Ehsan Karimi

Evaluation of Crocus sativus L. stigma phenolic and flavonoid compounds and its antioxidant activity.

Saffron (Crocus sativus L.) belongs to the Iridaceae family. The stigma of saffron has been widely used as spice, medicinal plant, and food additive in the Mediterranean and Subtropical countries. Recently, attention has been paid to the identification of new sources of safe natural antioxidants for the food industry. The antioxidant activities of spices are mainly attributed to their phenolic and flavonoid compounds. Saffron is one of the spices believed to possess antioxidant properties, but information on its antioxidant activity and phenolic, flavonoids compound are rather limited, therefore this research was carried out to evaluate the antioxidant activity of saffron stigmas extracted with different solvents. The phenolic and flavonoid compounds of saffron were also examined using reversed phase (RP)-HPLC. Results showed that saffron stigma possess antioxidant activity. The free radical scavenging and ferric reducing power activities were higher for the methanolic extract of saffron stigma at a concentration of 300 μg/mL, with values of 68.2% and 78.9%, respectively, as compared to the corresponding boiling water and ethanolic extracts, but the activities were lower than those of antioxidant standards such as BHT and α-tocopherol. The obtained total phenolics value for methanolic saffron extract was 6.54 ± 0.02 mg gallic acid equivalent (GAE)/g dry weight (DW), and for total flavonoids, 5.88 ± 0.12 mg rutin equivalent/g DW, which were also higher than values obtained from the ethanolic and boiling water extracts. In addition, the RP-HPLC analyses indicated the presence of gallic acid and pyrogallol as two bioactive compounds. In summary, saffron stigmas showed antioxidant activity and methanol appeared to be the best solvent to extract the active components, among which the presence of gallic acid and pyrogallol might contribute towards the stigma’s antioxidant properties. Hence, saffron stigma could be applied as a natural antioxidant source for industrial purposes.

Phytochemical Analysis and Antimicrobial Activities of Methanolic Extracts of Leaf, Stem and Root from Different Varieties of Labisa pumila Benth

A local herb, Kacip Fatimah, is famous amongst Malay women for its uses in parturition; however, its phytochemical contents have not been fully documented. Therefore, a study was performed to evaluate the phenolics, flavonoids, and total saponin contents, and antibacterial and antifungal properties of the leaf, stem and root of three varieties of Labisia pumila Benth. Total saponins were found to be higher in the leaves of all three varieties, compared to the roots and stems. Leaves of var. pumila exhibited significantly higher total saponin content than var. alata and lanceolata, with values of 56.4, 43.6 and 42.3 mg diosgenin equivalent/g dry weight, respectively. HPLC analyses of phenolics and flavonoids in all three varieties revealed the presence of gallic acid, caffeic acid, rutin, and myricetin in all plant parts. Higher levels of flavonoids (rutin, quercitin, kaempferol) were observed in var. pumila compared with alata and lanceolata, whereas higher accumulation of phenolics (gallic acid, pyrogallol) was recorded in var. alata, followed by pumila and lanceolata. Antibacterial activities of leaf, stem and root extracts of all varieties determined against both Gram positive (Micrococcus luteus, Bacillus subtilis B145, Bacillus cereus B43, Staphylococcus aureus S1431) and Gram negative (Enterobacter aerogenes, Klebsiella pneumonia K36, Escherichia coli E256, Pseudomonas aeruginosa PI96) pathogens showed that crude methanolic extracts are active against these bacteria at low concentrations, albeit with lower antibacterial activity compared to kanamycin used as the control. Antifungal activity of methanolic extracts of all plant parts against Fusarium sp., Candida sp. and Mucor using the agar diffusion disc exhibited moderate to appreciable antifungal activities compared to streptomycin used as positive control.

HPLC and GC-MS Determination of Bioactive Compounds in Microwave Obtained Extracts of Three Varieties of Labisia pumila Benth.

Microwave extraction of phytochemicals from medicinal plant materials has generated tremendous research interest and shown great potential. This research highlights the importance of microwave extraction in the analysis of flavonoids, isoflavonoid and phenolics and the antioxidant properties of extracts from three varieties of the Malaysian medicinal herb, Labisia pumila Benth. High and fast extraction performance ability, equal or higher extraction efficiencies than other methods, and the need for small samples and reagent volumes are some of the attractive features of this new promising microwave assisted extraction (MAE) technique. The aims of the present research were to determine the foliar phenolics and flavonoids contents of extracts of three varieties of L. pumila obtained by a microwave extraction method while flavonoid, isoflavonoid and phenolic compounds were analyzed using RP-HPLC. Furthermore, the antioxidant activities were measured by the DPPH and FRAP methods and finally, the chemical composition of the crude methanolic extracts of the leaves of all three varieties were analyzed by GS-MS.

