A gel filtration high-performance liquid chromatographic method for determination of hepatic and renal metallothionein of rat and in comparison with the cadmium-saturation method.

Abstract

Metallothionein (MT) is a low-mol-wt protein. The essential trace elements copper and zinc and the potentially toxic elements, such as cadmium, can induce the synthesis of and bind to MT. The major functions of MT are related to metal metabolism. This paper reported and evaluated a new method for determination of hepatic and renal MT of rat by using high performance liquid chromatography (HPLC) with a Superdex 75 gel filtration column. The tissue was homogenized and centrifuged, then the supernatant was pretreated simply by cadmium saturation and heating before HPLC determination. The MT was completely and clearly separated from other proteins in the rat tissues in a short time, and was quantitated directly as a function of UV absorbance at 250 nm. The recovery both for hepatic and renal MT of rat were exceed 90%. The coefficient of variation was 1.3% for hepatic MT of rat and 1.7% for renal MT of rat. The detection limit was 0.265 microgram for hepatic MT and 0.095 microgram for renal MT of rat. The present method was compared with the traditional cadmium-saturation method for determination of hepatic and renal MT of rat. A good correlation was found in these two methods.