|Budget Amount *help
¥3,200,000 (Direct Cost : ¥3,200,000)
Fiscal Year 2005 : ¥700,000 (Direct Cost : ¥700,000)
Fiscal Year 2004 : ¥700,000 (Direct Cost : ¥700,000)
Fiscal Year 2003 : ¥1,800,000 (Direct Cost : ¥1,800,000)
Radicals and paramagnetic intermediates are formed when medicinal and poisonous substances are affecting actively. In our present series of study from 2003 to 2005, simple, rapid and sensitive methods have been developed for the determination of these radicals and paramagnetic metal-chelate complexes using electron spin resonance (ESR).
1.Klear is an adulterant that conceal tetrahydrocannabinols and morphines present in urine. We produced NO radical from nitrite, the active component of Klear, and trapped NO with Fe-diethyldithiocarbamate(DDC) complex. Without sample pretretment, nitrite in plasma, urine or saliva was quantitated as NO-Fe-(DDC)_2 at the detection limit of 50 pg within 5 min.
2.Molybdenum (Mo) is an essential trace element for human and is contained 140 ng/kg. When it works as cofactor of enzymes it becomes paramagnetic Mo^<5+> and detectable by ESR, although Mo^<5+> was unstable in other reports. We have succeeded in producing a new stable Mo^<5+> complex and detected it
using ESR. Mo in 10 μl of urine could be quatitated at the detection limit of 50 pg within 5 min.
3.Urine Luck is an adulterant that conceal tetrahydrocannabinols and opiates, and its active component is chromate Cr^<6+>. Cr^<3+> does not work for that purpose. Cr^<6+> is known to induce cancer while Cr^<3+> is not. In this respect, speciation of Cr^<6+> and Cr^<3+> is required. We produced paramagnetic Cr^<6+>(DDC)_4 complex from only Cr^<6+> and quantitated Cr^<6+> sensitively.
4.To measure minute amounts of manganese (Mn), we applied ESR method because Mn^<2+> is paramagnetic. Tissue Mn levels were not affected by either dosing with paraquat at 125 ppm or restricting magnesium at half of recommended. When both treatments were applied together, Mn levels were lowered significantly from those of the control in liver, kidney and heart.
5.Thiocyanate (SCN) is the main metabolite of cyanide, a famous poison. The procedure for the determination of SCN was based on the formation of a paramagnetic complex Cu(SCN)_2(4-methylpyridine)_2 in benzene, followed by its detection using ESR. According to the present method, SCN at concentrations from 5μM to 2 mM in 0.5 ml solution could be determined. The recoveries were more than 95% from blood, plasma and urine and beverages.
6.Sodium azide (N_3) is used in industry as an explosive in automobile safety bags and an antifungal for diagnostic reagents. In 1998, several poisonous cases of azide occurred in Japan. The procedure was based on the formation of a paramagnetic complex Cu(N_3)_2(4-methylpyridine)_x in benzene, followed by its detection using ESR. Less