[Efficiency of supercritical fluid extraction for the production of non-volatile terpenoids from Taraxaci radix]. / Szuperkritikus extrakció hatékonysága a Taraxaci radix nem illó növényi terpenoidjainak elóállítására.

Supercritical fluid extraction (SFE) is an extraction technique which exploits the solvent properties of fluids above their critical point. As a result supercritical fluid extraction was used to gain various active substances from plants. The use of SFE to obtain bioactive substances from medicinal plants over the past twenty years has been proved to be a viable alternative for the extraction of natural compounds. Dandelion (Taraxacum officinale Wiggers et Webers, Asteraceae) is one of the best known European medicinal plants, not only as a traditional medicine but perspective raw material for modern phytopharmaceuticals. From the characteristic principles our attention has been directed to triterpenes and phytosterols with anti-inflammatory activity, which were extracted with supercritical carbon dioxide. Designed experiments were carried out to determine the optimal extraction parameters. The products obtained by supercritical fluids extraction were compared to extracts prepared by traditional extraction method (Soxhlet) with n-hexane and ethyl alcohol solvents. The content of triterpenes and sterols was monitored after saponification by thin layer chromatography-densitometry. The products gained by supercritical fluid extraction were different from the traditional ones both in their appearance and composition. Triterpenes and their esters were extracted quantitatively by supercritical fluid extraction using CO2 as solvent and the extraction dynamic for triterpenes and phytosterols was different. Triterpenes had a higher concentration in the SFE product then in the extracts prepared by traditional methods.

[New data on composition of esssential oil from inflorescence of everlasting (Helichrysum arenarium(L.) Moench.)]. / Uj adatok a homoki szalmagyopár virágzati illóolajának összetételéhez.

We investigated the composition of essential oil obtained from inflorescence of Helichrysum arenarium (everlasting) which is a perennial herb, native in our home and in Middle and South East Europe. We used essential oils obtained from Hungarian and Polish mercantile samples and a plant sample cultivated in Hungary (Soroksár) by steam-distillation. The GC and GC-MS investigation revealed the presence of more than 60 compounds from which 24 were identified. The identified compounds are as follows: linalool, alpha-terpineol, carvone monoterpenes; anethol, anisaldehyde, thymol, carvacrol, eugenol, beta-asarone, butylhydroxyanisole aromatic components; alpha-humulene, beta-caryophyllene, gamma-muurolene, delta-cadinene, copaene, alpha-gurjunene, caryophyllenol, delta-cadinol and globulol sesquiterpenes as well as caprilic, pelargonic, caprinic, laurinic acids and methyl palmitate as alkyl carbonic acid and their derivatives. We established that the main components of essential oil of Hungarian and Polish mercantile samples is methyl palmitate (28.5% and 21.7% respectively) but of cultivated sample is caprinic acid (19.8%). Comparing our results of essential oil composition with those obtained from other Helichrysum species it is surprising that the alkyl carbonic acids are the dominant essential oil constitutents of Helichrysum arenarium.

In vitro antioxidant properties of Helichrysum arenarium (L.) Moench.

The choleretic, hepatoprotective and detoxifying activities of the inflorescence of Helichrysum arenarium (L.) Moench (everlasting, immortelle: Asteraceae-Helichrysi flos syn. Stoechados flos) have been known for a long time from herbal medicine in Hungary. Antioxidant properties of its main phenolics, flavonoids, are supposed to be responsible for these effects. The aim of this study is to verify the antioxidant properties of the lyophilized water extracts from inflorescences, and to define the total polyphenol and flavonoid contents in Helichrysi flos water extracts as well as in lyophilized water extracts. The hydrogen-donating ability and the reducing power property of the lyophilizates were determined spectrophotometrically; their OH&z.rad; scavenging activity was measured, in the H(2)O(2)/OH&z.rad;-luminol-microperoxidase system, by a chemiluminometric method. Results were compared with the activity of the flavonoid silibinin, the main agent of the well-known milk thistle (Silybum marianum L.).

Free radical scavenging and membrane protective effects of methanol extracts from Anthriscus cerefolium L. (Hoffm.) and Petroselinum crispum(Mill.) nym. ex A.W. Hill.

The antilipoperoxidant activity of Anthriscus cerefolium L. (Hoffm.), chervil, Petroselinum crispum (Mill.) Nym. ex A. W. Hill., parsley extracts were evaluated with ascorbic acid induced lipid peroxidation on rat brain homogenates. These results are completed by the antiradical potential of these extracts against a solution of OH. radical. In all cases luteolin-7-O-glucoside was used as a reference material.

In vitro antioxidant activity of Anthriscus cerefolium L. (Hoffm.) extracts.

