines. Chen Jianmin et al [to further standardize the method for the determination of organochlorine pesticides in 11 kinds of Chinese medicines and 6 kinds of Chinese patent medicines, and put forward the recommendations of the limited standard according to the analysis results. The Pharmacopoeia published in 2000 has recorded the determination method of organochlorine pesticides, which indicates that the research on pesticide residues in medicinal materials in China has been further standardized and standardized.
2 Types of pesticide residues in Chinese medicines The types of low-toxicity pesticides that can be used in the production of Chinese herbal medicines are stipulated in the production of Chinese herbal medicines. However, due to the physical and chemical properties of the pesticides, the growers of medicinal herbs cannot influence the mechanism of the plant groups cultivated by the traditional Chinese medicinal herbs and their ecological environment. The impact and other factors, the use of some highly toxic and high-residue pesticides (sometimes mixed), to control pests and diseases on herbs. As a result, some highly toxic and highly-residual pesticides remain in Chinese herbal medicines. The types of pesticides that are forbidden to use in the production of Chinese medicinal materials are shown in Table 1.
Among them, organochlorine insecticides used to be the most widely used insecticides in various countries. Because of their large residue, high toxicity and strong pollution, they have received much attention in the research on pesticide residues. The detection of organochlorine pesticides in medicinal materials is mainly composed of BHC, DDT, and PCNB and Aldrin. The organophosphorus pesticide has many ester bonds in its chemical structure and is unstable, so there is less residue in the medicinal material.
3 Pretreatment methods for the detection of pesticide residues in traditional Chinese medicines Due to the wide variety of medicinal materials, the basic composition of various types of medicinal materials is complex, and the predicted pesticide composition is different, and the presence of pesticide homologues, isomers, degradation products, metabolites, etc. . Other chemical contaminants such as heavy metals that are exposed during the production, transportation, and processing of medicinal materials may also accumulate in the medicinal tissues, thereby increasing the difficulty in the analysis of pesticide residues in medicinal materials.
Before the determination of the pesticide residue, there must be a pretreatment method suitable for the Chinese medicinal materials, and the medicinal material is converted into a sample solution for measuring on the machine, which generally includes the pretreatment steps of extraction, purification, and concentration. According to the different types of pesticides tested, different pre-treatment techniques are required for medicinal samples. This requires the majority of scientific research workers to actively explore, and constantly improve the pre-treatment technology, the introduction of various types of pesticides to determine the best pre-treatment technology, such as the extraction method of organochlorine pesticides is mainly based on Steiwandter established sulfonation method.
4 Detection methods for pesticide residue of traditional Chinese medicine Due to the wide variety of pesticides, the chemical structures of various pesticides are different, and different pesticides need to adopt different detection methods. At the same time, if the nature of the chemical constituents in Chinese medicine is similar to that of pesticides, pesticides will The detection interfered. Residues of pesticides in Chinese medicine are in trace levels, all of which determine the difficulty in the detection of pesticides in Chinese medicines. Many scholars are committed to exploring methods for detecting pesticide residues in Chinese medicines. Many scientific and technological companies have also developed various types of pesticide residue detectors for the detection of pesticide residues in Chinese herbal medicines. According to reports in the literature, the existing detection methods for pesticides in traditional Chinese medicines mainly include:
4.1 Gas chromatography (GC) is the main analytical method for the determination of pesticide residues. The GC method still occupies an absolute advantage in the analysis of organochlorine and organophosphorus pesticides. In the past, it was mainly filled columns, and now it is mainly based on capillary columns.
4.2 The use of high performance liquid chromatography (HPLC) in pesticide residue analysis is becoming more and more widely used. It is suitable for the analysis of thermally unstable and highly polar pesticides and their metabolites, and can be used for pre-column extraction, purification and post-column fluorescence derivatization. The reaction is combined with MS and the like, which facilitates the automation of analysis. At the same time, the advent of some new detectors has improved the detection sensitivity of HPLC to some extent. The limitation is that it is mainly used for thermally unstable, volatile ion-type pesticides, and is now commonly used in analysis and determination of carbamate compounds.
