ABSTRACT
Objective:To quantitatively analyze the changes of Staphylococcus aureus in different processed products of Angelicae Sinensis Radix. Method:The real-time fluorescence quantitative polymerase chain reaction method (Real-time PCR) was established to quantitatively analyze S. aureus in Angelicae Sinensis Radix decoction pieces which bought from different producing areas, different enterprises and different storage time. The fluorescence quantitative reaction system was SYBR Premix Ex Taq Ⅱ of 10 μL, each of forward primer and reverse primer (10 μmol·L-1) of 0.8 μL, template/genome DNA of 1 μL, double distilled water of 7.4 μL. The reaction conditions of the fluorescence quantitative amplification curve were pre-denaturing for 30 s at 94 ℃, denaturing for 10 s at 94 ℃, annealing for 12 s at 60 ℃, extensing for 30 s at 72 ℃, cycling 45 times, single-point detection signal at 72 ℃. The melting curve was made from 72 ℃, and the step temperature of 0.5 ℃ was kept for 15 s to collect fluorescence. According to the results of Real-time PCR, representative samples were selected from Angelicae Sinensis Radix decoction pieces for comparison between plate counting method and Real-time PCR. Result:The content of S. aureus in different processed products was sorted by rank of raw Angelicae Sinensis Radix>soil-fried Angelicae Sinensis Radix>wine-processed Angelicae Sinensis Radix. The content of S. aureus was the lowest in the samples from Weiyuan area of Gansu province by comparing with other producing areas. Compared with the retail enterprises, the content of S. aureus in raw products and wine-processed products from production and sale enterprises was lower. Different storage time had certain effect on the content of S. aureus in raw products and wine-processed products, and the content of S. aureus increased with the increase of storage time. The detection results of plate counting method were 3-4 orders of magnitude lower than that of Real-time PCR. Conclusion:The established Real-time PCR is superior to plate counting method in specificity, sensitivity, reliability and reporting period, which can provide an effective method for rapid and accurate quantitative detection of S. aureus in different processed products of Angelicae Sinensis Radix.
ABSTRACT
Objective::Astragali Radix is an important medicinal and edible herb. To achieve standardized cultivation of Astragali Radix and improve cultivation results, Astragalus membranaceus var. mongholicus was cultivated with a row spacing of 30 cm and different plant spacing (8, 10, 12, 14, 16 cm) in a test base of Longxi County, Gansu Province. Method::The growth and development dynamics of green strains and the yield and quality of medicinal materials were measured, and the comprehensive evaluation of membership function was used to determine the rational transplanting density and explore the standard production technology of A. membranaceus var. mongholicus. Result::The transplanting topping had a significant effect on the growth and development of A. membranaceus var. mongholicus under the condition of the row spacing of 30 cm. With the increase of topping, the biomass of astragalus on the ground decreased, and the ratio of root to shoot increased, but the yield of medicinal material per unit area decreased. The appearance traits were improved as the transplant density decreased. Under the row spacing of 14 cm, the content of astragaloside and calycosin-7-glucoside were the highest. Under the row spacing of 8-16 cm, the content of root extract of Astragali Radix was better than the Chinese Pharmacopoeia standard. The comprehensive evaluation index of each plant spacing treatment was 14 cm>16 cm>10 cm>8 cm>12 cm in turn. Conclusion::Combined with the comprehensive evaluation of yield and economic benefit, the optimal transplanting plant spacing and row spacing were 30 cm×14 cm (with the density of 238 100 strains/hm2). Under this density, A. membranaceus var. mongholicus grows vigorously, with thick stems, big root crowns, a high yield and high quality.
ABSTRACT
Ten batches of Angelica sinensis from three producing areas( Tuoxiang,Minxian and Weiyuan of Gansu province) were selected as the research objects,and processed into raw A. sinensis,A. sinensis with alcohol,and A. sinensis with soil respectively through the standard processing methods. Ultra-high performance liquid chromatography( UPLC) was used to establish fingerprint for three processed products of A. sinensis,and determine the contents of 9 phenolic acids and phthalide compounds. The similarity was analyzed with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine,which showed that the chromatographic peaks of the same processed samples of A. sinensis were basically similar,with all similarities greater than 0. 950. The difference between different processed products and their control spectra was not obvious,with all similarities also higher than 0. 950.On the basis of using principal component analysis( PCA) and OPLS-DA to seek the difference components between groups,the improved distance coefficient method can be used to effectively distinguish the three processed products of A. sinensis by fingerprint similarity. At the same time,the determination method of nine phenolic acids and phthalide in A. sinensis was established by UPLC,and the comparison between different processed products was carried out. The results showed that the content of various components was changed as compared with the raw A. sinensis. The contents of coniferyl ferulate and ligustilide in the A. sinensis with alcohol were increased significantly,and the content of coniferyl ferulate was obviously increased in A. sinensis with soil. The method established in this paper can effectively distinguish different processed products of A. sinensis and determine the content of the main components in them.