Simultaneous quantification of three major bioactive triterpene acids in the leaves of Diospyros kaki by high-performance liquid chromatography method

The leaf of Diospyros kaki, which is a traditional Chinese medicine, has been used for the treatment of various diseases. In order to improve the quality assurance of the leaves of D. kaki, derived extracts and phytomedicines, a simple, rapid and accurate high-performance liquid chromatography (HPLC) method was developed to simultaneously assess the three bioactive triterpene acids: barbinervic acid (BA) and its epimer, rotungenic acid (RA), along with 24-hydroxy ursolic acid (HA). This HPLC assay was performed on a reversed-phase C18 column with methanol and aqueous H3PO4 as the mobile phase and using a monitoring wavelength at 210 nm. This method was successfully applied to quantify these three bioactive triterpene acids in five different solvent extracts of the leaves of D. kaki and in the leaves from six different locations in China. The results demonstrated the total content and quantity of each of the main bioactive compounds were strongly dependent on the extraction solvents and locations, indicating that the quality control of the bioactive ingredients in the leaves of D. kaki, derived extracts and phytomedicines is critical to ensure its clinical benefits. The content of the total triterpenoids was also determined by the less selective colorimetric method, and the comparison with the HPLC method was given.     

Metabolic conversion from co-existing ingredient leading to significant systemic exposure of Z-butylidenephthalide, a minor ingredient in Chuanxiong Rhizoma in rats

Pharmacokinetic (PK) study of medicinal herbs is a great challenge, because which component(s) is(are) the bioactive ingredients is largely unknown. Most of the reported PK studies of herbs focused on the major ingredients regardless of their in vivo bioactivities, while PK of components with low content in herbs is often ignored. The present study demonstrates how PK study can reveal potential importance of a low content ingredient to the herbal bioactivities using Z-butylidenephthalide (BuPh), a bioactive phthalide present in a significantly low quantity in medicinal herb Chuanxiong Rhizoma, as an example. PK of BuPh was investigated in rats using Chuanxiong extract, fraction containing BuPh and ligustilide, and pure BuPh, respectively. The results demonstrated that remarkable blood concentrations of BuPh were observed after administration of the herbal extract and its systemic exposure was significantly different between BuPh given in pure and mixed forms. More interestingly, AUC of BuPh via intake of fraction (9.3-fold) and extract (4.5-fold) was significantly greater than that obtained from pure BuPh, which was further evidenced to be mainly due to metabolic conversion from ligustilide, a major component in Chuanxiong. Our findings revealed that although it naturally occurred in low amount, BuPh reached significant systemic concentrations via metabolic conversion from ligustilide. Moreover, our results demonstrated that PK study is one of crucial and inevitable steps for revealing in vivo bioactive ingredients of herbal medicines, and such studies should be more appropriate to focus on in vivo profile of the ingredients co-existing in herbs rather than only studying them individually.     

Simultaneous determination of nine components in Qingkailing injection by HPLC/ELSD/DAD and its application to the quality control

High-performance liquid chromatography coupled with photo diode array detection and evaporative light scattering detection (HPLC/DAD/ELSD) was established to simultaneously determine nine ingredients in Qingkailing injection. Four wavelengths at 240, 254, 280 and 330 nm, respectively, were chosen as the monitoring wavelength to determine two nucleosides (uridine and adenosine), geniposide, baicalin and two organic acids (chlorogenic acid and caffeic acid), and an evaporative light scattering detector combined was employed to determine three steroids (cholic acid, ursodeoxycholic acid and hyodeoxycholic acid). This assay was fully validated in respect to precision, repeatability and accuracy. The proposed method was successfully applied to quantify the nine ingredients in 19 different Qingkailing injection samples and by principal component analysis (PCA) and hierarchical clustering analysis (HCA), it demonstrated significant variations in the content of these compounds in the samples from different manufacturers and preparation procedures. This method could be readily utilized as a quality control method for traditional Chinese medicine (TCM).     

