HPTLC determination of catechins in in-vitro cultures of two species of the genusPhyllanthus

2008 ◽  
Vol 21 (2) ◽  
pp. 103-106 ◽  
Author(s):  
Barbara Sparzak ◽  
Mirosława Krauze-Baranowska ◽  
Loretta Pobłocka-Olech
Keyword(s):  
In Vitro Cultures

scholarly journals Comparative Analysis of Therapeutically Important Indole Compounds in in vitro Cultures of Hypericum perforatum Cultivars by HPLC and TLC Analysis Coupled with Densitometric Detection

2014 ◽  
Vol 9 (10) ◽  
pp. 1934578X1400901 ◽  
Author(s):  
Bożena Muszyńska ◽  
Halina Ekiert ◽  
Inga Kwiecień ◽  
Anna Maślanka ◽  
Rawad Zodi ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Growth Regulators ◽  
Hypericum Perforatum ◽  
Dry Mass ◽  
In Vitro Cultures ◽  
Indole Compounds ◽  
Densitometric Detection ◽  
Tlc Analysis

Five indole compounds (5-hydroxy-L-tryptophan, L-tryptophan, indole-3-acetic acid, melatonin, serotonin) and hypericin were identified and quantified in methanolic extracts of shoot cultures of three Hypericum perforatum cultivars (Helos, Elixir, Topas) growing on two variants of Murashige -Skoog medium differing in concentrations of growth regulators (naphthalene-1-acetic acid and 6-benzylaminopurine). Extracts of the aboveground parts of field-grown plants ( Hyperici herba) were also analyzed by HPLC and TLC analysis coupled with densitometric detection. Determination of four compounds was based on our assay described earlier. The methods of determination of 5-hydroxy-L-tryptophan and hypericin were developed and validated in this study. The composition and contents of the metabolites under study differed between the cultivars cultured in vitro and between medium variants containing diverse contents of growth regulators. The contents of individual indole compounds in the biomass from in vitro cultures ranged from 39.6 to 343.2 mg/100 g dry mass. 5-Hydroxy-L-tryptophan was the dominating metabolite (from 78.2 to 343.2 mg/100 g dry mass). Extracts from shoots of the cultivar Helos also contained high contents of serotonin (319.9 and 197.4 mg/100 g dry mass). The contents of indole compounds in Hyperici herba were also diverse (from 7.1 to 55.3 mg/100 g dry mass). 5-Hydroxy-L-tryptophan was the dominating metabolite as well. Hypericin content of Hyperici herba, equaling 12.2 mg/100 g dry mass was from 3.3 to 10 times higher than in extracts from shoots cultured in vitro. The present report is the first analysis of endogenous accumulation of indole compounds in Hyperici herba which involves, apart from melatonin, four other compounds.

scholarly journals Galanthamine Content of Bulbs and in Vitro Cultures of Leucojum Aestivum L

2006 ◽  
Vol 1 (6) ◽  
pp. 1934578X0600100 ◽  
Author(s):  
Mamadou F. Diop ◽  
Agata Ptak ◽  
Françoise Chrétien ◽  
Max Henry ◽  
Yves Chapleur ◽  
...  
Keyword(s):  
Hplc Method ◽  
Growth Substance ◽  
In Vitro Cultures ◽  
Tissue Cultures ◽  
Qualitative And Quantitative ◽  
Leucojum Aestivum ◽  
Cholinesterase Inhibitory Activity

In vitro cultures, at different stages of morphogenesis, were established from leaves of Leucojum aestivum (Amaryllidaceae) for determination of their galanthamine content, an alkaloid that possesses cholinesterase inhibitory activity and which is used for the treatment of Alzheimer's disease. A suitable HPLC method for qualitative and quantitative determination of galanthamine in both in vitro and in vivo extracts has been developed. Confirmation of the identity of galanthamine in sample extracts was achieved using LC-MS-MS. A correlation was observed between the state of differentiation and the galanthamine content of the tissue cultures. No galanthamine was detected in the roots grown in vitro, while all bulblets grown in vitro showed the presence of this alkaloid, with dramatic variations in concentration levels, according to the growth substance balance. The best result (6.79 × 10−3 % of D.W.) was obtained with bulblets initiated with NAA (10 μM) combined with BA (0.5 μM).

scholarly journals High-Performance Thin-Layer Chromatography Densitometric Determination of β-Sitosterol in Phyllanthus Species

2009 ◽  
Vol 92 (5) ◽  
pp. 1343-1348 ◽  
Author(s):  
Barbara Sparzak ◽  
Mirosawa Krauze-Baranowska ◽  
Loretta Pobocka-Olech
Keyword(s):  
Plant Material ◽  
Mobile Phase ◽  
High Performance ◽  
Dry Weight ◽  
Phosphomolybdic Acid ◽  
In Vitro Cultures ◽  
Layer Chromatography ◽  
Interday Precision

Abstract An HPTLC densitometric method was established for the determination of -sitosterol in in vitro cultures of some species of the genus Phyllanthus. Two derivatization reagents commonly used in TLC for the visualization and detection of sterols, namely, anisaldehyde reagent and 5 phosphomolybdic acid, were compared with vanillin reagent. The densitometric quantification of -sitosterol was carried out on HPTLC Si60 F254 plates with the mobile phase chloroformhexanemethanol (65 + 30 + 5, v/v/v) at 525 nm. The method was validated for each derivatization reagent in terms of linearity, precision, repeatability, intra- and interday precision, LOD, and LOQ. The presence of -sitosterol was revealed in all analyzed plant material. The concentrations of -sitosterol determined ranged from 0.48 to 2.75 mg/g (dry weight). In addition, traces of -amyrin were detected in some plant samples.

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