A new two-step synthetic pathway developed for the transformation of arsenic trioxide [iAs (III); As2O3] into arsenobetaine (AB; Me3As+ CH2CO2−) involves treatment of iAs (III) with native B12 or biomimetic B12 in the presence of glutathione (GSH) to give TMAO with a high selectivity and a high conversion rate; subsequent treatment of TMAO with iodoacetic acid in the presence of GSH gives arsenobetaine.