The study of alkaloids from methanolic extract of Crucianella sintenisii from Rubiaceae family has not been studied yet. In this research, the methanolic extract was initially prepared from the aerial parts of Crucianella sintenisii, and then the resulting extract was acidified by acetic acid to remove neutral compounds. The neutral compounds were extracted by EtOAc (ethyl acetate). The resulting extract was alkalized with NH3 (25%), and then extracted with chloroform solvent. Finally, the crude alkaloid is extracted and ready to be injected into the MPLC device. One fraction was isolated, and the structure of 2-methyl-3-phenyl indole from the category of indole alkaloids was confirmed by using the spectroscopic techniques (NMR, ESI-MS, and IR). The therapeutic activities of the plant indole alkaloids are extensive, and most of these activities are related to the indole core in these structures.

Background: Zhumeria majdae showed antinociceptive and anti-inflammatory activity. The antinociceptive effect was inhibited by naloxone in hot-plate test.Objective: As Z. majdae antinociceptive activity was inhibited by an opioid antagonist, we decided to evaluate the effects of this plant on withdrawal syndrome.Methods: Dependence was induced using subcutaneous injections of morphine daily for three days (50, 50 and 75 mg/kg). On the fourth day, morphine was injected two hours prior to the intraperitoneal injection of naloxone. The number of jumps during the 30 minutes period after naloxone injection was considered as measure of withdrawal syndrome. The extracts and fractions were injected 30 min prior to morphine injection. The results indicated that methanolic extracts (0.28, 1.12 and 1.96 mg/kg) reduced the number of jumps (compared to control) up to %64, %85 and % 99.3, respectively. The methanolic-aqueous (3:2) extracted with chlorphorm also reduced withdrawal syndrome. Three MPLC fractions, 0.28 mg/kg, (A, B and C, similar Rf in each group) also reduced the number of jumping up to %97, %92 and %55, respectively in mice. In the open field test, on the contrary of MPLC fractions, the methanolic extract reduced locomotor activity.Conclusion: It is concluded that Z. majdae aerial parts methanolic extract and MPLC fractions reduced withdrawal syndrome via different mechanism of actions.

Background and purpose: Phytochemically, Allium species are a rich source of important secondary metabolites especially steroidal saponin and sapogenins, flavonoids, and sulfur compounds. As a member of this genus, Allium affine, which is locally known as “, tareh kouhi”, , is an endemic plant of middle Asian countries. Experimental approach: Bulbs of A. affine were collected and air-dried in the shade. The chloroformmethanol (9: 1) extract of the sample was subjected to purification by MPLC and HPLC. Structure elucidation of isolated compounds was done using comprehensive spectroscopic methods including 1D-NMR, 2D-NMR, and MS. Findings/Results: A steroidal saponin structurally related to parillin and a phenylpropanoid glycoside (coniferin) were isolated and identified from the plant chloroform-methanol extract. Conclusion and implication: To the best of our knowledge isolation of these potentially medicinal compounds from A. affine was reported for the first time in this study.

Stachys lavandulifolia Vahl is an herbaceous wild plant native to Iran which is traditionally used in Iranian folk medicine as a mild sedative tea for reducing anxiety and for treatment of gastrointestinal disorders. Our previous study on ethyl acetate extract of S. lavandulifolia proved anti-anxiolytic activity and so the present study was designed to determine chemical components of this biologically active fraction. The extract was prepared using maceration method. Column chromatography and medium pressure liquid chromatography (MPLC) was used respectively to separate the fractions. Finally, some evaluated fractions were used for high pressure liquid (HPLC) and peak shaving recycle technique to achieve more purification. Separated compounds were determined using NMR analysis and mass spectroscopy. Six compounds have been isolated from ethylacetate extract of aerial parts of S. lavandulifolia including four flavonoids (apigenin, kumatakenin, penduletin and 4’, 7-dihyroxy- 3, 5, 6-trimethoxy flavone), a labdan diterpenoid (labda-13-en-8, 15-diol), and an iridoid.