4.3 Gas Chromatography-Mass Spectrometry (GC-MS) The combination of GC-MS and GC-MS-MS has matured, and MS has become a common method in pesticide residue analysis. Since tandem mass spectrometry (MS-MS) can reduce the amount of Impact, improve the sensitivity of the instrument, with the development of mass spectrometry, mass spectrometry has become an effective means of analysis of pesticide residues, the use of compound mass spectrometry identification of components is superior to the retention time of the chromatogram, chromatographic retention time and mass fingerprint data on the compound Perform analysis to ensure maximum reliability of the analysis.
4.4 Thin Layer Chromatography (TLC) TLC requires no special equipment, is simple, can analyze multiple samples at the same time, and is often used for the separation and screening of complex mixtures. In addition to the use of special color reagents to observe spot color and characterize by Rf, TLC can be used not only qualitatively but also quantitatively analyze one or more components that are separated from the sample.
4.5 Enzyme linked immunosorbent assay (enzyme linked immunosorbent asaay, ELISA) use of enzyme-labeled antibodies, anti-antibodies or antigens, ELISA reaction between antigen and antibody, relying on colorimetry to determine the amount of pesticide residues. The advantages of this method are its high specificity, high sensitivity, and rapid and simple, which can be quasi-deterministic and quantitative. It is suitable for on-site analysis. The disadvantage is that it is difficult to prepare antibodies, and false positive or false negative phenomena may occur.
4.6 In vivo bioassay (ABBA) luciferin in luminescent bacteria is fluorescing by the action of luciferase when it is involved in aerobic, but when it is exposed to certain toxic compounds, luminescence is weakened, and its attenuation degree is consistent with the concentration of toxicants. The linear relationship can be quantified accordingly. The advantages of this method are simple, fast, cheap, sensitive, and suitable for on-site analysis. The disadvantage is that the linearity of pesticide concentration and luminescence intensity is not accurate enough, and it can only be semi-quantitative. The in-vivo bioassay has been successfully applied to pesticide residue testers.
4.7 Atomic absorption spectrometry is mainly used for the detection of inorganic salt pesticides.
In addition, neutron liquefaction analysis, spectroscopic techniques, biochemical methods, polarography can be used for pesticide determination.
In short, the following matters need to be paid attention to when producing green medicinal herbs. It is necessary to strengthen the publicity of 5 quality management practices for Chinese medicinal materials. The selection criteria for medicinal materials production bases are stipulated, and raw land is generally selected so as to avoid the influence of pesticides left over from years of planting on the quality of the planted medicinal materials. It stipulates the types of chemical pesticides that can be used for the production of traditional Chinese medicinal materials, the classification of toxicity, the time and method of pesticide application, the minimum effective amount of pesticides, and the interval between the last application and the harvesting.
For medicinal plant growers, the propaganda of plant protection knowledge should be strengthened to eliminate the use of highly toxic pesticides and counterfeit and inferior pesticides; education should be guided by ecology, and full use should be made of resistant varieties and natural enemies to minimize the use of pesticides, and gradually Transition to green Chinese herbal medicines that do not use pesticides. Improve equipment and storage conditions for Chinese medicine processing. Avoid the contamination of Chinese herbal medicines during processing and storage.
Gradually establish the detection methods, limit standards and monitoring agencies for pesticide residues in various types of Chinese medicines. With the implementation of the 5 regulations for quality management of Chinese medicinal materials (trial)6, it is believed that the production of Chinese medicinal herbs will surely enter an orderly and healthy development track, so that Chinese herbs will enter the international market with high quality and participate in market competition.
