hplc_dad_elsd法同时测定威灵仙药材中齐墩果酸和常春藤皂苷元的含量[1] - 副本（Simultaneous determination of oleanolic acid and hederagenin content of [1] - hplc_dad_elsd copy of Clematis）

hplc_dad_elsd法同时测定威灵仙药材中齐墩果酸和常春藤皂苷元的含量[1] - 副本（Simultaneous determination of oleanolic acid and hederagenin content of [1] - hplc_dad_elsd copy of Clematis） hplc_dad_elsd法同时测定威灵仙药材中齐墩果酸和常春藤皂苷元 的含量[1] - 副本(Simultaneous determination of oleanolic acid and hederagenin content of [1] - hplc_dad_elsd copy of Clematis) Simultaneous determination of Clematis herb by HPLC-DAD-ELSD Content of oleanolic acid and ivy saponins Qi Shuai, Hu Hua (Wuhan people's Liberation Army 61 hospital first 430063, Hubei Wuhan 430010) Abstract: Objective: to establish the evaporative light scattering detector (ELSD) and diode array detector (DAD) determination of oleanolic acid in Clematis HPLC method for the analysis of ivy saponins. Methods chromatographic column: M erck L ichro CA RT C18 column (4 mm @ 250 mm), 5 Lm); mobile phase: methanol water glacial acetic acid three ethylamine (volume ratio 87B13B0. 04B0. 02); flow rate: 1 mL# m in- 1; column temperature: 30 e; The detection wavelength of DAD is 210 nm; the temperature of ELSD evaporation tube is 80 e, and the gas pressure is 172.375 kPa. Results the saponins of ivy were homogeneous and homogeneous The linear range of oleanolic acid was 1. 43~ 14.3 Lg and 2. 1~ 21 Lg, respectively. The results of the determination of DAD and oleanolic acid were similar to those of oleanolic acid and oleanolic acid The recoveries were 98.02% and 98.94%, RSD were 2.61% and 2.34%, respectively, and the recoveries in ELSD detection were respectively For 98.72% and 100.03%, RSD were 2.51% and 2.22%, respectively. Conclusion the method is simple, reproducible and specific, for the improvement of wheeling The scientific basis for the quality control standard of Xian Xian herbs. Keywords: Weilingxian; oleanolic acid; hederagenin; HPLC; ELSD; diode array detector CLC number: R 284.1 document identification code: A do: I 10.3969 /.J issn. 1006- 8783. 2010. 1.020 Article number: 1006-8783 (2010) 01- 0067- 03 Determination of oleanolic acid and hederagenin from Rhizoma Clemat ID is by HPLCDAD- ELSD Q I Shua, iHU Hua (D epartment of Pharmacy, 161st Cen tra lH ospita l of PLA, Wuhan, H ubei 430010, China) Abstract: Objective To determ ine the con ten Rhizoma t of oleanolic acid and hederagen in from Rad IX Clematid is by RP-HPLC-ELSD-DAD at the same TMI E. Merck MethodsA L ICH rocartC18 column (4. 0mm) @ 250mm, 5 Lm) was used. The mob ile phasewas consisted ofmethano-lwater-acetic acid-tr iethyl am ine (87B13B0. 04B0. 02) w ith a flow-rate of 1 mL# m in- 1, and the column temperature was 30e. The DAD detection wavelengthw as 210 nm; The ELSD Dr if-t tube temperaturewas 80e, and gas p ressure was 172.375 kPa. Results The linear ranges of hederagen in and oleanolic acidw ere 1. 43- 14.3 Lg and 2. 1- 21 Lg respectively. The average recovery of hederagenin and oleanolic acid were 98.02% (RSD =) 2161%), 98.94% (RSD = 2134%) determ ined by DAD, and 98.72% (RSD = 2.51%), 100.03% (RSD) = 2.22%) determ ined by ELSD. Conclusion It. s a reliab of Le and accuratemethod for the quality con trol R. Clema tidis. Key words: Rhizoma Clematid is; oleanolic acid; hederagen in; HPLC; ELSD; DAD Received date: 2009- 11- 02 Brief introduction: Qi Shuai (1976-), male, in charge of pharmacists, Em a i:l Qishua i0101 @ yahoo. cn. Clematis from Ranunculaceae Clematis chinensis O sbeck, Clematis C lematis hexap etala Pa L.L or northeast China Dried roots and rhizomes of Clematis C lematismanshurica Rupr.i, For the Chinese Pharmacopoeia 2005 edition carried varieties (1). Expelling wind and removing dampness, Tongluozhitong achievements, for the treatment of rheumatic arthralgia, numbness of limbs, muscles