Rapid detection of aflatoxin B_1 in Chinese herbal medicines by indirect and direct competitive enzyme-linked immunosorbent assays: a comparative analysis / 中国中药杂志

The indirect competitive enzyme-linked immunosorbent assay(ic-ELISA) and direct competitive enzyme-linked immunosorbent assay(dc-ELISA) were performed for the rapid detection of aflatoxin B_1(AFB_1) in Astragali Radix, Zingiberis Rhizoma Recens, Jujubae Fructus, and Nelumbinis Semen with self-made antigens and antibodies. Different extraction methods were investigated to reduce the matrix effects of different medicinal parts in Chinese herbal medicines. The sensitivity of dc-ELISA method was improved by optimizing the molar ratio of AFB_1 to horseradish peroxidase(HRP). In this study, the sensitivity(IC_(50)) of ic-ELISA and dc-ELISA was 0.046 and 0.023 ng·mL~(-1), with the limit of detection(LOD) of 0.007 and 0.004 ng·mL~(-1), respectively. The detection time was 3 h and 50 min for ic-ELISA and dc-ELISA, respectively. The recovery rates were within the range of 62.96%-104.4%, with RSDs of less than 10%. Confirmed by LC-MS/MS, three positive samples of Nelumbinis Semen were detected from 53 samples. Two ELISA methods established in this study were accurate, rapid and sensitive, and can be used for rapid screening of AFB_1 in Chinese herbal medicines such as Astragali Radix, Zingiberis Rhizoma Recens, Jujubae Fructus, and Nelumbinis Semen. In addition, the advantages and limitations of the two methods were compared and discussed, which can provide a reference for the testing institutions to choose the proper method.

Overview of quality standards of Eucommiae Cortex in China and overseas / 中国中药杂志

Eucommiae Cortex is an authentic medicinal material with broad growing areas( such as Hunan and Sichuan provinces in China. It is well-known for its efficacy in tonifying liver and kidney,strengthening muscles and bones,and stabilizing fetus. It has also been proven in pharmacology to possess the functions such as lowering blood pressure and lipids. Hence,Eucommiae Cortex has attracted increasing attention. The current quality standards of Eucommiae Cortex vary in different countries or regions. The quality of Eucommiae Cortex products on the market is affected by mix-ups of non-medicinal parts and insufficient growth years. In view of these problems,this paper summarizes the current quality standards and research progress of Eucommiae Cortex in China and overseas,aiming to provide a reference for the establishment of the quality standards of Eucommiae Cortex.

Cloning of AhWRKY33 from Asarum heterotropoides and its genetic transformation in Arabidopsis / 中国中药杂志

The WRKY family genes, which play an important role in plant morphogenesis and stress response, were selected based on the data of the full-length transcriptome of Asarum heterotropoides. Using AtWRKY33, which regulates the synthesis of the camalexin in the model plant Arabidopsis to compare homologous genes in A. heterotropoides, primers were designed to amplify the open reading frame(ORF) fragment of AhWRKY33 gene by RT-PCR using total RNA of A. heterotropoides leaves as template. Real-time PCR results showed that there was a significant difference between the aerial part and the underground part of A. heterotropoides, the toxic aristolochic acid content is highly expressed in the leaves higher than the root. After verification, the WRKY33 gene of A. heterotropoides is ORF long 1 686 bp, encoding 561 amino acids.AhWRKY33 had two conserved WRKYGQK domains. According to the classical classification, it belongs to group Ⅰ WRKY transcription factor. A. heterotropoides WRKY33 had some homology with amino acids of other species. The study successfully constructed the plant eukaryotic expression vector PHG-AhWRKY33 and transformed Arabidopsis thaliana, the transgenic Arabidopsis was obtained by PCR detection and hygromycin resistant plate screening. It found that the germination of transgenic Arabidopsis seeds was accelerated and the stress resistance was increased. It laid a foundation for further analysis of WRKY transcription factor in the growth and development of A. heterotropoides and the synthesis of secondary metabolites.