日本川芎药材的HPLC指纹图谱分析方法研究

目的 建立日本川芎药材的指纹图谱分析方法。方法 用HPLC法建立日本川芎醇提取物的指纹图谱。结果和结论 所用方法可靠，简便，为提高日本川芎质量控制标准提供参考。     

RP-HPLC-DAD法同时测定吴茱萸汤中7个生物碱和2个黄酮苷含量(英文)

建立反相高效液相色谱-二极管阵列检测 (RP-HPLC-DAD) 法同时测定经典名方吴茱萸汤中7个生物碱 (去氢吴茱萸碱、10-羟基吴茱萸次碱、吴茱萸碱、吴茱萸次碱、1-甲基-2-正壬基-4(1H)喹诺酮、吴茱萸卡品碱和二氢吴茱萸卡品碱) 和2个黄酮苷 (异鼠李素-7-O-芸香糖苷和香叶木素-7-O-β-D-吡喃葡萄糖苷)的含量。RP-HPLC分离应用反相C18 色谱柱, 甲醇-1%醋酸水溶液梯度洗脱, 流速为1.0 mL/min, 检测波长为300 nm。上述9个分析物达到基线分离, 标准曲线线性关系良好(r2>0.9941), 日内和日间精密度和准确度符合分析方法学要求, 加样回收率为90.13%-102.48% (RSDs<3.6%)。经方法学验证, 所建立的方法适用于吴茱萸汤物质基础的质量控制。     

Simultaneous determination of protocatechuic acid, syringin, chlorogenic acid, caffeic acid, liriodendrin and isofraxidin in Acanthopanax senticosus Harms by HPLC-DAD

A high performance liquid chromatography (HPLC) method was developed for the first time to quantify simultaneously the six major active ingredients in Acanthopanax senticosus (Rupr. et Maxim.) Harms, namely protocatechuic acid, syringin, chlorogenic acid, caffeic acid, liriodendrin and isofraxidin. The analysis was performed by a reverse phase gradient elution with an aqueous mobile phase (containing 0.05% phosphoric acid) modified by acetonitrile and diode-array multiple-wavelength UV detector (DAD). Six regression equations showed good linear relationships between the peak area of each marker and concentration. The recoveries of the markers listed above were 92.3%, 93.9%, 90.3%, 93.1%, 94.3% and 90.7%, respectively. The relative standard deviation of intra-day and inter-day were less than 2.7% and 3.1%, respectively. This method was validated for specificity, accuracy, precision and limits of quantification. Medicinal materials of ten commercial brands were analyzed and found to contain different amounts of the six bioactive markers. The method developed can be used for the quality control of Acanthopanax senticosus (Rupr. et Maxim.) Harms.     

骨愈灵制剂质量标准统一的研究

目的：统一骨愈灵胶囊、片剂的质量标准。方法增加乳香、当归、川芎显微特征及没药薄层鉴别,建立梯度洗脱 HPLC法测定三七中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的含量。结果显微特征、薄层鉴别、含量测定方法专属性好。结论骨愈灵制剂质量标准的统一,能有效地控制该产品的质量。     

替代对照品法同时测定川芎中丁苯酞和藁本内酯的含量

目的:建立HPLC替代对照品法同时测定川芎中丁苯酞和藳本内酯。方法:采用Agilent Eclipse XDB-C18(4.6 mm×150 mm,5μm)色谱柱,以甲醇-水(52∶48)为流动相,流速1.0 mL.min-1,丁苯酞检测波长228 nm,藁本内酯检测波长330nm,柱温35℃。以丁苯酞作为藁本内酯的替代对照品,在不同条件下测定相对校正因子,利用相对校正因子和替代对照品同时测定川芎中丁苯酞和藁本内酯的含量。结果:丁苯酞和藁本内酯进样量分别在0.01～0.2μg(r=0.9999)和0.1～2.0μg(r=0.9999)范围内与峰面积呈良好的线性关系,测得相对校正因子f’为1.0806,不同条件下相对校正因子的重现性良好,利用相对校正因子计算川芎中待测成分含量与外标法实测值之间没有明显差异。结论:用丁苯酞替代藁本内酯作为对照品,同时测定川芎中丁苯酞及藁本内酯的含量,解决了藳本内酯对照品不稳定的难题,该方法可用于川芎的质量控制。     

Simultaneous quantification of active components in the herbs and products of Si-Wu-Tang by high performance liquid chromatography–mass spectrometry

Si-Wu-Tang (SWT), comprising Paeoniae, Angelicae, Chuanxiong and Rehmanniae, is one of the most popular Traditional Chinese Medicine (TCM) formulae for woman's health. Data mining from the available Chinese and English literatures indicated that the major bioactive components of SWT consist of paeoniflorin, paeonol, gallic acid, ferulic acid, Z-ligustilide, ligustrazine, butylphthalide, senkyunolide A and catalpol. Since content determination of the marker compounds is generally considered as an initial step for quality control of TCM product, a high performance liquid chromatography–mass spectrometric method employing both positive and negative electrospray ionization was developed for the simultaneous determination of the nine identified compounds in the raw herbs and products of SWT. The LOQ of the developed assay method for the tested components was 10 ng/ml for ligustrazine, 200 ng/ml for catalpol, and 100 ng/ml for the other seven compounds. The intra-day and inter-day variations of the current assay were within 17.5%. Paeoniflorin, ferulic acid, gallic acid, Z-ligustilide and senkyunolide A were found in all SWT products investigated. Variations in the contents of the studied compounds were observed among batches of raw herbs and SWT products. The currently developed method provides a sensitive and rapid quantification approach that can be useful in the quality control of raw herbs and products of SWT.     

