小弟做游离氨基酸,先用的梯度是:0.01min B 0% ,12min B 15% ,30min B 34% ,30.01min B 100% ,40min B 0% ,50min B 0%.这是经典的做氨基酸的梯度,但我用得不行,峰总有几个连在一起···· 已经排除了其他条件对峰的干扰可能(全部按标准做的) 后认为应该提高洗脱能力,对于反向柱,降低B的含量···· 于是采用这个梯度:0.01min B 0% ,13min B 7% ,23min B 23% ,29min B 35% ,35min B 40% ,40min B 100% ,45min B 100% ,47min B 0% 这个梯度已经降低了B的含量,但还是无法把峰分开····