QuEchERS pretreatment coupled to gas chromatography and tandem mass spectrometry to fast determination of 34 pesticide residues in Glycyrrhizae Radix et Rhizoma / 中国中药杂志

This study proposed a quantitative method for 34 pesticides including organochlorine,organophosphorus and pyrethroids in Glycyrrhizae Radix et Rhizoma herbs and medicinal slices,and analyzed the pesticide residues of collected Glycyrrhizae Radix et Rhizoma samples from different regions. With acetonitrile extraction and optimized Qu Ech ERS purification,the 32 batches of Glycyrrhizae Radix et Rhizoma herbs and medicinal slices were analyzed by matrix matching standard curve quantitative analysis under GC-MS/MS multi-response monitoring( MRM) mode. This study investigated the pretreatment of Glycyrrhizae Radix et Rhizoma samples based on the Qu Ech ERS method of Chinese Pharmacopoeia( 2015 edition,4),and the result showed that the recoveries of some pesticide was low and pigment has a strong interference in analysis,which result in worse purification effect. Therefore,this paper further optimized the Qu Ech ERS method and corrected the matrix matching standard curve method,and compensated the qualitative and quantitative effects of matrix effects on the detected target compounds in Glycyrrhizae Radix et Rhizoma. The results showed that 34 kinds of pesticide had good linear( R~2 of 0. 996 4 or higher) within a covering 0. 01-0. 2 mg·kg~(-1) concentration range. The limits of quantitation are less than 0. 01 mg·kg~(-1). This method was further applied to the simultaneous determination of 34 pesticide residues of typical organochlorine,organophosphorus and pyrethroids in 32 batches of Glycyrrhizae Radix et Rhizoma herbs and medicinal slices. Six batches containing beta-endosulfan,thiosulphate,o,p'-DDD and thrta-cypermethrin were detected,but none of them exceeded the limit of pesticide residues stipulated in the Chinese Pharmacopoeia and the EU Pharmacopoeia. This study indicates that the established method is rapid,convenient,accurate,and sensitive,which provides a rapid and efficient method for the simultaneous determination of typical organochlorine,organophosphorus and pyrethroids in Glycyrrhizae Radix et Rhizoma.

Survey on pesticide use in Crataegi Fructus and analysis of pesticide residues based on LC-ESI-MS/MS / 中国中药杂志

In order to study the pesticide residues of the medicinal Crataegi Fructus,this study aims to establish an analysis method for pesticide residues( mainly containing insecticides and fungicides) suitable for the actual situation of medicinal Crataegi Fructus based on the survey of the pesticides of the Crataegi Fructus base,combined with the blind screening results of the LC-ESI-MS/MS pesticide screening platform established by the research team in the early stage. Then,the pesticide residues in medicinal Crataegi Fructus from Shandong,Hebei,Henan,Shanxi,and Liaoning( main cultivation areas) were analyzed. The samples were pretreated by the modified Qu ECh ERS method,i.e.,extracted with acetonitrile-water( 9 ∶1),purified by PSA,C_(18),GCB,silica gel. The detection of pesticides was performed by LC-MS/MS. The ion source was ESI with positive scanning mode,and the linearity of 11 kinds of pesticides in the range of 5-300 μg·kg~(-1) was acceptable( R~2>0. 996 9). All the recoveries of pesticides were within 70. 02%~(-1)12. 0% in the low,medium and high levels,with RSD≤17%. The results showed that the detection rate of carbendazim,chlorpyrifos and difenoconazole is 79%,82%,56%,respectively. Besides,the prohibition pesticide carbofuran were detected in some of the batches,indicating the security risk. This study provides methodological references and basic data for risk assessment of Crataegi Fructus and government regulation.

