Ultrahigh pressure reversed-phase capillary liquid chromatography:: Isocratic and gradient elution using columns packed with 1.0-μm particles
Ultrahigh pressure reversed-phase capillary liquid chromatography:: Isocratic and gradient elution using columns packed with 1.0-μm particles
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DOI：
10.1021/ac9807013
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发表时间：
1999-02-01
影响因子：
7.4
通讯作者：
Jorgenson, JW
中科院分区：
文献类型：
--
作者：
MacNair, JE;Patel, KD;Jorgenson, JW
内径为33 μ m、长度为25-50 cm的熔融石英毛细管用1.0 μ m无孔十八烷基硅烷改性（C-18）二氧化硅球浆状填充。这些色谱柱用于在等度和梯度洗脱模式下进行超高压反相液相色谱分析。流动相压力高达5000巴（72 000 psi）施加到柱入口，以产生超过200 000理论塔板在6分钟（k'接近1）的小，有机分析物。分析物的平均容量因子被发现与所施加的压力线性增加。描述了一种能够产生高达9000巴（130 000 psi）的流动相压力的电驱动恒流注射泵。该泵与指数稀释法结合使用，用于在填充有1.0 μ m颗粒的27 cm长毛细管上从胰蛋白酶消化物中梯度分离肽。30分钟分析的峰值容量为300。
Fused-silica capillaries with inner diameters of 33 mu m and lengths of 25-50 cm are slurry-packed with 1.0-mu m nonporous octadecylsilane-modified (C-18) silica spheres. These columns are used to perform ultrahigh-pressure reversed-phase liquid chromatographic analyses in both isocratic and gradient elution modes. Mobile-phase pressures as high as 5000 bar (72 000 psi) are applied to column inlets to generate more than 200 000 theoretical plates in 6 min (k' approximate to 1) for small, organic analytes. Average capacity factors of analytes are found to increase linearly with applied pressure. An electrically driven constant-now syringe pump capable of generating mobile-phase pressures as high as 9000 bar (130 000 psi) is described. This pump is used in conjunction with an exponential dilution method for the gradient separation of peptides from a tryptic digest on a 27-cm-long capillary packed with 1.0-mu m particles. A peak capacity of 300 is demonstrated for a 30-min analysis.