RP-HPLC同时测定贝母药材中5种核苷类化合物
投稿时间:2009-07-03    责任编辑:  点此下载全文
引用本文:张建芝,宋昌慧,陈波,姚守拙.RP-HPLC同时测定贝母药材中5种核苷类化合物[J].中国中药杂志,2010,35(1):67.
DOI:10.4268/cjcmm20100114
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作者中文名作者英文名单位中文名单位英文名E-Mail
张建芝 ZHANG Jianzhi 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 Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research, Ministryof Education, Hunan Normal University, Changsha 410081, China  
宋昌慧 SONG Changhui 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 Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research, Ministryof Education, Hunan Normal University, Changsha 410081, China  
陈波 CHEN Bo 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 Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research, Ministryof Education, Hunan Normal University, Changsha 410081, China dr-chenpo@vip.sina.com 
姚守拙 YAO Shouzhuo 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 Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research, Ministryof Education, Hunan Normal University, Changsha 410081, China  
基金项目: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)
中文摘要: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 Materials XB-C18色谱柱(4.6 mm×250 mm, 5 μm),甲醇-5 mmol·L-1乙酸铵-乙酸缓冲盐(pH 4.30)作为流动相,以1 mL·min-1的流速进行梯度洗脱(洗脱梯度程序为: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);进样量20 μL;检测波长260 nm。 结果 :在0.24~13.60 mg·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>0.998 3;各待测物的日内精密度和日间精密度的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。
中文关键词:高效液相色谱法(HPLC)  核苷类化合物  贝母
 
Simultaneous determination of five nucleotides in BulbusFritillariae 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×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 1 mL·min-1 and monitored at 260 nm, the injection volume was 20 μL. 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. cirrhosaF. 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.
keywords:high performance liquid chromatography  nucleotides  Bulbus Fritillariae
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