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OBJECTIVE@#To evaluate the cytotoxic, apoptotic, mutagenic and immunomodulatory activities of Kaempferia galanga Linn. (KG) extract and ethyl-p-methoxycinnamate (EPMC) in vitro.@*METHODS@#The present study investigated the cytotoxic [using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide test], apoptotic (using a mitochondrial membrane potential assay), mutagenic (using a micronucleus test) and immunomodulatory (using flow cytometry) activities of the ethanolic extract of KG and its bioactive component, EPMC, against two cholangiocarcinoma (CCA) cell lines, CL-6 and HuCCT1, and one normal human cell line, OUMS-36T-1F.@*RESULTS@#Both KG extract and EPMC exhibited moderate cytotoxic activity against both CCA cells. The cytotoxic activity was supported by their concentration-dependent induction of apoptosis. CL-6 was most sensitive (3-4 fold) and selective to 5-fluorouracil (5-FU), compared with KG extract and EPMC [median half inhibiting concentration (IC) and selectivity index (SI) were 23.01 μg/mL and 17.32; 78.41 μg/mL and 4.44; 100.76 μg/mL and 2.20, respectively for 5-FU vs. KG extract vs. EPMC]. HuCCT1 was relatively more sensitive and selective to 5-FU and EPMC than KG extract [median IC and SI were 66.03 μg/mL and 6.04; 60.90 μg/mL and 3.65; 156.60 μg/mL and 2.23, respectively for 5-FU vs. EPMC vs. KG extract]. EPMC produced relatively potent cytotoxic activity against polymorphonuclear cells (IC = 92.20 μg/mL). KG extract and EPMC exhibited concentration-dependent mutagenic activity, as well as inhibition of tumor necrosis factor-α and interleukin-6.@*CONCLUSION@#Considering cytotoxic, apoptotic, immunomodulatory and mutagenic activities, further development of KG as a drug candidate is likely to focus on the oral pharmaceutical formulation of a standardized KG extract rather than isolated compounds.
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Objective: India has been a producer of a large number of aromatic medicinal plants which serves as a valuable genetic resource for future quality improvement to meet the ever-growing demand of human essential products. Thus, an urgent need arises for germplasm conservation of these high yielding varieties to help the pharmaceutical and other industries. For this understanding, the population structure is essential in order to explore their genetic identification by fingerprinting and molecular characterization. Methods: In the present study DNA was isolated using modified Cetyl Trimethyl Ammonium Bromide (CTAB) method and Polymerase Chain Reaction (PCR) was performed according to standardized method along with its data analysis. This study was undertaken to characterize the highly medicinal Kaempferia galanga collected from 4 different populations of Odisha using the molecular markers as Random Amplified Polymorphic DNA and Inter-Simple Sequence Repeats for the first time. Results: A dendrogram constructed through Sequential Agglomerative Hierarchical and Nested (SAHN) clustering and Unweighted Pair Group Method with Arithmetic mean (UPGMA) analysis showed an average similarity of 0.993 ranging between 0.967 to 1.000. Jaccard’s similarity coefficient of combined markers segregated the genotypes into two main clusters, 1 with six samples and the others at 0.98 similarity coefficient. Conclusion: Hence, the molecular analysis could be further used for the identification of important novel gene present in Kaempferia galanga which can be utilized for future crop improvement as well as pharmacological activities.
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Objective:To lay the material foundation for the research of subsequent pharmacological activities by the analysis of volatile components in Kaempferia galanga Linn. from different origins. Methods:It was the first time that headspace solid-phase microextraction and gas chromatography mass spectrometry (HS-SPME/GC-MS) technique was used to extract and analyze the volatile chemical components in Kaempferia galanga Linn. from Guangxi, Guangdong and Yunnan, and the area normalization method was used to calculate the mass fraction of each component. Results:Totally 42 chemical constituents were identified,mainly terpenoids,hydrocarbons,esters and aromatic compounds. A total of 41 chromatographic peaks were isolated from the volatile substances in Guangxi and 38 chemical constituents were identified,which accounted for 99.78% of the total of the volatile components. Totally 37 chromatographic peaks were isolated from the volatile substances in Guangdong, and 26 chemical constituents were identified, which accounted for 80.49% of the total of the volatile components. A total of 31 chromatographic peaks were isolated from the volatile compounds of Yunnan,and 24 chemical constituents were identified,which accounted for 64.72% of the total of the volatile components.Conclusion:The volatile components in Kaempferia galanga Linn. from Guangxi,Guangdong and Yunnan show little difference,and the characteristic components of the three habitats are methoxy cinnamate ethyl cinnamate, ethyl cinnamate and pentadecane, however, the relative contents of the three characteristic components from the three areas are much different,which are 45.02%,17.18% and 9.08% for Guangxi,41.08%,16.25% and 8.04% for Guangdong,and 30.78%,15.66% and 7.89% for Yunnan. One of the main active components in Kaempferia galanga Linn. is methoxy cinnamate ethyl cinnamate,which can be inferred that the quality of Kaempferia galanga Linn. from Guangxi and Guangdong is better than that from Yunnan. This experiment also provides evidence for the geoherbalism of Kaempferia galanga Linn.,and provides reference for the further development of the herb.
