Assessment of Copper Bioavailability in Spinach (Spinacia oleracea L) Leaves by Chemical Fractionation

An analytical fractionation scheme has been developed based on water, acetone, chloroform, diethyl ether, ethanol, n-hexane, or methanol extractions to identify free and/or copper bound complexes in spinach samples, sample extracts being analyzed by inductively coupled plasma-mass spectrometry. The total copper contents were determined after digestion of the samples in a microwave-assisted digestion system. Method validation parameters were defined in terms of the detection limits, accuracy, and precision. The limits of detection and quantification for copper were 0.07 and 0.23 mg kg(-1), respectively. The precision in terms of the repeatability and reproducibility, calculated from the relative standard deviations (%RSD), were 3.0 and 4.8 %, respectively. Solvent systems, which simulated gastric and intestinal or gastrointestinal digestion together with n-octanol extraction and activated carbon adsorption, were evaluated with respect to both sample-solvent compatibility and bioavailability.