Simultaneous determination of aflatoxin B_1,B_2,G_1,G_2,M_1,M_2 in Eupolyphaga Steleophaga by immunoaffinity column clean-up and HPLC-FLD with post-column photochemical derivatization / 中国中药杂志

The contamination of aflatoxin B_1,B_2,G_1,G_2,M_1 and M_2 in Eupolyphaga Steleophaga was determined by immunoaffinity column clean-up and HPLC-FLD with post-column photochemical derivatization. Chromatographic separations were carried out using a Cloversil C_(18) column( 4. 6 mm×250 mm,5 μm) that were eluted in isocratic with methanol-acetonitrile-water( 20 ∶ 20 ∶ 60) as the mobile phase. The excitation wavelength and the emission wavelength of fluorescence detector were maintained at 360 nm and 450 nm,respectively. The flow rate was 0. 8 m L·min~(-1),and the column temperature was 30 ℃ ． The sample was prepared using the immunoaffinity column,then the recovery was measured with 75. 47%-101. 8% with RSD values lower than 6. 7%. A total of 20 batches of Eupolyphaga Steleophaga samples were assayed. According to the Chinese Pharmacopoeia( 2015 edition,part 1),the aflatoxin B_1 limit should be less than 5 μg·kg~(-1),and the sum of aflatoxins( AFB_1,AFB2,AFG_1,AFG_2) should be less than 10 μg·kg-1. Therefore,the positive rate of the 20 samples was 50. 0%,and 7 batches of samples exceeded the standard,and the over-standard rate was as high as 70. 0%. Among them,aflatoxins B_1,B_2,G_1,G_2,M_1,and M_2 were detected in three batches( SD-1,AH-1,AH-3),and aflatoxins B_1,B2,G1,G2,and M1 were detected in one batch( AH-7). The results showed that the newly developed method in this work is suitable for the simultaneous determination of six aflatoxins in Eupolyphaga Steleophaga,and also suggested that it should be of high values to take the contamination with aflatoxins into concerns.

Safety evaluation of heavy metals contaminated Xiaochaihu Tang using health risk assessment model / 中国中药杂志

In order to further improve the quality and safety evaluation standards of traditional Chinese medicine compound preparation,Xiaochaihu Tang compound preparations were selected to analyze the pollution level of heavy metals deeply,and the potential health risks were evaluated under taking such kind of compound preparations. In this study,the contents of copper( Cu),arsenic( As),cadmium( Cd),mercury( Hg),and lead( Pb) in different Xiaochaihu Tang compound preparations were determinated by the method of inductively coupled plasma mass spectrometry( ICP-MS). Moreover,combined with target hazard coefficient method and in vitro artificial system,the bioaccessibility and health risk level was investigated in three main consumption ways including powder,decoction and granule. The result was showed that,under the maximum residual limit set by International Standard Organization,only one batch of raw herb was eight times exceeded the Hg MRL,however,in water decoctions and granules,the heavy metal residue rate was reduced to 2. 02%( Hg in granules)-42. 85%( Cd in granules). So,the heavy metal pollutions and health risks can be reduced to safe region in spite of the serious pollution in raw herbs. Besides,the THQ and CR values of the three consumption methods were lower than the standard values of non-carcinogenic and carcinogenic risks of each heavy metal. It can be seen that even if the heavy metals in the raw herbs exceed the standard,the use of Xiaochaihu Tang decoction and granules can reduce the harm of heavy metals to the human body. Above all,the establishment of this health risk assessment model can be provided experimental basis and reference value for improving the safety evaluation standard of other heavy metals contained traditional Chinese medicine( TCM) compound preparations,and further improving the quality control methods of other different toxic compounds in clinical use.

Current situation of heavy metal pollution,detection and removal techniques in medicinal marine organisms in China / 中国中药杂志

As an important branch of traditional medicines,medicinal marine organisms have many advantages,including biological diversity,remarkable biological activity,especial for the treatment of anti-cancer,anti-virus,anti-coagulation,analgesia,anti-bacterial,cardiovascular and cerebrovascular diseases. In recent years,with the continuous exploration of marine organisms by human beings,many marine organisms with specific biological activities and medicinal prospects have been found,which have attracted great attention around the world and thus called " new hope" to solve human health problems. However,due to the rapid development of modern industry,heavy metal pollution not only poses a great threat to medicinal marine living resources,but also hinders the development of marine biomedical industry and threatens human health. In view of this,this paper introduced the development trend of medicinal marine organisms and the current situation of heavy metal pollution and focusing on the analysis technology and chemical removal technology of heavy metals in medicinal marine organisms,which is to provide reference for the heavy metals control in marine medicines and the development and utilization of marine medicines.

