Pharmacognosy research and identification of Euphorbia prostrata Ait.

Background:Euphorbia prostrata Ait.is an annual herb widely distributed in the southern region of China with great medical values on Anti-inflammation,insect repellent,treatment of diarrhea.Despite its extensive uses as a traditional Chinese medicine,no systematic research on the identification of E.prostrata has been reported.Methods:The study aimed to establish an accurate identification system for E.prostrata through traditional pharmacognostical methods,including botanical origin,morphological characters,medicinal material characters,microscopic characters,physicochemical parameters determination,phytochemical screening,and DNA barcoding analysis.Results:Physicochemical results show that this plant likely contains flavonoids,anthraquinones,and other substances.The ITS loci of the nuclear genome and psbA-trnH loci of the chloroplast genome were selected and evaluated,which were the most variable loci.Conclusion:The findings of this study are expected to contribute to the development of species identification,as well as provide references for authenticity identification,genetic relationship analysis,and further utilization of E.prostrata.

Systematic Evaluation of Pharmacognostic Identification of Polygonum capitatum

[Objectives] To investigate the systematic evaluation of pharmacognostic identification of Polygonum capitatum . [Methods] 10 batches of P. capitatum cultivated in Guizhou were chosen for plant samples. Macroscopical identification was conducted on plant roots, stems, leaves, flowers and fruits. The P. capitatum powder was processed for physical and chemical distinction by FeCl 3 chromogenic reaction, hydrochloric acid magnesium powder reaction, AlCl 3 color development reaction and thin-layer chromatography.Microscope identification was carried out on the powder. Plant genome DNeasy Plant Kit was adopted for DNA molecular marker identification. [Results] The results showed that the stem of P. capitatum was tufted, the leaves were oval, 2 to 5 cm long, and 1 to 2 cm wide;the leaf apex was acute and cuneate at the base, the inflorescence was capitate, paired or solitary;the raceme was erect and nearly spherical, and the perianth was light red. Furthermore, for the chromogenic reaction of FeCl 3 ethanol extract of P. capitatum , appeared blue and turned to dark blue after long time storing at room temperature. For the reaction of hydrochloric acid magnesium powder, the alcohol extract of P. capitatum , exhibited deep red. In the color reaction of AlCl 3, the alcohol extract revealed yellow fluorescence under 360 nm UV lamp. Microscope identification of the powder displayed pollen grains, crystal sheath fibers, cellulose, vessels, starch grains, cork cells, and other characteristic fragments. In addition, DNA barcoding electrophoresis results showed that P. capitatum showed a clear and bright single band near 500 bp, and further sequencing results showed that the sequence differences were mainly concentrated in ITS1 and ITS2 region. [Conclusions] Systematic evaluation for the identification of P. capitatum is established, which combines with macroscopic identification, physicochemical identification, powder microscope identification, and DNA molecular identification. Finally, the original medicinal material is identified as P. capitatum Buch.-Ham. ex D. Don.

Identification of Mucilage Cavity as a Significant Microscopic Characteristic Existing in Phloem Instead of Pith of Radix et Rhizoma Rhei

The morphologic and microscopic features of Radix et Rhizoma Rhei were studied. The study verified that the mucilage cavities did exist in phloem of rhizomes and roots or abnormal vascular bundles. Also, they were in similar distribution in three species of Radix et Rhizoma Rhei. The diagnostic characteristic for microscopic identification was found to be the similar distribution of abnormal vascular bundles in pith of rhizomes in all three species. And the appearance of the crude drug varied more depending on the plants＇ geographical origin and different preliminary treatment on the spot of collection than on the species differences. Our findings, having not been delineated clearly so far in the previous reports, are helpful for clarifying current descriptions in different literatures or standards and make a full understanding on microscopic and macroscopic identification of Radix et Rhizoma Rhei.

Identification of Tibetan Medical Material Dracocephalum tanguticum Maxim.

[Objectives]To explore morphological identification,macroscopical identification,microscopic identification,thin layer chromatography(TLC)identification of Tibetan medical material Dracocephalum tanguticum Maxim.,and provide experimental data for its identification and application.[Methods]The Tibetan medical material was identified by means of original plant,characters,powder,paraffin section and thin layer chromatography(TLC).[Results]Tibetan medical material D.tanguticum Maxim.was obviously distinguished in character identification and microscopic identification,and the TLC method was simple and feasible.[Conclusions]The results will provide the source work foundation for the formulation of the quality standard of Sichuan Province(draft)for Tibetan medicinal material"D.tanguticum Maxim."and the development of pharmaceutical preparations for medical institutions.

