Dissertation
Dissertation > Medicine, health > Chinese Medicine > Of Pharmacy > Traditional Chinese medicine chemical > Chemical analysis and identification

Studies on Fingerprint Analysis and Total Flavone Extracts of Saussurea Involucrata ( Kar. Et Kir.) Sch.-Bip.

Author LvRuiHuang
Tutor XuJianGuo
School Xinjiang Medical University
Course Pharmacy
Keywords Saussurea involucrata ( Kar. et Kir.) Sch.-Bip fingerprints flavonoids extracts quality control pharmacodynamics
CLC R284.1
Type Master's thesis
Year 2011
Downloads 28
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Objective:①To establish chromatographic fingerprint of Saussurea involucrata (Kar. et Kir.) Sch.-Bip., and to provide the foundation for the quality control of this herb;②Through the studies on quality, stability and pharmacodynamics for flavonoids extracts from Saussurea involucrata, to provide the basis for the further development and utilization. Methods:①We collected ten batches herbs of Saussurea involucrata from different places in Xinjiang province. We established the HPLC chromatographic fingerprint using gradient elution method, and we applied the software "Evaluation Standard System to the similarity of fingerprints in Chinese traditional medicine (2004 version)" for analysis and evaluation of the ten fingerprints.②The contents of total flavone in the flavonoids extracts from Saussurea involucrata was determined by colorimetry.③Rutin and syringin in the flavonoids extract from Saussurea involucrata was determined by high performance liquid chromatography at the same time.④The stability of the flavonoids extracts from Saussurea involucrata was studied through the influence factors such as temperature, humidity, light.⑤Anti-inflammatory effect and analgesic effect of the flavonoids extracts of Saussurea involucrata were evaluated by mouse ear edema resulted from xylene, mouse writhing symptom caused by acetic acid, rat paw swelling induced by carrageenan, Freund’s complete adjuvant induced primary, secondary arthritis damage in rats and the weight of cotton granuloma in rat. The T and B lymphocyte proliferation of mouse were determined by MTT method.Results:①The chromatographic fingerprint of Saussurea involucrata was established. We found that there was not significant difference in the fingerprints although they came from different regions.②The colorimetric method to determine the content of total flavone in the flavonoids extracts of Saussurea involucrata was applied with NaNO2-A1(NO3)3-NaOH system. Rutin was used as reference substance. The method was validated to show convenient, stable and accurate. The standard curve equation was y=0.5251x-0.0223(r=0.9996). The linearity of total flavonoids was fine in the range of 0.2-1.2mg. The relative standard derivation(RSD) in the precision test was 0.46%. RSD in the stability test was 1.01%. That showed there was no influence on the determination for the sample solution placed 5 days. RSD in the replicate test was 0.70%. The average recovery rate was 102.32% and RSD was 0.96%. The contents of the flavonoids in 3 batches sample were 44.39%, 42.64% and 43.14% respectively.③Rutin and syringin in the flavonoids extracts from Saussurea involucrata were determined by high performance liquid chromatography at the same time. The method was validated to show convenient, stable and accurate. For syringin the standard curve equation was A = 25842626.81C-939.15(r=0.9999). The linearity of syringin was fine in the range of 0.00276-0.02208mg/ml. For rutin the standard curve equation was A= 17863886.50C– 28941.33(r=0.9996). The linearity of rutin was fine in the range of 0.02-0.16mg/ml. RSD of syringin and rutin in the precision test were 0.97% and 0.90% respectively. RSD of syringin and rutin in the replicate test were 1.16% and 1.13% respectively. RSD of syringin and rutin in the stability test were 1.78% and 1.89% respectively. The average recovery rate of syringin was 102.56% and RSD was 1.40%. The average recovery rate of rutin was 101.85% and RSD was 1.89%. The contents of syringin in 3 batches of flavonoids extracts were 0.88%, 0.85% and 0.97% respectively. The contents of rutin in 3 batches of flavonoids extracts were 7.22%, 6.34% and 6.82% respectively.④The temperature(60℃)and light(4500lx) had no influence to the appearance characters of flavonoids extracts from Saussurea involucrata. But the relative humidity (RH75%)caused moisture absorption of flavonoids extracts. The temperature(60℃) had no influence to the contents of total flavone, syringin and rutin in the flavonoids extracts from Saussurea involucrata. The relative humidity (RH75%) and light (4500lx) had no influence to the content of syringin in the flavonoids extracts, but they decreased the contents of total flavone and rutin in flavonoids extracts slightly. So the flavonoids extracts from Saussurea involucrata should be stored in dry and dark place.⑤The flavonoids extracts from Saussurea involucrata can protect the adjuvant arthritis damage of primary and secondary. They can inhibit the ear edema resulted from xylene and reduce writhing times caused by acetic acid significantly in mice. They can relieve rat paw swelling induced by carrageenan, too. They can reduce the weight of cotton granuloma in rat. The large doses of the flavonoids extracts were better than the positive control medicine wild papaya tablets in all models above. The flavonoids extracts can inhibited T and B lymphocyte proliferation significantly within 0.13-4mg/ml. The role of inhibition reduced obviously when the concentration below 0.06mg/ml. The flavonoids extracts from Saussurea involucrata had good anti-inflammatory effect, analgesia effect and immune modulation function. Conclusion:①We established the chromatographic fingerprint of Saussurea involucrata (Kar. et Kir.) Sch.-Bip. It can be the basis on the quality control for the herb.②The flavonoids extracts from Saussurea involucrata had good anti-inflammatory effect, analgesia effect and immune modulation function. They were stable and they can be control in quality easily. These provided the basic data for further development and utilization.

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