High-resolution method for regulatory control of Echinacea species in Nutraceuticals by CD-MEKC

One problem in the international regulatory control of Echinacea, a therapeutic Nutraceutical, is recognition of caffeoyl solutes and alkamides in different products. Cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) has been applied to Echinacea spp. in combination with pattern recognition of some caffeoyl solutes. A novel metric based on relative migration time (RMT) data has been developed in CE to address the problem of variable reported migration time.The CD-MEKC method of Gotti's group using hydroxypropyl-β-cyclodexrin (HP-β-CD; 100 mM) with sodium dodecyl sulphate (SDS; 110 mM), in a triacid background electrolyte (10 mM, pH 8) under 19 kV was adapted to identify two key hydrophilic solutes: chlorogenic acid and cichoric acid present in all commercial products. Two internal markers were taken as reference points to calculate the RMT of any target peak: RMT = t_{m (target)}/t_{m (marker)}.The RMT method was robust to temperature change from 20 to 40 °C, but sensitive to pH. The lateral shift and reproducibility of the target peak t_{m (target)} were significantly reduced by this novel transformation. In the worst cases migration time variability ranged up to 12% (n = 6); the RMT algorithm reduced this to less than 1%. In general, the RMT transformation reduced the variability of migration time data by a factor of 2–5.For systematic comparison of electrophoretic profiles for test sample and standard, a new pattern recognition algorithm permits sequential peak-by-peak comparison using specified segments of the electropherograms for comparison of test and Echinacea purpurea (root product) as a standard. This algorithm was capable of rapidly characterising the similarity of target peaks in a test sample relative to those in the reference standard. Combination of the RMT algorithm and pattern recognition in CE is expected to offer a robust approach to international regulatory characterisation and control of Nutraceuticals.     

Determination of heavy metals for the quality control in argentinian herbal medicines by ETAAS and ICP-OES

The determination of trace elements in Hypericum perforatum leaves and flowers, their teas, tinctures and tablets was carried out by Electrothermal Atomic Absorption Spectrometry (ETAAS) and Ultrasonic Nebulization System coupled to Inductively Coupled Plasma Optical Emission Spectrometry (USN-ICP-OES). Hypericum perforatum (St. John’s wort), is a phytomedicine used for the treatment of depression. Samples were collected from different sources in the argentinian market. Heavy metals contents in the investigated samples were found at different levels. Chromium and cobalt were undetectable above their limits of detection in both liquid and solid samples; while aluminum, cadmium, lead, iron and vanadium were present in the majority of samples.     

Evaluation of a method to determine the natural occurrence of aflatoxins in commercial traditional herbal medicines from Malaysia and Indonesia

Traditional herbal medicines, popularly known as ‘jamu’ and ‘makjun’ in Malaysia and Indonesia, are consumed regularly to promote health. In consideration of their frequent and prolonged consumption, the natural occurrence of aflatoxins (AF) in these products was determined using immunoaffinity column clean-up and high-performance liquid chromatography with pre-column derivatization. The evaluated method, which entails dilution of sample extracts with Tween 20–phosphate buffered saline (1:9, v/v) and a chromatographic system using isocratic mobile phase composed of water–methanol–acetonitrile (70:20:10, v/v/v), was effective in separating AFB₁, AFG₁ and AFG₂ from interference at their retention times. Results were confirmed using post-column derivatization with photochemical reactor. For 23 commercial samples analyzed, mean levels (incidence) of AFB₁, AFB₂ and AFG₁ in positive samples were 0.26 (70%), 0.07 (61%) and 0.10 (30%) μg/kg, respectively; one sample was positive for AFG₂ at a level of 0.03 (4%) μg/kg. In contrast to the high levels of AF in crude herbal drugs and medicinal plants reported previously by other researchers, the low contamination levels reported in this study may be attributed to the higher selectivity to AF of the method applied. Based on the AFB₁ levels and the daily consumption of positive samples, a mean probable daily intake of 0.022 ng/kg body weight was calculated.     

