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Title Comparison of Column Performance between Monolithic and Particulate Parking for Separation of Photosynthetic Pigments
Edition
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ISBN/ISSN
Author(s) Tatas H.P. Brotosudarmo
Leenawaty Limantara
Indriatmoko
Yuzo Shioi
Subject(s) particulate packing
Column
Monolithic
Photosynthetic pigment
Pigment analysis
Pleomele angustifolia
UFLC
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GMD Text
Language English
Publisher --
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Abstract/Notes Pleomele angustifolia has been used as a source of natural colorant for many years in Indonesia. By the wave of healthy life style campaign, it is reconsidered again the importance of this plant instead of use of artificial colorants and give us great potency in producing easy to use natural colorant-product through sophisticated Industries. This species is known that it contains highly concentrated green compound which is able to provide green chlorophyll-based colorant. Recently, we found that P. angustifolia contained some carotenoids, i.e., violaxanthin, zeaxanthin, and β-carotene in addition to chlorophylls a and b, whose have unique functional role in human health. High-performance and ultra- fast liquid chromatography (HPLC and UFLC) are the most common techniques for analyzing and quantifying photosynthetic pigments. One of the key roles in providing good separation in these methods is the column. In this study, we examined two different C18 columns in order to characterize their performance for the separation of pigments extracted from P. angustifolia. We used Chromolith (MERCK), as a representative of monolithic column which is no longer packed with small particles but instead consists of a single piece of high-purity silica gel with a bimodal pore structure and XR-ODS (Shimadzu), as a standard small particulate column using a simple isocratic system and photo diode array detector (430 nm detection). In addition, C8 column was also compared with C18 column to understand the effects of carbon chain length. We analyzed several chromatographic parameters such as eluent compositions, flow rates, and temperatures to determine the best system conditions in separating this leaf pigments. Separated factor and resolution were used as indicators of each peak quality as results of applied system. We also conducted a peak ratio analysis for three major pigments, violaxanthin (tR viol), chlorophyll a (tR Chl_a), and β-carotene (tR b-car) to understand column efficiency in separating polar and non-polar pigments under certain conditions. Column efficiency between two different structural properties will be compared and discussed
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