Imai K, Nishitani A, Tsukamoto Y, Wang W H, Kanda S, Hayakawa K, Miyazaki M
Branch Hospital Pharmacy, University of Tokyo, Japan.
Biomed Chromatogr. 1990 May;4(3):100-4. doi: 10.1002/bmc.1130040305.
The catalytic effect of bases (imidazole, pyridine, Tris and triethylamine) on the peroxyoxalate chemiluminescence (PO-CL) reaction for high performance liquid chromatography (HPLC) was investigated. Imidazole increased PO-CL intensity extraordinarily, whereas the other bases (pyridine, Tris and triethylamine) did not. The peak heights of dipyridamole (coronary vasodilator) obtained using the eluents containing buffers were largest at pH 7.0, a few times less at pH 6.0 and pH 5.0, 100 times less at pH 4.0 and a few hundred times less at pH 3.0. The eluents containing buffers at pH 3, 4, 5, 6 or 7 each with imidazole increased the peak heights by a few to ten times as compared with those without imidazole, and those peak heights were within one order of magnitude. On the other hand, the eluent containing buffer at pH 2 did not affect the peak heights with or without imidazole. Bis(4-nitro-2-(3,6,9-trioxadecyloxycarbonyl)phenyl) oxalate (TDPO) alone and bis(2,4-dinitrophenyl)oxalate (DNPO) plus TDPO were recommended to be used against eluents containing buffers of pH 5-7 and pH 3-4, respectively. Dipyridamole and benzydamine hydrochloride (anti-inflammatory drug) were separated on the ODS column and detected by the present system. The detection limits of dipyridamole and benzydamine hydrochloride were 40 amol and 270 fmol, respectively.
研究了碱（咪唑、吡啶、三羟甲基氨基甲烷和三乙胺）对用于高效液相色谱（HPLC）的过氧草酸酯化学发光（PO-CL）反应的催化作用。咪唑极大地提高了PO-CL强度，而其他碱（吡啶、三羟甲基氨基甲烷和三乙胺）则没有。使用含缓冲剂洗脱液获得的双嘧达莫（冠状血管扩张剂）的峰高在pH 7.0时最大，在pH 6.0和pH 5.0时减小几倍，在pH 4.0时减小100倍，在pH 3.0时减小几百倍。含pH 3、4、5、6或7缓冲剂且含有咪唑的洗脱液与不含咪唑的洗脱液相比，峰高增加了几倍到十倍，且这些峰高在一个数量级内。另一方面，含pH 2缓冲剂的洗脱液无论有无咪唑都不影响峰高。建议分别使用双（4-硝基-2-（3,6,9-三氧杂癸基氧基羰基）苯基）草酸酯（TDPO）和双（2,4-二硝基苯基）草酸酯（DNPO）加TDPO来对抗含pH 5 - 7和pH 3 - 4缓冲剂的洗脱液。在ODS柱上分离双嘧达莫和盐酸苄达明（抗炎药），并通过本系统进行检测。双嘧达莫和盐酸苄达明的检测限分别为40 amol和270 fmol。