Microscopic Identification of Pueraria phaseoloides,a Zhuang Medicine

This study aimed to investigate the structural characteristics of different tissues and tissue powder of Pueraria phaseoloides by microscopic identification. The main microscopic characteristics of roots, stems, leaves, root powder, stem powder and leaf powder of P. phaseoloides were determined by microscopic identifi- cation, which provided reference for the establishment of quality standards of P. phaseoloides.

Study on the Microscopic Identification of Chenopodium album L.

[Objectives]This study was conducted to identify the microscopic characteristics of Chenopodium album L.[Methods]The microscopic identification method was adopted.[Results]The xylem vessels and fiber bundles of the roots are arranged into 3-4 intermittent ring belts in a concave-convex pattern,alternately with the parenchymal cell ring belts;and the xylem rays vary in width.The cross section of the stems is polygonal;there are parenchyma cells at the corners of the cortex;there are many vascular bundles of varying sizes;and calcium oxalate clusters are common in the medulla and cortex.There are 2 to 4 vascular bundles in the main leaf vein,peltate;and fiber bundles exist below the phloem.The powder is characterized by numerous clusters of calcium oxalate,which are uniform in size,sharp at edges and corners.[Conclusions]This study can provide reference for the identification and quality standard of the crude drug.

Study on the Pharmacological Identification of Polygonum capitatum

[Objectives]This study was conducted to establish a microscopic identification method of Polygonum capitatum Buch.Ham.ex D.Don.[Methods]The cross sections were identified by microscopic identification.[Results]The stem cross section of P.capitatum is round-like,and shows pericyclic fibers forming a ring,strongly lignified,many vascular bundles,and a hollow pith part.There are many starch granules in the powder,and single granules are more common;and fibers are mostly bundled or scattered,lignified or non-lignified,and calcium oxalate cluster crystals are common.There are many pollen grains,obtuse triangular or round-like,and some of them have three germination apertures.They have fine thorn-like protrusions on the outer wall,the surface of which has reticulate carvings.[Conclusions]The results of microscopic identification are reliable and can be used as the basis for identification of P.capitatum.

Characters and Microscopic Identification of Aralia cordata

[Objectives]Characters and microscopic characteristics of Aralia cordata were studied,to improve identification method for Aralia cordata.[Methods]Using pharmacognostic identification method,original plant morphology of Aralia cordata was observed,and microscopic observation of its root cross section and powder was conducted.[Results]It is perennial herb,with rhizome recess,arranged in clusters of nodules.There are double or triple pinnate compound leaves.It is glabrous or sparsely pubescent,with umbel.Flowers are white,while fruits are spherical and purple black.In microstructure,the cork layer on the cross section of root consists of 6-10 rows of cells,and there is secondary cortex.The parenchyma cells are rich in starch granules,and the cambium is not obvious.The vessels are uniserial,and the wood fiber is less,and the primary xylem is bipartite.There are many starch granules in powder,mostly single granules,round or quasi round,oval,umbilicus herringbone or short slit,with many marginal pits and reticulate ducts.Calcium oxalate cluster crystals are mostly gathered.Cork cells are yellowish brown and arranged neatly.The fibrous wall is thin,and the pore groove is obvious,and stone cells are occasionally seen.[Conclusions]Using character and microscopic identification method,it could accurately identify medicinal material of Aralia cordata,and provide reference basis for development and utilization of its resources.

Establishment of Quality Standard for Mongolian Medicines Shoushen and Naishoushen

[Objectives] To systematically identify and analyze pharmacognostical features of Mongolian medicine Shoushen and Naishoushen( dairy tablets) to provide scientific basis for the establishment and identification of its quality standard. [Methods] According to relevant methods specified in Appendix to 2010 Chinese Pharmacopoeia,the water content,total ash,and extracts of Shoushen and Naishoushen were detected,and thin layer chromatography( TLC) was applied to make qualitative identification. Gastrodin was used as the reference substance,extracted with 70% methanol,and then sprayed with ethyl acetate-formic acid-water( 9∶ 1∶ 0. 2) as the developing solvent,and then sprayed with 10% phosphomolybdic acid ethanol solution and heated at 105℃ to clear spot color. UV-Visible spectrophotometry and high performance liquid chromatography( HPLC) were used to analyze the content of polysaccharides and gastrodins in Shoushen and Naishoushen. The chromatographic column was a ZOBB AX eclipse XDB-C_(18)( 4. 6 mm × 250 mm,5 cm) column with the methanol-0. 04% phosphoric acid solution( 8∶92) as the mobile phase,the flow rate of 1. 0mL/min,the detection wavelength of 222 nm,and the column temperature of 30℃.[Results]In the thin-layer chromatography test,spots of the same color appeared at the positions corresponding to the chromatogram of the reference substance. Gastrodin eluted under high pressure reached baseline separation. Gastrodin had a good linearity in the concentration range of 0. 009-0. 09 mg/m L. The regression equation was Y = 586 866 X + 425 821( R^2= 0. 999 6),and the average recovery rate was100. 1%. Precision test,reproducibility test,and stability test conformed to the requirements. The results of extracts of three batches of samples were 26. 13%-42. 58%,water content was 3. 47%-5. 31%,and total ash was 5. 43%-6. 33%. [Conclusions] The quality control method has high reliability,high sensitivity,high specificity,high accuracy,and high stability. The results are expected to provide a scientific basis for the identification,resource utilization,and improvement of quality standard for Shoushen and Naishoushen.