Rapid Immunoassay of Aflatoxin B1 and Its Application in Traditional Chinese Medicine / 中国实验方剂学杂志

Aflatoxin B1 (AFB1) is a toxic secondary metabolite produced by toxigenic Aspergillus spp. (such as A. flavus and A. parasiticus) with carcinogenic,teratogenic and mutagenic effects. Studies have shown that AFB1 is widely found in crops,food,feed and traditional Chinese medicine,which poses a serious safety hazards to humans healthy. The establishment of a rapid detection technique that is suitable for AFB1 in different matrices has a great significance in preventing contamination,controlling food and medicines safety and ensuring human health. With the continuous improvement of small-molecule immune technology,various rapid immunoassays of AFB1 have been developed and utilized in recent years. This review systematically summarized current relevant standards for the detection of AFB1 in China and the maximum recommended levels for the application of aflatoxins in Chinese herbal medicines in some regions and countries. These standards are mainly applicable for aflatoxins in food,feed and some easily contaminated samples of Chinese herbal medicines. Some studies have shown that except the Chinese herbal medicines specified with the maximum recommended levels,some medicinal herbs and their products were also contaminated by aflatoxins. In addition,this paper reviewed the preparation technology of antigen and antibody for AFB1,and the rapid detection methods based on the specific recognition ability between antigen and antibody, including enzyme-linked immunosorbent assay,immunochromatographic assay,fluorescence immunoassay,chemiluminescence immunoassay,and novel immunosensor method, were also summarized and compared. This review aims to provide the reference for rapid,accurate,and sensitive technical standards for the detection of AFB1 in traditional Chinese medicine during the agricultural planting,distribution, trade and quality supervision and for the market, pharmacies and hospitals,so as to ensure the quality and safety of the traditional Chinese medicines.

Recent research progress on natural medicines in treatment of cadmium toxicity / 中国中药杂志

Cadmium contamination of environment is a subject of serious international concern. Bioaccumulation of cadmium occurs primarily through ingestion of contaminated water and food. Cadmium poisoning came into prominence with the "itai-itai" disease event in Japan in the 1950s. It could also cause damages to liver, kidney, lung and other organs. Thus, the treatment of cadmium poisoning has become a research hotspot. Researchers are trying their best to explore prophylactic and therapeutic medicines for prevention and treatment of cadmium-induced poisoning. So far, chelation therapy, the conventional treatment for heavy metal toxicity, is reported to have a number of safety and efficacy issues. Natural medicines have a variety of advantages such as extensive sources, high safety, less adverse reactions, and thus have great potentials in treating cadmium poisoning. In this review, the progress in the antagonistic effects of natural drugs in cadmium poisoning and their therapeutic mechanisms were summarized in order to provide certain references for the future development and in-depth study of antagonistic substances.

Identification and quality evaluation of Tripterygium wilfordii / 中国中药杂志

With the extensive clinical application of Tripterygium wilfordii, there are many counterfeit products on the market. Traditional technology can not effectively identify the authenticity of the traditional Chinese medicine. Therefore, a strategy of accurate identification and quality evaluation of Tripterygium based on DNA barcode and chemical fingerprint spectrum was established. Based on DNA barcode technology, HMMer annotation method of hidden Markov model and K2P model were used to analyze genetic distance.BLAST1, nearest distance and phylogenetic tree (NJ-tree) methods were used to assess the identification efficiency of the ITS2 barcode. The fingerprint of 27 T. wilfordii was established by UPLC-PDA method, and the similarity of the fingerprint of different sources was evaluated. The main components of T. wilfordii were determined by LC-MS/MS. The results revealed that the intraspecific genetic distances of T. wilfordii were lower than the interspecific genetic distances between T. wilfordii and its adulterants. The results of similarity search showed that ITS2 sequence was used to identify T. wilfordii and its adulterants. The clustering of T. wilfordii and its adulterants was clear in the tree of NJ cluster, and 12 of 27 samples were identified as true T. wilfordii.The chemical fingerprint spectrum research indicates that the feature one region can distinguish the false product of tripterygium glycosides more intuitively. The cluster analysis of HCA-thermal map showed that the contents of six active components of T. wilfordii from different habitats were significantly different, which could be used to evaluate the quality of T. wilfordii. This paper is of guiding significance for the accurate identification and quality evaluation of Tripterygium medicinal plants.

