Summary
Three new methods of improving triglyceride separation by RP-HPLC using propionitrile as eluent and an RI detector are described. First the baseline was stabilized by indirect thermostating of the detector. Relatively high sensitivity adjustments (4×10−5 RI units full scale deflection) were possible allowing small fat samples to be injected (down to 200 μg). Second the increase of selectivity with decreasing temperature and the increasing sharpness of peaks at higher temperatures are shown. A high temperature is indispensible for saturated long chain compounds which tend to crystallize on the column. To fulfill this requirement and avoid loss of selectivity temperature programming has been investigated. Third at higher temperatures an additional axial temperature gradient up to 0.05 K/cm was applied to enhance peak symmetry. All chromatograms achieved by those temperature gradients show higher resolution than those obtained at constant temperatures. Butterfat could be separated into more peaks than has previously been possible. The chromatogram of soybean oil, besides showing excellent selectivity for more polar components shows a series of ten long chain compounds following POP.
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Abbreviations
- CN:
-
acyl carbon number
- ECN:
-
equivalent carbon number
- ND:
-
number of double bonds
- TG:
-
triglyceride
- La:
-
lauric acid
- M:
-
myristic acid
- P:
-
palmitic acid
- S:
-
stearic acid
- A:
-
arachidic acid
- O:
-
oleic acid
- L:
-
linoleic
- at:
-
attenuation
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Frede, E. Improved HPLC of triglycerides by special tempering procedures. Chromatographia 21, 29–36 (1986). https://doi.org/10.1007/BF02310591
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DOI: https://doi.org/10.1007/BF02310591