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Using high-performance liquid chromatography (HPLC), a quantitative method was developed to control the quality of ribonucleic acid for injection II (RA-II), which is used as an anti-cancer drug clinically in China. Using nuclease enzyme and under optimal hydrolysis conditions, the unstable RNA was hydrolyzed into stable nucleosides and nucleotides, which were easily detected by HPLC with diode array detection. Furthermore, by analyzing HPLC chromatograms of 10 batches of samples, six common peaks, namely, cytosine nucleotide, uracil nucleotide, adenine nucleotide, guanine nucleotide, guanosine and adenine, were identified by a HLPC coupled with electrospray ionization mass spectrometry. The similarity, the relative retention time and the relative peak area of each common peak, relative to the reference peak, were calculated to obtain the HPLC fingerprints, and their values were within the scope of the provisions. More importantly, all six components were simultaneously determined. Overall, the developed method in this investigation proved to be a useful tool for monitoring the quality of RA-II in terms of their batch variations and the raw material sources.

Fingerprint Analysis and Multi-Component Determination of Ribonucleic Acid for Injection II Recipe by HPLC–DAD and LC–ESI-MS Methods