RP-HPLC同时测定贝母药材中5种核苷类化合物  

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

作　　者：张建芝[1] 宋昌慧[1] 陈波[1] 姚守拙[1] 

机构地区：[1]湖南师范大学化学生物学及中药分析教育部重点实验室,湖南长沙410081

出　　处：《中国中药杂志》2010年第1期67-70,共4页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金(0875028)项目;国家重点基础研究发展计划(973)项目(2006CB504701)

摘　　要：目的：建立一种同时测定贝母药材中核5种苷类化合物尿苷、腺嘌呤、鸟苷、胸苷、腺苷的高效液相色谱法。方法：采用Welch MaterialsXB—C18色谱柱（4．6mm×250mm，5μm），甲醇-5mmol·L-1乙酸铵-乙酸缓冲盐（pH4．30）作为流动相，以1mL·min-1的流速进行梯度洗脱（洗脱梯度程序为：0～10min，0％～1％A；10～20min，1％～5％A：20～25min，5％A；25～35min，5％～30％A；35～37min，30％～0％A；37～40min，0％A）；进样量20μL；检测波长260nm。结果：在0．24～13．60mg·L-1，各核苷类化合物的响应峰面积与其相应的浓度呈现良好的线性关系，r〉0．9983；各待测物的日内精密度和日问精密度的RSD均小于2．1％；重复性良好（RSD〈5．5％）；回收率范围在93．55％～101．88％，RSD〈3．0％。结论：不同种贝母药材中核苷类化合物的含量大致顺序为湖北贝母〉浙贝〉川贝=平贝；本方法简便、可靠、准确，可用于贝母药材中核苷类化合物的含量测定，为全面开发利用贝母药材提供进一步依据。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-C,s column （4. 6 mm ×250 mm, 5 μm） was used and the chromatographic separation was achieved using 5 mmoL · 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 l mL ·min -1 and monitored at 260 nm, the injection volume was 20 p,L. The peak areas of nucleotides and the concentrations showed a good linear relation ranged from 0. 24 to 13.60 mg· L-1 , r 〉0. 998 3. 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 = F. ussuriensis. The method is simple, convenient and accurate. It can be used for the de- termination of nucleosides and supplying evidence for exploiting and applying of Bulbus Fritillariae.

关 键 词：高效液相色谱法(HPLC) 核苷类化合物 贝母 

分 类 号：R284.1[医药卫生—中药学]