北马兜铃的化学成分研究——Ⅱ、马兜铃酸E的化学结构

From the root of Aristolochia contorta Bunge six chemical constituents were isolated; one of them is a new phenanthrene compound containing nitro group for which the name aristolochic acid E is suggested. By means of spectral methods combined with chemical analysis the chemical structure of aristolochic acid E was determined to be 7-methoxy-8-hydroxy-aristolochic acid. The other components were identified as allantoin, aristolochic acid A, magnoflorine, β-sitosterol and daucosterol respectively.     

马兜铃酸肾不良反应及质量控制研究进展

马兜铃酸为硝基菲类有机酸类化合物,是马兜铃、关木通、细辛等植物的主要成分。马兜铃酸药理作用广泛,有抗感染、抗肿瘤、增强细胞免疫等功能。本文总结了马兜铃酸肾毒性研究进展及其分子结构,并对近5年不同中药材和中成药中马兜铃酸质量控制的研究进行综述,对比分析不同检测方法的优缺点,为今后对马兜铃酸的进一步研究及临床合理应用提供依据。     

Selectively preparative purification of aristolochic acids and aristololactams from Aristolochia plants

A novel method has been developed for the purification of aristolochic acids and aristololactams compounds from Aristolochia plants, a kind of typically toxic traditional Chinese medicine. In this method, Oligo (ethylene glycol) separation column which has “clustering function” for compounds in TCMs was used to produce the fractions containing the compounds with similar structures. A four-channel parallel preparative HPLC with C18 separation column was employed to purify the target compounds. The extraction sample of the blending of Radix Aristolochiae, Fructus Aristolochiae and Caulis Aristolochiae Manshuriensis was used to develop the method. Then, four aristolochic acids and three aristololactams were obtained using this method and the chemical identification was confirmed by Q-TOF-MS, ¹H NMR and ¹³C NMR. Thus, this method can deal with more than one traditional Chinese medicine simultaneously. Additionally, the results demonstrated that this method was an effective way to purify target compounds selectively from TCMs.     

马兜铃酸类成分检测与分析研究进展

马兜铃酸类成分是广泛存在于马兜铃科植物中的硝基菲类化合物,已被证实具有肾毒性、致癌和致基因突变等作用。我国自2003年以来采取一系列风险控制措施,其中马兜铃酸含量高的关木通、广防己和青木香等药材已被禁用。目前,一些马兜铃酸含量低的中药材与中成药仍在使用中,鉴于马兜铃酸成分对人体的严重危害性,有必要进一步加强相关药材与制剂的风险评估。在归纳马兜铃酸类成分结构等基本信息的基础上,对近年来的检测分析方法进展进行了总结,为其风险控制与安全使用提供了科学依据。     

关木通药材中马兜铃酸A的药动学研究

目的　采用高效液相色谱法测定血浆中马兜铃酸A的浓度,研究小鼠灌胃给予关木通提取液后体内马兜铃酸A的药动学特点。方法　采用DiamonsilTMC18色谱柱(250mm×4. 6mm, 5μm),以甲醇水冰乙酸(72∶27∶1)为流动相,流速1. 0mL·min-1,检测波长315nm。小鼠按马兜铃酸A4. 0mg·kg-1灌胃给予关木通提取液后,经HPLC法测定血浆药物浓度。结果　马兜铃酸A在0. 1098~3. 66mg·L-1的浓度范围内线性关系良好(r=0.9998,n=7),最低检测浓度为0. 0183mg·L-1。马兜铃酸A高、中、低3种不同浓度的平均回收率及相对标准偏差分别为97. 07% (1. 0% ), 97. 10% (1. 1% )和99. 36% (0. 8% ),该法的日内和日间精密度RSD值均小于9. 3%。小鼠给药后,药动学房室模型为一室模型,主要药动学参数T1 /2(ka),T1 /2(ke),Tmax, AUC, Cmax,CL/F(S),V/F(c)分别为: 3. 94, 16. 29, 10. 65min, 104. 88(mg/L)·min, 2. 84mg/L, 0. 038mg/kg/min/(mg/L), 0.89(mg/kg) /(mg/L)。结论　本实验建立了关木通药材中马兜铃酸A血药浓度的HPLC测定方法,阐明了马兜铃酸A的药动学特征。     

