Identification of Dalbergia odorifera and Its Counterfeits by HS-GC-MS / 中国实验方剂学杂志

ObjectiveTo screen the differential markers by analyzing volatile components in Dalbergia odorifera and its counterfeits， in order to provide reference for authentication of D. odorifera. MethodThe volatile components in D. odorifera and its counterfeits were detected by headspace gas chromatography-mass spectrometry（HS-GC-MS）， and the GC conditions were heated by procedure（the initial temperature of the column was 50 ℃， the retention time was 1 min， and then the temperature was raised to 300 ℃ at 10 ℃ for 10 min）， the carrier gas was helium， and the flow rate was 1.0 mL·min-1， the split ratio was 10∶1， and the injection volume was 1 mL. The MS conditions used electron bombardment ionization（EI） with the scanning range of m/z 35-550. The compound species were identified by database matching， the relative content of each component was calculated by the peak area normalization method， and principal component analysis（PCA）， orthogonal partial least squares-discrimination analysis（OPLS-DA） and cluster analysis were performed on the detection results by SIMCA 14.1 software， and the differential components of D. odorifera and its counterfeits were screened out according to the variable importance in the projection（VIP） value>2 and P<0.05. ResultA total of 26， 17， 8， 22， 24 and 7 volatile components were identified from D. odorifera， D. bariensis， D. latifolia， D. benthamii， D. pinnata and D. cochinchinensis， respectively. Among them， there were 11 unique volatile components of D. odorifera， 6 unique volatile components of D. bariensis， 3 unique volatile components of D. latifolia， 6 unique volatile components of D. benthamii， 8 unique volatile components of D. pinnata， 4 unique volatile components of D. cochinchinensis. The PCA results showed that， except for D. latifolia and D. cochinchinensis， which could not be clearly distinguished， D. odorifera and other counterfeits could be distributed in a certain area， respectively. The OPLS-DA results showed that D. odorifera and its five counterfeits were clustered into one group each， indicating significant differences in volatile components between D. odorifera and its counterfeits. Finally， a total of 31 differential markers of volatile components between D. odoriferae and its counterfeits were screened. ConclusionHS-GC-MS combined with SIMCA 14.1 software can systematically elucidate the volatile differential components between D. odorifera and its counterfeits， which is suitable for rapid identification of them.

Pharmacognostical Study of Leaf of Hamsapadi (Adiantum Lunulatum)

Pharmacognostical evaluation of leaves of Adiantum lunulatum is done for identification in field and for differentiation from other species of Adiantum. Methods: A detailed literary work of drug studied and its pharmacognostical, phytochemical and analytical study including powder microscopy and taste determination of leaves with suitable instruments. Results: Macroscopic evaluation reveals the pinnacles are half moon shaped, alternate, sub-opposite or opposite in apical portion of the frond, having lobes with minor incisions. In microscopic structure the two surfaces of pinna are covered by epidermis and epidermal cells are wavy in outline. The opening and closing of stomata will be regulated by guard cells, whereas there is absence of stomata on epidermis. The vascular cells are lined by an endodermal tissue. Transverse section of petiole showed that it is externally covered by an epidermis lined by cuticle on its outer wall which renders the petiole to be shining. Below the epidermis is a fairly thick hypodermis made up Sclerenchyma cells of 4-5 layers in thickness. They are dead cells compact in arrangement and look dark in colour. Powder microscopy of leaves reveals shows few Tracheids of the veins and the Parenchyma cells of the leaves. The Sori are very prominent structures occurring at the leaf margin, they are visible characteristically under the microscope. The Sori contain number of Sporangia each Sporangium is lined by cells with U shaped thickening having their outer wall thin in nature. Conclusion: This study will help in Authentication of Hamspadi plant.

