Reactive column HPLC: rapid analysis on RPC-5 of oligoadenylates that differ at the 3'-terminus.

Oligoadenylates can be analyzed according to the type of 3′-terminus (A_nA, A_nAp, and A_nA > p, oligoadenylates that have at the 3′-terminus no phosphate, a 2′(3′)-monophosphate, and a 2′,3′-cyclic phosphate respectively) by hplc on RPC-5 support using a novel dual-column technique. The first column separates A_nAp plus A_nA > p from A_nA, and at the start of the second column a layer of bacterial alkaline phosphatase enzyme converts the A_nAp into A_nA. Hence this A_nA emerges separately from the original A_nA and from the A_nA > p. The technique can be used to analyze a three-component mixture for a single chain length or a mixture of A_nAp and A_nA > p of mixed chain lengths (n = 3 to 7). The presence of poly(U) does not interfere with the analysis.