国产马兜铃属的植物和生药研究:资源利用

在全国调查的基础上,对我国马兜铃属18种药用植物的原植物,进行分类鉴定、薄层比较,并分析18种马兜铃属植物的根(或根茎)中马兜铃总酸的含量。     

反相离子对色谱法测定青木香中马兜铃酸A的含量

建立反相离子对色谱法测定青木香中马兜铃酸A含量。采用HypersilC18色谱柱(2 0 0mm× 4 6mm ,5 μm) ,以甲醇∶1 2 8%四丁基溴化氨∶pH 3 6醋酸 醋酸钠缓冲液 (体积比2∶1∶1)为流动相 ,流速 1 0mL/min ,检测波长 316nm。线性范围 2 0 8～ 10 4 0 μg/mL(r =0 9998) ,样品平均加样回收率为 97 0 % (n =6 ) ,精密度RSD为 1 5 % (n =6 )。该法简便、快速、准确 ,适用于青木香中马兜铃酸A的定量分析     

Development of HPLC fingerprint for species differentiation and quality assessment of Rhizoma Smilacis Glabrae

Rhizoma Smilacis Glabrae (RSG) is a commonly used herbal material in functional food and Traditional Chinese Medicine. A HPLC chromatographic fingerprint was developed for its quality control and species differentiation. Nine peaks were found in the chromatogram of RSG and all these peaks were identified by diode array detection and electrospray ionization–MS/MS: 5-O-caffeoylshikimic acid, taxifolin, engeletin, isoengeletin, trans-resveratrol, astilbin and its three stereoisomers. Six of these constituents were consistently found in 18 batches of samples. The standard fingerprint of RSG was generated by mean simulation of all tested samples. Using the standard fingerprint, RSG could be easily differentiated from Rhizoma Smilacis Chinae and Rhizoma Heterosmilacis, the two species that can be confused with RSG.     

马兜铃属植物的化学(Ⅱ)异马兜铃酸与马兜铃酸的关系

作者等前于土青木香(Aristolochia debilis, S. et Z.)中分出一种m.p. 275℃(分解)的黄色结晶(Ⅰ),从其元素分析、游离酸及其甲酯的熔点看来,均与異马兜铃酸(Ⅱ)一致.经继续研究,证明(Ⅰ)应与马兜铃酸(Ⅲ)相同,因此作这二种化合物的检查.按马兜铃酸是马兜铃属植物的一个共有成分,而異马兜铃酸则系1935年Krishnaswamy,Manjunath,Rao等氏从印度马兜铃(A. indica)中分出.Rosenmund,Reichstein等曾怀疑这二种成分可能是同一化合物,但没有充分的证据。     

十堰地区连翘药材高效液相指纹图谱研究

目的：建立十堰地区连翘药材化学成分的HPLC指纹特征图谱,为连翘药材的质量控制提供科学依据。方法：采用HPLC-梯度洗脱法,以Waters Sun Fire C18（250 mm×4.6 mm,5μm）为色谱柱,Diamonsil C18保护柱,以乙腈和0.4%醋酸溶液为流动相,梯度洗脱,流速为1 ml·min^-1,检测波长为277 nm,柱温为30℃。结果：不同产地的12批连翘药材化学成分特征图谱共检出14个共有峰。对不同产地的14个特征峰进行聚类分析,连翘药材产地与特征图谱相似度差异不大,样品中连翘酯苷A和连翘苷的含量有明显差别。结论：该方法简便、快速,可做为连翘药材的质量控制的有效手段。     

LC-MS法检测消风止痒颗粒中马兜铃酸A

目的:建立检测消风止痒颗粒中马兜铃酸A的液相色谱-质谱分析方法。方法:采用Venusil mp C18(250 mm×4.6mm,5μm)色谱柱,流动相为乙腈-0.1%甲酸水溶液(40∶60),流速1.0 mL.min-1。采用电喷雾电离源正离子质谱检测。结果:马兜铃酸A在0.005086～0.4069μg范围内线性关系良好(r=1.000),平均回收率(n=7)为97.1%(RSD=2.1%)。87批制剂均未检出马兜铃酸A。结论:本法快速、灵敏、准确,可用于消风止痒颗粒中是否混入关木通的确证检查,以防马兜铃酸A造成的用药安全,为消风止痒颗粒的质量控制提供了科学依据。     

反相高效液相色谱法测定关木通水煎剂中马兜铃酸A含量

目的 :建立反相高效液相色谱法测定关木通水煎剂中马兜铃酸A的含量。方法 :色谱柱 :SpherisorbODS2(4 6mm×250mm ,5μm ) ;流动相 :甲醇 -水 -冰醋酸 (72∶27∶1) ;流速 :1 0ml/min ;检测波长 :250nm。结果 :马兜铃酸A在0 0128μg～0 4096μg范围内线性关系良好 ,回归方程 :Y=4553 7 +5388319 3X ,r=0 9999 ;平均回收率为97 33 % ,RSD为2 34 %。结论 :本方法简便、灵敏、准确。     

