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OBJECTIVE：To establish the method for content determination of 6 active ingredients in Paeonia lactiflora during different harvest periods ，and to investigate its variation rules so as to determine the optimal harvesting period. METHODS ：HPLC method was adopted to determine the contents of gallic acid ， catechin， alibiflorin， paeoniflorin， benzoic acid and benzoylpaeoniflorin，and principal component analysis was conducted . The determination was performed on Thermo C 18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid aqueous solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was 230 nm，and column temperature was 35 ℃. The sample size was 10 µL. RESULTS ：The linear range of the above 6 ingredients were 0.013 2-0.25 mg/mL（r＝0.999 2），0.013 2-0.25 mg/mL（r＝0.999 9），0.026 8-0.51 mg/mL（r＝0.999 7）， 0.42-8.01 mg/mL（r＝0.999 2），0.016-0.31 mg/mL（r＝0.999 4），0.02-0.38 mg/mL（r＝0.999 8），respectively. The limits of quantification were 0.009 3，0.008 5，0.016 3，0.021 7，0.011 3，0.017 4 mg/mL，and the limits of detection were 0.003 3，0.002 7， 0.005 4，0.007 3，0.003 8，0.005 9 mg/mL. RSDs of precision ，stability and repeatability tests were less than 2％. The recoveries were 96.01％-99.43％（RSD＝1.23％，n＝9），97.95％-100.45％（RSD＝0.79％，n＝9），97.98％-100.11％（RSD＝0.68％，n＝ 9），98.83％ -100.09％ （RSD＝0.65％ ，n＝9），98.58％ - 100.95％（RSD＝1.35％，n＝9），96.28％-103.26％（RSD＝ 1.76％，n＝9）. The contents of above 6 ingredients in Radix Paeoniae Rubra （roots） were 0.016％ -0.057％ ，0-0.067％ ， 0.207％-0.640％，2.350％-5.887％，0.030％- 0.245％，0.054％- 0.381％，respectively. On May 30th，the drying rate of Radix 0451-87266873。E-mail：wangzhen_yue@163.com Paeoniae Rubra was the lowest （about 33％），and on Sept. 15th，the drying rate was the highest （about 49％）. The contents of gallic acid and paeoniflorin in the leaves of P. lactiflora were higher than the root during Jul.-Oct. Results of principal component analysis showed that the variance contribution rates of the first two principal components were 71.845％ and 18.170％，respectively；cumulative variance contribution rate was 90.015％. The months with higher comprehensive scores were May to Jun. and Sept. to Oct. CONCLUSIONS ：Established method is simple ， accurate，reproducible and precise. It can be used to determine the contents of 6 active ingredients in Radix Paeoniae Rubra during different harvest periods. Sept. 30th to Oct. 15th is the optimum harvesting periods for Radix Paeoniae Rubra ，and leaves can be harvested around Jul. 15th.
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OBJECTIVE： To establish a determination method for content of polysaccharide in Acanthopanax senticosus， and to conduct cluster analysis and ultrasonic extraction technology optimization. METHODS： The content of polysaccharide in A. senticosus was determined by phenol-sulfuric acid method. Cluster analysis was conducted by using SPSS 23.0 software of the polysaccharide in A. senticosus from 17 habitats. The ultrasonic extraction technology was optimized with L9（34）orthogonal test using extraction temperature， the ratio of material to liquid， extraction time as factors， the content of polysaccharides as index， and then validated. RESULTS： The linear range of glucose ranged from 0.007 75 to 0.151 mg/mL （r＝0.999 1）； the limits of quantification and detection were 2.854， 0.856 μg/mL； RSDs of precision， stability， and repeatability tests were less than 2％； recovery rates of the sample were 98.41％-101.58％（RSD＝1.23％，n＝6）. Cluster analysis results showed that 17 batches of samples could be clustered into three classes； S1， S6， S10， S12 and S13 were clustered into one class； S3-S5 and S7 were clustered into one class； and the rest samples were clustered into one class. The optimal ultrasonic extraction technology was extraction temperature 55 ℃， ratio of material to liquid 1 ∶ 10 （g/mL）， extraction time 35 min. Results of validation tests showed the content of the polysaccharide under the best process condition was 4.36％（RSD＝0.920％，n＝3）. CONCLUSIONS： Established method is simple，reproducible and suitable for determination of polysaccharide in A. senticosus； the optimized ultrasonic extraction technology is stable and feasible.