Background: Marine seaweeds synthesize different metabolites with various biological activities, including antimicrobial effects. Objectives: The main object of this study was to fractionate the seaweed Padina australis and analyze the antimicrobial activities of these fractions to isolate the bioactive compounds. Methods: Padina australis was collected from the Persian Gulf and extracted by maceration with methanol-ethyl acetate solvent. The extract was evaporated and partitioned by the Kupchan method to yield hexane, dichloromethane, water, and butanol partitions. Dichloromethane partition was further fractionated by MPLC, and antimicrobial activity of the crude extracts and all fractions were investigated using disc diffusion and agar plate count methods. The MIC values of hexane, dichloromethane, chloroform, and butanol partitions and all fractions were calculated. Results: Fraction numbers 11, 4, and 7 showed the highest inhibitory effect with MIC value of 87 µ g/mL, 156 µ g/mL, and 156 µ g/mL against E. coli, P. aeruginosa, and S. aureus, respectively. Conclusions: This study reveals that different fractions of P. australis have antimicrobial activities against various microorganisms.

Sea cucumbers are widely consumed in traditional medicine and food. These animals have a considerable secondary metabolite and also several potential biological activities. This study investigated the phytochemical and cytotoxic evaluation of Holothuria leucospilota (H. leucospilota), a sea cucumber from Persian Gulf. The saponin composition of H. leucospilota was studied by different partitioning and chromatography methods such as thin layer chromatography (TLC), medium pressure liquid chromatography (MPLC) and high performance LC (HPLC). The marine sea cucumber Holostane-type triterpenoids (1-3) were characterized by physical and spectroscopic examination (1 and 2 dimensional neuclear magnetic resonance and mass experiments) with data analysis. The structure of compounds 1-3 identified as echinoside A, holothurine A, and 24-dehydroechinoside A, showed moderate cytotoxic activity with IC50 values of 1. 9 ± 0. 07, 6. 8 ± 0. 23, and 2. 57 ± 0. 18 μ g/mL against HeLa and 10. 4 ± 0. 32, 8. 9 ± 0. 24, and 4. 4 ± 0. 13 on HUVEC cell line, respectively. In conclusion, the holostane-type triterpenoids showed moderate cytotoxic activity against HeLa cell line and have a prosperous future to be introduced as a lead structure.

The antiemetic effect of Mentha longifolia extracts and fractions was studied in young chickens in order to provide a basis for the traditional use of the plant. Emesis was induced with copper sulfate (60 mg/kg, orally) al1d ipecac suspension (600 mg/kg, orally). The LD 50 values of aqueous and methanolic extracts were 1561.7 mg/kg and 2161.6 mg/kg, respectively.The Maximum non-fatal doses of aqueous and methanolic extracts were 100 mg/kg and 800 mg/kg, respectively. The methanolic extract showed more efficacy, therefore the chloroformic and aqueous-methanolic fractions were prepared from it. Because of the better appearance and solubility of aqueous-methanolic fraction, other fractions were prepared from it by the method of MPLC. Among these, fractions C and F showed the most efficacy. In all groups, the positive control (metoclopramide) showed the most efficacy. In each step, more efficacy with lower dose showed that the extraction procedure was effective. Finally, fraction F showed more efficacy than fraction C and both of these fractions demonstrated more efficacy than the aqueous-methanolic fraction.According to these results, the aqueous and methanolic extracts as well as the fractions of Mentha longifolia showed antiemetic effects in young chickens with central and peripheral mechanisms.

