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Study on different parts and the best harvest time of Herba Sarcandrae

Author: Anonymous From: www.yourpaper.net Posted: 2009-08-30 13:09:26 Read:
Author: Li Chengren Wang Yuanyuan Li Xiaoyan Song Xiaohong Wei Qisheng Jiang Lin Ye Chuangxing
[Abstract] to determine the recovery position and the optimum harvest period of Herba sarcandrae.Different seasons of Herba Sarcandrae methods will be collected into root, stem, leaf of 3 parts, with 60% ethanol as solvent reflux extraction.The content of fumaric acid and isofraxidin by high performance liquid chromatography method for the determination of different seasons in different parts of Herba Sarcandrae; Determination of content of total flavonoids in the ultraviolet spectrophotometric method.Results the content of total flavonoids of Herba Sarcandrae leaves the highest content of fumaric acid, highest in stem, isofraxidin content is highest in roots, considering the 3 effective components, to June, the highest in August.Conclusion in June August, the best harvest time of Sarcandra glabra, to whole plant harvest is appropriate.
[keyword] sarcandra medicinal components harvest harvest parts
Sarcandra glabra as Chloranthaceae plants grass coral Sarcandra glabra (Thunb.) Nakai dry complete grass, also known as tea, elderberry, mainly distributed in Southern China, East China, southwest provinces, in Korea, Japan, Southeast Asia and India also have distribution."Chinese Pharmacopoeia" 2005 Edition ( Department) records, with Qingre Liangxue Huoxue Xiaoban, Qufeng Tongluo effect, can be used for treating blood-heat purpura, purpura, rheumatism, traumatic injury, disease (1).According to modern pharmacological experiments, glabrous sarcandra herb antibacterial anti-inflammatory, anti-tumor, has made various formulations for auxiliary treatment of various inflammatory diseases and cancer.The main efficacy components for fumaric acid, isofraxidin and flavonoids (2).The content of fumaric acid and isofraxidin in Sarcandra glabra in the HPLC method for the determination of (3, 4), flavonoid compounds through UV spectrophotometric method for the determination of (5).This experiment adopts the method of 3 kinds of medicinal components at the same time study, to evaluate the different seasons and different parts of Herba Sarcandrae medicinal value, provide the basis for the recovery of Herba sarcandrae.
1 instrument and reagent
1.1 Waters high performance liquid chromatograph (Waters515 pump, Waters 2487 UV detector), constant Austrian HT-230A column temperature box (Tianjin hengao company), YH3000 chromatography workstation (Guangzhou Yihai company).TU-1901 double beam UV-Vis spectrophotometer (Beijing Purkinje General Instrument Co., Ltd).
1.2 reagent fumaric acid reference substance (batch number: 1541-200001), isofraxidin control (batch number: 110837-200304), rutin reference substance (batch number: 100080-200) for the control of pharmaceutical and Biological Products Institute in china.Acetonitrile (HPLC, Merck), phosphate (HPLC, Fluka company), Mili-Q ultra-pure water, the rest of the reagents were analytically pure.