Standardised aqueous extracts of chervil (Anthriscus cerefolium L. Hoffm.) (Apiacae) were investigated for antioxidant effect. Numerous in vitro test methods were used to determine whether the extracts, from different vegetative parts (root, herb) had H-donor, metal binding, reductive, free radical scavenging and membrane protective activity. Apiin was used as a reference material. The herb extract showed better activity in all experiments than the root extract. The present results underline that the wateric chervil extracts have antioxidant and anti-lipoperoxidant activity.

Supercritical carbon dioxide extraction and fractionation of fennel oil.

Ground fennel seeds were extracted with supercritical carbon dioxide. Small-scale subsequent extractions of the same sample showed that the composition of volatile compounds was changed with the extension of extraction time and only principal volatile components (limonene, fenchone, methylchavicol, and anethole) were present in the last-extracted sample. Fennel oil was successfully fractionated into the essential oil rich and fatty oil rich products in pilot-scale apparatus using two separators in series. Designed experiments were carried out to map the effects of pressure and temperature in the first separator on the yields and compositions of the products. The minimum level of the total undesired components in both essential oil rich and fatty oil rich products appeared at a pressure of 80-84 bar and a temperature of 31-35 degrees C in the first separator. Supercritical CO(2) extraction of fennel seeds resulted in higher yield (10.0%) than steam distillation (3.0%), almost the same yield as hexane extraction (10.6%), and lower yield than alcohol extraction (15.4%). Analysis of the volatile compounds revealed the significant difference of the composition in distilled oil and oleoresins prepared by CO(2) and solvent extractions. Sensory evaluation showed that the CO(2) extraction product and distilled oil were more intense in odor and taste than alcohol and hexane extracts.

A study of the production of essential oils in chamomile hairy root cultures.

The active substances in chamomile (Matricaria recutita L.) belong to chemically different structural types. The largest group of medically important compounds forming the essential oils are primarily chamazulene, (-)-alpha-bisabolol, bisabololoxides, bisabolonoxide A, trans-beta-farnesene, alpha-farnesene, spathulenol and the cis/trans-en-in-dicycloethers. Flavonoids, coumarins, mucilages, mono- and oligosaccharides also have pharmacological effects. We studied the production of essential oils in genetically transformed cultures. Sterile juvenile chamomile plants were infected with A4-Y strains of Agrobacterium rhizogenes. They are known plant pathogens and are capable of inducing so-called hairy roots. The transfer DNA segment of the Ri-virulence plasmid of A. rhizogenes becomes integrated in the genome of the plant cells. The isolated hairy roots grow rapidly on hormone-free media. In order to obtain bacteria-free media, we cultured the transformed roots on Murashige-Skoog (MS) medium supplemented with carbenicillin (800 mg/l). To study the production of essential oils, the clones were propagated on liquid and solid MS and Gamborg (B5) media, respectively. According to gas chromatography, the composition of the essential oil of hairy root cultures on different media was found to be similar, but differing in proportion. The main component of the essential oil which was identified by gas chromatography and mass spectrometry was trans-beta-farnesene, as in the intact roots.

Biologically active compounds of Helichrysum arenarium (L.) Moench.

The biologically important components of Helichrysum arenarium (L.) Moench are the flavonoids, which may have choleretic, hepatoprotective and antimicrobial properties. During our study, we examined the flavonoids present in the lyophilizate of the inflorescence of Helichrysum arenarium (L.) Moench (Helichrysi flos, syn. Stoechados flos) and in the tea made from the inflorescence via qualitative and quantitative analysis, and determined the flavonoid content. We examined the properties of the lyophilizate antioxidant, and measured the H-donor activity, reducing power property and total scavenger capacity via spectrophotometric and chemiluminescent methods. We compared the results with those of the agent present in milk thistle (Silybum marianum L.), the silibinin flavonoid. Regarding the H-donor activity, our lyophilizate was determined to be more effective than the silibinin at the same amounts; on the other hand, its reducing power property and total scavenger capacity was lower than that of silibinin. The flavonoid content, which is responsible for the lyophilizate effect, was found to be 0.47% according to our measurements; it is, therefore, possible that an extract with such a concentration of flavonoids may be of therapeutic benefit.

[Phytochemical evaluation of essential oils, medicinal plants and their preparations]. / Illóolajok, illóolajtartalmú gyógynövények és készítményeik fitokémiai értékelése.

