CN101185663B - Method for obtaining the extraction from caper medicinal part and application of the extraction - Google Patents
Method for obtaining the extraction from caper medicinal part and application of the extraction Download PDFInfo
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- CN101185663B CN101185663B CN2006101183698A CN200610118369A CN101185663B CN 101185663 B CN101185663 B CN 101185663B CN 2006101183698 A CN2006101183698 A CN 2006101183698A CN 200610118369 A CN200610118369 A CN 200610118369A CN 101185663 B CN101185663 B CN 101185663B
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Abstract
The invention discloses a method of obtaining extract from the medical part of capparis spinosa L. This method includes the following steps: the medicinal part of capparis spinosa L. is pulverized; the crude extracts is obtained through heating reflux, cold soaking, percolate extraction and decompressed concentration. The crude extract is defatted by ligroin, and the invention is characterized inthat the residue after defatting is extracted by chloroform to get the extract. This extract is used for treating rheumatic arthritis, rheumatoid arthritis and periarthritis of shoulder. Pharmacodynamics study demonstrates that the extract obtained from the invention has obvious curative effect on multiple arthritis models (including type collagen II induced rheumatoid arthritis models).
Description
Technical field
The present invention relates to the Natural Medicine Chemistry field, specifically, relate to the method for acquisition extract from Cortex Capparis Spinosae Radicis and the application of this extract.
Background technology
Cortex Capparis Spinosae Radicis is Capparidaceae plant Cortex Capparis Spinosae Radicis Capparis spinoaL, has another name called in mallet fruit rattan, Cortex Capparis Spinosae Radicis, Herba hibisci trioni, the spinach to restrain fruit etc.Cortex Capparis Spinosae Radicis is a kind of fruticuli that is grown under the hot arid climatic conditions, and China mainly is distributed in the low mountain of Xinjiang Tianshan north and south stone matter, hilly upland, the foot of a mountain coombe or Gobi desert.In addition, on Gansu, Tibet and other places growth is arranged also.Cortex Capparis Spinosae Radicis mainly is distributed in area, gulf, Mediterranean abroad, and trophophase is that May is to October.The medicinal part of Cortex Capparis Spinosae Radicis is leaf, fruit and root bark, acrid in the mouth hardship, warm in nature, and among the people with diseases such as treating arthritis by using caper, ventilations, effect is remarkable.
(Agricultural and BiologicalChemistry 34 can only obtain cupreol, cupreol glucoside or 1-hydrogen-3-acetonitrile-base-4-methoxyl group-indole respectively in 1590-1592) from document " chemical constitution study of Herba hibisci trioni fruit " (Harbin University of Commerce's journal natural science edition 21 volume the 6th phase 684-686 page or leaf) and Isolation and identification of indolederivatives in clubroots of Chinese cabbage.And Chinese patent 200310113957.9 discloses a kind of method of extracting arthritis effective site from the plant Capparis fruit, this method comprises really to be pulverized Cortex Capparis Spinosae Radicis, reflux, merceration or percolation extract, concentrating under reduced pressure obtains crude extract, and crude extract is used earlier defat with petroleum ether, residue is used ethyl acetate and n-butyl alcohol system eluting successively, obtains ethyl acetate effective site and n-butyl alcohol effective site respectively.But test shows that ethyl acetate and n-butanol portion are invalid to the inductive rheumatoid arthritis model of II Collagen Type VI.
Summary of the invention
For solving the problems of the technologies described above, one of purpose of the present invention provides a kind of method that obtains extract from the Cortex Capparis Spinosae Radicis medicinal part, this method comprises pulverizes the Cortex Capparis Spinosae Radicis medicinal part, reflux, and merceration or percolation extract, concentrating under reduced pressure, obtain crude extract,, it is characterized in that the crude extract defat with petroleum ether, residue chloroform extraction after the defat obtains extract.
Two of purpose of the present invention provides the extract that is made by described method.
Described extract contains cupreol, cupreol glucoside and 1-hydrogen-3-acetonitrile-base-4-methoxyl group-indole.
Three of purpose of the present invention provides the application of described extract in the medicine of preparation treatment rheumatic arthritis, rheumatoid arthritis, scapulohumeral periarthritis.
Four of purpose of the present invention provides a kind of pharmaceutical composition, and this pharmaceutical composition contains described extract and acceptable accessories.