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Objective To establish an HPLC method for the simultaneous determination of quercetin and kaempferol inKaempferia galanga L..Methods ODS2 C18 (5μm, 4.6 mm×150 mm) was used as chromatographic column; methanol-0.4% phosphate (47:53) was the mobile phase; the flow rate was 1 mL/min; column temperature was 30℃; the detection wavelength was 367 nm; the injection volume was 10μL.Results Quercetin showed good linear relationship in the range of 0.016 5–1.65μg (r=0.999 7). The average recovery rate was 96.8%, RSD=2.02%; kaempferol showed good linear relationship in the range of 0.014 6–1.46μg (r=0.999 5). The average recovery rate was 97.3%, RSD=1.77%.Conclusion The method is simple, accurate, and with good reproducibility, which can be used for content determination of quercetin and kaempferol inKaempferia galanga L..
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OBJECTIVE:To establish a method for the content determination of kaempferol in Kaempferia galanga. METH-ODS:HPLC was performed on the column of Diamonsil ODS2 C18 with mobile phase of methanol-0.4% Phosphoric acid solution at a flow rate of 1 ml/min,detection wavelength was 367 nm,column temperature was 30℃,and injection volume was 10 μl. RE-SULTS:The linear range of kaempferol was 0.001 58-0.158 mg/ml;RSDs of precision,stability and reproducibility tests were low-er than 3%;recovery was 95.52%-99.32%(RSD=1.47%,n=6). CONCLUSIONS:The method is simple,accurate and reproduc-ible,and can be used for the content determination of kaempferol in K. galanga.
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Alcoholic extract of Kaempferia galanga was tested for analgesic and antiinflammatory activities in animal models. Three doses, 300 mg/kg, 600 mg/kg and 1200 mg/kg of the plant extract prepared as a suspension in 2 ml of 2% gum acacia were used. Acute and sub acute inflammatory activities were studied in rats by carrageenan induced paw edema and cotton pellet induced granuloma models respectively. In both models, the standard drug used was aspirin 100 mg/kg. Two doses 600 mg/kg and 1200 mg/kg of plant extract exhibited significant (P<0.001) antiinflammatory activity in carrageenan model and cotton pellet granuloma model in comparison to control. Analgesic activity was studied in rats using hot plate and tail-flick models. Codeine 5 mg/kg and vehicle served as standard and control respectively. The two doses of plant extract exhibited significant analgesic activity in tail flick model (P<0.001) and hot plate model (P<0.001) in comparison to control. In conclusion K. galanga possesses antiinflammatory and analgesic activities.
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Objective:To analyze and determine the main component in volatilization oil which is extracted from the Chinese herbal medicine Kaempferia galanga L.Methods:Volatilization oil is obtained by steam-distillation from commercial available Kaempferia galanga L..After re-crystallization,GC-MS,UV-Vis,NMR,IR were processed for analysis and structure determination.Results:The m/z=205 peak in MS represents the quasi-molecular ion peak.The strongest absorption peak at 304nm in UV-Vis shows that there is big conjugated system in the molecule.Along with the data of chemical shifts and coupling constants of 1H-NMR,13C-NMR and wave numbers of IR absorption peaks,the main ingredient of Kaempferia galanga L.volatilization oil is verified as ethyl 3-(4-methoxyphenyl)-(E)-acrylate.Conclusion:We have employed advanced methods in separation and analysis in the determination of structure of the main ingredient in volatilization oil extracted from Kaempferia galanga L.and get detailed data to support the results,the methods are reliable and the results are dependable.