Rapid authentication of different herbal medicines by heating online extraction electrospray ionization mass spectrometry / 药物分析学报

The rapid and accurate authentication of traditional Chinese medicines(TCMs)has always been a key scientific and technical problem in the field of pharmaceutical analysis.Herein,a novel heating online extraction electrospray ionization mass spectrometry(H-oEESI-MS)was developed for the rapid and direct analysis of extremely complex substances without the requirement for any sample pretreatment or pre-separation steps.The overall molecular profile and fragment structure features of various herbal medicines could be completely captured within 10-15 s,with minimal sample(＜0.5 mg)and solvent consumption(＜20 μL for one sample).Furthermore,a rapid differentiation and authentication strategy for TCMs based on H-oEESI-MS was proposed,including metabolic profile characterization,characteristic marker screening and identification,and multivariate statistical analysis model validation.In an analysis of 52 batches of seven types of Aconitum medicinal materials,20 and 21 key compounds were screened out as the characteristic markers of raw and processed Aconitum herbal medicines,respectively,and the possible structures of all the characteristic markers were comprehensively identified based on Com-pound Discoverer databases.Finally,multivariate statistical analysis showed that all the different types of herbal medicines were well differentiated and identified(R2X＞0.87,R2Y＞0.91,and Q2＞0.72),which further verified the feasibility and reliability of this comprehensive strategy for the rapid authentication of different TCMs based on H-oEESI-MS.In summary,this rapid authentication strategy realized the ultra-high-throughput,low-cost,and standardized detection of various complex TCMs for the first time,thereby demonstrating wide applicability and value for the development of quality standards for TCMs.

Padronização de uma PCR multiplex para a detecção de fraude em leite bubalino e caprino por adição de leite bovino / Standardization of a multiplex PCR for detection of fraud in buffalo and goat milk by addition of bovine milk

O consumo de leite de espécies como bubalino e caprino tem se popularizado por representarem uma alternativa para indivíduos que possuem restrições alimentares relacionadas ao leite bovino e em virtude das propriedades nutricionais desses alimentos. No entanto, fatores como a baixa produção e a sazonalidade predispõem a adulterações destes alimentos, principalmente pela adição de leite bovino, visando maior rendimento e lucratividade. Assim, o objetivo do estudo foi padronizar um método de PCR multiplex para autenticação de leites bubalino e caprino. Para isso, amostras de leite exclusivamente de cada espécie foram utilizados para a padronização da técnica. Em seguida, foi realizada a fraude pela adição de leite bovino ao caprino e ao bubalino, em proporções de 0,1% até 100%. A técnica foi eficaz, precisa, rápida e prática para a detecção do DNA de bovino, bubalino e caprino, separadamente e em conjunto. Na fraude experimental, o limite de detecção da técnica ocorreu a partir do menor percentual testado (0,1%) tanto no leite caprino quanto no bubalino. Dessa forma, a PCR multiplex testada mostrou ser uma importante ferramenta para a autenticação de leite, pendendo ser utilizada para fins de fiscalização por órgãos competentes.

Milk consumption of species such as buffalo and goat has become popular due to the nutritional properties of these foods and because they represent an alternative for individuals who have dietary restrictions related to bovine milk. However, factors such as low production and seasonality predispose to adulteration, mainly by the addition of bovine milk, aiming at higher yield and profitability. Thus, the aim of the present study was to standard a multiplex PCR method for buffalo and goat milks authentication. For this, the milks exclusively of each species were used to standardize the technique. Subsequently, fraud was performed by the addition of bovine milk to goat and buffalo in proportions from 0.1% to 100%. The technique was effective and accurate for detecting bovine, buffalo and goat DNA separately and together quickly and practically. In experimental fraud, the detection limit of the technique occurred from the lowest percentage tested (0.1%) in both goat and buffalo milk. Thus, the multiplex PCR tested proved to be an important tool for milk authentication, pending to be used for supervision by competent agencies.