Julian, flexion and extension, stuck throat. Chinese Pharmacopoeia only carries on this product There was no quantitative control for the TLC identification of oleanolic acid as the reference substance Item. [2]] the main effective component of this product is three terpene saponins The aglycone is oleanolic acid and ivy saponin. It is often used now The content of oleanolic acid was determined by TLC, 3], HPLC-UV[2 and 4] The detection wavelength was 220 or 215 nm; the TLC method was more stringent, The reproducibility was poor, while oleanolic acid belongs to the determination of oleanolic acid by HPLC-UV Journal of Guangdong Pharmaceutical University Journal of Guangdong Pharmaceu tical C ollege Feb. 2010, 26 (1) The three terpenoids of the five rings are absorbed at the end of ultraviolet 204 NM The influence of mobile phase is large, the separation is not ideal, and the detection sensitivity is low. It is also a five ring three terpenoids, but it has not been collected yet Determination of the content by HPLC-UV or HPLC-ELSD method Avenue。 The HPLC-ELSD-DAD method was used for the first time The contents of oleanolic acid and hederagenin in Clematis Chinensis Osbeck,. A comparative study on determination results of 2 kinds of detectors, for welling elixir Material quality control method provides experimental basis. 1 instrument and reagent W aters 2695 high performance liquid chromatography (2996 DAD and) 2420 ELSD detector); Empow Er chemical workstation; Sartorius Electronic analytical balance (Germany, d = 0.01 mg). Methanol is chromatography Pure, other reagents are analytical pure. Oleanolic acid reference substance (Chinese Medicine) Institute of biological products inspection, lot number 110709-200505), Ivy Reference substance of diosgenin (China Institute for drug and biological products, 111733- 200502). Clematis herbs purchased from Wuhan medicine company, tsunetake Hu Hua, chief pharmacist of the first 61 Hospital of the Chinese people's Liberation Army, was identified as Buttercup Dried root and rhizome of plant chinensis O sbeck weilingxian. 2 methods and results 2.1 chromatographic conditions and system suitability test column: Merck L ichrocart C18 column (4 mm @ 250 mm, 5 Lm), add pre Column: mobile phase: methanol water glacial acetic acid - three ethylamine (volume ratio 87B13B) 0. 04B0. 02); flow rate: 1 mL# m in- 1; column temperature: 30 E; violet The detection wavelength was 210 nm; the evaporative light scattering detector was used The evaporation tube temperature is 80 E and the gas pressure is 172.375 kPa. branch Do not accurately absorb the reference solution, the test solution 10 LL each, respectively Inject liquid chromatograph, record chromatogram, see figure 1. A. reference solution; B. solution solution; 1. Ivy saponin; 2. oleanolic acid Figure 1 H PLC map Weilingxian Figure 1 HPLC Chrom atogram s of Rhizoma C LEM atidis 2.2 standard solution preparation were accurately weighed olive The appropriate amount of acid and ivy saponins were prepared by adding methanol The concentration of ursolic acid was 2.100 mg# mL- 1, and the content of saponins in Ivy was the highest Reference substance solution with mass concentration of 1. 430mg# mL- 1. Pro When the same amount of reference solution was mixed with the same amount of precision, it was mixed pair Photographic solution. Preparation of Clematis medicinal powder of 2.3 tested solution (4 No.) 2 g, precision methanol, 50 mL, according to the 2005 Edition Chinese / Akebia assay Pharmacopoeia under item 0 [1] for sample preparation Preparation method, that is. 2.4 linear relationship was investigated for precision absorption of mixed reference solution 2, 5, 8, 10, 15, 20 LL, injected liquid chromatograph. DAD test junction The Yifeng area (Y) of sample quality control samples (X) were linear The regression equation was: Ivy saponin Y = 953094 X - 4227.9, r = 0.9999; 齐墩果酸y = 816930x - 326856, r = 0. 