Separation and direct detection of long chain fatty acids and their methylesters by the non -aqueous reversed phase HPLC and silver ion chromatography, combined with CO laser pumped thermal lens spectrometry
Bicanic, Dane Danijel, 1943- ...
The potential of the CO laser pumped dual beam thermal lens spectrometer (TLS)used as the detector of infrared (IR) absorbance in non-aqueous reversed-phase high pressure liquid chromatography ...(NARP-HPLC) and argentationchromatography (Ag-HPLC-TLS) has been investigated. The linoleic acid C18:2 (9,11) cis, cis, the conjugated linoleic acid C18:2 (9,12) cis, cis, and (9,11) cis, cis and oleic acid C18:1 (9) cis free fatty acids presentin the test mixture were separated and detected by NARP-HPLC-TLS directly, i.e., without a need for their prederivatization. The limit of detection (LOD) for the present NARP-HPLC-TLS set-up was determined using either oleic acid C18:1 (9) cis or linoleic acid C18:2 (9,12) cis, cis as the only analyte. The LOD obtained at 1750 cm -1 for C18;2 (9,12) cis, cis is 0.14% (vžv) with the standard deviation being 3.8% of the measured value. Thisis comparable to the LOD attainable by the bulk property refractive index detector (RID). For C18:1 (9) cis the LOD is 0.04% (v/v) with a standard deviation of 2.5%. The unique feature of the NARP-HPLC-TLS is its excellent selectivity; this was demonstrated experimentally by adding alcohols (octanol and decanol) that lack the C&dbnd;O carbonyl group to the aforementioned mixture of three fatty acids. In another experiment, the Ag-HPLC-TLS in IR wasused to directly detect two different mixtures of five fatty acid methyl esters (FAMEs) after separation on the column. The results were compared to those achieved by the conventional UV absorbance detector. Despite more favourable excitation cross sections of FAMEs in the IR, the concentration LODobtained for Ag-HPLC-TLS is inferior to that of the Ag-HPLC-UV. However, the mass LOD (11 g) achieved by Ag-HPLC-TLS in IR for C18:3 FAME is two timesbetter than that of the Ag-HPLC-UV absorbance detector. High IR absorption of eluent restricts, at present, the sensitivity of the Ag-HPLC-TLSand NARP-HPLC-TLS. Improved LOD's are anticipated by a...
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