Phytochemical Analysis of Podospermum and Scorzonera n-Hexane Extracts and the HPLC Quantitation of Triterpenes.

Previously tested n-hexane extracts of the Scorzonera latifolia showed promising bioactivity in vivo. Because triterpenes could account for this activity, n-hexane extracts were analyzed by HPLC to identify and quantify the triterpenes as the most abundant constituents. Other Scorzonera and Podospermum species, potentially containing triterpenic aglycones, were included in the study. An HPLC method for simultaneous determination of triterpene aglycones was therefore developed for analysis of Podospermum and Scorzonera species. n-Hexane extracts of root and aerial parts of S. latifolia, ten other Scorzonera species and two Podospermum species were studied to compare the content of triterpenes. HPLC was used for the qualitative and quantitative analysis of α-amyrin, lupeol, lupeol acetate, taraxasteryl acetate, 3-ß-hydroxy-fern-7-en-6-one acetate, urs-12-en-11-one-3-acetyl, 3-ß-hydroxy-fern-8-en-7-one acetate, and olean-12-en-11-one-3-acetyl. Limits of detection and quantification were determined for each compound. HPLC fingerprinting of n-hexane extracts of Podospermum and Scorzonera species revealed relatively large amounts of triterpenes in a majority of investigated taxa. Lupeol, lupeol acetate, and taraxasteryl acetate were found in a majority of the species, except S. acuminata. The presence of α-amyrin, 3ß-hydroxy-fern-7-en-6-one-acetate, urs-12-en-11-one-3-acetyl, 3ß-hydroxy-fern-8-en-7-one-acetate, and olean-12-en-11-one-3-acetyl was detected in varying amounts. The triterpene content could correlate with the analgesic and anti-inflammatory activity of Scorzonera, which was previously observed and Scorzonera species that have been determined to contain triterpenes in large amounts and have not yet been tested for their analgesic activity should be tested for their potential analgesic and anti-inflammatory potential. The presented HPLC method can be used for analysis of triterpene aglycones, for example dedicated to chemosystematic studies of the Scorzonerinae.

Turkish Scorzonera Species Extracts Attenuate Cytokine Secretion via Inhibition of NF-κB Activation, Showing Anti-Inflammatory Effect in Vitro.

Scorzonera species are used in different folk medicines to combat many diseases, including the illnesses connected with inflammation. Previous experiments showed anti-inflammatory activity of Scorzonera extracts in vivo. S. latifolia, S. cana var. jacquiniana, S. tomentosa, S. mollis ssp. szowitsii, S. eriophora, S. incisa, S. cinerea, and S. parviflora extracts were, therefore, evaluated for their inhibitory activities of TNF-α and IL-1ß production, and NF-κB nuclear translocation in THP-1 macrophages. The HPLC analysis was carried out to elucidate and to compare the composition of these extracts. Major compounds of the tested extracts have been isolated using different chromatographic techniques and further tested for their inhibitory activities on TNF-α and IL-1ß production. Several extracts showed promising anti-inflammatory activity in these in vitro tests. Results of HPLC analysis revealed chlorogenic acid as a compound present in all tested extracts. Hyperoside, quercetin-3-O-ß-d-glucoside and rutin were also present in varying amount in some Scorzonera species analyzed. Furthermore, eight phenolics which were identified as quercetin-3-O-ß-d-glucoside (1), hyperoside (2), hydrangenol-8-O-glucoside (3), swertisin (4), 7-methylisoorientin (5), 4,5-O-dicaffeoyl-quinic acid (6), 3,5-di-O-caffeoyl-quinic acid (7), and chlorogenic acid (8) have been isolated as major phenolic compounds of the tested extracts and, together with eight terpenoids (9-16) previously obtained from different Scorzonera species, have been tested for the inhibition of TNF-α production, unfortunately with no activity comparable with standard.

Antimicrobial and Anti-Inflammatory Activity of Some Lathyrus L. (Fabaceae) Species Growing in Turkey.

