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We developed a method for measuring blood coagulation using superparamagneticiron oxide nanoparticles (SPIONs) and an alternating magnetic field (AMF). The3rd and 5th harmonic signals from SPIONs mixed with blood induced by AMF weredetected using a gradiometer coil. Blood coagulation was induced artificiallyby adding CaCl2 solution to whole blood of sheep at various temperatures andhematocrits. We calculated the coagulation rate (k) and normalized signalintensity at infinite time (Sinf) by fitting the time course of the normalized3rd harmonic signal to S(t)=(1-Sinf)exp(-kt)+Sinf. The k values increasedsignificantly with increasing temperature and decreased significantly withincreasing hematocrit. The Sinf values decreased significantly with increasingtemperature and tended to increase with increasing hematocrit. Bloodanticoagulation was induced by adding heparin to the whole blood sampled frommice. There were significant differences in both the 3rd and 5th harmonicsignals between groups with and without heparin at 25 min or more after addingheparin. We also calculated the 3rd and 5th harmonic signals for viscositiesranging from 0.001 to 1 kg/m/s, with an assumption that the magnetization andparticle size distribution of SPIONs obey the Langevin theory of paramagnetismand log-normal distribution, respectively. The 3rd and 5th harmonic signalsincreased slowly with increasing viscosity and had peaks at approximately 0.015and 0.025 kg/m/s, respectively. After these peaks, they decreased monotonicallywith increasing viscosity. These results confirm the rationale of our method.In conclusion, our method will be useful for measuring blood coagulation andanticoagulation and for studying their processes.

Development of a method for measuring blood coagulation using superparamagnetic iron oxide nanoparticles and an alternating magnetic field