Optimization of TLC analysis of flavonoids and phenolic acids of Helleborus atrorubens Waldst. et Kit

Numerical methods for the evaluation of the separation power of thirteen thin-layer chromatographic systems for splitting a methanolic extract of leaves of Helleborus atrorubens Waldst. et Kit. into 15 compounds (flavonoids and phenolic acids) have been investigated. For this purpose, the following mathematical approaches have been applied: calculation of the information content (I), determination of discriminating power (DP) and formation of clusters and dendrogram. The most suitable chromatographic system for the separation of investigated compounds is ethyl acetate-formic acid-water (65:15:20, v/v/v).     

杭菊叶生长过程中绿原酸类和木犀草苷动态变化分析

目的：建立高效液相色谱（HPLC）法同时测定杭菊叶中5种绿原酸类成分及木犀草苷的方法,并分析不同生长过程杭菊叶中绿原酸类和黄酮类成分的动态变化,比较同期采收的杭菊叶与杭菊花中成分的差异,为杭菊叶的质量控制、合理利用提供参考。方法：采摘不同生长时期的杭菊叶样品及盛花期的杭菊花样品,采用HPLC法对杭菊叶及杭菊花中绿原酸类及黄酮类成分进行检测。结果：不同生长时期杭菊叶中绿原酸类及木犀草苷成分呈不同的规律。新绿原酸、隐绿原酸、异绿原酸B随着生长时间而逐渐累积,到9月含量达到峰值后逐步下降;绿原酸、异绿原酸A则随着成分累积,在9月达到峰值,下降后又在11月出现次峰值。木犀草苷则随着生长过程逐步增加,到11月达到含量峰值。同期采收的杭菊叶中绿原酸类及黄酮类成分高于杭菊花。结论：该研究结果为确定杭菊叶的采收时间及质量测定标准提供了参考和依据,为揭示杭菊叶生长过程中资源性化学成分的变化以及药材品质的控制奠定了一定基础。     

硫磺熏蒸前后金银花的HPLC指纹图谱比较和聚类分析

目的:建立硫磺熏蒸前后金银花HPLC指纹图谱,为评价硫磺熏蒸前后金银花样品的质量优劣提供依据。方法:采用RP-HPLC方法,色谱柱:Pursuit XRs C18(250 mm×4.6 mm,5μm);流动相:乙腈-0.1%磷酸水溶液,梯度洗脱;柱温30℃;检测波长238 nm。结果:建立了硫磺熏蒸前后金银花HPLC指纹图谱共有模式,并指认了其中4个色谱峰。通过与对照品的保留时间比较,4,5,13,14号峰的归属分别为绿原酸、咖啡酸、芦丁、木犀草苷。通过采用相似度计算法结合聚类分析对19批金银花进行比较分析,从而对其质量进行评价。结论:金银花在硫磺熏蒸前后,某些共有成分的含量会有不同程度的改变,硫磺熏蒸对金银花的质量影响较大。     

Development and validation of a dissolution method for warfarin sodium and aspirin combination tablets

A dissolution method for warfarin sodium-aspirin combination tablets was developed which utilizes USP Apparatus 1 (baskets) at 50 rpm with 900 ml of phosphate buffer (pH 6.8; 0.05 M) medium at 37°C. A reversed-phase liquid chromatographic method was also developed for the simultaneous determination of warfarin sodium, aspirin and salicylic acid on an octadecylsilica column using acetonitrile-tetrahydrofuran-glacial acetic acid-water (23:5:5:67, v/v/v/v) as the mobile phase with UV detection at 282 nm. Validation data were obtained which demonstrate that the dissolution methodology is accurate, precise, linear and rugged for the combination tablets.     

Analysis of some steroids by thin-layer chromatography using optimum mobile phases

The aim of this paper was the analysis of five androstane isomers by thin-layer chromatography (TLC). The choice of proper mobile phase and the optimization of the mobile phase composition are very important because the chromatographic separation is difficult to achieve. In the first step, the proper mobile phase system was chosen from seven elution systems presented in the literature using numerical taxonomy method. The proper solvent system was found to be the mixture of chloroform, acetone, and petroleum ether. In the second step the composition of mobile phase was optimized by "simplex" method and "prisma" method. The optimum TLC system can be applied for the separation of androstane isomers from real samples such as drug formulation, biological and natural sources.     