Pharmacological Effect of Glycyrrizae Radix et Rhizoma Compatibility and Its Mechanism / 中国实验方剂学杂志

Glycyrrizae Radix et Rhizoma has high medicinal value and is widely used in compatibility. It is used most frequently in the compatibility of Chinese medicine prescriptions，and is known as ''Guolao''（national medicine） and "master of all medicines". The characteristic active ingredients are mainly liquitin，glycyrrizic acid，glycyrrizin，and licochalcone. In different compatibilities，based on traditional and modern pharmacological theories，the corresponding effect of Glycyrrizae Radix et Rhizoma are brought into play through different mechanisms. Based on the traditional pharmacology of Glycyrrizae Radix et Rhizoma for tonifying spleen，replenishing Qi，clearing heat，removing toxin，dispelling phlegm，relieving cough and pain，and harmonizing various medicines，this paper used herbal authentication to analyze its compatibility application and mechanism. It was found that Glycyrrizae Radix et Rhizoma played corresponding effect in compatibilities through "tonification"，"harmonization"，and "regulation". For example，Glycyrrizae Radix et Rhizoma was combined with tonics including Ginseng Radix et Rhizoma and Atractylodis Macrocephalae Rhizoma to tonify the five Zang-organs through its strong tonifying effect，combined with Paeoniae Radix Alba and Aconiti Lateralis Radix Praeparata to relieve emergencies and pains through harmonizing medicine power and properties，and combined with Rhei Radix et Rhizoma and Natrii Sulfas to reduce medicine intensity through regulating medicine properties and body characteristics. The application law and mechanism of the modern pharmacological compatibility of Glycyrrizae Radix et Rhizoma were analyzed by data mining and network pharmacology. It was found that the modern clinical formula was often compatible with Glycyrrizae Radix et Rhizoma for anti-inflammation，cardiovascular and cerebrovascular protection，anti-virus，and anti-tumor，Ephedrae Herba and Scutellariae Radix for anti-inflammation，Bambusae Caulis in Taenias，Salviae Miltiorrhizae Radix et Rhizoma，and Aurantii Fructus Immaturus for cardiovascular and cerebrovascular protection，and Ophiopogonis Radix and Chuanxiong Rhizoma for nerves protection. Meanwhile，the important targets of the characteristic ingredients were protein kinase B1 （Akt1），interleukin-6 （IL-6），tumor necrosis factor （TNF），and epidermal growth factor receptor （EGFR）. The important characteristic pathways such as tyrosine kinase inhibitor resistance pathway and cyclic guanosine monophosphate （cGMP）/protein kinase G （PKG） signal pathway played the role of cardiovascular and cerebrovascular protection，and proteoglycan pathway in cancer played a neuroprotective role. This study is expected to provide references for the rational compatibility and application of Glycyrrizae Radix et Rhizoma，as well as the compatibility application of Chinese medicine prescriptions.

Identification of Armeniacae Semen Amarum and Persicae Semen from different origins based on near infrared hyperspectral imaging technology / 中国中药杂志

In order to establish a rapid and non-destructive evaluation method for the identification of Armeniacae Semen Amarum and Persicae Semen from different origins, the spectral information of Armeniacae Semen Amarum and Persicae Semen in the range of 898-1 751 nm was collected based on hyperspectral imaging technology. Armeniacae Semen Amarum and Persicae Semen from different origins were collected as research objects, and a total of 720 Armeniacae Semen Amarum samples and 600 Persicae Semen samples were used for authenticity discrimination. The region of interest(ROI) and the average reflection spectrum in the ROI were obtained, followed by comparing five pre-processing methods. Then, partial least squares discriminant analysis(PLS-DA), support vector machine(SVM), and random forest(RF) method were established for classification models, which were evaluated by the confusion matrix of prediction results and receiver operating characteristic curve(ROC). The results showed that in the three sample sets, the se-cond derivative pre-processing method and PLS-DA were the best model combinations. The classification accuracy of the test set under the 5-fold cross-va-lidation was 93.27%, 96.19%, and 100.0%, respectively. It was consistent with the confusion matrix of the predicted results. The area under the ROC curve obtained the highest values of 0.992 3, 0.999 6, and 1.000, respectively. The study revealed that the near-infrared hyperspectral imaging technology could accurately identify the medicinal materials of Armeniacae Semen Amarum and Persicae Semen from different origins and distinguish the authentication of these two varieties.