OBJECTIVES: The present study aimed to evaluate the antimicrobial and anti-inflammatory activities of methanol extracts and n-hexane, ethyl acetate, chloroform, and water fractions of five Lathyrus species, namely Lathyrus armenus, Lathyrus aureus, Lathyrus cilicicus, Lathyrus laxiflorus subsp. laxiflorus, and Lathyrus pratensis, growing in Turkey. MATERIALS AND METHODS: The antimicrobial activities were screened against Staphylococcus aureus ATCC 29213, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and Candida albicans ATCC 10231. Broth dilution was used to determine the antimicrobial activities of extracts and fractions. In vitro anti-inflammatory activity of these extracts and fractions was determined using human red blood cell membrane stabilization. RESULTS: The results demonstrated that ethyl acetate fractions of the tested species exhibited higher antimicrobial activity than the other extracts. Among all of the tested extracts and fractions, the highest anti-inflammatory activity was detected in water fractions. Furthermore, water fractions of L. pratensis showed better anti-inflammatory activity than acetylsalicylic acid and diclofenac sodium, which were used as standard drugs in this assay. CONCLUSION: The results indicate the membrane stabilizing effect of the various extracts and fractions of the Lathyrus species and could constitute preliminary work for in vivo anti-inflammatory activity experiments.

Evaluation of the Antidiabetic Activity of Alchemilla persica Rothm. in Mice with Diabetes Induced by Alloxan.

OBJECTIVES: Alchemilla species are used in Turkish folk medicine for the treatment of many diseases together with diabetes. Alchemilla persica, belonging to this genus, is widely distributed in Eastern Anatolia as well as in Caucasia, northern and northeastern Iran, and northern Iraq. MATERIALS AND METHODS: Methanolic-water extracts of the aerial parts and roots of A. persica were evaluated for their hypoglycemic activities in mice with alloxan-induced diabetes in order to verify its usage in folk medicine. RESULTS: None of the tested extracts exhibited a significant lowering effect on blood glucose levels. However, the aerial parts notably increased blood glucose levels at doses of 100 mg/kg and 200 mg/kg. CONCLUSION: A. persica usage as an antidiabetic is not confirmed in the present study.

Investigation of Antimicrobial Activities of Some Herbs Containing Essential Oils and Their Mouthwash Formulations.

OBJECTIVES: The aim of this study was to prepare pharmaceutical formulations of mouthwashes and to examine the antimicrobial activities of essential oils obtained from plants used traditionally in Turkey for oral infections. MATERIALS AND METHODS: Essential oils were obtained from herbal drugs using water distillation with Clevenger apparatus. The antimicrobial capacities of mouthwash formulations containing a mixture of essential oils with proportions of 4.5% and 9.0% were examined using disc diffusion and microbroth dilutions. RESULTS: The inhibition zone diameters were determined to vary between 7 and 59 mm. The static and cidal activity was generally 50% and greater than 50% when pure essential oil samples were applied on microorganism specimens. Formulation F2, which contained a mixture of essential oils with proportions of 4.5%, showed 6.25% minimum bactericidal effect on Staphylococcus aureus ATCC 25923, and 3.125% the minimum inhibitory concentration and minimum bactericidal concentration on all other microorganisms. The antimicrobial effect of pure essential oil samples applied on microorganisms was lower than of mouthwashes formulations; the antimicrobial effect of F2, which contained a mixture of essential oils with proportions of 4.5% was higher than formulation F1, which contained a mixture of essential oils with proportions of 9%. CONCLUSION: The results obtained by these methods allow us to conclude that the essential oils and the prepared F1 and F2 mouthwash formulations exerted activity against microorganisms affecting the oral cavity. The F2 formulation also had significant antimicrobial activity on the tested microorganisms.

Isoflavonoids as wound healing agents from Ononidis Radix.