A surway is presented on the authors' work in the field of volatile oil research. The gas chromatographic method previously used for analysis of essential oils was transformed to capillary gas chromatographic conditions. The method is also suitable for separation of compound-pairs frequently occurring in essential oils (peppermint, rosemary, lavender, sage, clary sage, thyme oils). Beside the gas chromatographic analysis of essential oils, which was necessary for their standardization and qualification, the influence of different extraction methods and some biological facts e.g. the ontogenesis on the change of essential oil composition are also discussed. It has been established that the water steam distillation from acidic medium can be more advantageous than the traditional one, if the volatile terpene derivatives were bound in form of glycosides or dimeric quajazolide lactons were present in plant (oregano, Sideritis, wormwood oils). Comparing the composition of essential oil obtained by water steam distillation and supercritical fluid extraction (SFE) it was found that the SFE fractions are richer in ester constituents because the possibility of hydrolysis is reduced, and the oils are more valuable than the classic oils. On the other hand, when the transformation processes are important (chamomile), the distillation is the better method. The change of essential oil composition of Ocimum basilicum L. and Anthriscus cerefolium L. was also studied during the vegetation period. It has been established that in budding and early flowering stages the basil oil was rich in monoterpenes; the quantity of sesquiterpenes and phenylpropane derivatives increased only in later stadiums. Finally the extraction and analytical processes are discussed which are used for standardization of complex plant preparations which contained essential oil as active agent and represented various medicinal forms.

[Investigation of the in vitro antioxidant effect of Petroselinum crispum (Mill.) Nym. ex A. W. Hill]. / A Petroselinum crispum (Mill.) Nym. ex A. W. Hill. in vitro antioxidáns hatásának vizsgálata.

Free radical reactions have excited excessive research in the past two decades. Since then it has been proven that these mechanisms may be important in the pathogenesis of certain diseases and aging. Many synthetic antioxidant components have shown toxic and/or mutagenic effects, which have directed most of the attention on the naturally occurring antioxidants. Their use has mainly centered around prevention, and the maintenance of health. Parsley, Petroselinum crispum (Mill.) Nym. ex A. W. Hill belonging to the Apiaceae family, is a well-known spice and vegetable. Its herb and root are widely known for their effects on digestion, stomach, kidney, blood, and liver. The essential oil obtained from the fruit has also strong action on the central nervous system. Characteristic constituents are: flavonoids (apiin, luteolin-, apigenin-glycosides), essential oil (apiol, miriszticin), cumarines, (bergapten, imperatorin) and vitamin C. In our experimental work, various extracts prepared from different vegetative organs of parsley have been investigated. The chemical composition of the extracts and fractions were analyzed by chromatographic (GC, HPLC) and spectroscopic (UV, UV-VIS) techniques. We intended to provide evidence for the antioxidant activity of vegetable drugs and also studied the free radical scavenger activities by means of spectrophotometry (H-donor activity, reducing capability, chelat formation) and chemiluminometry. To prove the free radical scavenging effect of the extracts, the reduced intensity of the H2O2/.OH-isoluminol, microperoxidase system was studied in vitro. The highest correlation was found between the chemical property and the antioxidant effect of the flavonoid rich samples. At the same time it was also observed that the essential oil plays a significant role in the scavenging effect as well. More experiments are in progress to study the most promising compounds in the vegetative and generative organs of parsley.

[Plant anatomical and phytochemical evaluation of Salvia species]. / Zsálya-fajok anatómiai és fitokémiai vizsgálata.

Plant-anatomical and phytochemical investigations were carried out on three Salvia species: S.officinalis L., S.sclarea L., S.pratensis L. It was established that the structure of the glandular hairs of the three species doesn't differ from each other but the characteristics of the covering hairs are different. The covering hairs of sage (Salvia officinalis) consist of 1-4 cells and have protective function. The hairs of the other two Salvia species are bristle hairs. The highest essential oil content was found in sage and the lowest one in S. pratensis. The qualitative and quantitative composition of the essential oils of Salvia species was also various but the qualitative composition of leaf, calix and petal of the same Salvia species was characteristical standard; significant differences were found only in their quantitative composition. Sclareol diterpene alcohol was the main component of the hexane extract obtained from clary sage flowering herb. In polyphenol ingredients Salvia officinalis was the richest. The results demonstrated that the 20% ethanol is the best of the 20, 40, 70% alcoholic solvents, for the extraction of polyphenol compounds. It was established that the rosmarinic acid depside was the main component of polyphenols. Mineral elements were also analysed in the Salvia species leaves as well in the alcoholic and wateric extracts of sage. The magnesium content was considerable in S. pratensis, the zinc content was the highest in S. officinalis.

[Instrumental analysis of medicinal plants and their drug products]. / Gyógynövények és drogjaik müszeres vizsgálatának lehetöségei.

The experiences obtained during the development of gas chromatographic and other (GC, TLC and infrared spectrophotometric) methods for the 7th edition of the Hungarian Pharmacopoeia for essential oils and drugs containing essential oils are summarized with emphasis on the selection of suitable internal standard for the gas chromatographic assays. The qualitative and quantitative estimation of bitter compounds and polyphenoles e.g. flavonoids and procyanidines by means of ultraviolet and infrared spectrophotometry and HPLC is also described. Some HPLC methods for the determination of anthocyan and carotinoid derivatives are also presented. These are not yet included in the pharmacopoeia but are successfully used for the analytical investigation of commercially available medicinal plants and drug products made thereof.