Evaluation of quality control strategies in Scutellaria herbal medicines

The statutory regulation of herbal medicines is under review within the United Kingdom (UK) and by 2011 all herbal medicines will require either a Product Licence or a Traditional Herbal Registration. The species Scutellaria baicalensis has been shown to possess anti-inflammatory, anti-viral and anti-tumor properties and is one of the most widely used Chinese herbal extracts in Eastern and Western medicines. The bioactivity of this herbal medicine is due to the radical scavenging activities of the flavone components of which there are more than 60. This research has characterised 5 key flavones in 18 extracts of Scutellaria using a combination of HPLC with DAD and MS detection. Employing an internal standard approach, the validated HPLC method afforded good sensitivity and excellent assay precision. Assays for the ferric reducing antioxidant power (FRAP) and total phenol determinations enabled determination of the antioxidant coefficient (PAC) of each Scutellaria extract. The potential usefulness of employing multivariate statistical analysis using a combination of the key parameters collected namely, FRAP activity, total phenol content, levels of 5 flavone biomarkers and the PAC as a means of quality evaluation of the Scutellaria herbal extracts was investigated. The PAC value was predicted by soft independent modelling of class analogy (SIMCA) as being the most discriminatory parameter and applying this ranking the herbal extracts were grouped into 3 clusters. The second most influential parameter in determining the clustering of the samples was the level of baicalin in each extract. It is proposed that the PAC value alone or in combination with a chromatographic fingerprint of key biomarkers [e.g. baicalin or (baicalin+baicalein)] may be useful indicators to adopt for the quality control of S. baicalensis.     

一测多评中药质量控制研究进展

一测多评(QAMS)是近年来新兴的中药质量控制方法,能够在缺少对照品的情况下实现多成分质量控制。迄今,QAMS报道的中药材、饮片以及中药制剂已达数十种。植物次级代谢产物是多数中药发挥药效作用的物质基础,也是评价相应中药质量优劣的关键。本文从植物次级代谢产物的角度,综述了近年来QAMS在中药质量评价中的研究进展,为该方法进一步的研究和应用提供了参考。     

中药开拓欧盟植物药市场的机会与威胁分析

目的:帮助我国中药企业加深对欧盟植物药市场的了解,推动中药国际化。方法:在认识欧盟植物药市场的市场容量、细分市场、消费方式和发展趋势等的基础上,分析中药开拓欧盟植物药市场所面临的机会与威胁。结果与结论:中药开拓欧盟植物药市场,机会与威胁并存,但总的来说机会大于威胁。我国企业应加大对欧盟植物药市场的开拓力度。     

Development and validation of an improved HPLC method for the control of potentially counterfeit isometamidium products

Isometamidium, a mixture of related substances of which 8-(3-m-amidinophenyl-2-triazeno)-3-amino-5-ethyl-6-phenylphenanthridinium chloride hydrochloride (M&B4180A) is the principal active component, is the only chemical agent available for prophylaxis of veterinary trypanosomiasis. A method for the simultaneous quantitation of the major constituents M&B4180A, 3-(3-m-amidinophenyl-2-triazeno)-8-amino-5-ethyl-6-phenylphenanthridinium chloride hydrochloride (M&B38897), 7-(m-amidinophenyldiazo)-3,8-diamino-5-ethyl-6-phenylphenanthridinium chloride hydrochloride (M&B4250) and 3,8-di(3-m-amidinophenyltriazeno)-5-ethyl-6-phenylphenanthridinium chloride dihydrochloride (M&B4596) is described. The related substances are resolved on a Gemini C18 column (150 mm x 4.6 mm, 5 microm) using a mobile phase composed of a mixture of acetonitrile and 50 mM ammonium formate buffer pH 2.8 (25:75 v/v) at a flow rate of 1 ml/min with UV detection at 320 nm. The method is compatible with electrospray ionisation mass spectrometry and provides a tool for the control of substandard and counterfeit commercial preparations of isometamidium.     

红外光谱法测定中药掺杂化学药品的方法改进

目的:改进专利中"用红外光谱检测中药中化学药物的方法",提高其准确性。方法:考察求导计算方法和随机采样范围对结果的影响,优选条件。结果:采用斜率法或平滑窗口小的拟合法进行求导,以及扩大至负值区域的随机采样方法,得到了更理想的结果。结论:经改进以后,方法准确率提高,进一步完善了该专利方法。     

振动法对外敷七厘散动物模型骨折愈合的病理研究

目的探讨振动法对中药七厘散外敷动物骨折愈合的促进作用,为临床提高骨折康复质量提供依据。方法将家兔随机分为4组,1组为对照组,2-4组分别在骨折后1天、2天、3天给予中药七厘散外敷包扎,配合低频率间歇振动,骨折前及骨折后1周、2周、3周拍X线片,观察骨痂形成情况,3周末处死,行弯曲试验及病理切片检查。结果3、4组经振动后最大载荷平均增加了20％;最大弯矩平均增加了39％;组织学愈合良好,软组织血供丰富;三周末骨痂形成率平均较对照组增加了21．4％。结论采用中药七厘散外敷包扎配合小型振动仪间歇振动可有效促进骨折愈合速度和质量;振动介入以骨折后2天、3天为佳。     