1 Problems with pesticide residues in medicinal herbs The quality of Chinese herbal medicines directly affects the quality of Chinese herbal medicines. In addition to the active ingredients of Chinese herbs, the quality standards of Chinese herbal medicines also include the pollution of chemical pesticides and heavy metals. The issue of pesticide residues in medicinal herbs is increasingly attracting the attention of countries in the world. Since 1970, the issue of pesticide residues in medicinal herbs has been studied internationally. In 1980, the World Health Organization listed the pesticide residue determination as a test item, which has become a worldwide research hotspot in recent years. Egyptian scholars analyzed organochlorine and organophosphorus pesticides in 303 samples of 20 species of plants. Since 1986, researchers in our country have gradually carried out research on pesticide residue testing, and have gradually stipulated the limit standards for pesticide residues in various Chinese medicines. Li Qingmin et al measured the content of 666 pesticides in ginseng. In addition to determining the content of 666 ginseng, the content of DDT was also determined. The residues of pesticides (BHC, DDT) in five medicinal materials such as safflower were analyzed. Han Guiru et al [analyzed residues of organochlorine pesticides and organophosphorus pesticides in Chinese medicinal materials such as honeysuckle, gardenia, Daqingye, Zhimu and Codonopsis pilosula; established gas chromatograms for determination of organochlorine pesticides in American ginseng, astragalus and licorice law. The contents of various organochlorine pesticides in Chinese medicinal materials such as licorice and astragalus were analyzed by capillary gas chromatography. Tian Jinjiao et al. analyzed the residues of organochlorine pesticides in proprietary Chinese medicines. Chen Jianmin et al [to further standardize the method for the determination of organochlorine pesticides in 11 kinds of Chinese medicines and 6 kinds of Chinese patent medicines, and put forward the recommendations of the limited standard according to the analysis results. The Pharmacopoeia published in 2000 has recorded the determination method of organochlorine pesticides, which indicates that the research on pesticide residues in medicinal materials in China has been further standardized and standardized.
2 Types of pesticide residues in Chinese medicines The types of low-toxicity pesticides that can be used in the production of Chinese herbal medicines are stipulated in the production of Chinese herbal medicines. However, due to the physical and chemical properties of the pesticides, the growers of medicinal herbs cannot influence the mechanism of the plant groups cultivated by the traditional Chinese medicinal herbs and their ecological environment. The impact and other factors, the use of some highly toxic and high-residue pesticides (sometimes mixed), to control pests and diseases on herbs. As a result, some highly toxic and highly-residual pesticides remain in Chinese herbal medicines. The types of pesticides that are forbidden to use in the production of Chinese medicinal materials are shown in Table 1.
Among them, organochlorine insecticides used to be the most widely used insecticides in various countries. Because of their large residue, high toxicity and strong pollution, they have received much attention in the research on pesticide residues. The detection of organochlorine pesticides in medicinal materials is mainly composed of BHC, DDT, and PCNB and Aldrin. The organophosphorus pesticide has many ester bonds in its chemical structure and is unstable, so there is less residue in the medicinal material.
3 Pretreatment methods for the detection of pesticide residues in traditional Chinese medicines Due to the wide variety of medicinal materials, the basic composition of various types of medicinal materials is complex, and the predicted pesticide composition is different, and the presence of pesticide homologues, isomers, degradation products, metabolites, etc. . Other chemical contaminants such as heavy metals that are exposed during the production, transportation, and processing of medicinal materials may also accumulate in the medicinal tissues, thereby increasing the difficulty in the analysis of pesticide residues in medicinal materials.
Before the determination of the pesticide residue, there must be a pretreatment method suitable for the Chinese medicinal materials, and the medicinal material is converted into a sample solution for measuring on the machine, which generally includes the pretreatment steps of extraction, purification, and concentration. According to the different types of pesticides tested, different pre-treatment techniques are required for medicinal samples. This requires the majority of scientific research workers to actively explore, and constantly improve the pre-treatment technology, the introduction of various types of pesticides to determine the best pre-treatment technology, such as the extraction method of organochlorine pesticides is mainly based on Steiwandter established sulfonation method.
4 Detection methods for pesticide residue of traditional Chinese medicine Due to the wide variety of pesticides, the chemical structures of various pesticides are different, and different pesticides need to adopt different detection methods. At the same time, if the nature of the chemical constituents in Chinese medicine is similar to that of pesticides, pesticides will The detection interfered. Residues of pesticides in Chinese medicine are in trace levels, all of which determine the difficulty in the detection of pesticides in Chinese medicines. Many scholars are committed to exploring methods for detecting pesticide residues in Chinese medicines. Many scientific and technological companies have also developed various types of pesticide residue detectors for the detection of pesticide residues in Chinese herbal medicines. According to reports in the literature, the existing detection methods for pesticides in traditional Chinese medicines mainly include:
4.1 Gas chromatography (GC) is the main analytical method for the determination of pesticide residues. The GC method still occupies an absolute advantage in the analysis of organochlorine and organophosphorus pesticides. In the past, it was mainly filled columns, and now it is mainly based on capillary columns.
