Factorial design of essential oil extraction from Fagraea fragrans Roxb. flowers and evaluation of its biological activities for perfumery and cosmetic applications.

OBJECTIVE: To optimize the extraction yields of essential oil from Fagraea fragrans Roxb. flowers in hydro-distillation using a central composite design (CCD) and to evaluate its biological activities for perfumery and cosmetic applications. METHODS: Central composite design was applied to study the influences of operational parameters [water to flower weight (X(1)) and distillation time (X(2))] on the yields of essential oil (Y). Chemical compositions of the essential oil extracted from the optimized condition were identified by gas chromatography-mass spectrometry. Antioxidant activities of the essential oil were determined against ABTS(•+) and DPPH(•) radicals, and the cytotoxic effects were assessed on human embryonic kidney (HEK293) cells by the use of the MTT assay. Also, the aromatic properties of the essential oil were evaluated by five healthy trained volunteers. RESULTS: The best conditions to obtain the maximum essential oil yield were 7.5 mL g(-1) (X(1)) and 215 min (X(2)). The experimental yield of the essential oil (0.35 ± 0.02% v/w) was close to the value predicted by a mathematical model (0.35 ± 0.01% v/w). 3-Octadecyne, Z,Z,Z-7,10,13-hexadecatrienal, E-nerolidol, pentadecanal and linalool were the major constituents of the essential oil. The essential oil showed moderate antioxidant capacities with no toxic effects on HEK293 cells at 1-250 µg mL(-1). Also, the essential oil exhibited a very strong aroma and was classified to be top- to middle-notes. CONCLUSION: The results offer the effectively operational conditions in the extraction of essential oil from F. fragrans using hydro-distillation. The essential oil could be used as a natural fragrance, having antioxidant activity with slight cytotoxicity, for perfumery and cosmetic applications.

Crown gall -- a plant tumour with biological activities.

Petroleum ether, acetone, 80% MeOH and water extracts of crown gall, a plant tumour, obtained from Eucalyptus globulus tree were screened for cytotoxic, antioxidant, antiinflammatory, embryotoxic, antitumour-promoting and antimicrobial activities. In terms of bioactivity the 80% MeOH extract was most effective followed by the acetone extract. The petroleum ether extract showed weak to moderate cytotoxic activity in dose-dependent manner against PC12 cells, mouse L fibroblasts and 1321N1 glia cells, whereas the hydroalcohol extract had no or a weak cytotoxic effect. The 80% MeOH extract exhibited strong antioxidant activity. Based on the in vitro HET-CAM assay all the extracts were effective against inflammation. The extracts did not show any embryotoxic effect at the concentrations tested. Antitumour-promoting activity (100% inhibition; 100 microg/mL) was observed in the 80% MeOH and acetone extracts. In the antimicrobial screening all extracts displayed predominantly antifungal activity against Candida sp. The extracts also showed various levels of antibacterial activity against E. faecalis, Ps. aeruginosa, Bac. subtilis and Staph. epidermidis. From the results of the investigations it can be concluded that crown gall is a valuable plant tumour tissue having interesting biological activities.

Study of alkaloids from Adhatoda vasica Nees on their antiinflammatory activity.

Adhatoda vasica Nees is a shrub widespread throughout the tropical regions of southeast Asia. It possesses a wide spectrum of medicinal properties including positive effects on inflammatory diseases. The antiinflammatory activity of the methanol extract, the non-alkaloid fraction, the saponins and the alkaloids was evaluated by the modified hen's egg chorioallantoic membrane test. The alkaloid fraction showed potent activity at a dose of 50 microg/pellet equivalent to that of hydrocortisone while the MeOH extract and the other fractions showed less activity.

Quality assessment of Paliurus spina-christi extracts.

Because the flavonoid glycosides can be considered as suitable compounds for the quality assessment of Paliurus spina-christi Mill., a high-performance liquid-chromatographic method for the separation and quantification of these compounds in methanolic extracts of the different plant parts (leaves, flowers, fruits) is described. The system used is a reversed-phase column and gradient elution with water-phosphoric acid-acetonitrile. The analysis showed the flavonoid contents of the different plant parts and the different composition of the flavonoid pattern. The highest flavonoid content was found in the leaves in June and July. No significant influence of growing site or year of harvesting on the flavonoid content was observed. As quercetin 3-O-rhamnoglucoside 7-O-rhamnoside and rutin are the main flavonoid compounds present in all plant parts investigated, it is suggested that these compounds should be used for the quality assessment of Paliurus spina-christi.

Antibacterial steroid alkaloids from the stem bark of Holarrhena pubescens.

Holarrhena pubescens (syn. H. antidysenterica) (L.) WALL. stem bark was tested for antibacterial efficacy against Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus faecalis, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa using the microdilution broth method as well as the disc diffusion method. The crude methanolic extract was active against all tested bacteria. Further chemical fractionation indicated that the antibacterial activity was mainly associated with the alkaloids. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined for the crude extract, the total alkaloids and the neutral fraction using microdilution broth method. The results were compared with reference antibiotics. The total alkaloids showed remarkable activity against S. aureus (MIC = 95 microg/ml).