脑伤泰颗粒的质量标准研究

目的 :建立脑伤泰颗粒的质量标准。方法 :采用薄层色谱法对脑伤泰颗粒中4味中药进行定性鉴别 ;用薄层扫描法对黄芪甲苷进行含量测定。结果 :脑伤泰颗粒剂中4味中药的薄层色谱鉴别呈阳性 ,黄芪甲苷的平均含量为0 0294 % ,方法平均回收率为98 18 % ,RSD=1 85 %。结论 :本方法简便、重现性好 ,可用于脑伤泰颗粒的质量控制。     

Determination of five components in Ixeris sonchifolia by high performance liquid chromatography

A high performance liquid chromatography (HPLC) method was developed for the simultaneous quantification of five major active ingredients (markers) in Ixeris sonchifolia (Bge.) Hance, namely chlorogenic acid, caffeic acid, luteolin-7-O-beta-D-glucuronide, luteolin-7-O-beta-D-glucoside and luteolin. Samples were extracted with 70% methanol. The chromatographic separation was performed on a Hypersil ODS(2) column (250 mm x 4.6 mm i.d.; 5 microm) with a gradient of acetonitrile and 0.5% (v/v) aqueous acetic acid, at a flow rate of 1.0 ml/min, detected at 335 nm. Five regression equations showed good linear relationships (r(2)>0.999) between the peak area of each marker and concentration. The assay was reproducible with overall intra- and inter-day variation of less than 3.2%. The recoveries, measured at three concentration levels, varied from 94.1% to 100.7%. This assay was successfully applied to the determination of the 5 bioactive compounds in 18 samples. The results indicated that the developed assay method was rapid, accurate, reliable and could be readily utilized as a quality control method for I. sonchifolia (Bge.) Hance.     

脑得生片的薄层色谱鉴别

目的：建立脑得生片中川芎的质量标准。方法：采用薄层色谱法对片剂中的川芎进行了鉴别。结果：薄层色谱鉴别可检出样品中相应的薄层斑点。结论：方法简便，灵敏，专属性强，重现性好，可有效地控制制剂的质量。     

妇科调经胶囊的显微特征及薄层色谱鉴别

目的建立妇科调经胶囊的质量控制方法。方法对处方中的当归、川芎、白术和延胡索进行了显微鉴别;并用薄层色谱法对妇科调经胶囊中的当归、川芎、香附、延胡索4味药材进行定性鉴别。结果该处方所含的当归、川芎、白术和延胡索4味药材的显微特征明显,且在薄层色谱中分别检出当归、川芎、香附、延胡索的特征斑点,阴性对照无干扰。结论该方法简便可行,专属性强,重现性好,可有效控制妇科调经胶囊的质量。     

Simultaneous determination of eight components in Radix Tinosporae by high-performance liquid chromatography coupled with diode array detector and electrospray tandem mass spectrometry

High-performance liquid chromatography (HPLC) coupled with electrospray tandem mass spectrometry (ESI-MS-MS) and diode array detection (DAD) was used to identify and simultaneously determine eight major ingredients in Radix Tinosporae. The assay was performed on a Diamonsil C(18) analytical column with a gradient solvent system of A (water containing 0.2% formic acid, 20mM ammonium acetate) and B (methanol/acetonitrile=1/1, v/v). The 217, 248, 270 and 347 nm, respectively, were chosen as the monitoring wavelengths to determine four structural types of components, say columbin, phytoecdysteroids (including 20-hydroxyecdysone, 2-deoxy-20-hydroxyecdysone 3-O-beta-d-glucopyranoside and 2-deoxy-20-hydroxyecdysone), menisperine and protoberberine alkaloids (including columbamine, jatrorrhizine and palmatine). This method was validated in respect to precision, repeatability and accuracy, and was successfully applied to quantify the eight components in 39 batches of R. Tinosporae for quality control purpose. The results indicated that the proposed method could be readily utilized as a quality control method for traditional Chinese medicine (TCM).     