Quality Standard of Zijinbiao

[Objectives]To establish the quality standard for Zijinbiao.[Methods]Microscopic identification,physical and chemical identification,and thin-layer chromatography(TLC)were used to qualitatively identify Zijinbiao.The moisture,total ash,acid-insoluble ash,and alcohol-soluble extract content were determined.The content of Plumbagin was determined by high performance liquid chromatography(HPLC).[Results]Microscopic identification,physical and chemical identification and thin layer identification features were remarkable.The moisture,total ash,acid-insoluble ash,and extract content of the 15 batches of samples were 7.49%-11.84%,2.43%-5.81%,0.59%-3.18%and 13.80%-20.45%,respectively.The linear equation of Plumbagin was Y=38094X,R^(2)=0.9996.Plumbagin had a good linear relationship in the range of 0.01-0.53 mg/mL.[Conclusions]This method is specific and reproducible,and can be used for quality control of Zijinbiao.

Study on Quality Standard of Tibetan Medicine Corydalis dasyptera Maxim.

[Objectives]To establish the quality standard of Tibetan medicine Corydalis dasyptera Maxim.[Methods]According to the research method of drug quality standard in the appendix of 2020 edition of Chinese Pharmacopoeia,8 batches of C.dasyptera Maxim.from different habitats were studied by character identification,microscopic identification and TLC identification.The content of water,total ash,acid-insoluble ash and alcohol-soluble extract was determined,and the content of protopine in medicinal materials was determined by high performance liquid chromatography(HPLC).[Results]The properties and microscopic characteristics of C.dasyptera Maxim.were determined.The TLC characteristic spots of the medicinal materials were clear,the degree of separation was good,and the specificity was strong.Both the test sample and the control sample showed the same yellow-green spots in the corresponding position.It was tentatively determined that the water content of C.dasyptera Maxim.should not exceed 14.0%,the total ash content should not exceed 14.0%,the acid-insoluble ash content should not exceed 3.0%,and the alcohol-soluble extract content should not be less than 18.0%.There was a good linear relationship between the concentration of protopine and the peak area in the range of 16.64-166.40μg·10-3(r=0.9996).The average recovery rate was 98.47%and the RSD was 1.21%(n=6).The content of protopine in 8 batches of C.dasyptera Maxim.was 0.023%-0.093%.[Conclusions]The established quality research method is simple,stable and reliable,and can be used for the quality control of C.dasyptera Maxim.

Pharmacognostic Study on Adenanthera pavonina Linn.var.microsperma

[Objectives]To study the pharmacognosy of the stems and leaves of Adenanthera pavonina Linn.var.microsperma and to provide reference materials for clinical application and further development and utilization.[Methods]The stems and leaves were sliced by paraffin section method,and the tissue structure and microscopic characteristics were observed by routine microscopic technique.[Results]It was found that the epidermal cell wall was red with unicellular non-glandular hairs,the phloem fiber bundles were arranged in rings,and the xylem vessels were large and few,arranged in the shape of"V".The non-glandular hairs in the cross section of leaves were mostly unicellular,and the palisade tissues were in 2-3 rows.Most of the vessels in the powder were with marginal pores and crystal sheath fibers,and there were a lot of non-glandular hairs in single cells.[Conclusions]The above characteristics can be used as the main basis for the identification of Adenanthera pavonina Linn.var.microsperma.

