菊花中绿原酸的含量测定

菊花中绿原酸的含量测定3李宗　陈在敏　廖雷生33　林少山33　(福建省药品检验所　福州350001)摘要　目的:测定菊花中绿原酸的含量。

方法:HP LC法,色谱柱为Nova2Pak C18柱(4μm,416mm×250mm),流动相为甲醇20.1mol・L-1磷酸盐缓冲液(pH2.65)30∶70,检测波长为328nm。

结果:20批菊花样品的绿原酸含量,以干燥品计为0.060%～0.467%。

结论:绿原酸可作为菊花的质量控制指标成分。

关键词　菊花　绿原酸　含量测定 菊花为菊科植物菊Chrysanthemum mori2 folium Ramat.的干燥头状花序,含挥发油、菊甙、氨基酸、黄酮类及绿原酸等多种成分[1]。

菊花为常用中药,使用量大,产地广,加工方法各异,虽有关于菊花挥发油、木犀草素的含量测定的报道[2],但尚无稳定、可靠的质量控制方法。

绿原酸已广泛作为金银花及其制剂的质控指标,测定的方法有分光光度法[3]、薄层扫描法[4]、HP LC法[5]等,但关于菊花中绿原酸的含量测定尚未见报道,因此,我们建立了HP LC法测定菊花中绿原酸的含量,报告如下。

1　仪器和材料111　仪器和试药　P E Integral4000高效液相色谱仪(带二极管阵列检测器),美国P E公司产品;Nova2Pak C18色谱柱(4μm,416mm×250mm),大连依利特公司产品;绿原酸对照品由中国药品生物制品检定所提供。

112　材料　菊花药材经鉴定均为菊科植物菊花Chrysanthemum morifolium Ramat1的头状花序。

鉴定人李宗。

2　方法和结果211　色谱条件　使用大连依利特公司的Nova2 Pak C18色谱柱(4μm,416mm×250mm)。

流动相的配制:取磷酸二氢钠适量制成0.1mol・L-1的溶液,按2ml・L-1的比例加入磷酸适量,调整p H为2.65,超滤后与甲醇按70∶30的比例混合。

检测波长为328nm,柱温为室温,流速0. 8ml・min21,进样量10μl。

2.2　对照品溶液的制备及线性关系考察　精密称取绿原酸对照品适量,加水配制成每1ml 约含0.1mg绿原酸的对照品溶液。

取不同体积的对照品溶液注入液相色谱仪,测定峰面积值,以进样量为横坐标(X),峰面积为纵坐标(Y),得回归方程:Y=277382+24027460X,r =0.9999。

绿原酸浓度在0.2072～2.072μg内与峰面积呈良好的线性关系。

213　供试品溶液的制备　取菊花粉末(过1号筛)1g,精密称定,置干燥烧瓶中,(同时按《中国药典》1995年版一部附录水分测定一法测定水分),准确加入50.0ml甲醇,称定重量,水浴加热回流提取2h,冷却,用甲醇调至原重,摇匀,滤过,精密吸取续滤液10ml于烧杯中,水浴蒸干,残渣加入5ml氯仿浸泡3min,弃去氯仿液,残渣挥干氯仿,准确加入5.0ml蒸馏水使绿原酸充分溶解,溶液用微孔滤膜超滤,滤液作为供试品溶液备用。

214　精密度考察　取同一浓度对照品溶液,连续进样5次,进样量均为10μl,测得峰面积值为11898229,12303678,12032046,11843952, 11740327,RSD=1182%。

215　准确度考察　取同一批样品,按213项下方法制备6份供试品溶液,测得绿原酸含量分别为0.236%,0.238%,0.239%,0.250%, 0.225%,0.232%,平均值为0.237%,RSD= 3.5%。

3福建省卫生厅科研基金资助项目　No.Q972233福建中医学院中药系97届实习生216　回收率考察　精密称取6份菊花样品,分别准确加入一定量的绿原酸对照品溶液,按213项下方法制备供试品溶液,测定并计算回收率。

结果加样提取回收率分别为104.9%, 10018%,10019%,9516%,9615%,9717%平均回收率为9914%,RSD=315%。

217　样品测定结果　先后测定了20批菊花样品的绿原酸含量和水分含量,各批菊花的含量测定结果如表1及图1。

表1　不同产地菊花绿原酸及水分含量测定编号商品名产地水分%绿原酸%I II1杭菊　福建福州16.60.1570.1882杭菊　福建福州16.40.1860.2223杭菊　福建福州17.80.1940.2364杭菊　福建福州19.60.2150.2675杭菊　福建福州14.20.2210.2586杭菊　浙江杭州14.80.2330.2737杭菊　福建福州12.40.1220.1368杭菊　浙江桐乡13.00.2520.2849贡菊　安徽 13.00.2610.32210贡菊　安徽黄山17.10.3880.46711滁菊　安徽滁县21.10.3060.38212亳菊　安徽亳州12.90.1020.11713黄菊　云南昆明13.40.2290.26414菊花　四川 7.60.3550.38415菊花　河南禹州8.90.2430.26716菊花　河南新乡7.10.0810.08717菊花　广西南宁12.80.2800.32018祁菊　河北安国14.80.2440.28619甘菊　福建福州13.60.1400.16220甘菊　福建福州12.40.0530.060 注:样品均为市售品,拉丁学名均为C1morifolium;I为干燥前商品,II以干燥品计结果显示菊花中绿原酸的含量在0.053%～0.388%,以干燥品计则为0.060%～0.467%,菊花中绿原酸含量因产地和商品规格不同而有一定差异,含水量亦有较大差异。