Simple and robust differentiation of Ganoderma species by high performance thin-layer chromatography coupled with single quadrupole mass spectrometry QDa / 中国天然药物

In this study, a high performance thin-layer chromatography/single quadrupole mass spectrometry QDa (HPTLC-QDa) method for robust authentication of Ganoderma lucidum, a popular and valuable herbal medicine, has been developed. This method is simple and practical, which allows direct generation of characteristic mass spectra from the HPTLC plates automatically with the application of in situ solvent desorption interface. The HPTLC silica gel plates were developed with toluene-ethyl formate-formic acid (5 : 5 : 0.2, V/V) and all bands were transferred to QDa system directly in situ using 80% methanol with 0.1% formic acid as desorption solvent. The acquired HPTLC-QDa spectra showed that luminous yellow band b3, containing ganoderic acid B/G/H and ganodeneric acid B, the major active components of Ganoderma, could be found only in G. lucidum and G. lucidum (Antler-shaped), but not in G. sinense and G. applanatum. Moreover, bands b13 and b14 with m/z 475/477 and m/z 475/491/495, respectively, could be detected in G. lucidum (Antler-shaped), but not in G. lucidum, thus allowing simple and robust authentication of G. lucidum with confused species. This method is proved to be simple, practical and reproducible, which can be extended to analyze other herbal medicines.

Development Of Bioanalytical Standardization Parameters Of Alocasia Indica Tuber By Hptlc Technique

Alocasia indica is perennial herb growing widely and used as traditional medicine in India, China and Bangladesh. The divine herb has potent medicinal values for the treatment of different type of illnesses. The HPTLC techniques were used to separate active components from ethanolic extract of tuber part of A. indica. This examination was intended to designed a HPTLC fingerprint profile of crude extract of the plant in ethanol. A HPTLC method for the isolation of various active constituents in A. indica ethanolic extract have been developed and solvent system for quercetin the mobile phase used was toluene: ethyl acetate: formic acid (5:2:1) and for analysis of β-sitosterol the mobile phase used was chloroform: ethyl acetate: formic acid (6:4:1) . In the present investigation, HPTLC fingerprint of extract of dried tuber part of A. indica have been performed and the results demonstrated that important information for standardization. The HPTLC system for routine quality control of present species can be used for ethanolic extract and serve in qualitative, quantitative and was appropriate for standardization of the plant.

Identificación molecular y relaciones evolutivas de Pomacea nobilis, base para la autenticación específica del churo negro de la Amazonia peruana / Molecular identification and evolutionary relationships of Pomacea nobilis, basis for the specific authentication of the black apple snail from Peruvian Amazonia

Resumen En la Amazonia Peruana los caracoles dulceacuícolas de la familia Ampullariidae son conocidos como churos y originalmente han sido descritas para Perú alrededor de 20 especies. Aunque son muy usadas para alimentación, medicina tradicional y objeto de muchos estudios para su cultivo e industrialización, solamente es mencionada en la literatura la especie Pomacea maculata. Se llevó a cabo la identificación molecular sobre la base del marcador mitocondrial COI, de individuos de churos negros (Pomacea) comercializados en los mercados de Iquitos, así como los usados en platos a la carta en la ciudad de Lima, contrastados con otros individuos de procedencia de su hábitat natural. Se encontró que estos especímenes expendidos corresponden a la especie Pomacea nobilis (Reeve, 1856). El análisis filogenético molecular mostró que P. nobilis es especie hermana de P. guyanensis, en el grupo de P. glauca, distantemente relacionada de P. maculata. Las distancias no corregidas encontradas entre ellas, para el marcador mitocondrial COI, fueron de 11.33% a 13.17%, mientras que con P. maculata fueron de 13.67% a 15.33%. Estos resultados demostraron la eficacia del código de barras de ADN para la identificación y autenticación de la especie, lo que le da un valor agregado para su eventual comercio de exportación.