Recent advances in analysis of endogenous toxic components in traditional Chinese medicines / 中国中药杂志

Endogenous toxic components have become an important topic in the field of traditional Chinese medicines (TCMs). Since the endogenous toxic components in TCMs are often used as clinical effective components, the safety and effectiveness of endogenous toxic substances has become an important part of the research of TCMs. In this paper, the classification and evaluation criteria of toxic Chinese medicinal materials are described, and the analytical methods of endogenous components in TCMs are summarized and expounded base on with the techniques of chromatography, spectroscopy, immunoassay, and so on. On this basis, the problems in terms of endogenous toxic components are analyzed and discussed. This paper could provide ideas and methods for the evaluation of the validity and safety of TCMs containing endogenous toxic components.

Research progress of rapid detection of ochratoxin A and its application prospect in traditional Chinese medicine / 中国中药杂志

Ochratoxin A (OTA) is a toxic secondary metabolite mainly produced by Aspergillus and Penicillium species, with strong renal toxicity, teratogenic, carcinogenic, mutagenic effect. Studies have shown that OTA is not only widely contaminated in food and feed crops, but also has been widely contaminated in Chinese herbal medicines such as spices, licorice and so on. In view of OTA's universality and harmfulness, this paper summarizes the flow visualization test strip, microsphere, electrochemical sensor, surface enhanced Raman spectroscopy technology in OTA rapid detection, which provides reference for the research and application of high throughout detection instrument miniaturization in order to achieve OTA quick detection and simple operation.

Application of natural plant pigments in enlarged health industry / 中国中药杂志

Natural plant pigment is rich in resources, with the features of natural color and environment friendly, which has a broad space for development and market prospects. In order to further develop and utilize of natural plant pigment, this paper mainly introduces the natural plant dyes in the domains of food, cosmetics and health care products on the historical development process and their application. In addition,this paper summarizes the application of representative natural plant pigment, dyes, and prospects the market of natural plant pigment, so as to provide reference for the development of natural plant pigment in the enlarged health industry of China.

Study on essential oils of medicinal plants in insect repellent / 中国中药杂志

Mosquitoes are seriously harmful to human health for transmitting some mortal diseases. Among the methods of mosquito control, synthetical insecticides are the most popular. However, as a result of longterm use of these insecticides, high resistant mosquitos and heavy environmental pollution appear. Thus, eco-friendly prevention measures are taken into the agenda. Essential oils extracted from medicinal plants have repellent and smoked killing effects on mosquitoes. With abundant medical plants resources and low toxicity, they have the potential of being developed as a new type of mosquito and insect repellent agent. The recent application advances of essential oils of medicinal plants in insect repellent and its application limitations are overviewed. This review will provide references for the future development and in-depth study of essential oils.

Application of natural plant pigment in hair dyes / 中国中药杂志

With the development of living condition, more and more people tend to show unique personality, thus hair dyes as hair cosmetics are highly favored. By the year 2012, the global sales of hair dye had exceeded $15 billion, with a sustained growth at a rate of 8%-10% annually. However, the harm caused by long-term use of hair dyes has aroused widespread public concern, so people begin to seek non-toxic or low toxic natural plant hair dyes. The types of commonly used hair dyes and the corresponding dyeing mechanisms were summarized in this manuscript, and the representative natural botanic dyes were listed. Thereafter, their effective fractions, constituents and application status were described. In addition, the values of botanic hair dyes and their broad market prospect were discussed. Finally, the problems that exist in the research and development of plant hair dyes were issued. This review may help to provide thought for developing novel, green and ecological natural plant hair dyes.