穆坪马兜铃化学成分的研究

自穆坪马兜铃(Aristolochia moupinensis Franch)根、茎中分得十三个化合物,其中化合物Ⅰ～Ⅻ经鉴定分別为马兜铃酸Ⅰ(aristolochic acid Ⅰ)(Ⅰ),尿囊素(allantoin)(Ⅱ),紫丁香酸(syringic acid)(Ⅲ),香豆酸(P-coumaric acid)(Ⅳ),马兜铃酸Ⅳ(aristolochic acid Ⅳ)(Ⅴ),β-谷甾醇(Ⅵ),木兰碱(magnoflorine)(Ⅶ),马兜铃酸Ⅳ甲醚aristolochic acid Ⅳmethyl ether(Ⅵ),棕榈酸(Ⅸ),马兜铃酸Ⅱ(aristolochic acid Ⅱ)(Ⅹ),N(Phydroxyphenethy1)P-coumaramide(Ⅺ),马兜铃酸Ⅳ甲醚甲酯(aristolochic acid Ⅳmethyl ether methyl ester)(Ⅻ),化合物ⅩⅢ为新化合物,经光谱分析和化学合成等方法证明其结构为N(P-hydroxyphenethyl)-ferulamide,命名为穆坪马兜铃酰胺(moupinamide)。初步药理试验表明穆坪马兜铃酰胺体外有抑制血小板聚集和影响血小板内前列腺素合成的作用。     

酒萸肉超高效液相色谱指纹图谱的建立及化学计量分析

目的建立酒萸肉的超高效液相色谱指纹图谱,并进行聚类分析和主成分分析。方法色谱柱为ACE3 C18-AR柱(100 mm×2.1 mm,3μm),流动相为乙腈-0.3%磷酸溶液(梯度洗脱),流速为0.2 m L/min,检测波长为254 nm,柱温为25℃,进样量为1μL。以马钱苷为参照峰绘制76批酒萸肉药材样品的指纹图谱,并与10批山萸肉药材样品的指纹图谱进行比较。采用Chempattern化学计量学软件进行相似度评价,并进行聚类分析和主成分分析。结果76批酒萸肉药材指纹图谱共确定7个共有峰、6种成分,分别为没食子酸、5-羟甲基糠醛、莫诺苷、马钱苷、獐牙菜苷、山茱萸新苷;76批酒萸肉药材样品中有60批次的相似度大于0.8;76批酒萸肉药材样品和10批山萸肉药材样品聚为两大类,两大类的5-羟甲基糠醛含量差异明显;经主成分分析,峰6和峰2均是导致药材样品质量差异的重要因素。结论所建立的超高效液相色谱指纹图谱可用于酒萸肉的质量评价。     

基于指纹图谱和网络药理学预测经典名方温胆汤中枳实的质量标志物

目的 基于指纹图谱和网络药理学预测经典名方温胆汤(Wendan decoction，WDD)中药材枳实的质量标志物(Q-Marker)。方法 建立枳实药材和WDD基准样品的指纹图谱，使用中药色谱指纹图谱相似度评价软件(2012年A版)进行相似度分析和共有成分指认；采用网络药理学筛选共有成分的相关靶点和关键通路，并建立成分-靶点-通路网络，预测枳实发挥祛痰止呕，清胆和胃功效潜在的Q-Marker，并测定其含量。结果 建立了15批枳实药材和15批WDD基准样品的指纹图谱，15批枳实药材和WDD基准样品指纹图谱相似度>0.9，指认出5个化学成分，分别为柚皮苷、橙皮苷、新橙皮苷、川陈皮素、桔皮素；网络药理学筛选分析得到5个化合物的TP53、MAPK8、EGFR、JUN、MMP9等32个核心靶点和癌症通路、内分泌抵抗通路等10条关键通路，构建了“成分-靶点-通路”网络图，预测柚皮苷、橙皮苷、新橙皮苷、川陈皮素、桔皮素为WDD中枳实的Q-Marker。WDD中柚皮苷、橙皮苷、新橙皮苷、川陈皮素、桔皮素的含量≥0.011 7%，0.025 3%，0.025 7%，0.010 4%，0.010 7%。结论 WDD中枳实的Q-Marker预测分析为建立完整的WDD质量评价体系提供参考，也为阐明其药效物质基础的作用机制奠定了基础。     