Phytochemistry,biological properties and quality control of Chuanxiong Rhizoma: a review

Chuanxiong Rhizoma is a widely used Chinese herbal medicine in the past 2000 years. Chuanxiong Rhizoma is composed of volatile oils, phthalide lactones, phenolic acids, polysaccharides and other compounds. To date, more than 149 compounds in Chuanxiong Rhizoma have been isolated and identified, and some of them have been reported to possess promising biological properties on cardiovascular and central nervous system disorders besides their anti-cancer and antioxidant effects. Modulation of inflammatory mediators and apoptotic factors are believed to contribute to its bioactivities. Analytical methods, such as HPLC, GC and UPLC, are employed for qualitative evaluation of Chuanxiong Rhizoma. In this work, harvest period, growing habitat, processing method and storage, which can affect the quality of Chuanxiong Rhizoma, were also discussed. Comprehensive quality control methods should be developed to ensure the safety, quality and efficacy use of Chuanxiong Rhizoma. Herein, we collected and analyzed the literature of Chuanxiong Rhizoma published on CNKI, ScienceDirect, Springer link, Wiley and PubMed in past two decades, and up-to-date information of Chuanxiong Rhizoma was provided in this paper. We suggested ligustilide, butylidenephthalide and total senkyunolides as the chemical markers to evaluate the quality of Chuanxiong Rhizoma. Additionally, the influences of soil conditions and processing methods on Chuanxiong Rhizoma as future research perspectives should also be further assessed.     

复方维肤乳的制备及临床应用

目的 :制备复方维肤乳 ,观察其对婴、幼儿湿疹的疗效。方法 :以黄芩、当归、川芎、二甲基硅油等为处方制备复方维肤乳 ,并建立质量控制方法。结果与结论 :该制剂性质稳定 ,质控方法可行 ;治疗婴、幼儿湿疹总有效率达90 % ,未发现明显不良反应 ,值得临床推荐应用。     

十珍香附丸质量标准的研究

目的：建立十珍香附丸的质量标准。方法：采用TLC法对方中的香附、当归、川芎、黄芪进行了定性鉴别；采用HPLC法测定制剂中的芍药苷含量。结果：定性定量方法简便，准确，专属性强，重现性好。结论：提高后的质量标准更有效的控制该产品的质量。     

柴白安神合剂的质量控制研究

目的建立柴白安神合剂的质量标准,有效控制柴白安神合剂的质量。方法采用薄层色谱法（TLC）对柴胡、百合、白芍、五味子、川芎进行定性鉴别,高效液相色谱法（HPLC）对黄芩苷进行含量测定。结果TLC可定性鉴别柴胡、百合、白芍、五味子、川芎5味药材,且Rf值合理,重复性好;HPLC中黄芩苷在0.1032～1.032μg内线性关系良好（r=0.9998）,平均回收率为99.5%,RSD为1.4%（n=6）。结论该质量控制方法准确,简单、专属性强,可有效控制柴白安神合剂的质量。     

不同方法提取川芎挥发油的GC-MS分析

目的:对川芎饮片不同提取方法所得挥发油的化学成分进行比较分析。方法:采用气相色谱-质谱联用仪对提取物进行比较分析。结果:4种方法提取的挥发油有9个化合物相同。其中水蒸气蒸馏法提取的挥发油样品中组分种类最多,主要是烯萜类成分。结论:4种提取方法均能提取出川芎挥发油中的主要成分,主要成分为内酯类,相对含量最高的为反式-藁本内酯。     

HPLC-DAD-ELSD法同时测定复方贞术调脂胶囊15个活性成分(英文)

复方贞术调脂胶囊(FTZc)是一种用于治疗高脂血症的专利药物,由八味中药组成,含有多活性成分的复方制剂。本文建立了可行的HPLC-DAD-ELSD方法用于FTZc的质量控制,同时测定15个活性成分(红景天苷、特女贞苷、木兰碱、迷迭香酸、丹酚酸B、非洲防己碱、表小檗碱、药根碱、黄连碱、巴马汀、小檗碱、5,7-二甲氧基香豆素、人参皂苷Rg1、人参皂苷Rb1和齐墩果酸)。色谱柱采用Ultimate XB C18,流动相为乙腈–水(0.25%乙酸,0.13%三乙胺)(梯度洗脱),流速为0.8mL/min,检测方法采用光电二极管阵列检测器的多波长检测模式和蒸发光散射检测器联合应用。方法学验证表明该方法精密度、准确度良好,成功用于不同批次FTZc的测定。结果表明,本文建立的测定多种生物活性成分HPLC-DAD-ELSD方法切实可行、方法可靠,特别适用于复方贞术调脂胶囊的质量评价。