Benton Christopher M, Lim Chang Kee, Moniz Caje, Jones Donald J L
Clinical Biochemistry, King's College Hospital, Denmark Hill, London, SE5 9RS, UK.
Biomed Chromatogr. 2012 Jun;26(6):714-9. doi: 10.1002/bmc.1720. Epub 2011 Oct 27.
Ultra high-performance liquid chromatographic (UHPLC) systems on columns packed with materials ranging from 1.9 to 2.7 µm average particle size were assessed for the fast and sensitive analysis of porphyrins in clinical materials. The fastest separation was achieved on an Agilent Poroshell C(18) column (2.7 µm particle size, 50 × 4.6 mm i.d.), followed by a Thermo Hypersil Gold C(18) column (1.9 µm particle size, 50 × 2.1 mm i.d.) and the Thermo Hypersil BDS C(18) column (2.4 µm particle size, 100 × 2.1 mm i.d.). All columns required a mobile phase containing 1 m ammonium acetate buffer, pH 5.16, with a mixture of acetonitrile and methanol as the organic modifiers for optimum resolution of the type I and III isomers, particularly for uroporphyrin I and III isomers. All UHPLC columns were suitable and superior to conventional HPLC columns packed with 5 µm average particle size materials for clinical sample analysis.
评估了平均粒径在1.9至2.7 µm范围内的填充材料的超高效液相色谱（UHPLC）系统，用于临床材料中卟啉的快速灵敏分析。在安捷伦Poroshell C(18)柱（粒径2.7 µm，内径50×4.6 mm）上实现了最快的分离，其次是赛默飞Hypersil Gold C(18)柱（粒径1.9 µm，内径50×2.1 mm）和赛默飞Hypersil BDS C(18)柱（粒径2.4 µm，内径100×2.1 mm）。所有色谱柱都需要一种流动相，该流动相包含1 m醋酸铵缓冲液（pH 5.16），并以乙腈和甲醇的混合物作为有机改性剂，以实现I型和III型异构体的最佳分离，特别是对于尿卟啉I和III异构体。所有UHPLC色谱柱都适用于临床样品分析，并且优于填充平均粒径为5 µm材料的传统HPLC色谱柱。