Sarcandra glabra mining in Donglan County of Guangxi GAP base of Sarcandra glabra, from 2007-05 to 2007-12 every month from whole plant, root, stem and leaf, 60 drying.
2 methods and results of
2.1 standard solution preparation
2.1.1 total flavonoids rutin standard solution weigh accurately control in 20 mg, 50 ml volumetric flask, add distilled water dissolved and diluted to the scale, shake up, into the concentration of 0.4 mg/ml rutin solution (5).
2.1.2 fumaric acid and isofraxidin control solution weigh accurately fumaric acid was 7.5 mg, isofraxidin was 1.8 mg, 25 ml volumetric flask, add distilled water dissolved and diluted to the scale, shake up, made of mixed standard solution containing fumaric acid (two to 0.3 mg/ml, isofraxidin 0.072 mg/ml).
2.2 the preparation of sample solution weigh accurately drying, crushing, over 3 sieve Sarcandra glabra root, stem, leaf powder 0.5 g, 250 ml flask, by 1 30 were added to liquid ratio of 60% ethanol reflux extraction 1 h, after filtering 2 times, merging filtrate repeat extraction, decompressing concentration to dry, with distilled water to dissolve the wreckage, set the volume to 25 ml, spare.
2.3  determination of total flavonoids; wavelength selection and rutin solution 0.5 ml, 10 ml volumetric flask, with 30% ethanol added to 5 ml, add 5%NaNO2 solution for 0.3 ml, placed 5 min, adding 10%Al (NO3) 3 solution for 0.3 ml, 6 min after adding 4%NaOH solution for 2 ml, the solution color red, mixed with 30% ethanol diluted to the scale, placed 10 min spectrophotometry in the ultraviolet meter under full wavelength spectrum scanning, the maximum absorption at 500 nm to 500 nm, so as the detection wavelength of total flavonoids.
2.4 HPLC conditions of the chromatographic column is Agilent Hypersil ODS C18 column (4.6 mm 250 mm, 5 m); the mobile phase A 0.05% water solution of phosphoric acid; mobile phase B of acetonitrile; the detection wavelength was 210 nm; column temperature of 35 .Gradient conditions of 0 ~ 8 min mobile phase B 5%; 8 ~ 30 min B5% ~ 12%; 30 ~ 45 min B12% to 19%.After the test, the samples were separated in 1.5 above, fumaric acid, the number of theoretical plates in 5000 above, isofraxidin theoretical plate number at more than 10000.
2.5 drawing standard curve
2.5.1 total flavonoids rutin standard curve were obtained from the reference solution of 0, 0.25, 0.5, 0.75, 1.25, 1.5 ml to 10 ml volumetric flask, according to the "2.3 methods" under, in the UV absorbance meter, the detection wavelength was 500 nm.The standard sample concentration as the abscissa, the absorbance value of standard curve is drawn as a vertical coordinate, and obtained the regression equation of Y=9.497 5X 0.0413, r=0.999 9, linear range: 0.01 ~ 0.06 mg/ml.
2.5.2 fumaric acid and isofraxidin standard curve accurately draw volume was 0.04, 0.2, 0.4, 2, 4, 6, 8 ml of fumaric acid and isofraxidin hybrid control solution to a 25ml volumetric flask, dilute with water to the mark, a definite volume with 0.45 m microporous membrane filter accurate lessons, 20 L, injected into the HPLC determination.The sample quality as abscissa, the peak area as a vertical coordinate drawing standard curve regression equation, respectively, fumaric acid and isofraxidin was: Y=6 966848.636, 2X 702 737.2867, r=0.999 6; Y=14 391526.525 4X 221 2.0133, r=0.999 8; the linear range: Fumaric acid is 0.024 4.8 g; differences in skin with 0.006 1.152 G.Study on the method of 2.6
2.6.1
science investigation methods Flavonids


Precision test: Sarcandra glabra plant samples, according to the "2.2" under the preparation of sample solution, according to the "2.3" under the method of processing and measuring 500 nm absorbance, measured 5 times, in terms of total flavonoid content, RSD 2.49%, show that the precision of the method is good.


Repeatability: take the same batch of medicinal plant samples of 6, according to the "law of 2.2" under the method of preparing the sample solution was determined, according to the "2.4" under the method, in terms of total flavonoid content, RSD =2.53%, repetitive method show good.


Stability test: take the same number of medicinal plant samples, according to the "law of 2.2" under the method of preparing the sample solution
, 3 h every 30 min were measured by "2.3" under the method, the results of RSD =1.58%, show the stable sample within 3 h.


The recovery experiment: Sarcandra glabra samples of known content, adding a certain amount of control, according to the "2.2" under the method of preparation of sample solution, according to the "2.3" after the determination of the light absorption value of total flavone, average recovery rate was 101.8%, RSD=3.44%.
2.6.2
science investigation of fumaric acid and isofraxidin method


Precision test: Sarcandra glabra plant samples, according to the "2.2" under the preparation of sample solution, repeat 5 samples, record the peak area ratio, fumaric acid and isofraxidin RSD values were 1.43% and 2.35%, show that the precision of the instrument.


Repeatability: take the same number of Sarcandra glabra samples of 6, according to the "2.2" under the preparation of sample solution, were injected into the HPLC analysis, fumaric acid, RSD = 4.64%; isofraxidin RSD = 4.13%, better repeatability that method.


Stability test: Sarcandra glabra sample solution at 0, 2, 4, 8, 24, 48 h respectively sampling analysis, fumaric acid and isofraxidin in RSD were 1.93% and 3.02%, showed that the samples were stable in 48 h.


The recovery experiment: Sarcandra glabra samples of known content, adding a certain amount of control, processing method according to the "2.2" under preparation of the sample solution, were analyzed by high performance liquid chromatography, the average recovery measured fumaric acid was 101.8%, RSD = 3.53%; the average recovery of isofraxidin the rate is 99.9%, RSD=3.44%.
2.7 content determination of effective components in a sample
2.7.1 Determination of content of total flavone from the precise 0.5 ml sample solution, according to the "2.3" under the method of order determination.The light absorption value was calculated by regression equation.The content of total flavonoids in different parts of each month is shown in table 1.
Table 1 from 5 to December, total flavonoids in different parts of Herba Sarcandrae (omitted)
2.7.2 Determination of fumaric acid and isofraxidin content accurately extracted sample solution 20 l injected into the liquid chromatograph, were determined according to the "2.4" under the chromatographic conditions (control sample and sample chromatogram is shown in Figure 1 to 2), calculated by peak area into the regression equation.Different positions of each month of fumaric acid and isofraxidin content table 2 ~ 3.
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