Abstract In the Peruvian Amazon, freshwater snails of the Ampullariidae family are known as churos, and around 20 species have originally been described for Peru. Although they are widely used for food, traditional medicine and the object of many studies for their cultivation and industrialization, only the species Pomacea maculata is mentioned in the literature. Molecular identification was carried out based on the mitochondrial marker COI of individuals of "churo negro" apple snails (Pomacea) commercialized in the markets of Iquitos, as well as those used in restaurant dishes in the city of Lima, and contrasted with specimens from their natural habitat. It was found that these specimens, correspond to the species Pomacea nobilis (Reeve, 1856). The molecular phylogenetic analysis showed P. nobilis as the sister species of P. guyanensis, in the P. glauca group, distantly related to P. maculata. The uncorrected distances found between them, for the mitochondrial marker COI, were from 11.33% to 13.17%, while with P. maculate were from 13.67% to 15.33%. These results demonstrated the effectiveness of the DNA barcode for the identification and authentication of the species, which gives it added value for its eventual export trade.

Genetic authentication of Eclipta prostrate (Asteraceae) from Penthorum chinense (Penthoraceae) by Sequence Characterized Amplified Region (SCAR) markers

Introduction: For the rapid and accurate genetic identification and authentication of living organisms, improved random amplified polymorphic DNA (RAPD) fragment based development of sequence-characterized amplified region (SCAR) markers is an important genetic technique. Objective: This study aimed to develop SCAR markers for perennial herb Eclipta prostrate (E. prostrate). Methods: Here the RAPD fragments by improved RAPD amplification with primers A11 and N-7 for E. prostrate were cloned into pGEX-T vector, and PCR amplification identified the positive clones. After the enzymatic digestion, they were sequenced with Sanger sequencing. Results: Two SCAR markers were developed, which were very specific to E. prostrate, not found in Penthorum chinense Pursh(P. chinense). The nucleotide sequence search by BLAST GenBank database showed that they are novel in E. prostrate, therefore they were deposited in Genbank with accession number KX671034, KX671035. The markers did not show any identity to other species. Conclusions: Thus, in this study two specific SCAR markers were developed for genetically distinguishing and identifying the plant species E. prostrate from herb P. chinense and others.

Introducción: Verificación genética del arbusto Eclipta prostrate (Asteraceae) (Para la identificación y verificación genética rápida y precisa de organismos vivos, el uso de fragmentos de ADN polimórfico amplificado aleatoriamente (RAPD) mejorado de marcadores de región amplificada caracterizada por secuencia (SCAR) es una técnica genética importante. Objetivo: Este estudio tuvo como objetivo desarrollar marcadores SCAR para la hierba perenne Eclipta postrate (E. postrate). Métodos: En este estudio os fragmentos RAPD mediante amplificación RAPD mejorada con los cebadores A11 y N-7 para E. postrate se clonaron en el vector pGEX-T, y la amplificación por PCR identificó los clones positivos. Después de la digestión enzimática, se realizó una secuenciación Sanger. Resultados: Se desarrollaron dos marcadores SCAR, muy específicos para E. postrate, que no se encuentran en Penthorum chinense Pursh (P. chinense). La búsqueda de las secuencias de nucleótidos con BLAST en GenBank mostró que son nuevos en E. postrate, por lo que fueron depositados en Genbank con los números de acceso: KX671034 y KX671035. Los marcadores no mostraron ninguna identidad a otras especies. Conclusiones: En este estudio se desarrollaron dos marcadores SCAR específicos para distinguir e identificar genéticamente la especie de planta E. postrate de la hierba P. chinense y otras.