HPLC检查非布司他原料药中的有关物质

目的 采用HPLC法测定Febuxostat原料药中的有关物质.方法 采用Agilent Eclipse plus C18色谱柱,以0.05％磷酸溶液-乙腈(40∶60)为流动相,检测波长220 nm,流速1.0 mL· min-1,柱温40℃.结果 在建立的色谱条件下,Febuxostat与其有关物质的分离良好(R＞1.5),杂质A、B、C的相对校正因子分别为1.344、1.588、2.157.结论 所用方法操作简便、灵敏度高、专属性强,可用于Febuxostat中有关物质的检查.     

国际植物药监管合作组织（IRCH）的发展及对我国植物药监管的启示

目的：介绍国际植物药监管合作组织（IRCH）发展情况,为进一步做好我国植物药的监督和管理提供参考。方法：对该组织的发展历程、成员和构架、工作形式及最新一届年会的情况进行介绍,提出对我国植物药监管的启示。结果与结论：中国是该组织的第一批成员国,一直积极参与国际植物药监管合作组织相关工作。参与IRCH活动,不仅可展示国家食品药品监督管理总局在完善中药注册管理法规体系方面取得的成绩,促进中国食品药品检定研究院领导的第二工作组的交流与合作;还可了解各国植物药掺假的现状和监管手段,更好地推动未来的植物药监管工作。     

中药化学成分体内代谢产物的制备方法

传统中药多为口服用药,其在体内发挥药效的物质可能是中药中的化学成分,还可能是中药成分在体内的代谢产物,中药产生毒副作用和不良反应的原因也可能与中药成分的体内代谢产物有关。而迄今中药成分代谢产物的研究多局限在分析和结构推测鉴定水平,仅对少数可通过简单方法间接获取的代谢产物进行了较深入的生物学研究。因此,代谢产物的获取是深入研究其生物活性,进而明确中药药效和毒效的关键物质基础。本文结合本课题组近几年来的研究成果,对目前中药活性成分体内代谢产物的制备方法进行了总结和综述,希望能够为中药成分的体内代谢研究提供借鉴。     

多次进样循环使用流动相HPLC-DAD法的建立及在快速筛查消渴类中成药中非法添加化学成分中的应用

目的:建立无需更换流动相而可用于中成药中非法添加化学成分现场快速筛查的HPLC-DAD法。方法:采用岛津Shim-pack VP-ODS色谱柱,使用2种等组成的流动相:Ⅰ为乙腈-0.02 mol.L-1甲酸铵缓冲液(pH3.8)(50∶50);Ⅱ为甲醇-0.01 mol.L-1乙酸铵(35∶65)。检测波长229 nm,参比波长550 nm,DAD波长扫描范围190~600 nm。测定时先将样品检出峰的保留时间与对照品比较,找出保留时间一致的疑似物;再将疑似物的光谱与相应的对照品比较。结果:建立的色谱条件可以分离消渴类中成药中常见的10种非法化学添加成分;循环使用流动相进样150次后,除基线稍有提高外,色谱峰分离度和理论塔板数均未明显变化;采用循环使用150次后的流动相成功筛查出未知样品中的非法添加物格列本脲,并得到了LC-MS的确认。结论:将流动相循环使用检测多个样本是可行的,适合现场采用车载或便携式HPLC仪进行快速药品筛查的需要;本文建立的色谱条件可以用于快速筛查消渴类中成药中的非法添加化学成分。     

HPLC analysis of coumarins in Turkish <Emphasis Type="Italic">Seseli</Emphasis> species (Umbelliferae)

The n-hexane extracts of aerial and underground parts of three Seseli species, S. gummiferum subsp. corymbosum, S. resinosum, and S. hartvigii growing in Turkey were investigated concerning the presence of coumarins by normal-phase HPLC (high-performance liquid chromatography). In this respect, two coumarins, osthol (1) and corymbocoumarin (2) [(−)-(3′S, 4′S)-3′-acetoxy-4′-isovaleryloxy-3′, 4′-dihydroseselin] isolated from aerial parts of S. gummiferum subsp. corymbosum, were used as standard samples. The amount of these compounds were discussed and evaluated in this study.     