Morphological and microscopic identification studies of Cordyceps and its counterfeits

Macroscopic and microscopic studies were applied to distinguish Cordyceps sinensis(Berk.)Sacc.and its 5 common counterfeits.Transverse sections of stroma and larvae and surface sections of stroma of C.sinensis,Cordyceps gunnii,Cordyceps barnesii,Cordyceps gracilis,Cordyceps liangshanensis and Cordyceps militaris were examined and their morphological and microscopic features photographed.The main morphological and microscopic features of the 6 species of Cordyceps were basically similar except for certain diagnostic differences.These included macroscopic differences from C.sinensis as follows:the stroma of C.gunnii is stout and rough with sterile bulgy or branched apex;the larvae of C.barnesii has a pair of teeth on the head;the stroma of C.liangshanensi is thread-like;C.gracilis is without stroma;and C.militaris is without larvae.There were also microscopic differences:from C.sinensis as follows:the stroma of C.barnesii is without perithecia;C.gunnii,C.liangshanensis and C.gracilis are without bristles on the larva body.These differences allow C.sinensis and its counterfeits to be easily distinguished.

Combined application of extended depth of field imaging, image stitching and polarized microscopy techniques in identification of Spatholobus suberectus

Objective:As traditional techniques for microscopic identification of Chinese medicines currently lack objective and high-quality reference images,here we developed a systemic procedure to be used in microscopic identification of Chinese medicines,which would lead to more objective,effective and accurate identification process.Methods:Spatholobi Caulis(Jixueteng in Chinese)was used as the specimen in the development of such procedure.Jixueteng samples were microscopically examined in bright-and dark-field microscopy.Microscopic images were obtained by regular,EDF,and image stitching techniques.Results:The microscopic images of the characteristics in pulverized Jixueteng were captured,thanks to EDF imaging and image stitching techniques which allowed the detailed and full sighting of each characteristic to be obtained simultaneously.Different layers in anatomical transverse section,including cork,phelloderm,cortex,phloem,cambium,xylem and pith,were distinctively observed.Moreover,by comparing images of bright-and dark-field microscopy,birefringent and non-birefringent components could readily be distinguished.Conclusion:With application of the developed procedure,high-definition,panoramic and microscopic images were acquired,which could be used as the reference images for microscopic identification of Chinese medicines.

Microscopic identification of the remnant hair or feather of five animal drug components in Shenrongbian pill

A comparative study was performed to identify the microscopic characteristics of hair or feather in the five animal drug components contained in Shenrongbian pill.Penis et Testis Canis is 40±0.07 in the medulla index,with long circular,banana or triangular circular shaped medulla cells arranged in one line or network,and the hair cuticle is in imbrication(d,m)and flat wave(p)shape.Penis et Testis Equi is 66±0.10 in the medulla index,with ellipse,spindle or long strip-shaped medulla cells arranged in network,and the hair cuticle is in flat wave shape.Penis et Testis.Bovis is 67±0.05 in the medulla index,with rectangle,spindle or polygon-shaped medulla cells arranged in ladder or network form,and the hair cuticle is in flat wave shape.Penis et Testis Mustelae is 29±0.05 in the medulla index,with ellipse-like,square-like or circular shaped medulla cells arranged in one line generally,and the hair cuticle is in acuminate(d,m),imbrication(m,p)and slightly flat wave(p)shape.Musculus et Bonis Passeris is 24±0.05 in the medulla index,with bam boo joint-shaped barbs and unclear medulla cells,without hair cuticle.

Pharmacognostical study of Cynanchum stauntonii and Cynanchum glaucescens,botanical sources of TCM Baiqian

Objective： Cynanchum stauntonii and Cynanchum glaucescens are botanical species of Baiqian（Cynanchi Stauntonii Rhizoma et Radix） in Chinese Pharmacopoeia, in which, however, there are no microscopic identification. Therefore, we provided the morphological and microscopic identification of the crude drug for updating Chinese Pharmacopoeia.Methods： Twelve batches of C. stauntonii and three batches of C. glaucescens and their crude drugs were taxonomically, morphologically, and microscopically examined.Results： Taxonomically, C. stauntonii had narrowly lanceolate leaves with acuminate apex and 5 mm long petiole; Whereas C. glaucescens was oblong-lanceolate or oblong with rounded or acute apex in leaves,and had very short or no petiole. Morphologically, rhizomes of C. stauntonii and C. glaucescens both had hollow pith, but the hollow pith occupied about a half of the rhizome＇s diameter in C. stauntonii, whereas only a very small proportion of the overall diameter in C. glaucescens. Moreover, microscopic observation showed the difference in the proportion of xylem and in rhizome transverse-sections of the two species along with the difference in the size of the pith. Finally, laticifers and rhizome epidermal secretory cells were present in the powders of C. stauntonii, but absent from C. glaucescens.Conclusion： Based on observation of morphological and microscopic characteristics, the two species can be distinguished by the size of the pith, proportion of xylem of rhizomes, and crude drug powder characters such as laticifers and secretory cells.