[Simultaneous determination of five nucleotides in Bulbus Fritillariae by RP-HPLC]

A high-performance liquid chromatography method was developed for determination of five nucleotides in Bulbus Fritillariae. The five nucleotides were uridine, adenine, guanosine, thymidine, adenosine, respectively. A Welch materials XB-C18 column (4.6 mm x 250 mm, 5 microm) was used and the chromatographic separation was achieved using 5 mmoL x L(-1) ammonium acetate-acetic acid buffer solution (pH 4.30, B) and methanol (A) as mobile phases, the gradient elution program: 0-10 min, 0%-1% A, 10-20 min, 1%-5% A, 20-25 min, 5% A, 25-35 min, 5%-30% A, 35-37 min, 30%-0% A, 37-40 min, 0% A with a flow rate of 1 mL x min(-1) and monitored at 260 nm, the injection volume was 20 microL. The peak areas of nucleotides and the concentrations showed a good linear relation ranged from 0.24 to 13.60 mg x L(-1), r > 0.9983. The intra- and inter-day pecision results were adequate with the RSDs of 2.1% or below. The repeatability was good and the RSD were smaller than 5.5%. The recoveries of nucleosides were in the range of 93.55% and 101.9%, RSD < 3.0%; The order of nucleotides contents in different Bulbus Fritillariae was F. hupehensis > F. thunberqii > F. cirrhosa approximately F. ussuriensis. The method is simple, convenient and accurate. It can be used for the determination of nucleosides and supplying evidence for exploiting and applying of Bulbus Fritillariae.