Anthocyanins paper chromatography

Paper chromatography (PC) and thin layer chromatography (TLC) have been used since the 1940s. Preparative PC on Whatman 3 paper, analytical PC on Whatman 1 paper, and analytical TLC on microcrystalline cellulose, silica gel, or polyamide have been applied with a variety of solvents and the behaviors of anthocyanins have been similar in all media. Two-dimensional TLC allows the separation of several compounds and has been nsed to clarify the anthocyanin compositions of different commodities. ... 

The anthocyanin profile of the flowers of Vanda (Orchidaceae) was investigated with a similar technique. Flowers (2 kg) were extracted with 101 of methanol-acetic acid-water (9 l 10,v/v) at ambient temperature for 24 h. The extract was purified by column chromatography, paper chromatography, TLC and preparative RP-HPLC. Analytical HPLC was carried out in an ODS column (250 X 4.6 mm, i.d.) at 40°C. Gradient conditions were from 40 per cent to 85 per cent B in 30 min (solvent A 1.5 per cent H3P04 in water solvent B 1.5 per cent H3P04, 20 per cent acetic acid and 25 per cent ACN in water). The flow rate was 1 ml/min and analytes were detected at 530 nm. The chemical structures of acylated anthocyanins present in the flowers are compiled in Table 2.90. The relative concentrations of anthocyanins in the flower extracts are listed in Table 2.91. It can be concluded from the results that the complex separation and identification methods (TLC, HPLC, UV-vis and II NMR spectroscopy, FAB-MS) allow the separation, quantitative determination and identification of anthocyanins in orchid flowers [262],... 

Bate-Smith, E.G. (1948) Paper chromatography of anthocyanins and related substances in petal extracts. Nature, 161,835-8. 

Paper and thin layer chromatography have been used since decades for flavonoid analysis. As anthocyanins are colored substances, separation can be directly monitored and may serve as a quick, qualitative test for the anthocyanin distribution in a given sample. Classical paper chromatography involves two-dimensional development followed by treatment with selective spraying reagents or exposure to ammonia vapor and allocation of spots in visible or UV-light [90]. 

Paper chromatography may be used as one step for separation and/or isolation of anthocyanins from complex samples. Thin-layer chromatography, performed on cellulose or silica gel plates, was in the first period of use a convenient extension of paper chromatography. The introduction of new stationary phases and the development of instrumentation offering the possibility of quantitative analysis provided new possibilities for method developments. The advantages of TLC are the simplicity in terms of instrumentation, sample preparation and data evaluation. 

Rf Values of Common Anthocyanins and Anthocyanidins by Paper Chromatography... 

Although neither paper nor high performance- or thin-layer chromatography plays a major role for routine analysis of beny anthocyanins, the value of these methods for research oriented work should never be underestimated. 

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