New aristolochic acid, aristololactam and renal cytotoxic constituents from the stem and leaves of Aristolochia contorta

Two novel phenanthrene derivatives, aristololactam IVa (1) and 9-hydroxy aristolochic acid I (2) were isolated from the stem and leaves of Anstolochia contorta Bunge, together with 17 known compounds (3-19). The structures of these compounds were determined by spectroscopic analysis. The phenanthrenes obtained were tested for cytotoxicity against renal proximal tubular epithelial cell line (HK-2). Aristololactam IVa and 7-methoxy aristololactam IV were found to have strong cytotoxic activity against HK-2 cells with a potency similar to or even stronger than those of aristolochic acid I and aristololactam I.     

Rapid resolution liquid chromatography (RRLC) analysis and studies on the stability of Shuang-Huang-Lian preparations

Shuang-Huang-Lian (SHL) is a traditional Chinese formula which comprises of three medicinal herbs: Flos Lonicerae, Radix Scutellariae and Fructus Forsythiae, and is commonly used to treat acute upper respiratory tract infection, acute bronchitis and light pneumonia. A simple, reliable and reproducible rapid resolution liquid chromatography (RRLC) method was developed for the quality control of SHL preparations, which baseline separates the major bioactive compounds within 6 min. The method uses a C18-HST column (2.5 μm, 100 mm × 3.0 mm) kept at 40 °C. The mobile phases consist of 0.1% phosphoric acid aqueous solution and acetonitrile. Flow rate is 1.0 ml/min and UV detection is performed at 327 nm from 0 to 4 min and 229 nm from 4 to 7 min. This method was further validated according to the ICH guidelines. Eight batches of commercial SHL preparations obtained from different pharmaceutical manufacturers as well as individual herbs were examined and their chromatographic profiles were compared. The stability test revealed that chlorogenic acid is stable only at acidic pH, and hence it is necessary to further evaluate and optimize the preparatory procedures and storage conditions for commercial SHL preparations.     

女贞子标准汤剂含量转移率异常原因分析

目的 探讨分析11批次女贞子标准汤剂中特女贞苷含量转移率异常的原因。方法 通过肉眼观察和显微鉴别比较不同产地、不同批次女贞子在性状和显微鉴别中的差异,采用高效液相色谱法对11批次女贞子粉末和11批次女贞子壳的指纹图谱进行比对,并比较同批次女贞子的壳和粉末中指纹图谱情况。结果 不同产地、不同批次的女贞子的外观形状存在差异,在显微鉴别上均与药典描述一致;11批女贞子粉末和11批壳指纹图谱与对照指纹图谱的相似度均>0.88;同批次女贞子壳中特女贞苷含量是粉末的1.15~3.5倍。结论 女贞子壳和粉末中特女贞苷含量差异大,在进行标准汤剂研究中因捣碎破壁程度不一,因而导致标准汤剂中特女贞苷含量转移率异常。建议在生产中对壳和粉末比例或对捣碎程度进行规定,以保证批间含量稳定。     

马兜铃属植物的化学Ⅲ青木香酸结构(1)

我们于前报中报告了从土青木香(Aristolochia debilis, S. et Z.)里分得一个过去文献上没有的新化合物:青木香酸(Ⅰ),现从实验说明它是和马兜铃酸(Ⅱ)属于同一类型的天然含硝基化合物。青木香酸分子式是C_(18)H_(13)O_7N,m.p.>350°(在300°左右渐渐变黑),R_A~D=0.87.其甲酯C_(19)H_(15)O_7N(Ⅲ), m.p. 260°.(Ⅰ)遇浓硫酸也呈绿色,在鹼性溶液中呈红色,与其他试剂     

Alkaloids from the Roots of Aristolochia Triangularis(I)

从马兜铃科植物Ａｒｉｓｔｏｌｏｃｈｉａｔｒｉａｎｇｕｌａｒｉｓ的根部分离出十五个生物碱成分。其中两种成分ｔｒｉａｎｇｕｌａｒｉｎｅ－Ａ和ｔｒｉａｎｇｕｌａｒｉｎｅ－Ｂ系新生物碱，三种系阿卟啡类季铵碱。这些化合物的结构经各种波谱（ＩＲ，ＵＶ，ＭＳ，２ＤＮＭＲ）解析和化学转化技术得以确定。