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Title Separation of Photosynthetic Pigments by High-Performance Chromatography: Comparison of Column Performance, Mobile Phase, and Temperature
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ISBN/ISSN
Author(s) Tatas H.P. Brotosudarmo
Leenawaty Limantara
Indriatmoko
Yuzo Shioi
Subject(s) HPLC
chromatography
Photosynthetic pigmets
Analitical detection
Classification
Series Title
GMD Prosiding Ilmiah Internasional
Language English
Publisher Sams
Publishing Year 2014
Publishing Place Malang
Collation
Abstract/Notes High-performance liquid chromatography (HPLC) has been commonly used as method of separating and identifying photosynthetic pigments such as chlorophylls and carotenoids because of such advantages as speed, high resolution and sensitivity. In this technique, high separation relies largely on the type of column material. Previously, we analyzed two silica particulate packed columns (C18 and C8) as the standard in addition to C18 monolithic type column to understand the effect of carbon chain length and the difference between particulate and monolithic types on the pigment separation. Our results showed that monolithic column provided better resolution and faster analysis, although each column have showed their characteristic features. In the present study, we examined two new types of column manufactured by Cosmosil: π-NAP and Cholester columns. π-NAP column is a reverse phase column with stationary phase materials of napthylethyl. Napthylethyl group has stronger π-π interactions than phenyl column and has improved selectivity for structural isomers. On the other hand, Cholester column is a reverse column with cholesteryl bonded silica packing material which has strong stereoselectivity compared to usual ODS column. The experiment conditions were the same as previously used, i.e., a simple isocratic solvent system composed of methanol and acetonitrile as a mobile phase at a flow rate of 0.5 ml per min and column temperatures of 30°C and 40°C. The pigments were detected by Photodiode array detector (200-800 nm) (SPD-M20A) and analyzed by LC workstation, Class LC20/M20A (Shimadzu). Here, we used pigments extracted from leaves of Pleomele angustifolia, an indigenous source of natural colorants. It contains common 6 major pigments such as chlorophylls a and b, violaxanthin, zeaxanthin, α-carotene, and β-carotene. The efficiency of the columns will be compared and discussed in this study. The results provide recommendation for the column selection in separating photosynthetic pigments, furthermore, it can help the optimization of rapid time analysis by using a simple, safe, and low cost mobile phase system
Specific Detail Info Humbolt Kolleg II in Conjuction with The 2nd International Conference on Natural Sciences (ICONS) 2014, Malang, 25-28, September 2014
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