title: [Simultaneous determination of five nucleotides in Bulbus Fritillariae by RP-HPLC].
abstract: 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.
authors.affiliation: Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research, Ministry of Education, Hunan Normal University, Changsha 410081, China.
authors.forename: Jianzhi
authors.initials: J
authors.lastname: Zhang
authors.forename: Changhui
authors.initials: C
authors.lastname: Song
authors.forename: Bo
authors.initials: B
authors.lastname: Chen
authors.forename: Shouzhuo
authors.initials: S
authors.lastname: Yao
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D031410:Fritillaria / Q000737:chemistry*; D009711:Nucleotides / Q000737:chemistry*; D015203:Reproducibility of Results
chemical_list: D004365:Drugs, Chinese Herbal; D009711:Nucleotides