Advances of toxicity evaluation of cinnabar and compatibility necessity analysis / 中国中药杂志

Cinnabars are widely used in the clinic and shows unique efficacy, and it has been used in treating diseases for thousands of years either single-use or combination with other traditional Chinese medicines. Cinnabar mainly contains mercury, which displays obvious hepatotoxicity and nephrotoxicity. The safety of cinnabar and its rational use caused extensive attention. However, there are some misunderstanding that cinnabar shows equivalent toxicity to the toxicity of mercury compounds such as mercuric chloride and methylmercury in safety evaluation of cinnabar, which in our view exaggerates the toxicity. For traditional Chinese medicine containing cinnabar, some researchers questioned the medicinal value of cinnabar, and even proposed to remove cinnabar in order to avoid possible poisoning. This review reported adverse events, summarized its toxicity characteristics, mechanism and the medicinal value in preparations of traditional Chinese medicine. The significance of this paper is to further understand the toxicity of cinnabar and reduce its side effect in clinical application, and to provide a scientific basis for reasonable clinical application and a scientific understanding of cinnabar.

Study on species and valence state of heavy metals and deleterious elements of mineral medicine / 中国中药杂志

As an important part of traditional Chinese medicine (TCM), mineral medicine plays an irreplaceable role. However, little has been reported on its species and valence state of heavy metals and deleterious elements, and also the relevance to pharmacological effect and toxicology. The present paper, in a new perspective, summarized the determination of the species and valence state of heavy metals and deleterious elements in recent years, discussed the progress of the pharmacological effect and toxicology, and prospected for future study which might provide reference for mineral medicine.

Research progress of Chinese herbal medicine raw materials in cosmetics / 中国中药杂志

Advocating green, nature, environmental protection, safety and the pursuit of efficacy are the trends of cosmetics in the world. In recent years, more and more Chinese herbal extracts with mild, high safety and small irritation are applied to cosmetics as the natural additives. This has become a new hot spot. The recent application advances of Chinese medicine raw materials in cosmetics are overviewed according to their main functions. This review will provide useful references for the future development and application of Chinese medicinal herbs cosmetics.

Sangxingtang inhibits the inflammation of LPS-induced acute lung injury in mice by down-regulating the MAPK/NF-κB pathway / 中国天然药物

In the present study, we investigated anti-inflammatory effects of Sangxingtang (SXT) on acute lung injury using a lipopolysaccharide (LPS)-induced acute lung injury (ALI) mouse model. The cell counting in the bronchoalveolar lavage fluid (BALF) was performed. The degree of lung edema was evaluated by measuring the wet/dry weight (W/D) ratio. The superoxidase dismutase (SOD) and myeloperoxidase (MPO) activities were assayed by SOD and MPO kits, respectively. The levels of inflammatory mediators, including tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), were assayed by the enzyme-linked immunosorbent assay methods. Pathological changes of lung tissues were observed by Hematoxylin and eosin (HE) staining. The inflammatory signaling pathway-related proteins nuclear factor mitogen activated protein kinases (P38MAPK), extracellular regulated protein kinases (Erk), c-Jun N-terminal kinase (Jnk) and nuclear transcription factor (NF-κB) p65 expressions were measured by Western blotting. Our results showed that the treatment with the SXT markedly attenuated the inflammatory cell numbers in the BALF, decreased the levels of P-P38MAPK, P-Erk, P-Jnk and P-NF-κB p65 and the total protein levels in lungs, improved the SOD activity and inhibited the MPO activity. Histological studies demonstrated that SXT substantially reduced the LPS-induced neutrophils in lung tissues, compared with the untreated LPS group. In conclusion, our results indicated that SXT had protective effects on LPS-induced ALI in mice.