[Evaluation of pharmacopoeia methods for determination of antimicrobial agents, especially of natural substances]. / Bewertung von Arzneibuchmethoden zur Bestimmung der antimikrobiellen Wirksamkeit unter besonderer Berücksichtigung von Naturstoffen.

Microbiological assays referring to antibiotics were first mentioned in 1955 in the US Pharmacopedia XV and in the Pharmacopedia of India I. In the pharmacopedias two general methods are employed: The first group of methods is grounded on diffusion (disc assay, cylinder-plate and hole-plate assay), the second one is based on the determination of optical density (turbidimetric method). Both methods involve certain problems, particularly with respect to the testing of low-active natural substances. In the course of our investigations those test methods were evaluated and the advantages and disadvantages discussed. The methods were compared to test systems not being described in the pharmacopedias (microdilution test, bioautographic TLC assay). In addition to that we examined spectrophotometrically (OD580nm) the influence of an antimicrobial substance and of a solutizer on the bacterial growth. Tetracycline hydrochloride was used as reference and naringenin as test substance. The results were analyzed with statistical methods cited in the pharmacopedias, e.g. straight-line method and compared to other common methods, e.g. analysis of variance.

Influence of physical parameters on the germ-reducing effect of microwave irradiation on medicinal plants.

Plant material is to a different extent contaminated by microorganisms. As DAB 10 demands limiting values for aerobic bacteria, for yeasts and moulds, a reduction of microorganisms of the crude herbal drug is necessary. An effective method is the application of microwave irradiation. The microorganisms isolated and identified on different herbal drugs (leaves, flowers, herbs) were mainly spore-forming aerobic bacteria, Micrococcus luteus, and moulds of the genera Aspergillus and Penicillium. Various parameters such as the use of different microwave powers and the influence of humidity on the total count of microorganisms were tested according to the method described in DAB 10. It can be concluded from the results that microwave treatment allows careful drying with a germ-reducing effect, provided testing takes place under optimized and accurately defined conditions.

Antibacterial activity of plant extracts used externally in traditional medicine.

Plants of 28 families, selected on the basis of medicinal folklore reports and literature data, were studied in a screening program. Aqueous extracts of plants used externally for the treatment of infected skin lesions were tested for their antibacterial potential. The results indicated that about 60% of the plant extracts tested exhibited some level of antibacterial action.

Applicability of diffusion methods required by the pharmacopoeias for testing antibacterial activity of natural compounds.

The diffusion methods required by various pharmacopoeias were investigated in comparison to the bioautographic TLC assay in regard to the applicability and the statistical evaluation for testing the antibacterial activity of natural substances. Naringenin as an example for a weakly effective lipophilic natural compound and tetracycline as standard were used. Data were analysed by linear regression and ANOVA. The comparison of the disk diffusion method, the hole-plate diffusion method, and the bioautographic TLC assay showed that nature and amount of substance influence to a great extent the selection of the test method. The evaluation of parallelism demanded by the pharmacopoeias may provoke problems in comparing natural substances with strongly effective antibiotics. The simultaneous influence of different variables was examined with the help of ANOVA. The bioautographic TLC assay showed for spotted compounds a comparable reproducibility and accuracy to the diffusion methods mentioned above.

[Ion exchange interactions on silica gel layers. II. Halogen salts of compounds containing organic tertiary and quaternary nitrogen]. / Szilikagél rétegen végbemenö ioncserés kölcsönhatások. II. Szerves tercier és kvaterner nitrogént vegyületek haloidsói.

Primary and secondary ion exchanges--of hydrochloric acid and hydrobromic acid salts of well hydrolyzing organic bases as well as quaternary ammonium bromide which are important drug substance--taking place on silica gel using methanol as mobile phase have been investigated by thin-layer chromatographic and spectrophotometric methods. In case of tertiary ammonium salts (hydrolyzing salts) basis linked to silanate ion and halogen acid have been formed by primary ion exchange. During secondary ion exchange halogen acid has exchanged metal ions linked to silanate ions on the layer. In case of non hydrolyzing salts, the quaternary ammonium bromide salts it could not surely be proved by the applied methods whether primary ion exchange had been followed by secondary ion exchange or only primary ion exchange had occurred.

[Ion exchange interactions on silica gel layers. III. Salts of organic bases with organic acids. Organic acids and sodium salts]. / Szilikagél rétegen vébemenö ioncserés kölcsönhatások. III. Szerves bázisok szerves savakkal alkotott sói. Szerves savak és nátriumsók.

By thin-layer chromatographic and spectroscopic (UV, IR) methods it has been proved that bass and acid are formed from salts of organic bases with organic acids during primary interaction because of the ion exchange behaviour of the silica gel layer. The organic acids formed have interacted with metals on the layer during secondary ion exchange and they migrate further as salts again. It has also been established that the extent of secondary ion exchange between metal silanate groups and acids depends on pKS values of acids. On the basis of investigations it can be ascertained with great probability that organic acids of pKS greater than 7 do not interact with metal silanate groups of silica gel layer. Sodium salts of organic acids of pKS less than 3 values similarly do not interact on the other hand sodium salts of organic acids of pKS greater than 3 values are able to interact with silanol groups. It has been established by flame atomic emission method that salicylic acid interacts with sodium ion and 86% of it migrates further as sodium salicylate.