Discussion on high quality development and authentication system construction of Dao-di herbs based on blockchain / 中国中药杂志

Dao-di herbs are an important part of clinical medicine in traditional Chinese medicine. They are also precious wealth left to human beings from history, which contain deep traditional Chinese cultural connotations and play an important role in supporting and serving the Chinese medicine business. The relevant policy documents introduced by various national ministries and commissions have many contents and requirements related to the promotion of Dao-di herbs protection and industrial development. Due to the Dao-di herbs industry has a series of characteristics, such as a long chain, many involved links, long cycles, multiple production entities, multiple locations, and various types, the high-quality development of the industry has put forward higher requirements on the linkage between upstream and downstream, production entities, traceability of the whole process and information sharing. This article takes Dao-di herbs certification work as an application scenario and entry point, and discusses it from the perspective of block chain and information technology. It proposes the following work ideas: establish multi-party consensus from the macro-organizational management, business, and operational technical levels, and unblock channels for data and information, to achieve institutionalization of certification; establish certification-related standards and specifications to achieve certification standardization; build a certification hardware system to achieve certification networking; build a certification software system to develop functions for specific information content such as identity, origin, production, production process, quality, product and brand of authentic medicinal material production interactively, and realize certification programmatic; data security and sharing of related production activities to achieve socialization of certification. Make full use of modern technologies such as blockchain, the internet of things, big data and information technology, and through the joint participation of management, production, use and the public, the whole process information of Daodi herbs is integrated to form an interconnected information sharing application mode, thus, to serve and promote the high-quality development of Dao-di herbs industry.

Systematic Identification and Quality Evaluation of Anemonis Flaccidae Rhizoma from 16 Different Places of Origin / 中国实验方剂学杂志

Objective:To construct a systematic identification system of Anemonis Flaccidae Rhizoma, and to evaluate the comprehensive quality of Anemonis Flaccidae Rhizoma from 16 regions in China, so as to lay a foundation for its origin selection and clinical medication safety. Method:The authenticity of Anemonis Flaccidae Rhizoma was quickly identified by traditional identification method and DNA barcode molecular identification technology, and HPLC-UV was used to determine the contents of 5 active ingredients in Anemonis Flaccidae Rhizoma. All high pressure chromatographic separations were performed with a Welch Ultimate XB-C18 column (4.6 mm×250 mm, 5 μm), the mobile phase consisted of acetonitrile-0.01% trifluoroacetic acid aqueous solution (30∶70) at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 210 nm and the column temperature was maintained at 30 ℃. Result:The authenticity of Anemonis Flaccidae Rhizoma could be precisely and rapidly identified by ribosomal DNA internal transcribed spacer 2 (ITS2) sequence and traditional identification methods. BLAST comparative analysis found that medicinal materials from 16 areas were all Anemone flaccida. Based on the contents of multi-index components, it was shown that the total content of 5 triterpenoid saponins in Anemonis Flaccidae Rhizoma from Banqiao, Enshi, Hubei was the highest (10.59%), followed by Hezhang, Bijie, Guizhou (6.28%) and Duzhenwan, Changyang, Hubei (5.64%). Conclusion:DNA barcoding can be used as an effective supplement to the traditional identification technology, it can ensure the authenticity of Anemonis Flaccidae Rhizoma and the safety of clinical use. The comprehensive evaluation of multi-index components of HPLC and cluster analysis show that the quality of medicinal materials in Enshi, Changyang, Wufeng of Hubei, Bijie of Guizhou and Jinfoshan of Chongqing is superior, which can be considered as important origin of Anemonis Flaccidae Rhizoma.

Application of near- and mid-infrared spectroscopy combined with chemometrics for discrimination and authentication of herbal products: A review

Herbal medicines along with its preparations have been commonly used as preventive and promotive agents around theworld, especially in developing countries. Motivated by economic profits, the high-priced value of herbal medicinesmay be substituted or adulterated with less expensive ones; therefore, the authentication methods must be developedto overcome the adulteration practices. Due to their properties as fingerprint analytical techniques, near-infrared (NIR)and mid-infrared (MIR) spectroscopies offered fast and reliable techniques for authentication of herbal medicine.The data generated during authentication of herbal medicines were complex and difficult to be interpreted; therefore,the statistical approach called chemometrics has been used to treat data. The objective of the present review was tohighlight the updates on the application of NIR and MIR spectroscopies and chemometrics techniques (discrimination,classification, and quantification) for discrimination and authentication of herbal medicine.