进样量分别在1 999 9. 43 ~ 14. 3 lg及2. 1 ~ 21. lg 范围内线性关系良好 0. elsd 检测 工程第三方检测合同工程防雷检测合同植筋拉拔检测方案传感器技术课后答案检测机构通用要求培训 结果以峰面积对数 (y) 对对照品进样质 量的对数 (x) 进行回归, 得回归方程分别为: 常春藤 皂苷元y = 1. 4438 x + 5. 8851, r = 0. 齐墩果 999 1; 酸y = 1. 4438 x + 6. 1566, r = 0. 进样量分别 999 2; 在1. 43 ~ 14. 3 lg及2. 1 ~ 21. 0 lg范围内线性关 系良好. 2. 5 精密度试验 精密吸取对照品溶液10 yy, 连 广东药学院学报 第26卷 68 续进样6次, 测定峰面积, 结果dad 检测常春藤皂 苷元和齐墩果酸峰面积的rsd 分别为1. 06% 、 0. elsd检测常春藤皂苷元和齐墩果酸峰面积 32%; 的rsd 分别为3. 49% 、4. 01%. 2. 6 重复性试验 精密称取同一批样品6 份, 同 / 2. 30项下方法平行制备, 测定, 结果dad 检测常 春藤皂苷元和齐墩果酸含量的rsd 分别为0. 82% 、 0. elsd检测常春藤皂苷元和齐墩果酸含量的 41%; 分别为2 rsd. 97% 、3. 93%. 2. 7 稳定性试验 取供试品溶液分别于0、1、2、4、 8 h进样10 yy, 测定峰面积, 常春藤皂苷元和齐墩 果酸的rsd 分别为1. 15% 、0. elsd检测常春 47%; 藤皂苷元和齐墩果酸的rsd 分别为3164% 、 4. 26%.表明在8h内溶液基本稳定. 2. 8 加样回收试验 取已知含量的威灵仙药材粉 末 (过4号筛) 1 g, 共6份, 精密称定, 分别精密加入 常春藤皂苷元对照品溶液1 ml 与齐墩果酸对照品 溶液3ml, 同 / 2. 30项下方法处理, 测定, 计算加样 回收率.结果常春藤皂苷元和齐墩果酸在以dad 检测时的加样回收率分别为98. 02% 、98. 94%, rsd 分别为2. 61% 、2. 34% (n = 6), 在以elsd 检测时 的加样回收率分别为98. 72% 、100. 03%, rsd 分别 为2. 51% 、2. 22% (n = 6). 2. 9 样品测定 取不同批号样品, 制备供试品溶 液, 精密吸取10 yy, 注入高效液相色谱仪, 测定. 结果常春藤皂苷元在以dad 检测时的平均含量为 0. 186%, 在以elsd检测时的平均含量为0. 143%; 齐墩果酸在以dad检测时的平均含量为0. 791% 在以elsd 检测时的平均含量为0. 627% (n = 3). 3 讨论 3. 1 波长选择 分析系统适应性试验时dad 检测 器收集的信息可知, 齐墩果酸和常春藤皂苷元的最 大吸收波长为 (210? 2) nm, 故选择210 nm 作为 uv测定波长. 3. 2 二极管阵列检测器 (dad) 是测定具有紫外吸 收的化学成分含量的常用检测器, 而蒸发光散射检 测器 (elsd) 广泛用于医药、化工等多种领域 [5, 6]. 分析威灵仙药材的dad 及elsd图谱可知, dad检 测因在近紫外区, 干扰较大, 峰形不好; 而elsd 图 谱几无干扰, 峰形好, 故宜使用elsd对威灵仙中齐 墩果酸和常春藤皂苷元进行含量测定. 3. 3 采用hplc - dad - elsd 法同时测定威灵仙药 材中齐墩果酸及常春藤皂苷元的含量, 方法简便、重 现性好、专属性强, 为完善威灵仙药材质量控制标准 提供了科学依据. 参考文献: 国家药典委员会 [1]. 中华人民共和国药典: 2005年版一部 [m]. 北京: 化学工业出版社, 2005: 175, 43. [2] 杨林, 裴瑾, 万德光. Containing three terpene aglycones of oleanolic acid in Clematis Chinensis Osbeck Determination of the amount of H PLC [J]. natural products research and development, 2006, 18 (2): 313- 315. 3] [Xiong Huilin, Zheng Maoxin. Determination of oleanolic acid in Radix Clematidis by TLC scanning Content [J]. Chinese Journal of traditional Chinese medicine, 2007, 32 (4): 357- 359. [4] Xie Le, Ma Xiaoli, Liu Lifang, et al. Determination of Weilingxian from different H PLC Oleanolic acid content [J]. Chinese patent medicine, 2009, 31 (1): 100- 103. [5] LI W K, JOHN F. Dete RM ina tion o f 24 (R) -pseudog inseno Side F11 in North Am Er Ican g inseng using H ighper Fo rm ance liqu ID chrom atography w ith evapo rative light SCA tter ing detection [J]. J P harm B IOM ed Anal, 2001, 25 (2): 257- 265. [6] BONNIE A, KRISHNA K, M ITCHELL A. Rap ID determ ination Of Artem isinin and re lated ana logues using H igh-perform ance Liquid chrom atography and an evapo RA tive light scatter ing De tector[J]. J Chroma togr A, 1999, 730 (1): 71- 80. (editor in chief: Chen Xiang) Studies show red fruits and vegetables reduce heart disease risk Experts at the University of Michigan cardiovascular disease research center found that the pigment in red fruits and vegetables such as cherry and tomato can be greatly reduced and moved The risk of hardening related diseases, therefore, helps reduce the risk of heart disease. A recently completed animal study at University of Michigan found that cholesterol and triglyceride levels were greatly reduced in Cherry fed animals, So that the risk of heart disease is greatly reduced. From February 4th to February 10th, it was designated by the American Heart Association / national red food week 0 to encourage people to eat more Red fruits and vegetables deepen understanding of heart health benefits of red fruits and vegetables. (Library feeds) First Qi Shuai, et al. HPLC-DAD-ELSD method for simultaneous determination of oleanolic acid and hederagenin Clematis in 69