Phenolic Compounds Characterization and Biological Activities of Citrus aurantium Bloom

Citrus plants are known to possess beneficial biological activities for human health. In addition, ethnopharmacological application of plants is a good tool to explore their bioactivities and active compounds. This research was carried out to evaluate the phenolic and flavonoid analysis, antioxidant properties, anti inflammatory and anti cancer activity of Citrus aurantium bloom. The total phenolics and flavonoids results revealed that methanolic extract contained high total phenolics and flavonoids compared to ethanolic and boiling water extracts. The obtained total phenolics value for methanolic Citrus aurantium bloom extract was 4.55 ± 0.05 mg gallic acid equivalent (GAE)/g dry weight (DW), and for total flavonoids it was 3.83 ± 0.05 mg rutin equivalent/g DW. In addition, the RP-HPLC analyses of phenolics and flavonoids indicated the presence of gallic acid, pyrogallol, syringic acid, caffeic acid, rutin, quercetin and naringin as bioactive compounds. The antioxidant activity of Citrus aurantium bloom were examined by the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay and the ferric reducing/antioxidant potential (FRAP). The free radical scavenging and ferric reducing power activities were higher for the methanolic extract of Citrus aurantium bloom at a concentration of 300 μg/mL, with values of 55.3% and 51.7%, respectively, as compared to the corresponding boiling water and ethanolic extracts, but the activities were lower than those of antioxidant standards such as BHT and α-tocopherol. Furthermore, the anti-inflammatory result of methanolic extract showed appreciable reduction in nitric oxide production of stimulated RAW 264.7 cells at the presence of plant extract. Apart from that, the anticancer activity of the methanolic extract was investigated in vitro against human cancer cell lines (MCF-7; MDA-MB-231), human colon adenocarcinoma (HT-29) and Chang cell as a normal human hepatocyte. The obtained result demonstrated the moderate to appreciable activities against all cell line tested and the compounds present in the extracts are non-toxic which make them suitable as potential therapeutics.

Phenolics and Flavonoids Compounds, Phenylanine Ammonia Lyase and Antioxidant Activity Responses to Elevated CO2 in Labisia pumila (Myrisinaceae)

A split plot 3 × 3 experiment was designed to examine the impact of three concentrations of CO2 (400, 800 and 1,200 µmol·mol−1) on the phenolic and flavonoid compound profiles, phenylalanine ammonia lyase (PAL) and antioxidant activity in three varieties of Labisia pumila Benth. (var. alata, pumila and lanceolata) after 15 weeks of exposure. HPLC analysis revealed a strong influence of increased CO2 concentration on the modification of phenolic and flavonoid profiles, whose intensity depended on the interaction between CO2 levels and L. pumila varieties. Gallic acid and quercetin were the most abundant phenolics and flavonoids commonly present in all the varieties. With elevated CO2 (1,200 µmol·mol−1) exposure, gallic acid increased tremendously, especially in var. alata and pumila (101–111%), whilst a large quercetin increase was noted in var. lanceolata (260%), followed closely by alata (201%). Kaempferol, although detected under ambient CO2 conditions, was undetected in all varieties after exposure. Instead, caffeic acid was enhanced tremendously in var. alata (338~1,100%) and pumila (298~433%). Meanwhile, pyragallol and rutin were only seen in var. alata (810 µg·g−1 DW) and pumila (25 µg·g−1 DW), respectively, under ambient conditions; but the former compound went undetected in all varieties while rutin continued to increase by 262% after CO2 enrichment. Interestingly, naringenin that was present in all varieties under ambient conditions went undetected under enrichment, except for var. pumila where it was enhanced by 1,100%. PAL activity, DPPH and FRAP also increased with increasing CO2 levels implying the possible improvement of health-promoting quality of Malaysian L. pumila under high CO2 enrichment conditions.

Antifungal, anti-inflammatory and cytotoxicity activities of three varieties of Labisia pumila benth : from microwave obtained extracts.

