We present a new simple thin-layer chromatographic method designed for determination of the main alkaloids of Chelidonium majus L. In this study, we used roots and herb of the plant collected in spring and autumn. The alkaloid fractions were prepared according to modified pharmacopeial procedure .
In our method, we performed two-step elution onto silica gel plates. The first eluent consisted of chloroform, methanol, and water mixed with 70:30:4 proportion. The second eluent comprised of toluene, ethyl acetate, and methanol with 83:15:2 proportion. The described thin-layer chromatography (TLC) system allows qualitative and quantitative determination of the following alkaloids: sanguinarine, chelerythrine, chelidonine, coptisine, and berberine. For determination of protopine, eluent with n-buthanol, acetic acid, and water in 15:1.5:4 proportion was investigated.
The dominant alkaloids observed in studied fractions were coptisine (1027.096 ± 13.367–287.474 ± 3.069 mg/100 g dry matter ± sdv) and chelidonine (1780.667 ± 263.522– 115.929 ± 14.694 mg/100 g dry matter ± sdv). The alkaloid detected in the least amount was chelerythrine (30.74 ± 7.526–1.143 ± 0.0651 mg/100 g dry matter ± sdv). The highest total amount of all alkaloids was determined in the fractions obtained from herbs in spring, and the lowest amount was detected in herbs autumn.
Additionally, we compared amounts of studied alkaloids in different parts of plants (aerial parts and roots). The plants were collected in spring and autumn.
Authors concluded that the presented method can be used as a valuable tool for screening studies on C. majus L.
Authors:Marek Och, Anna Och, Łukasz Cieśla, Janusz Kocki and Anna Bogucka-Kocka
Eleven Juniperus species and a total of 62 different varieties collected at different localities in Poland were extracted with methanol in order to determine the level of umbelliferone present in the extracts. Thin-layer chromatography (TLC) analysis was performed on glass silica gel 60 F254 plates first developed with n-hexane-ethyl acetate (70:30, 7p/7p) and redeveloped after drying with dichloromethane and diethyl ether (80:20, 7p/7p). Densitometric determinations were performed in fluorescence mode by using a deuterium lamp (D2) for excitation at 326 nm and a K400 filter for measurement of the emitted light using a CAMAG TLC Scanner 3. A linear regression equation was used to determine the amount of umbelliferone in the extracts. The mobile phases used for TLC separated well umbelliferone from other compounds present in the extracts. A good linear regression model was obtained in the range of 10-80 ng with a correlation coefficient (r) of 0.99844. Juniperus communis L. “Pendula” contained the highest (88.3 mg/100 g DW) UMB content. Umbelliferone quantitative screening in so many Juniperus species has been performed for the first time. There are no reports dealing with the quantitative assessment of umbelliferone in the Juniperus species. The described method has been validated and it is simple, fast, reliable, specific, precise, and useful for routine assays of Juniperus extracts containing umbelliferone.
Authors:Łukasz Cieśla, Anna Petruczynik, Michał Hajnos, Anna Bogucka-Kocka and Monika Waksmundzka-Hajnos
Coumarins are natural, biologically active substances, normally found in complex mixtures. Unfortunately their separation causes many difficulties, because of to their similar chemical structure and physicochemical properties. A new, reliable method has been established for analysis of coumarin fractions present in selected fruit extracts. The substances were separated in chromatographic systems that enabled use of orthogonal separation mechanisms (i.e. characterized by different selectivity). The greatest selectivity differences were obtained by use of two chromatographic systems — first dimension CN-silica with 30% ACN in H
O as mobile phase (triple developed) and second dimension SiO
with 35% AcOEt in
-heptane as mobile phase (triple developed), or first dimension SiO
with 35% AcOEt in
-heptane as mobile phase (triple developed) and second dimension RP-18 with 55% MeOH in H
O as mobile phase. The aforementioned two-dimensional TLC systems were used for separation of coumarin fractions present in extracts from
Archangelica officinalis, Pastinaca sativa and Heracleum sphondylium fruits
Authors:Lukasz Cieśla, Anna Petruczynik, Michał Hajnos, Anna Bogucka-Kocka and Monika Waksmundzka-Hajnos
Coumarins are interesting group of natural compounds, because of their biological and pharmacological activity, and are widely investigated. They are normally found in complex mixtures, e.g. plant extracts, and are difficult to separate in one chromatographic run. Mixtures of coumarins have been separated by two-dimensional thin-layer chromatography on CN-silica plates by use of aqueous and nonaqueous mobile phases. Complete separation was also achieved by use of graft thin-layer chromatography on connected layers — silica with RP-18W or CN-silica with silica. The systems characterized by the best efficiency and selectivity were used for separation of coumarin fractions from extracts of selected
plants. These orthogonal systems were used for quantitative analysis of selected coumarins. The results obtained show two dimensional thin-layer chromatography is useful tool for quantification of some furanocoumarins in plant extracts. The best results were obtained on connected silica and octadecyl silica layers.
Authors:Anna Hawrył, Anna Bogucka-Kocka, Ryszard Świeboda, Mirosław Hawrył, Adam Stebel and Monika Waksmundzka-Hajnos
Twenty land mosses were extracted by double maceration and ultrasonic extraction techniques using the mixture of 80% ethanol and water. The obtained extracts were analyzed using thin-layer chromatography (TLC) with silica gel (the mobile phase was consisted of ethyl acetate–formic acid–acetic acid–water, 14.0:1.5:1.5:2.0, v/v) and RP-18 (the composition of eluent methanol–water–formic acid, 7.0:2.5:0.5, v/v) chromatographic plates. After developing and drying, the plates were sprayed using the Naturstoff reagent, and after drying, the chromatograms were photographed and the obtained images were processed using the ImageJ program. Principal component analysis was performed to confirm the chemical similarity between the studied moss extracts. The cytotoxic activity of the ethanolic extracts of Bryophyta species was studied using the cell lines CCRF/CEM and CEM. For most of the moss samples, the cytotoxic activity was confirmed.