High-performance Thin Layer Chromatographic
HIGH-PERFORMANCE THIN LAYER CHROMATOGRAPHIC
AND DENSITOMETRIC DETERMINATION OF PERSILBEN
IN SPECIES OF POLYGONUM L
H.D. Smolarz1, G. Matysik2
1Department of Pharmaceutical Botany, Medical University,
Staszica st. 4, 20-081 Lublin, Poland
2Department of Inorganic and Analitycal Chemistry, Medical University,
Staszica st. 4, 20-081 Lublin, Poland
It has been reported in many works that TLC is a method appropriate to investigate multicomponental samples of natural origin. The compounds predominating in a sample do not cause any problems in their determination. Chromatography of minor components requires some preliminary work with more concentrated extracts. Some raw extracts are very sticky due to the presence of accompanying components. That is the cause of some complicating phenomena occurring in HPLC column like, e.g., lailing. In this case, HPTLC combined with densitometry is a method appropriate for analysis of such samples because it eliminates preliminary purification of drug extract. In HPTLC it is possible to get rid of ballast matrix choosing suitable solvent systems such that their values RF are higher than 0.9 or quite low RF
Stilbene derivatives are phytoalexin-like compounds which have antifungal and antimicrobial properties. A wide array of these phenolic substances in several studies have been also reported to possess substantial estrogenic, anticarcinogenic and cardioprotective activities. A constituent of the skin grapes and red wine resveratrol (3,4’,5-tetrahydroxystilbene) was reported to be one of the most potent chemopreventive agents able to block all three phases of tumor development, that is initiation, promotion, and progresion. This compound has been shown to inhibit the growth of wide variety of tumor cells, including leukemic, prostate, breast and endotherial cells .An other stilbene derivatives, tamoxifen currently used in the standard hormonal treatment of breast cancer, is an antiestrogen in intensive clinical use.
In this work, persilben (3.5-hydroxy 2-methoxystilbene) was determined in drug extracts from the following taxons of Polygonum: P. lapathifolium ssp.
tomentosum (Schrank) Dans., P. lapathifolium ssp. nodosum (Pers.) Dans., P. amphibium L., P. hydropiper L., P. bistorta L., P.convolvulus L. (Bilderdykia convolvulus (L.) Dumort.
The separation of components in drug extracts from plant material was performed on 10 cm · 20 cm glass HPTLC plates precoated with silica gel Si60 with the fluorescence indicator F254 (E. Merck, Darmstadt, Germany) and 10 · 10 cm Diol F254 HPTLC precoated plates (E. Merck, Darmstadt, Germany). The sample of standard and samples of extracts were spotted by AS30 automatic applicator (Desaga, Heidelberg, Germany). Mobile phases were prepared from mixtures of hexane, chloroform and methanol. Chromatograms were developed in horizontal Teflon DS. chamber (Chromdes, Lublin, Poland).
Videodocumentation of the results was achieved by the use of videoscanning equipment comprising an Hitachi 3 CCD video camera and UV Reprostar 3 (Camag, Muttenz, Switzerland). The documentation was processed with Videostore 2 software, running under Microsoft Windows (minimum 95), adapted to work with the camera.
Densitograms of the extracts were obtained by the use of densitometer Desaga CD 60 (Germany) under the control of a Compaq computer. Illumination with UV light (λ = 360 nm) was used for the measurments. For quantitative determinations the chromatograms were scanned with a zig-zag meander 9 mm (2 · 4.5mm) wide with slit dimension of 0.2 mm · 2 mm. The location of the spots was determined under UV light (λ = 360nm).
Among the eight samples obtained from herbs and rhizomes of different of species of Polygonum L. persilben was detected in five. The content of the detected compound ranges between 46.3 μg and 174.2 μg per gram of dry herb.
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