LCGC Asia Pacific September 2016

September 2016 | Volume 19, Issue 3
Cover Story
Carotenoids are a class of natural pigments, widely distributed in vegetables and fruits. A comprehensive two-dimensional liquid chromatography (LC×LC) method, based on the use of a cyano and an octodecylsilica column, placed in the first and second dimension, respectively, was applied to evaluate carotenoid composition and stability in selected overripe fruits representing the waste generated by a local food market. This research also evaluates if post-climacteric biochemical changes are linked to carotenoid degradation in the investigated fruits. A total of 22 compounds was separated into seven different chemical classes in the two-dimensional space, and identified by photodiode array (PDA) and mass spectrometry (MS) detection. The results prove that the waste generated by the large distribution of food still represent an important source of bioactives that could be used for other purposes.
Features
Ultrahigh-pressure liquid chromatography (UHPLC) instruments from different manufacturers and instruments with different configurations can produce significant variations in chromatographic separation. The variety in instrument configuration increases the complexity of the method development process, which now requires a more thorough evaluation of the effect of instrument variations on the method. The studies presented here determined the typical interinstrument variations in dwell volume, extracolumn dispersion, and mixing efficiency as measured by mobile-phase compositional accuracy. Additionally, the dwell volume and extracolumn dispersion were independently and systematically varied to evaluate the resulting impact on resolution for a small-molecule test mixture during gradient elution. To account for these interinstrument variations, dwell volume and wash-out volume method translation and adjustment techniques were evaluated.
GC Connections
While gas chromatographers may take their septa for granted, in fact these small and seemingly unremarkable polymer disks keep air out of the carrier-gas stream when used in an inlet and keep samples intact and uncontaminated when used in sample vials. Choosing the wrong septa can compromise method accuracy and repeatability as well as reduce column life in extreme cases. This instalment addresses septa for inlets and sample vials.
Sample Preparation Perspectives
The past couple instalments of “Sample Preparation Perspectives” have looked at current trends in the field. Another recent trend is dried blood spot analysis and other analysis methods using minute sample amounts. This month we take a quick look at the role of sample homogeneity and the determination of sample size. Microsampling approaches, including dried blood spots, are discussed.
Perspectives In Modern HPLC
Significant benefits can be obtained by standardizing high performance liquid chromatography (HPLC) columns in a pharmaceutical development laboratory. Here is a story of how one organization attempted to encourage its staff to develop HPLC methods using fewer column brands and dimensions to reduce waste and efforts in method transfers downstream.
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