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  • Title: A simplified method to determine [99mTc]TRODAT-1 in human plasma.
    Author: Mu M, Kung MP, Plössl K, Acton PD, Mozley PD, Kung HF.
    Journal: Nucl Med Biol; 1999 Oct; 26(7):821-5. PubMed ID: 10628563.
    Abstract:
    [99mTc]TRODAT-1 is a useful imaging agent in evaluating changes in presynaptic dopamine transporters (DAT) for Parkinson's disease and other central nervous system (CNS) neurodegenerative diseases, for which a reduction of dopamine neurons is indicated. As part of an effort to establish a quantitative single-photon emission tomography (SPECT) procedure for imaging CNS DAT, measurement of nonmetabolized [99mTc]TRODAT-1 in human plasma was investigated. After an intravenous injection of [99mTc]TRODAT-1, there are three possible radioactive components in human plasma: hydrophilic compounds (pertechnetate, etc.), lipophilic metabolite(s), and unchanged [99mTc]TRODAT-1. Based on the differences in lipophilicity of [99mTc]TRODAT-1 and its lipophilic metabolite [99mTc]BAT, a new quantitative method for measuring [99mTc]TRODAT-1 with a simple solvent extraction method was developed. Various organic solvents or mixtures of solvents were tested, among which cyclohexane gave the best extraction yield for [99mTc]TRODAT-1 (76.06 +/- 3.32%) with a low extraction for [99mTc]BAT (2.43 +/- 0.82%). Extractions of [99mTc]TRODAT-1 and [99mTc]BAT mixtures in different predetermined ratios to simulate the actual human plasma samples with cyclohexane from phosphate buffer (5 mM, pH 8.0) were evaluated. The experimentally obtained ratios were in good agreement with the theoretical ratios. To investigate further the possibility of replacing the previously established high performance liquid chromatography (HPLC) method with the new solvent extraction method for the clinical application, both HPLC and extraction methods were used side by side to determine the unchanged [99mTc]TRODAT-1 in human plasma samples during [99mTc]TRODAT-1/SPECT imaging studies. The results from four human subjects showed that both methods consistently produced similar values for unchanged [99mTc]TRODAT-1 in the plasma samples. This improved solvent extraction method provides an easy and reliable technique to quantify unchanged [99mTc]TRODAT-1 in human plasma, thus making the clinical application of this agent readily available for quantitation of the DAT binding sites in the brain by SPECT imaging.
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