14 research outputs found
HPLC analysis of alizarin and purpurin produced by Rubia tinctorum L. hairy root cultures
We have determined the quantities of the anthraquinones alizarin and
purpurin, with especial regard to their effective antigenotoxic
activity, in genetically transformed hairy root cultures of Rubia
tinctorum L. Hairy roots were cultured on solid and in liquid Gamborg
B5 and 1/2 NMS media in a shaking cabinet and in a bioreactor.
Methanolic extracts of lyophilized hairy roots were hydrolysed, and
then purified by solid phase extraction with good recovery. A new HPLC
method was developed for the determination of alizarin and purpurin.
The analysis was performed on a Luna C8 RP column using a 45:55 (v/v)
mixture of acetonitrile:20 mM ammonium formate-formic acid buffer (pH
3.00) as eluent. Peaks were identified by addition of standards and by
diode-array detection. External standardization allowed the
determination of alizarin and purpurin with good sensitivity and
reliability. The maximum purpurin content was observed in cultures
cultivated on solid B5 medium (5.94 mg g(-1)). The highest alizarin
content was measured in cultures cultivated on solid 1/2 NMS medium
(2.14 mg g(-1))
Flavones in genetically transformed Scutellaria baicalensis roots and induction of their synthesis by elicitation with methyl jasmonate
The composition and content of flavones were estimated in pRi
T-DNA-transformed skullcap (Scutellaria baicalensis Georgi) roots
obtained by the inoculation of axenically grown seedlings with a wild
A4 strain of the soil bacterium Agrobacterium rhizogenes. It was
elucidated by analytical and preparative HPLC of phenolic compounds in
the extracts from the pRi T-DNA-transformed roots and also by
ultraviolet spectroscopy and H-1 and C-13 NMR that cultured skullcap
roots contained similar basic flavones as intact roots of this plant
species. i.e., baicalein and wogonin and corresponding glucuronides,
baicalin and wogonoside. The content of these flavones in cultured
roots was threefold lower than in the roots of intact five-year-old
plants. When skullcap roots were cultured on B5 or Murashige and Skoog
medium, the ratios between major flavones changed but their total
content remained unchanged. The treatment of three-week-old cultured
roots with methyl ether of jasmonic acid (MeJa) doubled the total
concentration of major flavones in roots; the content of aglycons,
baicalein and wogonin, increased to a greater degree, e.g., by 2.3 and
3.3 times, respectively. The induction of flavone production by
elicitors indicates that flavones behave as phytoanticipins because
major flavones of skullcap manifest a distinct antimicrobial activity.
The results of the short-term treatment of skullcap roots with MeJa
show that stress biotic factors can considerably increase the content
of physiologically active flavones
New terpenoids in cultivated and wild chamomile (in vivo and in vitro)
The effect of Chamomilla recutita (L.) Rauschert is made up by several
groups of active substances, among which terpenoids in the
inflorescences are of greatest importance. Among cultivated species,
the Hungarian BK-2 contains more chamazulene in its essential oil than
the German Degumil type, which is mainly cultivated for its
(-)-alpha-bisabolol. Both components have important antiinflammatory
activities. Among wild chamomile populations in Hungary, a population
was found in the area of Szabadkigyos containing significant amounts-on
average 48%-of (-)-alpha-bisabolol in its inflorescence oil. In vitro
cultures were made from this population to obtain propagation material
containing a high number of active substances. The intact roots
contained no (-)-alpha-bisabolol but the sesquiterpene alcohol
beta-eudesmol as new compound was identified by our group. Sterile
plantlets, cultured in vitro, were multiplied for phytochernical
investigations. Pharmacologically important compounds of the essential
oils were followed in great detail. The amount of in vitro cultured
terpenoids and polyin compounds was compared with that of in vivo
plants. These volatile compounds were identified by comparing their
retention times with those of authentic standards, essential oils of
known composition and peak enrichment. The confirmation of identity was
done by comparison of their mass spectra with those reported in the
literature and reference compounds. The percentage evaluation of each
component was made by area normalisation. Gas chromatography (GC) and
mass spectrometry (MS) showed that sterile chamomile cultures generated
the most important terpenoid and polyin compounds characteristic of the
parent plant. We identified germacrene-D, berkheyaradulene, 4-(2', 4,
4'-trimethyl-bicyclo[4.1.0]hept-2'-en-3'-yl)-3-buten-2-one,
geranyl-isovalerate and cedrol as new components in these sterile
cultures. (C) 2003 Elsevier B.V. All rights reserved
GC and GC-MS studies on the essential oil and thiophenes from Tagetes patula L.
The production and the composition of the compounds (mono-,
sesquiterpenes and acetylenic thiophenes) obtained by the steam
distillation of Tagetes patula L. have been investigated. The volatile
oil was produced by steam distillation. GC was carried out on three
types of stationary phase using flame ionization and mass selective
detection. Percentage data were calculated by the area normalization
method with very good repeatability (RSD below 5%). Oils from
flowerheads were rich in beta-caryophyllene (53.5%) and the leaves
contained terpinolene in high concentration (21.1%). The main volatile
component of the hairy roots and intact roots was
5(3-buten-1-ynyl)-2,2'-bithienyl (BBT) yielding 28.5% and 44.0% in the
oils. Three new minor constituents were identified as alpha-guriunene,
beta-caryophyllene and (E)-beta-farnesene. A flash chromatographic
method was developed for the isolation of thiophenes from a solvent
extract of intact roots. The collected fractions were screened by TLC
and analyzed by GC-MS. Three thiophene fractions were obtained
containing BBT, 5-(4-acetoxy-1-butynyl)-2,2'-bithienyl (BBTOAc) and
5-(4-hydroxy-1-butynyl)-2,2'-bithienyl (BBTOH)