The concrete steps of the method that obtains extract from Cortex Capparis Spinosae Radicis of the present invention are as follows:
1) Cortex Capparis Spinosae Radicis fruit, stem and leaf and root bark are pulverized, carried out reflux with 60-90% ethanol, merceration or percolation extract, and extract 1-3 time, and extracting solution obtains the crude extract of Cortex Capparis Spinosae Radicis fruit, stem and leaf and root bark at 40-60 ℃ of concentrating under reduced pressure;
2) crude extract is earlier used defat with petroleum ether, the residue chloroform extraction obtains containing the extract of cupreol, cupreol glucoside and 1-hydrogen-3-acetonitrile-base-4-methoxyl group-indole.
By method of the present invention, can obtain containing the Cortex Capparis Spinosae Radicis chloroform extract of cupreol, cupreol glucoside and 1-hydrogen-3-acetonitrile-base-4-methoxyl group-indole.The test of pesticide effectiveness proves, by the extract that contains cupreol, cupreol glucoside and 1-hydrogen-3-acetonitrile-base-4-methoxyl group-indole of chloroform extraction treatment rheumatic arthritis, rheumatoid arthritis and scapulohumeral periarthritis shown good effect.This extract all shows good effect to multiple arthritis model (comprising the inductive rheumatoid arthritis model of II Collagen Type VI), and toxicity is very low, and is based on the little chemical constituent of polarity.
Description of drawings
The arthritic influence of chmice acute that Fig. 1 causes for oral Cortex Capparis Spinosae Radicis chloroform extract on Carrageenan;
Fig. 2 is the preventive effect of oral Cortex Capparis Spinosae Radicis chloroform extract to the inductive rat kind rheumatic arthritis of II Collagen Type VI;
Fig. 3 is the therapeutical effect of oral Cortex Capparis Spinosae Radicis chloroform extract to the inductive rat kind rheumatic arthritis of II Collagen Type VI;
Fig. 4 is the preventive effect of oral Cortex Capparis Spinosae Radicis chloroform extract to rat assist agent arthritis;
Fig. 5 is the therapeutical effect of oral Cortex Capparis Spinosae Radicis chloroform extract to rat assist agent arthritis;
Fig. 6 is the TLC collection of illustrative plates of the cupreol of Cortex Capparis Spinosae Radicis chloroform extract, and wherein 1 is the Cortex Capparis Spinosae Radicis chloroform extract, and 2,3 is cupreol, and developing solvent is a chloroform: methanol=20: 1, developer are vanillin-concentrated sulphuric acid;
Fig. 7 is the TLC collection of illustrative plates of the cupreol glucoside of Cortex Capparis Spinosae Radicis chloroform extract, and wherein 1 is the Cortex Capparis Spinosae Radicis chloroform extract, and 2,3 is the cupreol glucoside, and developing solvent is a chloroform: methanol=15: 1, developer are vanillin-concentrated sulphuric acid;
Fig. 8 a and 8b are respectively the HPLC collection of illustrative plates of Cortex Capparis Spinosae Radicis chloroform extract and 1-hydrogen-3-acetonitrile-base-4-methoxyl group-indole, and wherein high-efficient liquid phase chromatogram condition is as follows: the HP1100 chromatographic column be DiamonsilC18 (250 * 4.6mm, 5 μ, Dikma).(acetonitrile Merck) changes: 0 → 20min 5%B → 30%B, 21 → 50min 31%B → 70%B in gradient for mobile phase A (ultrafiltration water) and B; Detect wavelength 254nm, flow velocity 1.0mL/min, 25 ℃ of column temperatures.
The specific embodiment
Embodiment 1:
2 kilograms of Cortex Capparis Spinosae Radicis fruits, stem and leaf and root bark are pulverized, with 20 liter of 75% ethanol carry out reflux, merceration is carried, and gets extraction 2 times, extraction time is 2 hours, merge extractive liquid,, extracting solution can obtain Cortex Capparis Spinosae Radicis arthritis crude extract extractum at 40 ℃ of concentrating under reduced pressure;
Crude extract extractum is successively used defat with petroleum ether, and the residue chloroform extraction obtains containing the extract of cupreol, cupreol glucoside and 1-hydrogen-3-acetonitrile-base-chemical compounds such as 4-methoxyl group-indole.
