ABSTRACT
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.
ABSTRACT
In this study,DNA molecular identification technology and chemical fingerprint method were adopted to evaluate the quality system of precise powder decoction pieces (PPDP) of S.suberectus dunn (SSD).ITS2 sequence was taken as DNA barcode to indentify SSD.Different specifications of PPDP were prepared,their dry extract contents were quantified in contrast with that of original slices.Three batches of SSD original slices were gleaned and the content uniformity,fingerprint and similarity evaluation before and after the mixing and pulverization were valued by HPLC-DAD.As a result,ITS2 successfully and accurately identified the SSD in this study.The extract rate of PPDP was 15.5％,1.11 times as much as the original slices.RSD of inter-assay dissolution of cepicatechin from the original slices was 11.0％,which was reduced to 1.0％ after mixing and preparing into PPDP.The relative peak area of the 14 common peaks identified by fingerpringts were larger,while the RSD values significantly decreased.It was concluded that the PPDP of SSD improved the extraction efficiency and uniformity of the original slices,featuring quite prospective in more reasonable and scientific clinical use.
ABSTRACT
Objective To establish an HPLC method for the fingerprint analysis of Spatholobus suberectus, so as to provide evidence for the quality control of it. Methods Separation was performed on a ZORBAX SB-C18 column (150 mm × 4.6 mm, 5 μm) and the mobile phase was acetonitrile-0.2% phosphoric acid with gradient elution. The flow rate was 1 m/min and the wavelength was 260 nm. The column temperature was 30 ℃. The cluster analysis (CA) and principal component analysis (PCA) were applied to studying the HPLC fingerprint and chemical pattern recognition. Results The HPLC fingerprint of S. suberectus has been established with 35 common peaks, and six of common peaks were identified. The values of similarity evaluation mostly agreed with the result of CA and PCA. Conclusion The application of HPLC combined with CA and PCA could quickly and objectively evaluate the quality of S. suberectus from different regions.
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Objective The dynamic accumulation trend of secondary metabolite in cultivated Spatholobus suberectus Dunn at different growth time was studied to provide scientific basis for confirming its reasonable harvesting time. Methods Ethanol-soluble extractives were measured by the method recorded in Chinese Pharmacopoeia ( A) Appendix XA ( 2010 edition). The content of total flavonoids was tested by ultraviolet spectrophotometer. The contents of protocatechuic acid, catechin, and epicatechin in Spatholobus suberectus Dunn were determined by high performance liquid chromatography ( HPLC). Results The contents of ethanol-soluble extractives, total flavonoids and the major components such as protocatechui cacid, catechin and epicatechin in Spatholobus suberectus Dunn showed increasing trend with the prolongation of growth time. The changes of contents of alcohol-soluble extract, total flavonoids and epicatechin in samples collecting at different months in one year showed a parabolic trend, and the contents reached the peak values from July to September. Catechin content kept increasing continually. The content of ethanol -soluble extractives of Spatholobus suberectus Dunn aged 4-6 years met the requirements of the current edition of Chinese Pharmacopoeia when harvested from July to September. Conclusion It is preliminarily recommended that Spatholobus suberectus Dunn cultivated for more than 7 years is suitable for harvesting, but the optimal harvest period and the harvest month still need to be further studied.
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Objective To investigate the status of soil fertility of Good Agricultural Practices ( GAP) base for Spatholobus suberectus Dunn. (SSD) in Pingyuan county of Guangdong province, thus to provide reference for GAP research and the subsequent fertilization for SSD. Methods The deep layer and superficial layer of GAP soil were collected for the physiochemical detection and nutrient assay. Compared with the classification standard of the second national general soil investigation, single base soil fertility index was diagnosed and the comprehensive soil fertility was evaluated with modified Nemoro Index. Results The soil pH value and the contents of exchangeable calcium and magnesium were unbalanced, and the contents of macroelements of nitrogen and phosphonium, microelements, and organic matter were low. Therefore, the measures for improving the base soil fertility should be as follows: ( 1) soil amendments of bentonite, gypsum and slaked lime should be used to adjust the soil pH value; ( 2) each plant should be given 10 kg of slaked organic fertilizer as base fertilizer; ( 3) in the process of nurturing, some special micro-fertilizer solution should be used to treat the cut slips, and 5 kg of urea should be used for every 667 meter square of land; ( 4) besides compound fertilizer, every 667 meter square of land should be fertilized with 15 kg of ammonium dihydrogen phosphate for the supplement of nitrogen and phosphorus, and slaked lime and magnesium carbonate should be used for the supplement of soil moderate-quantity elements after transplantation. Conclusion The comprehensive fertility of Pingyuan GAP base for Spatholobus suberectus Dunn. is at low level, and should be improved in combination with GAP requirements.
ABSTRACT
Objective To determine the content of protocatechuic acid in Spatholobus suberectus Dunn.Methods HPLC method was developed for the determination of protocatechuic acid in Spatholobus suberectus Dunn.The separation was performed on C18 column with acetonitrile-water-orthophosphoric acid(8 ∶92 ∶0.1) as a mobile phase with the flow rate being 1.0 mL/min.The detection wavelength was set at 260 nm.Results A satisfactory separation between protocatechuic acid and impurity was obtained.The calibration curve was linear in the range of 0.020～2.008 ug,r=0.999 9.The average recovery was 98 %,and RSD was 0.1 %(n=5).Conclusion The method is accurate and can be used for the detemrmination of protocatechuic acid in Spatholobus suberectus Dunn.
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Aim To study effects of nine compounds extracted from Spatholobus suberectus Dunn (SSD) on proliferation of hematopoietic progenitor cell (HPC) in marrow-depressed mice. Methods Serum pharmacology experiment was used to observe the influence of nine compounds on growth of CFU-E、BFU-E、CFU-GM、CFU-Meg in marrow-depressed mice. Results Compared with the control, all compounds except pyromucic acid and ononin could significantly stimulate the growth of CFU-GM (P