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OBJECTIVE:To determine the content of 17 amino acid in sarcocarp of northeast Empetrum nigrum simultaneous-ly. METHODS:HPLC-ELSD method was adopted. The determination was performed on Waters Symmetry C18 column with mobile phase consisted of 5.0 mmol/L heptafluorobutyric acid solution(containing 0.7% trifluoroacetic acid)-acetonitrile(gradient elution) at the flow rate of 2.0 L/min,the column temperature was 37 ℃,the drift tube temperature was set at 60 ℃. RESULTS:The line-ar ranges of glycine,serine,aminosuccinic acid,aminopropionic acid,threonine,glutamic acid,cystinol,diaminocaproic acid, histidine,arginine,proline,valine,methionine,oxyphenylaminopropionic acid,isoleucine,aminocaproic acid,phenylalanine were 0.0059-0.1877,0.0082-0.2628,0.0104-0.3328,0.0070-0.2227,0.0093-0.2978,0.0115-0.3678,0.0094-0.3004,0.0114-0.3655,0.0121-0.3880,0.0136-0.4355,0.0090-0.2878,0.0092-0.2930,0.0117-0.3730,0.0142-0.4530,0.0103-0.3280, 0.0103-0.3280 and 0.0129-0.4130 mg/mL(all r>0.9990). RSDs of precision,stability and repeatability tests were not all less than 2.0%. Recoveries were 98.0%-101.2%(RSD<2.0%,n=6). CONCLUSIONS:This method is simple,precision,stable and repeatable,and can be used for simultaneous determination of 17 amino acid in sarcocarp of northeast E. nigrum.
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Objective:To obtain AP2/ERF genes from Brassica oleracea by using in silico cloning. Methods: AP2/ERF genes were cloned by retnieving the EST database and using the bioinformatics software with the Arabidopsis thaliana AP2/ERF as a querying probe. Results:Two AP2/ERF family transcriptional regulators (BoAP2/ERF1 and BoAP2/ERF2) were isolated from Brassica olera-cea by the in silico cloning method. Some characters of AP2/ERF protein were analyzed and predicted by the tools of bioinformatics in the following aspects including the composition of amino acid sequence, hydrophilicity and hydrophobicity, subcellular localization, secondary and tertiary structure of protein and function. Conclusion:Bioinformatical analysis shows BoAP2/ERF1 and BoAP2/ERF2 gene encode 40. 85kDa and 39. 44kDa protein with 371 and 352 amino acids. The domain is predicted to locate on nucleus. Sequence analysis indicates the protein may be involved in signaling transducer and stressing response roles in plantbiotic stresses.
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The numerous noxious reactions caused by misusing medicinal plant ingredients have been widely concerned at home and abroad. The problems of safe drug use needed to be solved. DNA barcode is a powerful molecular identification method, which can cover the shortage of tradditional morphological and chemical identification. In recent years, DNA barcode provided new aspects for the identification of Chinese herbal medicines and have obtained outstanding achievement. Several popular DNA regions ( ITS, ITS2, ps-bA-trnH, rbcL and matK) have been used for the identification of Chinese herbs.
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Objective To investigate the effects of spine region of interest (ROI),tracking range,and real-time image contrast ratio on the positioning errors of cyberknife.Methods The LTT dynamic phantom was used to develop a spine tracking plan and perform treatment,and the target positioning system was used to preset the phantom and obtain real-time preset image and positioning error.Based on the realtime preset image,spine ROI,tracking range,and real-time image contrast ratio were changed to observe the changes in positioning error and related parameters.Pearson correlation analysis was performed.Results The change in tracking range did not change the positioning error in spine tracking,and tracking range was not correlated with positioning error (R =0,P =1).The changes in ROI and image contrast ratio did not affect the translation error,but affected the rotation error,especially the rotation error in left-right direction (r =0.533 and 0.693,P=0.002 and 0.026).The image contrast ratio had the most obvious effect,with an amplitude of variation up to 2.2°.Conclusions The change in tracking range does not affect the positioning errors in spinal tracking,but the changes in ROI and image contrast ratio can cause varying degrees of changes in positioning errors,which should be taken seriously in cyberknife treatment.
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Objective To explore the relationship of X-ray exposure parameters and false-node rate during image-guidance treatment with CyberKnife spine tracking.Methods Using spine tracking planning on a chest phantom,several combinations of X-ray exposure parameters were used to locate.The false-node ratio and the surface absorbed dose were investigated and the radiation dose was optimized.Results The false-node ratio and surface absorbed radiation dose decreased when the X-ray exposure parameters increased until they saturated.In the range of ≤5.0％ false-node rate,the surface absorbed radiation dose was 0.11,0.26 mGy,and 0.31-0.46 mGy,when the false-node rate was 2.77％,1.07％and 1.0％,respectively.Conclusions In image-guided treatment of CyberKnife,the radiation dose would be optimized,and the patient's radiation dose would be reduced greatly,which is important to protect the patients.
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Objective To measure and analyze Varian Clinac 21EX accelerator induced radioactivity,and to provide specific recommendations and ways of radiation protection for radiotherapy technicians.Methods To simulate the working environment of radiotherapy technician,and to detect induced radioactivity variation of Varian Clinac 21EX accelerator that induced by 15 MV X-rays under different conditions of beam field area,dose,time,distance and by high energy electron beam (12,16,20 MeV)at the different time.Results The induced radioactivity level was not influenced by different beam field area,and was increased with increasing dose (r =0.930,P ＜ 0.05),decreased with time increase (r =-0.84,P ＜ 0.05),decreased with distance increase(r =-0.975,P ＜ 0.05).The induced radioactivity attenuation levels of the different doses at the same time are different,and have the common characteristic that the induced radioactivity attenuation rate is faster in the initial times.The induced radioactivity levels of high energy electron beams were significantly lower than those of high-energy X-rays.Conclusions During radiotherapy positioning,it is necessary to take measures to protect against the induced radioactivity when high-energy rays with energy greater than 10 MeV will be used.The radiotherapy technician should take the different time and energy interval steps for the different ray type and energy and dose in order to meet the radiation protection principle of optimization.