Embodiment 2:
5 kilograms of Cortex Capparis Spinosae Radicis fruits, stem and leaf and root bark are pulverized, carried out reflux with 100 liter of 70% ethanol, percolation extracts, extract 2 times, extraction time is 5 hours, merge extractive liquid,, extracting solution can obtain Cortex Capparis Spinosae Radicis arthritis crude extract extractum at 45 ℃ of concentrating under reduced pressure;
Crude extract extractum is successively used defat with petroleum ether, and the residue chloroform extraction obtains containing the extract of cupreol, cupreol glucoside and 1-hydrogen-3-acetonitrile-base-chemical compounds such as 4-methoxyl group-indole.
3 kilograms of Cortex Capparis Spinosae Radicis fruits, stem and leaf and root bark are pulverized, carried out reflux with 45 liter of 80% ethanol, merceration extracts, extract 1 time, extraction time is 10 hours, merge extractive liquid,, extracting solution can obtain Cortex Capparis Spinosae Radicis arthritis crude extract extractum at 50 ℃ of concentrating under reduced pressure;
Crude extract extractum is successively used defat with petroleum ether, and the residue chloroform extraction obtains containing the extract of cupreol, cupreol glucoside and 1-hydrogen-3-acetonitrile-base-chemical compounds such as 4-methoxyl group-indole.
5 kilograms of Cortex Capparis Spinosae Radicis fruits, stem and leaf and root bark are pulverized, carried out reflux with 40 liter of 90% ethanol, percolation extracts, extract 1 time, extraction time is 8 hours, merge extractive liquid,, extracting solution can obtain Cortex Capparis Spinosae Radicis arthritis crude extract extractum at 60 ℃ of concentrating under reduced pressure;
Crude extract extractum is successively used defat with petroleum ether, and the residue chloroform extraction obtains containing the extract of cupreol, cupreol glucoside and 1-hydrogen-3-acetonitrile-base-chemical compounds such as 4-methoxyl group-indole.
Embodiment 5
2 kilograms of Cortex Capparis Spinosae Radicis fruits, stem and leaf and root bark are pulverized, with 30 liter of 65% ethanol carry out reflux, merceration is carried, and gets extraction 2 times, extraction time is 2 hours, merge extractive liquid,, extracting solution can obtain Cortex Capparis Spinosae Radicis arthritis crude extract extractum at 40 ℃ of concentrating under reduced pressure;
Crude extract extractum is successively used defat with petroleum ether, and the residue chloroform extraction obtains containing the extract of cupreol, cupreol glucoside and 1-hydrogen-3-acetonitrile-base-chemical compounds such as 4-methoxyl group-indole.
The arthritis drug effect experimental example of the chloroform extract that from Cortex Capparis Spinosae Radicis, obtains
One, the chloroform extract on Carrageenan is brought out the influence of mice pedal swelling
The 20g kunming mice is divided into administration group: 500mg/kg, po earlier; Positive control indometacin group: 10mg/kg, po; The blank group; Administration is three days as stated above, cause survey earlier before scorching respectively organize the right back sufficient sole of the foot of rat volume once, the last administration caused inflammation at the right back sufficient sole of the foot middle part subcutaneous injection 1% carrageenin 0.1ml of rat after one hour, surveyed sufficient sole of the foot volume 1 time every one hour later on, calculate swelling rate and suppression ratio, group difference compares with the t check.The result is as follows.
Table 1 chloroform extract on Carrageenan causes influence (%, n=10, the x ± s) of chmice acute inflammation
In () for swelling rate (%)＜in be suppression ratio (%)
By table 1 as seen, chloroform extract can suppress carrageenin significantly and cause mice foot sole of the foot acute inflammation, and is higher than the effect of indometacin group.
Two, chloroform extract is to the influence of the inductive rat arthritis of II type glue unit (CII)
Proinflammatory agent: II type glue unit is dissolved in the 0.1mol/L acetic acid, making final concentration is 2mg/ml glue unit solution, 4 ℃ are spent the night, add to II type glue unit drips of solution in the cold incomplete Freund's adjuvant next day, it is fully emulsified that (II type glue unit liquid: incomplete Freund=1: 1), making II type glue unit final concentration is 1mg/ml.