Mass spectrometry-based approaches for the detection of proteins of Staphylococcus species

Mass spectrometry (MS) has become a powerful and popular method to analyze macromolecules from biological systems towards the application of clinical chemistry. Disease markers related to infections can be identified with MS analysis in combination with electrophoresis or chromatographic separations. Further, direct analysis of whole pathogenic bacterial cells (taken directly from a colony) by MS can reveal specific biomarkers that can be used for taxonomy. A brief introduction to the two advanced ionization techniques, electrospray ionization and matrix-assisted laser desorption/ionization, for MS is provided in this review. Sample preparation, separation and MS-related techniques for staphylococcal proteins analysis are summarized. The review is concluded with some current clinical applications of mass spectrometry in the area of biomarker research, vaccine development, diagnosis and strain typing of infectious Staphylococci.     

正交试验优选滁菊总黄酮的提取工艺

目的优选滁菊总黄酮的最佳提取工艺。方法以滁菊总黄酮含量为考察指标,采用正交试验法,对提取过程中乙醇浓度、加醇量、提取时间和提取次数四个影响因素进行研究。结果影响滁菊总黄酮提取的显著因素是加醇量和乙醇浓度。结论以总黄酮含量为考察指标的滁菊最佳提取工艺为用12倍75%乙醇提取2次,每次1 h。     

表面活性剂用于黄酮药物的薄层色谱分离鉴定的研究

The micellar thin layer chromatographic behaviours of rutin, quercetin and morin were studied. The methods of separation and identification were established: Polyamide thin coating was used as the stationary phase and 4% Zeph--acetylacetone--H2O (5:1.5:1.5) used as the mobile phase. The method was applied to the separation and identification of quercetin and rutin in samples of Sophorae flos and satisfactory result was obtained.     

表面活性剂用于黄酮药物的薄层色谱分离鉴定的研究

以表面活性剂的胶束溶液为流动相的胶束色谱技术已引人重视,并已成功地应用于药物的薄层色谱分离鉴定。黄酮类化合物是中草药中广泛存在的一类有效成分。本文以十余种表面活性剂的胶束溶液为流动相研究了槲皮素、桑色素和芦丁的胶束薄层色谱行为,拟出了以4％Zeph—乙酰丙酮—水(5:1.5:1.5)溶液为流动相,聚酰胺薄膜为固定相分离此三种黄酮的层析体系,并对槐花进行分离和鉴定,得到了良好效果。     

中药路路通化学成分的研究

目的研究中药路路通(Fructus liquidambaris)的化学成分。方法用色谱法分离路路通的化学成分,根据理化性质和波谱数据鉴定其结构。结果分离得到11个化合物,分别鉴定为:β-谷甾醇(1),3-oxo-11α,12α-epoxyoleanan-28,13β-olide (2),3-oxo-12α-hydroxy-oleanan-28,13β-olide (3),3α-acetoxyl-25-hydroxyolean-12-en-28-oic acid (4),齐墩果酸(5),熊果酸(6),胡萝卜苷(7)桦木酮酸(8),没食子酸(9),正二十九烷(10),正三十烷酸(11)。结论4为新化合物,3为首次从天然产物中分离得到,5,6,7,9,10和11为首次从路路通中分离得到。     

Iberis amara L. and Iberogast--results of a systematic review concerning functional dyspepsia

A systematic review referring to efficacy and tolerability of the herbal combination Iberogast (Iberis amara planta totalis, Chelidonii herba, Cardui mariae fructus, Melissae folium, Carvi fructus, Liquiritiae radix, Angelicae radix, Matricariae flos, Menthae piperitae folium) was performed in patients with functional dyspepsia. Three placebo-controlled trials and a reference-controlled trial showed a statistical significant and therapeutical relevant reduction of the gastrointestinal symptom-scores in 595 patients. The therapeutic efficacy was also found in one observational study (2267 patients). In accordance with the available evidence Iberogast seems to be an effective phytotherapeutic preparation to reduce the symptoms of dyspepsia yet, without central nervous side effects.     

Simultaneous reversed-phase high-performance liquid chromatographic method for the determination of diosmin,hesperidin and naringin in different citrus fruit juices and pharmaceutical formulations

Diosmin, hesperidin and naringin are flavonoid glycosides that occur naturally in citrus fruits. They exert a variety of pharmacological properties such as anti-inflammatory, antioxidant and free radical scavenging and antiulcer effects and also inhibit selected cytochrome P-450 enzymes resulting in drug interactions. A reversed-phase high-performance liquid chromatographic method has been developed for the simultaneous determination of diosmin, hesperidin and naringin in different citrus fruit juices and pharmaceutical preparations. Diosmin, hesperidin, naringin and the internal standard rhoifolin were separated using tetrahydrofuran/water/acetic acid (21:77:2, v/v/v) as the mobile phase at 34 degrees C, using a C8 reversed-phase column. The method was linear in the 0.25-20.0 microg/ml concentration range for all three flavonoid glycosides (r>0.999). The method has been successfully applied to the determination of all three flavonoid glycosides in several samples of different citrus fruit juices sold in Greece and for the determination of diosmin and hesperidin in pharmaceutical preparations.     