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OBJECTIVE:To conduct pharmacognostical study of Caulopyhllum robustum Diels so as to offer evidences for exploiting and utilization of this herbal plant. METHODS:A pharmacognostical study of C.robustum was performed by carrying out identification of characteristics,microscopic identification and physicochemical identification. RESULTS:The pharmacognostical features of C.robustum have been established in our study. CONCLUSIONS:This study is suitable for the identification of the origin of C.robustum and it provides approach and evidence for the establishment of the quality standard and sustainable development of resources of C.robustum.
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Objective To study the chemical constituents from the root of Rumex gmelini.MethodsThe compounds were isolated and purified by chromatography,polyamide column chromatography,and preparation HPLC etc.Their structures were elucidated by physicochemical and spectroscopic evidences.Results Ten compounds were identified as:nepodin (Ⅰ),emodin (Ⅱ),citreorosein (Ⅲ),chrysophanol 8-O-?-(6'-acetyl) glucopyranoside (Ⅳ),chrysophanol 8-O-?-D-glucopyranoside (Ⅴ),resveratrol (Ⅵ),9,9'-dianthranone-2,2'-dimethyl-5,5'-bis (?-D-glucopyranose)-9,9',10,10'-tetrahydro-4,4'-dihydroxy-10,10'-dioxo (trivial name:rumoside A) (Ⅶ),emodin-8-O-?-D-glucopyranoside (Ⅷ),resveratrol-3-O-?-D-glucoside (Ⅸ),and rutin (Ⅹ).Conclusion Compound Ⅶ is a new compound,named rumoside A.Compounds Ⅴ,Ⅷ,and Ⅹ are separated from R.gmelini for the first time.Compounds Ⅲ and Ⅳ are the compounds which have been found in the plants of Rumex L.for the first time.
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Object To provide a scientific basis for identification and rational use of the herbal medicine by the pharmacognostic study on Cortex Fraxini. Methods Microscopic identification, TLC, HPLC were used. Results There were remarkable differences in the genuine product, confusing product and fakements of Cortex Fraxini in characteristics of microscopic identification, fluorescence, TLC and HPLC. Conclusion The scientific basis has been established and provided for differentiation of confusing product and fakements.
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Objective To study the effect of three fungi of Helminthosporium on the seven kinds of bioactive constituents in root of seedling Rumex gmelini. Methods Let the R. gmelini infect the fungi of Helminthosporium by dealing the leave surface of R. gmelini with fungal spore suspension. The content of seven kinds of bioactive constituents was determined by HPLC and the results were analyzed by variance analysis. Results During the treatment time, little effect of three fungi of Helminthosporium on content of polydatin, chrysophanol 1-glucoside, and physcion, but the obvious effect on resveratrol, musizin, emodin, and chrysophanol was found. Conclusion The effects of H. turcicum 001 on yield and content of resveratrol are significant in early-middle stage (0—40 d), but treatment time over 40 d is not suitable. The results of this study could provide the new idea and theoretical basis for the exploiting of natural medicines and the planting of Chinese herbal medicine.
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Objective To compare the HPLC fingerprint of Rumex gmelini from different habitats by RP-HPLC (DAD). Methods In this paper, 12 different samples were studied. Separation was performed on an Planetsil C_ 18 column, with mobile phase consisting of methanol and 0.1% phosphoric acid-water and with gradient elution at the flow rate of 1.0 mL/min. The UV detection wavelength was 254 nm, column temperature was 35 ℃, and the analysis time was 50 min. Results The results showed that this method has a good repeatability and the ratio of common peaksarea of different samples had some difference. Conclusion This method can be used to establish the chromatographic fingerprint of R. gmelini with high specificity.
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Objective: To determine the contents of aesculin and aesculetin in periderms and leaves of Cortex Fraxini among differnet provenances. Methods: HPLC was used with acetonitrile-water(15∶85) as the mobile phase and detected wavelength at 348nm. C 18 column was adopted. Results: The calibration curves were linear. The value of correlation coefficient were 0.9992 and 0.9994, respectively. The average recovery of aesculin was 98.2% and aesculetin was 99.2%. RSD were 2.24% and 2.15%, respectively. The quality differences of Cortex Fraxini among different provenances were remarked. The quality of Cortex Fraxini from Shanxi province was the best. Conclusion: The method is applied in determination and analytics of content of Cortex Fraxini and is rapid, simple and easy to carry out. The method is with the feature of accuracy, repetition and stability.