Van Beek H, Baars A J
Central Veterinary Institute, Department of Analytical Chemistry & Toxicology, Lelystad, The Netherlands.
J Chromatogr. 1988 Jun 17;442:345-52. doi: 10.1016/s0021-9673(00)94482-0.
Metallothioneins (MTs) were separated and quantitated by reversed-phase high-performance liquid chromatography (RP-HPLC), in direct combination with atomic absorption spectrometry (AAS) for quantitation of the metal contents in MTs. MTs were eluted from an RP-8 column with a gradient of Tris buffer pH 7.0 and methanol, and were detected by UV absorbance (220 nm). Commercially available purified MTs from horse kidney and rabbit liver were analyzed for purity and metal composition. One lot of horse kidney yielded only 50% of the estimated value. In some cases, the certified metal content differed considerably from the values found. The method was also tested with rat liver and fetal bovine liver. The metal contents found in MTs by use of graphite-furnace atomic absorption spectrometry corresponded well with the values found by RP-HPLC-AAS. Molar ratios of cadmium, zinc and copper were calculated in MT-1, MT-2 and total MT.
通过反相高效液相色谱法（RP-HPLC）分离并定量金属硫蛋白（MTs），并直接结合原子吸收光谱法（AAS）来定量MTs中的金属含量。MTs用pH 7.0的Tris缓冲液和甲醇梯度从RP-8柱上洗脱下来，并通过紫外吸光度（220 nm）进行检测。对市售的来自马肾和兔肝的纯化MTs进行纯度和金属组成分析。一批马肾MTs的产量仅为估计值的50%。在某些情况下，认证的金属含量与实际测得的值有很大差异。该方法也用大鼠肝脏和胎牛肝脏进行了测试。使用石墨炉原子吸收光谱法在MTs中测得的金属含量与RP-HPLC-AAS法测得的值非常吻合。计算了MT-1、MT-2和总MT中镉、锌和铜的摩尔比。