3　讨论311　比较了多种流动相对绿原酸的分离效果,结果表明用0.1mol・L-1,pH2.65的磷酸盐缓冲液与甲醇混合洗脱,可较好地抑制色谱峰的拖尾现象,使用了4μm粒径的Nova2Pak C18色谱柱,使绿原酸与杂质得到良好的分离。

312　由于菊花产地多而分散,加工方法各异,如阴干法(亳菊)、硫黄蒸晒法(滁菊)、烘干法(贡菊)、蒸透法(杭菊)等[6]。

从实验结果来看,贡菊含量最高,滁菊、川菊次之,杭菊普遍较低,甘菊仅为0.060%。

提示加工方法对绿原酸的含量有一定的影响。

有必要就加工方法对菊花的影响作进一步的考察。

图1　菊花(滁菊)供试液HP LC色谱图C1绿原酸313　由于菊花成分复杂,木犀草素含量仅0.031%～0.066%,挥发油含量仅为0.1074%～～0.2144%[2]。

其他有效成分的含量分析尚未见报道,而绿原酸的药理活性与菊花的功用是相符的[7],根据所收集样品的性状分析,绿原酸含量相对较低者,香气一般偏弱,花序多破碎,色泽偏暗,多呈浅棕色或浅棕褐色。

可能为加工方法不佳或储存条件不良所致,因此用绿原酸含量作为菊花的质量控制指标是可行的。

4　参考文献1　黄泰康.常用中药成分与药理手册1北京:中国医药科技出版社,1994.15652　陈发奎.常用中草药有效成分含量测定1北京:人民卫生出版社,1997.6063　张广强,粱生旺,姜玉凤,等.三波长分光光度法测定清热解毒注射剂中绿原酸的含量.中国中药杂志,1990,15(8): 334　刘伟,杜天信,郑松波,等.小儿清热解毒口服液中绿原酸的薄层扫描测定.中成药,1989,11(8):145　潘扬,马国祥,郭立玮,等.RP.HP LC法测定紫金解毒饮中绿原酸的含量.中国药科大学学报,1995,26(4):2176　徐国均,徐珞珊,何宏贤,等.中国药材学(下册).北京:中国医药科技出版社,1996.9937　季宇彬.中药有效成分药理与应用.哈尔滨:黑龙江科学技术出版社,1995.110致谢　部分菊花样品由浙江、安徽、昆明等地药检所和其它有关单位提供。

1998208226收稿ABSTRACT OF ORIGINAL ARTICL ESPhamacognostical Identification of Medicinal Plants of PaederiaHe K aijia　(Phamacology Department,G uangxi Health Staff C ollege,Nanning530021) Abstract　Objective:T o distinguish accurately the dried medicinal plants of Paederia in G uangxi Province.Method: Using regular methods for identification of crude drugs.R esults:Obvious distinctions were found among leaf quality,pre2 sence of hair characteristics,inflorescence morphology,presence of non2glandular hair and scleroid cells along with their sizes,forms and superficial characteristics.Obvious distinctions were also found among cuticular veins of leaf cuticles,pro2 jecting of cells’outer walls,development of metaxylem,shapes and separation of crystal cells and wood fibers under micro2 scope.Conchusion:All the above2mentioned characteristics can be used for species identification.K ey words　plants of Paederia,pharmacognostical identification;G uangxi Province(original article on page323)IR Spectral Identification of Rhizoma Corydalisand Rhizoma Corydalis R epentisT ian Jinguo,Chen Y onglin,Ren Jian,Lou H ongxiang and G ao Y anhui(Faculty of Pharmaceutical Sciences,Shandong Medical University,Jinan250012) Abstract　Objective:T o study the IR spectra of Rhizoma C orydalis and Rhizoma C orydalis Repentis.Method:The petroleum ether,ethyl ether and water extracts obtained by the same isolating procedure from Rhizoma C orydalis and Rhi2 zoma C orydalis Repentis were determined by IR spectrophotometry.R esults:The IR spectral evidences of the two samples showed distinct characteristics and good repeatability.Conclusion:The IR spectral evidences can be used to differentiate Rhizoma C orydalis and Rhizoma C orydalis Repentis.K ey words　IR spectra;identification;Rhizoma C orydalis;Rhizoma C orydalis Repentis(original article on page327)Determination Chlorogenic Acid in Chrysanthemum morif olium R amat.Flow erLi Z ong,Chen Zaimin,Liao Leisheng and Lin Shaoshan(Fujian Provincial Insititute for Drug C ontrol,Fuzhou350001)Abstracts　Objective:T o determine the contents of chlorogenic acid in flower of Chrysanthemum morifolium. Method:C olumn:Nova2Pak C18,4μm,4.6mm×250mm,mobile phase:MeOH20.1mol・L-1NaH2PO4(pH2.65)30∶70,de2 tection wavelength:328nm.R esults:Flowers of20batches bought from the market were analysed.C ontents of clorogenic acid were0.060%～0.467%.Conclusion:Chlorogenic acid is a suitable component for the quality control of C.morifolium.K ey words　Chrysanthemum morifolium;chlorogenic acid;content determination(original article on page329)E ffect of Space Flight on U ltrastructure in Medicinal PlantDatura innoxia Mill.G ao Wenyuan,Zhao Shuping,Xue Lan,Quan Lihui,Fu Rongzhao and Xiao Peigen(Institute of Medicinal Plants,Chinese Academy of Medical Sciences and Peking Union Medical C ollege,Beijing100094) Abstract　Objective:T o substantiate the effect of space environment on medicinal plants.Method:The seeds of Datura innoxia were set up in retrievable satellites.After returning to earth,the ultrastructural changes were observed by means of electron microscope.R esult:In cells of weightless group and hit group,some changes were found in chloroplast。