4 ℃ of refrigerators of II type glue unit Emulsion are preserved standby.
Immunization method: every rat is divided 5 back intradermal injection immunities, every some 0.1ml, 5ml (contain II type glue unit amount and be 0.5mg) altogether, after 7 days with same method supplementary immunization once.Matched group injection 0.1mol/L acetic acid and incomplete Freund's adjuvant emulsion reach immunity and reinstated the swelling degree that drainage is measured right back ankle and foot joint in back 20 days before the injection.
1. chloroform extract is to the inductive arthritic preventive effect of II type glue unit (CII)
150～180gWistar rat is divided into the administration group: chloroform extract 50mg/kg, po; Indometacin group 1mg/kg, po; The blank group.The beginning administration is 8 days after the administration treated animal first immunisation.Measure before the immunity and 20,22,24,27 days ankle arthroncus value after the first immunisation with drainage, calculate swelling rate and suppression ratio, group difference compares with the t check.
Table 2 chloroform extract is to the inductive arthritic preventive effect of II type glue unit (CII) (%, n=8, x ± s)
In () for swelling rate (%)＜in be suppression ratio (%)
By table 2 as seen, chloroform extract can prevent the inductive arthritis of II type glue unit significantly, and it acts on all apparently higher than the indometacin group.
2. chloroform extract is to the inductive arthritic therapeutical effect of II type glue unit (CII)
150～180gWistar rat is divided into the administration group: chloroform extract 50mg/kg, po; Indometacin group 1mg/kg, po; The blank group.The 19th day began successive administration 8 days after the administration treated animal first immunisation.Measure before the immunity and 19,21,23,26,28 days ankle arthroncus value after the first immunisation with drainage, it is the same to calculate swelling rate and suppression ratio, and group difference compares with the t check.
Table 3 chloroform extract is to the inductive arthritic therapeutical effect of II type glue unit (CII) (%, n=8, x ± s)
In () for swelling rate (%)＜in be suppression ratio (%)
As shown in Table 3, chloroform extract can be treated the inductive arthritis of II type glue unit, and its effect all is higher than the indometacin group.
Three, chloroform extract is to the influence of adjuvanticity rat arthritis
1.Zhp-I to the inductive rat assist agent arthritis preventive effect of adjuvant
130～150gSD rat is divided into heavy dose of group: chloroform extract 250mg/kg, iv; Middle dosage group: chloroform extract 125mg/kg, iv; Small dose group: chloroform extract 62.5mg/kg, iv; Indometacin group: 1mg/kg, po; The blank group.Animal causes scorching preceding elder generation with the volume of surveying the right back sufficient sole of the foot once, cause inflammation at the right back sufficient sole of the foot of rat middle part subcutaneous injection Freund's complete adjuvant 0.1ml then, begin by above-mentioned grouping after causing scorching 7 days, and beginning administration 8 days, survey sufficient sole of the foot volume at regular intervals, calculate swelling rate and suppression ratio, group difference compares with the t check.
Table 4 chloroform extract is to the preventive effect of rat assist agent arthritis (%, n=8, x ± s)
| 0day | 2day | 4day | 7day | 9day | 11day | 13day | 15day | |
| Blank | 1.22±0.09 | 1.28±0.07 | 1.29±0.04 | 1.29±0.07 | 1.29±0.06 | 1.27±0.05 | 1.28±0.04 | 1.29±0.03 |
| Cause scorching contrast | 1.35±0.14 | 2.53±0.30 (113.98±11.07) | 2.53±0.30 (87.91±12.40) | 2.32±0.32 (72.64±17.36) | 2.85±0.31 (113.39±30.89) | 2.92±0.35 (118.42±31.98) | 2.84±0.40 (111.57±29.78) | 2.99±0.37 (122.38±15.07) |
