Differential Thermal Inactivation Enables Simultaneous Quantitation of Ricin and Abrin

Ricin and abrin are highly lethal Type II ribosome-inactivating proteins. They depurinate the same site of the 28S rRNA to inhibit protein synthesis. Consequently, standard molecular-level activity assays used to detect the toxic activity of ricin or abrin do not distinguish between the two in mixed samples without prior physical separation or specially designed substrates. This study proposes a novel, cost-effective method to separately and simultaneously quantify the activities of ricin and abrin in mixtures by exploiting their distinct thermal stabilities. Thermal inactivation was used to demonstrate that heating samples at 80 °C for 5 min maximized the difference in their activities; while ricin retained most of its activity, abrin activity dropped to 20% after thermal treatment. This thermal treatment yielded 4 standard curves-ricin or abrin, thermally treated or not treated-in the 0.3 to 50 µg/mL range. By applying Cramer's rule, the individual concentrations of active ricin and abrin in mixed samples were successfully calculated. However, this method should be used with a method detecting presence of ricin/abrin, to avoid unexpected reactivity due to contaminating RIPs.