1.Study on effect of different extraction methods on anti-inflammatory and analgesic activity of Anemone hupehensis
Xiang LI ; Yihong XU ; Xiaohui SUN ; Zhongyao HAN ; Yunyan FU ; Junke WU
Chongqing Medicine 2018;47(15):1990-1992,1996
Objective To investigate the effect of different extraction methods on anti-inflammatory and analgesic activity of Anemone hupehensis.Methods The different abstracts were prepared from the whole herb of Anemone hupehensis.The analgesic effect was observed by adopting the mouse torsion and electric heating plate method,and the anti-inflammatory activity was comprehensively evaluated by using the mouse ear tumefaction,toe tumefaction and tampon granulation tumefaction exprements.Results Compared with the blank model group,the anti-inflammatory action difference of low dose in the water layer parts of mouse ear tumefaction,toe tumefaction and tampon granulation tumefaction had no statistical significance(P>0.05),and the extracting parts of rest doses all had significant anti-inflammatory and analgesic effect (P<0.05).Ethyl acetate part had strongest activity in the electric heating plate experiment.N-butanol part had strongest activity in the ear tumefaction,toe tumefaction,tampon granulation tumefaction experiments and torsion method.Conclusion The whole herb of Anemone hupehensis has prominent anti-inflammatory and analgesic effect,and the ethyl acetate part E and N-butanol part are main effective parts.
2.Optimization of the Extraction Technology of Senecio scandens by Orthogonal Design Combined with Informa- tion Entropy Weighting Method
Yan LI ; Yanfan PAN ; Tao ZHANG ; Zhongyao HAN
China Pharmacy 2020;31(12):1470-1474
OBJECTIVE:To optimize the extraction technology of Senecio scandens ,and to provide reference for the further development and utilization of the medicinal material. METHODS :The method of reflux extraction with 80 ℃ water bath was used to extract S. scandens . HPLC method was adopted to determine the contents of chlorogenic acid and hyperoside. The determination was performed on Diamonsil C 18 with mobile phase consisted of 0.2% glacial acetic acid water solution-methanol (82∶18,V/V)at the flow rate of 1.0 mL/min;the column temperature was set at 35 ℃;the detection wavelength was 327 nm,and the sample size was 5 μL. UV-Vis spectrophotometry was used to determine the contents of total flavonoids(by rutin )and total alkaloids (by matrine)and the detection wavelengths were set at 509 nm and 208 nm,respectively. Based on single factor tests ,using ethanol volume fraction (A,%),solvent folds (B,mL/g),extraction time (C,h),extraction times (D,times)as factors ,using the contents of chlorogenic acid ,hyperoside,total flavonoids and total alkaloids as indexes ,and t he weight coefficients of above indicator components were calculated based on information entropy weighting method so as to calculate their comprehensive scores , then L 9(34)orthogonal design was used to further optimize the extraction technology. The validation tests were also performed. RESULTS:The optimal extraction technology of S. scandens included that 8-fold 50% ethanol,extracting for 3 times,lasting for 1.5 h each time. Results of 3 times of validation tests showed that RSDs of the contents of chlorogenic acid ,hyperoside,total flavonoids and total alkaloids were all lower than 1.5%(n=3). CONCLUSIONS :The optimized technology is reproducible , stable and feasible ,and can be used for the extraction of S. scandens .
