Amperometric Titration Largely Overestimates Chloride Concentrations in Chloroplast Extracts

Determination of chloride contents in aqueously isolated chloroplasts from spinach by amperometric titration indicated chloride concentrations of 60 to 100 mmol 1-1 (based on a chloroplast volume of 25 μl mg-1 chlorophyll). However, when analyzed by anion chromatography, chloride contents in chloroplasts were much lower (1-8 mmol 1-1). In leaf extracts chloride concentrations obtained with both methods were rather similar. Boiling of chloroplast extracts prior to chloride titration reduced indicated chloride contents by a factor of three or four, but did not change results obtained with anion chromatography. It is concluded that chloroplasts contain large amounts of silver complexing agents giving rise to a drastic overestima­tion of chloroplast chloride concentrations when measured by amperometric titration. Boiling and centrifugation of extracts apparently precipitates these compounds only insufficiently.