Determination of ambroxol hydrochloride, methylparaben and benzoic acid in pharmaceutical preparations based on sequential injection technique coupled with monolithic column

The porous monolithic columns show high performance at relatively low pressure. The coupling of short monoliths with sequential injection technique (SIA) results in a new approach to implementation of separation step to non-separation low-pressure method. In this contribution, a new separation method for simultaneous determination of ambroxol, methylparaben and benzoic acid was developed based on a novel reversed-phase sequential injection chromatography (SIC) technique with UV detection. A Chromolith SpeedROD RP-18e, 50-4.6 mm column with 10 mm precolumn and a FIAlab 3000 system with a six-port selection valve and 5 ml syringe were used for sequential injection chromatographic separations in our study. The mobile phase used was acetonitrile-tetrahydrofuran-0.05M acetic acid (10:10:90, v/v/v), pH 3.75 adjusted with triethylamine, flow rate 0.48 mlmin(-1), UV-detection was at 245 nm. The analysis time was <11 min. A new SIC method was validated and compared with HPLC. The method was found to be useful for the routine analysis of the active compounds ambroxol and preservatives (methylparaben or benzoic acid) in various pharmaceutical syrups and drops.     

手性流动相HPLC法拆分氟比洛芬对映体研究

目的:建立氟比洛芬手性药物的高效液相色谱拆分方法.方法:利用C18柱,以β-环糊精及其衍生物作为手性流动相添加剂,调节有机修饰剂比例和添加不同量的峰型修饰剂对氟比洛芬对映体进行拆分.结果:采用单6-L-脯氨酸-β-环糊精作为手性流动相添加剂,利用C18柱可直接拆分S,R-氟比洛芬对映体.最佳色谱分离条件为:流动相为1.4％的6 -L - proline -β - CD甲醇溶液(w/v):pH为4.0,体积分数1.0％的三乙胺水溶液(V/V) =25:75(V/V);柱温t=25℃;流速1.0ml/min;进样量10ul,检测波长254nm.S,R-氟比洛芬对映体获得了良好分离,分离度为1.65.结论:建立的手性流动相添加剂法能有效拆分氟比洛芬对映体.     

Preparative separation of high-purity troxerutin and related substances from mother liquor of troxerutin by silica gel column chromatography and semi-preparative liquid chromatography

Troxerutin (TRO) is a mixture of semi-synthetic flavonoids prepared by hydroxyethylation of rutin, and it is commonly used for the treatment of cerebrovascular diseases. The main active ingredient is trishydroxyethyl rutin. The mother liquor of TROcontains a lot of TRO and other derivatives of hydroxyethylated rutin. In order to make full use of the mother liquor of TRO, an efficient method was developed for recovering high-purity TRO from mother liquor of TRO by combining silica gel column chromatography with semi-preparative liquid chromatography. In the silica gel column chromatographic separation, the ratio of silica gel to sample and eluent composition were investigated to obtain optimum separation effect. The results showed that when the ratio of silica gel to sample was 50, and acetone–ethyl acetate–water–glacial acetic acid (10:10:3:1, v/v/v/v) was used as the eluent, the separation effect of TRO and adjacent impurities was good. Moreover, 150 g ofTRO with a purity of 80% could be obtained from 1 kg of mother liquor of TRO by the silica gel column chromatographic separation, and the results were consistent with the quality standard of TRO raw material. Subsequently, the semi-preparative HPLC was performed, and 100 g TRO with a purity of up to 98% (w/w) was obtained. Meanwhile, tetrahydroxyethylrutin and tetrahydroxyethylquercetin with purity greater than 98% were obtained. This work proposed the separation and preparation of TRO with high-purity from the production waste of TRO for the first time, which had certain environmental benefits and economic benefits.     

巴戟天不同炮制品的高效薄层色谱指纹图谱研究

目的以巴戟天药材生品为基础,建立巴戟天不同炮制品的高效薄层色谱(HPTLC)指纹图谱,并比较分析生品的指纹图谱。方法硅胶GF254高效预制薄层板,乙酸乙酯部位以石油醚-乙酸乙酯(5∶5.5)为展开剂,上行展开8 cm,10%硫酸乙醇溶液显色;正丁醇部位以正丁醇-冰醋酸-水(4∶1∶5)上层溶液为展开剂,上行展开8 cm,α-萘酚浓硫酸试剂显色。显色后置紫外光灯365 nm或自然光下检视,得薄层色谱指纹图谱,并进行相关分析。结果巴戟天生品的薄层荧光色谱指纹图谱与巴戟肉(蒸制)、盐巴戟天、制巴戟天(甘草制)图谱比较分析,彼此具有明显的区别,可用于生品与巴戟天炮制品的鉴别。结论通过HPTLC指纹图谱考察,生品巴戟天与巴戟肉、盐巴戟天、制巴戟天的区别较大,应分别制定质量评价的标准。     