BackgroundLabisia pumila, locally known as Kacip Fatimah, is a forest-floor plant that has tremendous potential in the herbal industry. It is one of the five herbal plants identified by the government as one of the national key economic areas to be developed for commercial purposes. There are three varieties of L. pumila namely, L. pumila var. pumila, L. pumila var. alata and L. pumila var. lanceolata and each has its own use.MethodsThe leaves and roots of the three varieties of L. pumila Benth. were extracted using microwave assisted extraction (MAE). Antifungal activity of all plant extracts were characterized against Fusarium sp., Candida sp. and Mucor using the agar diffusion disc. Anti-inflammatory assays were performed using NO production by macrophage RAW 264.7 cell lines induced by LPS/IFN-g and cytotoxic activity was determined using several cancer cell lines and one normal cell line.ResultsThe overall result demonstrated that leaf and root extracts of all three varieties of L. pumila exhibited moderate to appreciable antifungal activity against Fusarium sp., Candida sp. and Mucor compared to streptomycin used as positive control. Leaf and root extracts of all varieties significantly decreased NO release. However, the root extracts showed higher activity compared to the leaf extracts. Cytotoxic activity against MCF-7, MDA-MB-231 and Chang cell lines were observed with all extracts.ConclusionsThese findings suggest the potential use of L. pumila Benth. as a natural medicine and indicated the possible application of this medicinal plant such anti inflammatory activity and cytotoxic agents.

Involvement of Salicylic Acid on Antioxidant and Anticancer Properties, Anthocyanin Production and Chalcone Synthase Activity in Ginger (Zingiber officinale Roscoe) Varieties

The effect of foliar application of salicylic acid (SA) at different concentrations (10−3 M and 10−5 M) was investigated on the production of secondary metabolites (flavonoids), chalcone synthase (CHS) activity, antioxidant activity and anticancer activity (against breast cancer cell lines MCF-7 and MDA-MB-231) in two varieties of Malaysian ginger, namely Halia Bentong and Halia Bara. The results of high performance liquid chromatography (HPLC) analysis showed that application of SA induced the synthesis of anthocyanin and fisetin in both varieties. Anthocyanin and fisetin were not detected in the control plants. Accordingly, the concentrations of some flavonoids (rutin and apigenin) decreased significantly in plants treated with different concentrations of SA. The present study showed that SA enhanced the chalcone synthase (CHS) enzyme activity (involving flavonoid synthesis) and recorded the highest activity value of 5.77 nkat /mg protein in Halia Bara with the 10−5 M SA treatment. As the SA concentration was decreased from 10−3 M to 10−5 M, the free radical scavenging power (FRAP) increased about 23% in Halia Bentong and 10.6% in Halia Bara. At a concentration of 350 μg mL−1, the DPPH antioxidant activity recorded the highest value of 58.30%–72.90% with the 10−5 M SA treatment followed by the 10−3 M SA (52.14%–63.66%) treatment. The lowest value was recorded in the untreated control plants (42.5%–46.7%). These results indicate that SA can act not only as an inducer but also as an inhibitor of secondary metabolites. Meanwhile, the highest anticancer activity against MCF-7 and MDA-MB-231 cell lines was observed for H. Bara extracts treated with 10−5 M SA with values of 61.53 and 59.88%, respectively. The results suggest that the high anticancer activity in these varieties may be related to the high concentration of potent anticancer components including fisetin and anthocyanin. The results thus indicate that the synthesis of flavonoids in ginger can be increased by foliar application of SA in a controlled environment and that the anticancer activity in young ginger extracts could be improved.

Bioactive Compounds, Antioxidant, Xanthine Oxidase Inhibitory, Tyrosinase Inhibitory and Anti-Inflammatory Activities of Selected Agro-Industrial By-products