Application of near- and mid-infrared spectroscopy combined with chemometrics for discrimination and authentication of herbal products: A review

Herbal medicines along with its preparations have been commonly used as preventive and promotive agents around theworld, especially in developing countries. Motivated by economic profits, the high-priced value of herbal medicinesmay be substituted or adulterated with less expensive ones; therefore, the authentication methods must be developedto overcome the adulteration practices. Due to their properties as fingerprint analytical techniques, near-infrared (NIR)and mid-infrared (MIR) spectroscopies offered fast and reliable techniques for authentication of herbal medicine.The data generated during authentication of herbal medicines were complex and difficult to be interpreted; therefore,the statistical approach called chemometrics has been used to treat data. The objective of the present review was tohighlight the updates on the application of NIR and MIR spectroscopies and chemometrics techniques (discrimination,classification, and quantification) for discrimination and authentication of herbal medicine

Research progress of Alismatis Rhizoma and prediction analysis on its Q-marker / 中草药

Alismatis Rhizoma was first published in the “Shennong’s Classic of Materia Medica”, listed as top grade, and most of the ancient herbals have been collected. Alismatis Rhizoma is mainly distributed in Fujian, Sichuan, and Jiangxi provinces. Alismatis Rhizoma has the effect of diffusing water and dampness, releasing heat, removing turbidity, and reducing lipid. Its modern pharmacological activity is extensive, and its clinical research is deepening gradually. This paper summarizes the research status of Alismatis Rhizoma from the aspects of herbal textual research, chemical composition, and pharmacological action. On this basis, the differences of different radicals of Alismatis Rhizoma are clarified. Based on the concept of quality marker (Q-marker), the Q-markers of Alismatis Rhizoma are predicted from the point of origin, combined with the research of pharmacodynamics, new medicinal uses and processing, in order to perform qualitative and quantitative analysis of the effective components of Alismatis Rhizoma and provide scientific basis for formulation of new standards.

SEM and XRD Identification of Mineral Chinese Medicine Alumen, Calcined Alumen, and Their Processed Products / 中国实验方剂学杂志

Objective: Ammonium alum is a common counterfeit of Alumen,and the processed product of ammonium alum is a common counterfeits of calcined Alumen. This paper aims to establish a method for identifying Alumen,calcined Alumen,ammonium alum and their processed products. Method: The samples were analyzed by scanning electron microscope (SEM) and X ray diffraction (XRD) in this paper. Result: Ammonium alum and Alumen showed obvious changes in morphology after processing. Both Alumen and ammonium alum showed obvious differences in morphology at×250 and×1 000 times microscope. Alumen presented irregular fragments,clear edge corners,smooth surface,scattered irregular small particles,occasional holes and longitudinal edges. Ammonium alum presented irregular clumps,blunt edges,not obvious edges and corners,uneven surface,scattered smaller and round-like particles. The difference in morphology was not obvious at×250 times microscope between Alumen and ammonium alum processed products. While at×1 000 times,the surface of calcined Alumen was uneven with coarse particles; the surface of counterfeit calcined Alumen was flat,and the coarse particle characteristics were not obvious. XRD can be used to rapidly and accurately identify the primary phase of Alumen,calcined Alumen,ammonium alum and ammonium alum processed products:KAl(SO4)2·12H2O,NH4Al(SO4)2·12H2O,KAl(SO4)2,and NH4Al(SO4)2 respectively, with 2θ angle characteristic value of 23,12,22 and 5 respectively for XRD peak. Conclusion: SEM and XRD techniques can be used for the identification of Alumen,calcined Alumen,ammonium alum and their counterfeit products.