| High dose 250mg/kg | 1.31±0.07 | 2.41±0.22 (111.60±19.15) | 2.41±0.22 (84.55±14.08) | 2.16±0.23 (65.19±16.75) | 2.60±0.48 (97.94±28.68) <13.62> | 2.35±0.38 (79.26±24.08) <33.07> | 2.41±0.24 (84.80±16.06) <23.99> | 2.48±0.36 (89.65±28.27) <26.74> |
| Middle dosage 125mg/kg | 1.37±0.13 | 2.54±0.37 (107.86±16.96) | 2.54±0.37 (85.72±16.83) | 2.30±0.18 (68.64±16.35) | 2.80±0.35 (104.99±22.49) <7.40> | 2.57±0.36 (88.04±21.38) <25.66> | 2.59±0.23 (89.89±17.77) <19.43> | 2.74±0.25 (101.37±20.33) <17.17> |
| Low dosage 62.5mg/kg | 1.22±0.09 | 2.22±0.19 (111.27±22.18) | 2.22±0.19 (82.69±19.94) | 2.06±0.14 (68.94±16.34) | 2.34±0.24 (92.53±23.44) <18.39> | 2.37±0.20 (94.65±23.29) <20.07> | 2.38±0.22 (95.31±21.47) <14.57> | 2.41±0.18 (98.27±18.91) <19.70> |
| Radix Tripterygii Wilfordii 1.5mg/kg | 1.34±0.07 | 2.75±0.22 (105.26±11.59) | 2.26±0.16 (68.82±8.54) | 2.49±0.20 (86.83±20.05) | 2.76±0.43 (107.77±41.79) <4.96> | 2.45±0.26 (84.23±25.14) <28.87> | 2.80±0.06 (109.57±12.18) <1.79> | 2.55±0.20 (90.99±17.93) <25.65> |
| Indometacin 1mg/kg | 1.24±0.12 | 2.54±0.27 (105.18±8.84) | 2.32±0.24 (86.94±2.86) | 2.24±0.45 (81.92±34.54) | 2.20±0.25 (78.82±23.89) <30.49> | 2.19±0.15 (77.85±16.90) <34.26> | 2.21±0.14 (79.93±16.58) <28.36> | 2.30±0.26 (87.37±26.38) <28.61> |
In () for swelling rate (%)＜in be suppression ratio (%)
As shown in Table 4, the high dose chloroform extract is equivalent to the indometacin group to rat assist agent arthritis, organizes a little more than Radix Tripterygii Wilfordii.
2.Zhp-I to the inductive rat assist agent arthritis therapeutical effect of adjuvant
130～150gSD rat is divided into heavy dose of group: chloroform extract 250mg/kg, iv; Middle dosage group: chloroform extract 125mg/kg, iv; Small dose group: chloroform extract 62.5mg/kg, iv; Indometacin group: 1mg/kg, po; Radix Tripterygii Wilfordii group: 1.5mg/kg, po.Animal causes scorching preceding elder generation with the volume of surveying the right back sufficient sole of the foot once, cause inflammation at the right back sufficient sole of the foot of rat middle part subcutaneous injection Freund's complete adjuvant 0.1ml then, begin by above-mentioned grouping after causing scorching 19 days, and beginning administration 8 days, survey sufficient sole of the foot volume at regular intervals, calculate swelling rate and suppression ratio, group difference compares with the t check.
Table 5 chloroform extract is to the therapeutical effect of rat assist agent arthritis (%, n=8, x ± s)
| 0day | 19day | 22day | 24day | 27day | 29day | |
| Cause scorching contrast high dose 250mg/kg | 1.13±0.04 1.16±0.07 | 2.63±0.25 (140.13±26.67) 2.52±0.3 (107.58±26.43) <23.23> | 2.63±0.25 (133.06±20.43) 2.52±0.3 (117.80±29.28) <11.47> | 2.79±0.31 (146.74±25.52) 2.62±0.3 (127.03±28.00) <13.43> | 2.71±0.33 (139.71±26.77) 2.58±0.28 (123.66±30.09) <11.48> | 2.72±0.26 (140.85±24.60) 2.55±0.23 (120.61±23.69) <14.37> |
| Middle dosage 125mg/kg | 1.07±0.04 | 2.31±0.26 (122.32±19.81) <12.71> | 2.31±0.26 (116.88±21.06) ＜12.16 | 2.33±0.27 (118.10±22.30) <19.52> | 2.29±0.29 (114.44±20.40) <18.09> | 2.26±0.3 (111.99±23.40) <20.49> |
| Low dosage 62.5mg/kg | 1.11±0.07 | 2.48±0.22 (123.55±20.96) <11.83> | 2.48±0.22 (125.24±22.53) ＜5.88 | 2.54±0.19 (130.14±22.33) <11.31> | 2.51±0.2 (127.87±2206) <8.48> | 2.57±0.2 (133.25±24.24) <5.40> |
| Radix Tripterygii Wilfordii 1.5mg/kg | 1.13±0.05 | 2.5±0.22 (122.82±23.38) <12.35> | 2.49±0.32 (121.22±24.54) <8.90> | 2.61±0.17 (131.55±10.05) <10.35> | 2.48±0.19 (121.02±23.92) <13.38> | 2.4±0.21 (113.12±18.05) <19.69> |
| Indometacin 1mg/kg | 1.14±0.08 | 2.6±0.36 (127.69±15.64) <8.88> | 2.58±0.38 (125.53±19.50) <5.66> | 2.48±0.34 (117.38±19.70) <20.01> | 2.4±0.13 (112.42±20.71) <19.53> | 2.34±0.12 (106.99±16.97) <24.04> |
In () for swelling rate (%)＜in be suppression ratio (%)
As shown in Table 5, the high dose chloroform extract is equivalent to the indometacin group to rat assist agent arthritis, organizes a little more than Radix Tripterygii Wilfordii.