Background: Our previous studies have shown that the aqueous and ethanolic extracts of Rosmarinus officinalis L. aerial parts reduce morphine withdrawal syndrome in mice.Objective: As R. officinalis L. aerial parts total extracts reduced morphine withdrawal syndrome, we decided to evaluate the effects of rosemary extract fractions on the withdrawal syndrome.Method: Dependence was induced using subcutaneous injections of morphine daily for three days (50, 50 and 75 mg/kg). On the fourth day, morphine was injected two hours prior to the intraperitoneal injection of naloxone. The number of jumps during the 30 minute period after naloxone injection was considered as measure of the withdrawal syndrome.Results: The results indicate that the aqueous, methanolic-aqueous and chlorformic fractions (0.96 g/kg and 1.68 g/kg, i.p.) and also clonidine (0.3 mg/kg) and diazepam (5 mg/kg) reduce the number of jumps when they are injecte 1 h prior the last dose of morphine. Two evaluable MPLC chloroformic fractions also reduce the number of jumping in mice. In the open field activity, test, on the contrary of fraction 2, the fraction 1 and clonidine reduce locomotor activity.Conclusion: It is concluded that the reduction of withdrawal morphine syndrome by R. officinalis aerial parts fraction 1 may be due to the reduction of movement by this fraction, but the fraction 2 may reduce withdrawal syndrome via an interaction with opioid system.

The genusAchillea (Asteraceae) consisting of important medicinal species, growing wildly in Iran, of which A. tenuifolia is found in Iran-o-Turan regions. Regarding the traditional use of Achillea species for treatment of diabetes and also lack of information on phyto-constituents of A. tenuifolia underground parts, in this study anti-diabetic activity of the plant have been reported. In order to find the main active components, underground parts of the plant were extracted with water and fractioned by hexane, ethyl acetate, and methanol and the separation of the main compounds were carried out via medium pressure liquid chromatography (MPLC). Also, anti-diabetic effects of the extract were investigated on rat pancreatic islets. The root extract of the plant as well as the compound β-sitosterol showed moderate α-amylase inhibitory activity, however prangol did not suppress the enzyme activity. The results of islet cells’ bio-function assays revealed that the herb root extract was able to increase the secretion of insulin in high concentration (10 mg/mL) and improved the cell viability with no toxicity in all doses. Furthermore, the herbal extract could reduce the levels of reactive oxygen species (ROS) and lipid peroxidation (LPO). The plant extract also significantly decreased the enzyme activity for both caspase-3 and -9 and increased the antioxidant capacity of the isolated cells. Taking together, preparations or extracts from the underground parts of the plant are good candidates for further anti-diabetic investigation and clinical trials.

Background: This study was designed to investigate the various extracts of Allium giganteum in term of their cytotoxic effects on gastric and colon cancer cell lines and then fractionation of extract with the most desired effects on cancer cells in comparison to the normal cell line. Materials and Methods: The A. giganteum air-dried bulbs were extracted in four solvents including aqueous, butanol, chloroform: methanol (9: 1), and dichloromethan extracts. Evaluating the cytotoxicity of extracts were done by MTT assay against HT-29, KATO-III, and HUVEC cell lines. The extract with The most cytotoxicity on cancer cells and lower toxicity on normal cell line was subjected to the fractionation by MPLC using a RP-18 silica gel column and in the next stage and the cytotoxicity of resulted fractions were determined by MTT assay. Results: Chloroform: methanol extract was the safest extract on HUVEC cells with great cytotoxic effects against cancer cell lines (IC50 17. 3 and 23 µg/mL, for KATO-III and HT-29, respectively). The results of MTT assay for 7 fractions of the chloroform: methanol extract showed that 5th fraction (E) was the most cytotoxic fraction (IC50 9 and 11. 2 µg/mL for HT-29 and KATO-III, respectively). TLC and NMR analysis of the fractions suggested the presence of steroidal saponins as the main constituents. Conclusion: Significant cytotoxic effects of Allium giganteum was established against GI cancer cell lines especially fractions rich in steroidal saponins. So, these fractions are attractive candidate for the isolation and identification of effective ingredients.