HPLC流动相的缓冲液的配制1.docVIP

HPLC流动相的缓冲液的配制 HPLC流相的冲液动动动动动动 动动动动动动冲液流相可以根据下面的操作配制:步 将冲液品溶解在水中制成冲液，溶液动动动动动动动动动动动动动动动动动动A,。 用pH动动动动动确定或溶液A的pH。 将一定体，如动动动200ml,的有机溶动B与由步动2配制的溶液A，如800ml,混合,制成最动 流相，此的有机相动动动动动动动动动动20%,。 动动动动动动动最流相的pH，此,。动动动动动动 由于定含有有机的流相动动动动动动动动动动动动动pH动动动动生漂移,使其定pH不很可靠,所以上面的步动4 一般只有在考察差的主要来源采用,大多数室皆略去此动动动动动动动动动动动动动动动动动动。步 步动1一般是将配制冲液所用的两，动动动动动动动动动动动A1和A2,分按算量先溶解在动动动动动动动动动 水中制成水溶 液,然后再将两溶液按一定的比例混合得到期动动动动动动动动动动动动动动动动动pH的溶液A。若在此步动2中动动pH, 动动可以采用溶A滴定溶动B,也可反来滴定,并通动动动动动动动动pH动动量得到所需pH的最冲溶动动动动 液。在大多数室中,动动动动动pH的量精度通常不会达到动动动动动动动动动动?0.05—0.1个pH动动动动动位,会著 影响一些品的分离。当动动动动动动动HPLC方法动pH非常敏感,动动动动步2只能大致确定pH动。 将准确称重的冲液配制与精确体的脱气蒸水混合,使动动动动动动动动动动动动动动动动动动动动动动 其成均一的溶液，不一动动动动动动整步pH,,如此配得得冲溶液得动动动动动pH可以控制在非常窄得范动 内，?0.02动位,,已知准确pH动动动动动动动动动动动动得冲液溶液已有市售。 动动动动动动动动动动动动动动动动了改善峰形或塔板数,有在流相中加入酸或碱，如三乙 胺、乙酸等,,如果些添加并非配制冲溶液所采用得动动动动动动动动动动动动动动动动动动动动动 那将些加入到流相中后,需重新滴加酸或碱,使流相达到所需的动动动动动动动动动动动动动动动动动动动动动动动动动动动动动动动动动动pH动。 常用冲液的配制动动动动动动 当冲液被稀或,动动动动动动动动动动动pH也不。其动动动动动pH动动动动动动动动近似恒定不,或当其中 的离解的阳离子，K+、Na+,或离子，动动动动Cl-、Br-,被其它的离子替,表动动动动动动动动动动1—4中 动动动动动动动动动动动动动动出了响的配制方法,平衡阳离子Na+,冲液的最度动动动动动动动动动0.1mol/l,于其它动动动动动 度的冲溶液或阳离子，通常动动动动动动动动动动动动动K+,更好,表中的数据也可供参考。稀的或动动动动动动 的冲溶液,及包含有不同阳离子的溶液,其动动动动动动动动动动动动动动动动动动动pH动会与表中的数据稍有出入。在方法建立 动动动程中,精确的流相pH动动动动往往并不重要,重要的是次新配制的流相的每pH动与第一次 基本相同，最好控制在?0.02范内,。注意,只有当冲溶液的动动动动动动动动动动动动动动动pH在冲解离常数动动动动动动动动 pKIa?1范内,冲溶液才具有有效的冲容量，如乙酸的动动动动动动动动动动动动动动动动动动动动动动动动pKa=4.6,其适宜的pH范 动动3.6—5.6,。 表1 低pH动，25?,磷酸冲溶液的配制方法动动动动动动动动动动 溶液A1:0.1M磷酸。配制精确pH动动动动动动动动动动动冲液,所用磷酸需滴定H3PO4的含量。 溶液A2:0.1M磷酸二溶液,称量动动动动动动动13.8g NaH2PO4?H2O溶解于水中,定容于1L容量瓶 中。 pH A1体，动动ml, A2体，动动ml, pH A1体，动动ml, A2体动 ，ml, 2.0 565 435 2.8 175 825 2.2 455 545 3.0 110 890 2.4 345 655 3.2 55 945 2.6 250 750 表2 不同pH动，25?,醋酸冲溶液的配制方法动动动动动动动动动 溶液A1:0.1M醋酸溶液。6.0g，5.8ml,冰醋酸溶于水中,定容到1L容量瓶中。 溶液A2:0.1M醋酸溶液。动动动动8.2g C2H3O2Na，或13.6g C2H3O2Na?3H2O,溶解于水中定 容到1L容量瓶中。 pH A1体，动动ml, A2体，动动ml, pH A1体，动动ml, A2体，动动ml, 3.6 926 74 4.8 400 600 3.8 880 120 5.0 296 704 4.0 820 180 5.2 210 790 4.2 736 264 5.4 176 824 4.4 610 390 5.6 96 904 4.6 510 490 表3 不同pH动，25?,檬酸冲溶液的配制方法动动动动动动动动动动动动 溶液A1:0.1M动檬酸溶液。21.0g