Molecular identification of Armillaria gallica by PCR-RFLP analysis / 中国中药杂志

Armillaria gallica is a symbiotic fungus in the cultivation process of Gastrodia elata and Polyporus.The rhizomorph of A. gallica invades the stalk of the G. elata or the Sclerotium of the Polyporus,and is digested and utilized by the latter,becoming their important source of nutrition. Different nature of A. gallica affects the growth of G. elata and Polyporus. The authors collected A. gallica from 13 commercially available regions and screened two A. gallica,A and B,at the genetic and metabolic levels,in order to distinguish between the two A. gallica market. We have established convenient and effective DNA molecular identification method.By comparing the sequence differences between the A. gallica type A and type B invertase genes,PCR-RFLP primers were designed based on differential fragment. Primer ZTM.F/ZTM.R can amplified A. gallica type A and B,producing a band of about 304 bp in length. The restriction endonuclease EcoR V could recognize the difference sequence of A and B types of A. gallica. The type B was digested to form two fragments,thereby specifically identifying the A. gallica as type B. The established methods of PCR-RFLP is an accurate identification method for A. gallica. Therefore,in the cultivation process of G. elata and Polyporus,suitable strains can be selected according to different needs of variety,growth stage and ecological environment,and the yield and quality can be improved according to local conditions.

Microscopic characterization and HPTLC of the leaves, stems and roots of Fadogia agrestis - an African folk medicinal plant

ABSTRACT Fadogia agrestis Schweinf. ex Hiern (Vangueria agrestis (Schweinf. ex Hiern) Lantz), Rubiaceae, is an African traditional medicinal plant also used as a dietary supplement in the US. The present paper is the first report of the pharmacognostic study of the leaf, stem and root of F. agrestis by microscopy, HPTLC and total phenolic/flavonoid content analyses. Noteworthy microscopic features that can help in identification and quality control are septate and lignified non-glandular trichomes on leaf and stem epidermises, paracytic stomata on leaf abaxial epidermis, numerous cells containing yellow substances of presumably phenolic compounds in leaf and stem, calcium oxalate druses and prismatic crystals in leaf and styloids in stem, primary phloem fibers in stem, brachysclereids in stem and root, spherical starch grains in root, and vessels with vestured pits and simple perforated end walls. In addition to microscopy, a total phenolic/flavonoid content determination and an HPTLC method were also developed for rapid chemical fingerprint analyses of Fadogia samples and dietary supplements.

HPLC fingerprint and simultaneous quantitative analysis of phyllanthin and hypophyllanthin for identification and authentication of Phyllanthus niruri from related species

Abstract A precise and accurate method for the identification and authentication of Phyllanthus niruri L. from P. debilis Klein ex Willd. and P. urinaria L., Phyllanthaceae, was developed using high-performance liquid chromatography. Chromatographic fingerprint analysis was combined with simultaneous quantification of phyllanthin and hypophyllanthin for the developed method. Phyllanthin and hypophyllanthin were successfully separated and quantified under this proposed method. The highest amount of phyllanthin and hypophyllanthin was found in P. niruri compared to P. debilis and P. urinaria. Fingerprint chromatogram of the three Phyllanthus species showed distinct profiles that these may be used to identify and authenticate each Phyllanthus species, which improved by marker compounds present in each species. The combination of chromatographic fingerprint analysis and discriminant analysis was successfully discriminated all three species, including P. niruri adulterated with P. debilis or P. urinaria. The method can be used for the identification and authentication of P. niruri from related species, such as P. debilis and P. urinaria.

Electrochemical fingerprint of traditional Chinese medicines based on BrO⁻₃-Ce⁴⁺-H⁺-malonic acid/tartaric acid chemical oscillating system / 中国中药杂志

A new composite organic oscillating reaction system based on BrO₃-Ce(SO₄)₂-H₂SO₄-malonic acid/tartaric acid was proposed in this paper. On the basis of the influence of the concentration of each component on the stability and characteristic parameters of the blank system, the electrochemical fingerprints of 30 kinds of traditional Chinese medicines (TCM) were obtained. The results showed that the electrochemical fingerprint can be used for the identification of TCMs, the distinguishment of different parts and the appraisal of genuineness, which is fast, sensitive and accurate. At the same time, we explored and verified the mechanism of oscillation and the formation mechanism of TCM fingerprint.