Evaluation of abundantly available agro-industrial by-products for their bioactive compounds and biological activities is beneficial in particular for the food and pharmaceutical industries. In this study, rapeseed meal, cottonseed meal and soybean meal were investigated for the presence of bioactive compounds and antioxidant, anti-inflammatory, xanthine oxidase and tyrosinase inhibitory activities. Methanolic extracts of rapeseed meal showed significantly (P < 0.01) higher phenolics and flavonoids contents; and significantly (P < 0.01) higher DPPH and nitric oxide free radical scavenging activities when compared to that of cottonseed meal and soybean meal extracts. Ferric thiocyanate and thiobarbituric acid tests results showed rapeseed meal with the highest antioxidant activity (P < 0.01) followed by BHT, cotton seed meal and soybean meal. Rapeseed meal extract in xanthine oxidase and tyrosinase inhibitory assays showed the lowest IC50 values followed by cottonseed and soybean meals. Anti-inflammatory assay using IFN-γ/LPS stimulated RAW 264.7 cells indicated rapeseed meal is a potent source of anti-inflammatory agent. Correlation analysis showed that phenolics and flavonoids were highly correlated to both antioxidant and anti-inflammatory activities. Rapeseed meal was found to be promising as a natural source of bioactive compounds with high antioxidant, anti-inflammatory, xanthine oxidase and tyrosinase inhibitory activities in contrast to cotton and soybean meals.

Impact of Organic and Inorganic Fertilizers Application on the Phytochemical and Antioxidant Activity of Kacip Fatimah (Labisia pumila Benth)

A study was conducted to compare secondary metabolites and antioxidant activity of Labisia pumila Benth (Kacip Fatimah) in response to two sources of fertilizer [i.e., organic (chicken dung; 10% N:10% P2O5:10% K2O) and inorganic fertilizer (NPK green; 15% N, 15% P2O5, 15% K2O)] under different N rates of 0, 90, 180 and 270 kg N/ha. The experiment was arranged in a randomized complete block design replicated three times. At the end of 15 weeks, it was observed that the application of organic fertilizer enhanced the production of total phenolics, flavonoids, ascorbic acid, saponin and gluthathione content in L. pumila, compared to the use of inorganic fertilizer. The nitrate content was also reduced under organic fertilization. The application of nitrogen at 90 kg N/ha improved the production of secondary metabolites in Labisia pumila. Higher rates in excess of 90 kg N/ha reduced the level of secondary metabolites and antioxidant activity of this herb. The DPPH and FRAP activity was also highest at 90 kg N/ha. The results indicated that the use of chicken dung can enhance the production of secondary metabolites and improve antioxidant activity of this herb.

Combined effect of CO 2 enrichment and foliar application of salicylic acid on the production and antioxidant activities of anthocyanin, flavonoids and isoflavonoids from ginger

BackgroundThe increase in atmospheric CO2 concentration caused by climate change and agricultural practices is likely to affect biota by producing changes in plant growth, allocation and chemical composition. This study was conducted to evaluate the combined effect of the application of salicylic acid (SA, at two levels: 0 and 10-3 M) and CO2 enrichment (at two levels: 400 and 800 μmol·mol−1) on the production and antioxidant activities of anthocyanin, flavonoids and isoflavonoids from two Malaysian ginger varieties, namely Halia Bentong and Halia Bara.MethodsHigh-performance liquid chromatography (HPLC) with photodiode array detection and mass spectrometry was employed to identify and quantify the flavonoids and anthocyanins in the ginger extracts. The antioxidant activity of the leaf extracts was determined by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and thiobarbituric acid (TBA) assays. The substrate specificity of chalcone synthase, the key enzyme for flavonoid biosynthesis, was investigated using the chalcone synthase (CHS) assay.ResultsCO2 levels of 800 μmol·mol−1 significantly increased anthocyanin, rutin, naringenin, myricetin, apigenin, fisetin and morin contents in ginger leaves. Meanwhile, the combined effect of SA and CO2 enrichment enhanced anthocyanin and flavonoid production compared with single treatment effects. High anthocyanin content was observed in H Bara leaves treated with elevated CO2 and SA. The highest chalcone synthase (CHS) activity was observed in plants treated with SA and CO2 enrichment. Plants not treated with SA and kept under ambient CO2 conditions showed the lowest CHS activity. The highest free radical scavenging activity corresponded to H Bara treated with SA under high CO2 conditions, while the lowest activity corresponded to H Bentong without SA treatment and under atmospheric CO2 levels. As the level of CO2 increased, the DPPH activity increased. Higher TBA activity was also recorded in the extracts of H Bara treated with SA and grown under high CO2 conditions.ConclusionsThe biological activities of both ginger varieties were enhanced when the plants were treated with SA and grown under elevated CO2 concentration. The increase in the production of anthocyanin and flavonoids in plants treated with SA could be attributed to the increase in CHS activity under high CO2 levels.