高效液相色谱法测定2种獐牙菜中龙胆苦苷的含量

目的建立测定川东獐牙菜、显脉獐牙菜中龙胆苦苷含量的高效液相色谱方法。方法HYPERSILBDSC18（4.6mm×200mm,5μm）色谱柱,流动相为甲醇-水梯度洗脱（MeOH%：0min,15%;40min,45%）,流速为1.0mL·min^-1,检测波长为276nm,柱温为25℃。结果龙胆苦苷在0.22～6.50μg线性良好,r=0.99996,平均回收率为96.30%,RSD〈3%（n=6）。结论本方法操作简便,分离效果好,灵敏度高,重现性好,为控制川东獐牙菜和显脉獐牙菜质量提供参考。     

运用Excel建立卡马西平TDM室内质控方法

目的应用Excel电子表格建立HPLC法卡马西平TDM室内质控方法。方法用本实验室建立的卡马西平血药浓度测定方法进行血样及质控品中卡马西平浓度测定,并用Excel电子表格处理质控数据并自动创建室内质控图。结果常规条件下卡马西平已知值质控血清的平均值分别为5.94 mg.L-1,标准差(s)为0.104,相对标准差(RSD)1.8%,3倍RSD值均小于10%,以质控结果超过2S为警告限,超过3S为失控限。结论该方法简单、方便、易于管理,适用于用HPLC法卡马西平TDM的室内质控。     

洛索洛芬钠片中间控制快速检测方法研究

目的建立洛索洛芬钠片中间控制中颗粒含量的快速检测方法。方法采用紫外分光光度法与高效液相色谱法比较。结果在223nm处有最大吸收,其浓度在4.0～24.0μg.mL-1与吸收度线性关系良好;回归方程为C0=30.59A-0.2631;相关系数r=0.999。将紫外检测结果乘以校正因子（K=0.9615）,即C=29.41A-0.2530,与高效液相色谱法比较,两种检测方法结果基本一致。结论紫外分光光度法简便易行、快速、准确、重现性好,可用于洛索洛芬钠片中间控制中颗粒含量的快速检测。     

Combinative method using HPLC fingerprint and quantitative analyses for quality consistency evaluation of an herbal medicinal preparation produced by different manufacturers

A combinative method using HPLC-DAD fingerprint and quantitative analysis was developed and validated for manufacturer-to-manufacturer quality consistency evaluation of Yiqing preparations. For fingerprint analysis, 22 peaks were selected as the characteristic peaks to evaluate the similarities of different samples collected from different manufacturers. The similarities of 12 Yiqing samples were beyond 0.90, indicating that samples from different manufacturers were, to some extent, consistent. Additionally, simultaneous quantification of nine markers including berberine, aloe-emodin, rhein, emodin, chrysophanol, baicalin, baicalein, wogonoside, and wogonin in Yiqing was performed to interpret the quality consistency. The results from the quantitative data showed that the contents of these nine marker compounds were quite consistent for batches produced within one manufacturer and significantly different from manufacturer-to-manufacturer. This study demonstrated that a combination of the chromatographic fingerprint and quantitative analysis offers an efficient way to quality consistency evaluation of herbal preparation.     

Simultaneous determination of 11 active components in two well-known traditional Chinese medicines by HPLC coupled with diode array detection for quality control

A simple and sensitive high-performance liquid chromatography coupled with diode array detection (HPLC-DAD) method was investigated for simultaneous determination of 11 components (chlorogenic acid, coptisine, epiberberine, jatrorrhizine, berberine, palmatine, baicalin, wogonoside, baicalein, wogonin and chrysin) in Qinhuanghouzheng (QHHZ) capsule and Xiaoerqingre (XEQR) tablet, for quality control of these two well-known traditional Chinese medicines (TCMs). The method was established using an Eclipse Plus C₁₈ (150 mm × 4.6 mm i.d., 5 μm) column. The mobile phase comprising methanol (A) 3% phosphoric acid (B) (pH 2.0, adjusted by triethylamine) was used to elute the targets in gradient elution mode. Flow rate and detection wavelength were set at 0.8 mL/min and 270 nm, respectively. All calibration curves showed good linearity with R² > 0.9995. Inter- and intra-day precisions for all investigated components expressed as relative standard deviation (R.S.D.) ranged from 0.26% to 1.77%. Recoveries measured at three concentrations were in the range of 95.0–103.0% with R.S.D. ≤ 3%. The validated method is simple, reliable, and successfully applied to determine the contents of the selected compounds in QHHZ capsule and XEQR tablet for quality evaluation and control. The 11 main active marker compounds measured occur only in 2 or 3 plant species out of 7–10 species comprising the two TCMs. Additional procedures need to be developed for the quality control of plant materials other than Coptis chinensis Franch, Scutellaria baicalensis Georgi and Phellodendron amurense Rupr.