ETHNOPHARMACOLOGICAL RELEVANCE: Dried roots of Ononis spinosa L. are traditionally used for their diuretic, anti-inflammatory and wound healing effects. AIM OF THE STUDY: Isolation of the bioactive compounds of Ononis spinosa L. subsp. leiosperma (Boiss.) Sirj. MATERIALS AND METHODS: Ethyl acetate extract prepared from the roots of Ononis spinosa L. subsp. leiosperma (Boiss.) Sirj. was subjected to silica gel column. The fractions were tested for their wound healing and anti-inflammatory activities. Linear incision and circular excision wound models and hydroxypyroline estimation assay were used for the wound healing activity. Carrageenan-induced hind paw edema, TPA-induced ear edema and acetic acid-induced increase in capillary permeability tests as acute inflammation; FCA-induced arthritis as chronic inflammation models were used for the assessment of anti-inflammatory activity. Antioxidant capacities of the fractions were tested using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) scavenging activity assay, reducing power assay and hydroxyl radical (OH-) scavenging assay. The isolation procedure was continued with the active fraction (Fr-E5). RESULTS: Fr-E5 exhibited remarkable wound healing activity with the 33.4% tensile strength value on the linear incision wound model and 51.4% reduction of the wound area at the day 12 on the circular excision wound model. Hydroxyproline content of the tissue treated by Fr-E5 was found to be 30.9 ± 0.72µg/mg. Acetic acid induced increase in capillary permeability test results revealed that Fr-E5 inhibited inflammation by the value of 40.3%. Fr-E5 showed 28.1-32.2% inhibition in carrageenan-induced hind paw edema test while did not possess activity on TPA-induced ear edema and FCA-induced arthritis models. Trifolirhizin, ononin, medicarpin-3-O-glucoside, onogenin-7-O-glucoside and sativanone-7-O-glucoside were isolated from Fr-E5 and tested for their wound healing activities using by measuring their inhibition of hyaluronidase, collagenase and elastase enzymes. Ononin and sativanone-7-O-glucoside inhibited hyaluronidase and elastase enzymes by 31.66% and 41.75%; 45.58% and 46.88% values respectively at the dose of 100µg/mL. CONCLUSION: Among five isolated compounds, ononin and sativanone-7-O-glucoside were found to inhibit hyaluronidase and elastase enzymes. According to the results, these compounds may majorly be responsible for the wound healing activity of the extract.

Wound healing and anti-inflammatory activity of some Ononis taxons.

Ononis species are used for their laxative, diuretic, analgesic, anti-inflammatory, antiviral, cytotoxic and antifungal effects as well as against skin diseases for wound healing activity. In the light of this information n-hexane, ethylacetate and methanol extracts prepared from Ononis spinosa L. subsp. leiosperma (Boiss.) Sirj., Ononis variegata L., Ononis viscosa L. subsp. brevifolia (DC) Nym. and Ononis natrix L. subsp. natrix L. were tested for their wound healing, anti-inflammatory and antioxidant activities. Linear incision and circular excision wound models and hydroxypyroline estimation assay were used for the wound healing activity. For the assessment of chronic inflammation FCA-induced arthritis and for acute inflammation carrageenan-induced hind paw edema, TPA-induced ear edema and acetic acid-induced increase in capillary permeability tests were conducted. 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) scavenging activity assay, reducing power assay and hydroxyl radical (OH-) scavenging assay were used for determining antioxidant activities of the extracts. Results showed that O. spinosa subsp. leiosperma roots ethyl acetate extract exhibited remarkable wound healing activity with the 42.6% tensile strength value on the linear incision wound model and 60.1% reduction of the wound area at the day 12 on the circular excision wound model. Hydroxyproline content of the tissue treated by O. spinosa subsp. leiosperma roots ethyl acetate extract was found to be 41.3µg/mg. Acetic acid induced increase in capillary permeability test results revealed that O. spinosa subsp. leiosperma roots ethyl acetate extract and O. spinosa subsp. leiosperma roots methanol extract inhibited inflammation by 40.4% and 35.4% values respectively. O. spinosa subsp. leiosperma roots ethyl acetate extract showed 21.2-27.2% inhibition in carrageenan-induced hind paw edema test while did not posses activity on TPA-induced ear edema and FCA-induced arthritis models.