Recent years have seen widespread use of ultra-high-pressure liquid chromatography (UHPLC) in biological fluids. Most commonly the emphasis is on developing high throughput assay methods to reduce the analysis time and cost. Particle size and temperature are chromatographic parameters that can be changed to improve efficiency and obtain rapid separations. UHPLC and high-temperature liquid chromatography (HTLC) are two techniques that reduce the analysis time by decreasing the size of the column packing material and increasing the column temperature, respectively. Both of these techniques have advantages and limitations. In this article we have summarized the history, theoretical background of UHPLC and HTLC and the various advantages and limitations of sample preparation techniques and the detection systems (mass spectrometry and ultraviolet) used for the bioanalytical assays. In addition, selected bioanalytical applications of the two techniques have been reviewed and tabulated.
Ordinary linear regression shows that the methods are well correlated for all compounds. The Deming regression, which assumes error in both the methods, suggests the existence of a proportional and constant bias for 11-dehydro TXB₂ and only proportional bias for 8-isoprostane, PGF(2α), PGD₂ and 15-deoxy Δ(12,14) PGJ₂ between the two methods. According to Deming regression, the two methods are statistically similar for 6-keto PGF(1α) and PGE₂. The Bland-Altman analyses indicate the two methods are commutable.
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