Claims (6)
1. obtain the method for extract from Cortex Capparis Spinosae Radicis fruit, stem and leaf and root bark, this method comprises:
(1) Cortex Capparis Spinosae Radicis fruit, stem and leaf and root bark are pulverized, carried out reflux with 60-90% ethanol, merceration or percolation extract, concentrating under reduced pressure, obtain Cortex Capparis Spinosae Radicis really, the crude extract of stem and leaf and root bark;
(2) crude extract is used earlier defat with petroleum ether, the residue after the defat obtains extract after with chloroform extraction.
2. the method for claim 1, wherein merceration or percolation extract and carry out 1-3 time, and the temperature of concentrating under reduced pressure is 40-60 ℃.
3. the extract that makes by the described method of claim 1.
4. extract as claimed in claim 3 is characterized in that, this extract contains cupreol, cupreol glucoside and 1-hydrogen-3-acetonitrile-base-4-methoxyl group-indole.
5. claim 3 or the 4 described extracts application in the medicine of preparation treatment rheumatic arthritis, rheumatoid arthritis, scapulohumeral periarthritis.
6. pharmaceutical composition contains claim 3 or 4 described extract and acceptable accessories.
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| CN102423323B (en) * | 2011-12-12 | 2013-03-13 | 中国科学院新疆理化技术研究所 | Compound medicine for treating rheumatism and preparation method thereof |
| CN106667990A (en) * | 2016-11-17 | 2017-05-17 | 郑州郑先医药科技有限公司 | Medicine for treating rheumatism |
| CN106620315A (en) * | 2016-11-17 | 2017-05-10 | 郑州郑先医药科技有限公司 | Medicinal composition for treating rheumatism |
| CN109303790B (en) * | 2018-12-04 | 2021-07-20 | 上海中医药大学附属曙光医院 | Medical application of caper or caper extract |
| CN111690603B (en) * | 2020-06-22 | 2021-09-24 | 青岛赛尔斯干细胞库有限公司 | Biological agent for promoting proliferation and migration of dental pulp stem cells |
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| CN1541700A (en) * | 2003-11-05 | 2004-11-03 | 中国科学院新疆理化技术研究所 | Method for extracting active arthritis resisting element from Capparis fruit |
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| CN1541700A (en) * | 2003-11-05 | 2004-11-03 | 中国科学院新疆理化技术研究所 | Method for extracting active arthritis resisting element from Capparis fruit |
Non-Patent Citations (4)
| Title |
|---|
| 姜薇，林文翰，郭守东.野西瓜果实的化学成分研究.哈尔滨商业大学学报（自然科学版）21卷 6期.2005,21(6),684-689. |
| 姜薇,林文翰,郭守东.野西瓜果实的化学成分研究.哈尔滨商业大学学报(自然科学版)21卷 6期.2005,21(6),684-689. * |
| 季宇彬，郭守东，汲晨锋.野西瓜的化学和药理研究.哈尔滨商业大学学报（自然科学版）22卷 1期.2006,22(1),5-10. |
| 季宇彬,郭守东,汲晨锋.野西瓜的化学和药理研究.哈尔滨商业大学学报(自然科学版)22卷 1期.2006,22(1),5-10. * |
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