Application of first-derivative UV-spectrophotometry, TLC-densitometry and liquid chromatography for the simultaneous determination of mebeverine hydrochloride and sulpiride

Three methods are described for the simultaneous determination of mebeverine hydrochloride (MB) and sulpiride (SU) in combined pharmaceutical tablets. The first method depends on first-derivative ultraviolet spectrophotometry, with zero-crossing measurement method. The first derivative amplitudes at 214.2 and 221.6 nm were selected for the assay of MB and SU, respectively. Calibration graphs follow Beer's law in the range of 10-30 and 2-8 microg/ml(-1), and the linearity was satisfactory (r = 0.9999), for MB and SU, respectively. The second method was based on the application of the thin layer chromatographic separation of both drugs followed by the densitometric measurements of their spot areas. After separation on silica gel GF254 plates, using ethanol: diethyl ether: triethylamine (70:30:1 v/v) as the mobile phase, the chromatographic zones corresponding to the spots of MB and SU were scanned at 262 and 240 nm, respectively. The calibration function was established in the ranges of 4-12 microg for MB and 2-8 microg for SU. The third method was an internal standard procedure based on high performance liquid chromatographic separation of the two drugs on a reversed-phase, Bondapak CN column. The detection was done at 243 nm using buclizine hydrochloride as internal standard. All chromatographic methods showed good linearity, precision and reproducibility. No spectral or chromatographic interference from the tablet excipients were found. The proposed methods were successfully applied to the assay of commercial tablets and content uniformity test. The procedures were rapid, simple and suitable for quality control application.     

Chiral resolution of some piperidine 2, 6-dione drugs by high performance liquid chromatography on Kromasil CHI-DMB column

Kromasil CHI-DMB chiral stationary phase (CSP) is used to resolve some piperidine-2, 6-diones under normal mobile phase mode. Base-line separation was achieved using hexane and dioxane (90 : 10 v/v) for aminoglutethimide (AG), pyridoglutethimide (PG), cyclohexylaminoglutethimide (ChAG), thioglutethimide (TG), and N-acetyl aminoglutethimide (AAG). The chromatographic parameters are reported. The chiral recognition mechanisms between the analytes and the CSP depend primarily on hydrogen bonding and/or (pi-pi) interactions.     

多腺悬钩子黄酮类化学成分研究

目的研究藏药多腺悬钩子的黄酮类成分。方法采用各种柱色谱技术对其成分进行分离纯化,根据理化性质和波谱分析鉴定化合物的结构。结果从中分离得到8个黄酮类化合物,经鉴定分别为:槲皮素(1),5,7,4′-三羟基双氢黄酮(2),木犀草素(3),木犀草素-7-O-β-D-葡萄糖苷(4),5,7,3′-三羟基-8,4′,5′-三甲氧基黄酮(5),香叶木素-7-O-β-D-葡萄糖苷(6),槲皮素-3-O-β-D-葡萄糖基-7-O-α-L-鼠李糖苷(7)和槲皮素3-O-β-D-葡萄糖苷(8)。结论所有化合物均为首次从该植物中分离得到。     

HPLC法拆分氟比洛芬对映体

建立氟比洛芬手性药物的高效液相色谱拆分方法。方法:手性流动相添加剂HPLC法:利用C18柱,以羟丙基-β-环糊精作为手性流动相添加剂,调节有机修饰剂甲醇的比例和添加不同量的三乙胺对氟比洛芬进行拆分;手性固定相HPLC法:利用Chiral-pakAD手性柱,以正己烷-乙腈为流动相基本成分,调整两者不同比例和添加不同量的三乙胺,对氟比洛芬进行拆分。结果:手性流动相添加剂法:使用C18柱对氟比洛芬对映异构体进行拆分,调节流动相中有机修饰剂甲醇浓度、手性流动相添加剂羟丙基环糊精浓度、峰型修饰剂三乙胺的浓度等都不能使氟比洛芬对映体达到基线分离,只能部分分离。手性固定相法:氟比洛芬对映体在Chiral-pakAD手性柱上能达到较好的分离。在正己烷-乙腈流动相系统中,正己烷体积含量为90%,三乙胺体积含量为0.05%的条件下,氟比洛芬对映体得到了较好的分离,分离度为10.0。结论:建立的手性固定相法能有效拆分氟比洛芬对映体而手性流动相添加剂法不能拆分氟比洛芬对映体。