4.2 The use of high performance liquid chromatography (HPLC) in pesticide residue analysis is becoming more and more widely used. It is suitable for the analysis of thermally unstable and highly polar pesticides and their metabolites, and can be used for pre-column extraction, purification and post-column fluorescence derivatization. The reaction is combined with MS and the like, which facilitates the automation of analysis. At the same time, the advent of some new detectors has improved the detection sensitivity of HPLC to some extent. The limitation is that it is mainly used for thermally unstable, volatile ion-type pesticides, and is now commonly used in analysis and determination of carbamate compounds.
4.3 Gas Chromatography-Mass Spectrometry (GC-MS) The combination of GC-MS and GC-MS-MS has matured, and MS has become a common method in pesticide residue analysis. Since tandem mass spectrometry (MS-MS) can reduce the amount of Impact, improve the sensitivity of the instrument, with the development of mass spectrometry, mass spectrometry has become an effective means of analysis of pesticide residues, the use of compound mass spectrometry identification of components is superior to the retention time of the chromatogram, chromatographic retention time and mass fingerprint data on the compound Perform analysis to ensure maximum reliability of the analysis.
4.4 Thin Layer Chromatography (TLC) TLC requires no special equipment, is simple, can analyze multiple samples at the same time, and is often used for the separation and screening of complex mixtures. In addition to the use of special color reagents to observe spot color and characterize by Rf, TLC can be used not only qualitatively but also quantitatively analyze one or more components that are separated from the sample.
4.5 Enzyme linked immunosorbent assay (enzyme linked immunosorbent asaay, ELISA) use of enzyme-labeled antibodies, anti-antibodies or antigens, ELISA reaction between antigen and antibody, relying on colorimetry to determine the amount of pesticide residues. The advantages of this method are its high specificity, high sensitivity, and rapid and simple, which can be quasi-deterministic and quantitative. It is suitable for on-site analysis. The disadvantage is that it is difficult to prepare antibodies, and false positive or false negative phenomena may occur.
4.6 In vivo bioassay (ABBA) luciferin in luminescent bacteria is fluorescing by the action of luciferase when it is involved in aerobic, but when it is exposed to certain toxic compounds, luminescence is weakened, and its attenuation degree is consistent with the concentration of toxicants. The linear relationship can be quantified accordingly. The advantages of this method are simple, fast, cheap, sensitive, and suitable for on-site analysis. The disadvantage is that the linearity of pesticide concentration and luminescence intensity is not accurate enough, and it can only be semi-quantitative. The in-vivo bioassay has been successfully applied to pesticide residue testers.
4.7 Atomic absorption spectrometry is mainly used for the detection of inorganic salt pesticides.
In addition, neutron liquefaction analysis, spectroscopic techniques, biochemical methods, polarography can be used for pesticide determination.
In short, the following matters need to be paid attention to when producing green medicinal herbs. It is necessary to strengthen the publicity of 5 quality management practices for Chinese medicinal materials. The selection criteria for medicinal materials production bases are stipulated, and raw land is generally selected so as to avoid the influence of pesticides left over from years of planting on the quality of the planted medicinal materials. It stipulates the types of chemical pesticides that can be used for the production of traditional Chinese medicinal materials, the classification of toxicity, the time and method of pesticide application, the minimum effective amount of pesticides, and the interval between the last application and the harvesting.
For medicinal plant growers, the propaganda of plant protection knowledge should be strengthened to eliminate the use of highly toxic pesticides and counterfeit and inferior pesticides; education should be guided by ecology, and full use should be made of resistant varieties and natural enemies to minimize the use of pesticides, and gradually Transition to green Chinese herbal medicines that do not use pesticides. Improve equipment and storage conditions for Chinese medicine processing. Avoid the contamination of Chinese herbal medicines during processing and storage.
Gradually establish the detection methods, limit standards and monitoring agencies for pesticide residues in various types of Chinese medicines. With the implementation of the 5 regulations for quality management of Chinese medicinal materials (trial)6, it is believed that the production of Chinese medicinal herbs will surely enter an orderly and healthy development track, so that Chinese herbs will enter the international market with high quality and participate in market competition.
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