[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.
Zhang
Jianzhi
J
Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research, Ministry of Education, Hunan Normal University, Changsha 410081, China.
Song
Changhui
C
Chen
Bo
B
Yao
Shouzhuo
S
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
D004365:Drugs, Chinese Herbal; D009711:Nucleotides