Doehl J, Greibrokk T
Department of Chemistry, University of Oslo, Norway.
J Chromatogr. 1990 Jul 13;529(1):21-32. doi: 10.1016/s0378-4347(00)83804-9.
Prostaglandins of the E type (PGEs) in human seminal fluid have been determined by reversed-phase high-performance chromatography on a C18 column and ultraviolet detection at 230 nm after solid-phase extraction (C18) and oxidation to the corresponding 15-oxoprostaglandin derivatives by pyridinium dichromate in acetonitrile. Under optimized conditions, PGEs from 10-ml seminal samples were extracted into 4 ml of methyl formate with high recoveries (estimated at greater than 95%) and subsequently separated under mild chromatographic conditions (0.5 mM formic acid-acetonitrile, apparent pH 3.8). Comparable analytical sensitivities were obtained with detection at 230 nm with a conventional deuterium lamp spectrophotometer and a photometer equipped with a cadmium emission source, while with a diode-array spectrophotometer, signal-to-noise ratios were reduced with factors between 4.4 and 3.1, depending on the spectral bandwidth of the instrument. Theoretical aspects of signal-to-noise optimization of ultraviolet detectors are discussed. The stability of dilute standard solutions of PGE2 and PGD2 was measured, showing solutions in dichloromethane at 20 degrees C to be as stable as acetonitrile solutions at 5 degrees C over a period of thirty days. Absolute ethanol and acetonitrile were equally suited as solvents.
采用C18柱反相高效液相色谱法，并在固相萃取（C18）后于230 nm处进行紫外检测，通过在乙腈中用重铬酸吡啶将其氧化为相应的15 - 氧代前列腺素衍生物，测定了人类精液中的E型前列腺素（PGEs）。在优化条件下，将10 ml精液样本中的PGEs萃取到4 ml甲酸甲酯中，回收率很高（估计大于95%），随后在温和的色谱条件下（0.5 mM甲酸 - 乙腈，表观pH 3.8）进行分离。使用传统的氘灯分光光度计和配备镉发射源的光度计在230 nm处检测可获得相当的分析灵敏度，而使用二极管阵列分光光度计，根据仪器的光谱带宽，信噪比会降低4.4至3.1倍。讨论了紫外检测器信噪比优化的理论方面。测定了PGE2和PGD2稀标准溶液的稳定性，结果表明，在30天的时间内，20℃二氯甲烷溶液中的稳定性与5℃乙腈溶液中的稳定性相当。无水乙醇和乙腈同样适合作为溶剂。