3.Analysis of Ongoing Change Characteristics of the Contents of Syringin and Total Flavonoids in Different Medicinal Parts of Toricellia angulata from Guizhou
Zhongyao HAN ; Wei SONG ; Yan LI ; Tao ZHANG ; Wenshuang TANG ; Fujun ZHOU
China Pharmacy 2020;31(17):2124-2128
OBJECTIVE:To study ongoing change characteristics of the contents of syringin and total flavonoids in different medicinal parts (root bark ,tree bark ,leaf)of Toricellia angulata from Guizhou ,and to provide reference for the development and application of T. angulata . METHODS :The root bark ,tree bark and leaf parts of T. angulata during different harvesting periods (Jan.-Dec.) were taken as the research samples. The content of syringin was determined by HPLC. The determination was performed on Agela Promosil C 18 column with mobile phase consisted of 0.5% phosphoric acid solution-acetonitrile (gradient elution)at the flow rate of 1.0 mL/min. The detection wavelength was set as 210 nm,and column temperature was 35 ℃. The sample size was 5 μL. The content of total flavonoids was determined by UV-visible spectrophotometry under detection wavelength of 510 nm. RESULTS :The linear range of syringin and total flavonoids were 0.095 9-1.150 8 mg/mL(r=0.999 6)and 0.072 2- 1.083 0 mg/mL(r=0.999 9),respectively. RSDs of precision ,stability and repeatability tests were all less than 3%(n=6). The average recoveries were 101.74%(RSD=2.36% ,n=6)and 99.63%(RSD=2.19% ,n=6),respectively. During different harvesting periods ,the contents of syringin in root bark ,tree bark ,leaf of T. angulata collected on Aug. ,May and Sept. were the highest,and the contents of total flavonoids in samples collected on Feb. ,Dec. and Sept. were the highest. The contents of syringin in different medicinal parts of T. angulata were in descending order as follows as tree bark >root bark >leaf,and the content of syringin was commonly relatively high in tree bark part ;the content of total flavonoids in different medicinal parts of T. angulata were in descending order as follows as root bark >tree bark >leaf,and the contents of total flavonoids in three medicinal parts was generally low. The content of total flavonoids in root bark was the highest in Feb. of that year ,and the content of syringin in root bark at same month was second only to Aug. of that year ;the content of syringin in tree bark was the highest in May ,and the content of total flavonoids in tree bark at same month was second only to Oct. and Dec. of that year ;the contents of total flavonoids and syringin in leaf were the highest in Sept. of that year. CONCLUSIONS :It is suggested that Feb. is the best time for harvesting root bark ,May for tree bark and Sept. for leaf of T. angulata .
4.Establishment of HPLC Fingerprint ,Chemical Pattern Recognition Analysis and Content Determination of the Leaves of Toricellia angulata from Different Regions
Zhongyao HAN ; Jun XIANG ; Jianyu CHEN ; Yiyong SONG ; Shiwai LI ; Wenshuang TANG ; Zujun YE ; Linsu ZHANG ; Hao TIAN ; Wanle WANG
China Pharmacy 2021;32(10):1224-1229
OBJECTIVE:To provide reference for the quality control of the leaves of Toricellia angulata . METHODS :HPLC method was adopted. The determination was performed on Agela Promosil C 18 column with 0.2% phosphoric acid solution-acetonitrile(gradient elution )as mobile phase at the flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm,and column temperature was 35 ℃. The sample size was 10 μL. HPLC fingerprint of 10 batches of the leaves of T. angulata was established and similarity evaluation was conducted by using Similarity Evaluation System of TCM Chromatographic Fingerprint(2004 edition). The chromatographic peak was identified by comparing with the chromatogram of reference substance. Cluster analysis ,PCA and PLS-DA were used to identify chemical patterns ,and the quality differential markers were screened. The contents of hyperoside and isoquercitrin were determined by the same HPLC. RESULTS :The similarities of HPLC fingerprint of 10 batches of the leaves of T. angulata with control fingerprint were 0.923-0.983. A total of 11 common peaks were identified ,and the peaks 4 and 5 were hyperoside and isoquercitrin ,respectively. Results of cluster analysis ,PCA and PLS-DA showed that 10 batches of leaves of T. angulata could be divided into two categories ,Y10 was clustered into one category ,and others were clustered into one category. PLS-DA analysis showed that 6 common peaks (peaks 4,3,10,2,6 and 11) with variable importance projection (VIP)greater than 1 were selected. Average contents of hyperoside and isoquercitrin in 10 batches of the leaves of T. angulata were 0.47-6.97,0.21-1.87 mg/g,respectively. CONCLUSIONS :Established HPLC fingerprint and the method for content determination are stable and reliable ,and can be used for the quality control of the leaves of T. angulata from different areas. Six quality differential markers including hyperoside in the leaves of T. angulata from different areas are qnyz202034) preliminarily screened.