Use of Capillary Zone Electrophoresis and Micellar Electrokinetic Chromatography for Separations of Anthraquinone Derivatives

The behavior of seven hydroxy anthraquinone derivatives was studied by capillary zone electrophoresis and micellar electrokinetic chromatography. The effects of buffer pH (6.5–10.8) and sodium dodecyl sulfate concentration (10–20 mmol L^?1) on the effective mobilities of the analytes and their separation were tested. A comparison of the two optimized separation systems showed that micellar electrokinetic chromatography was superior as it permits separation of all the seven analytes within 15 min, using 15 mmol L^?1 sodium dodecyl sulfate in 10 mmol L^?1 tetraborate buffer, pH 8.5, at a voltage of 20 kV. The calibration curves were linear in the concentration range from 5.0 · 10^?7 to 5.0 · 10^?4 mol L^?1 for most of the analytes, at a detection wavelength of 254 nm. LOD and LOQ values of the analytes were in the ranges of 2.10 · 10^?7–1.28 · 10^?6 mol L^?1and 6.99 · 10^?7–4.25 · 10^?6 mol L^?1, respectively. The proposed separation conditions were applied to determination of 1,2-dihydroxy anthraquinone (alizarin) and 1,2,4-trihydroxy anthraquinone (purpurin) in Rubia tinctorum aglycone and of the recently described 1-acetyl-2,4,5,7-tetrahydroxy-9,10-anthraquinone and 1-acetyl-2,4,5,7,8-pentahydroxy-9,10-anthraquinone in the mycelium of fungi Geosmithia lavendula.     

Propolis Integration Methods into Solutions for Highly Loaded Propolis Fibers by Needleless Electrospinning

A load-bearing matrix filled with biologically active compounds is an efficient method for transporting them to the target location. Bee-made propolis has long been known as a natural product with antibacterial and antiviral, anti-inflammatory, antifungal properties, and anti-oxidative activity. The aim of the research is to obtain stable propolis/PVA solutions and produce fibers by electrospinning. To increase propolis content in fibers as much as possible, various types of propolis extracts were used. As a result of the research, micro- and nano-fiber webs were obtained, the possible use of which have biomedical and bioprotective applications. All used materials are edible and safe for humans, and fiber webs were prepared without using any toxic agent. This strategy overcomes propolis processing problems due to limitations to its solubility. The integration of different combinations of extracts allows more than 73 wt% of propolis to be incorporated into the fibers. The spinning solution preparation method was adapted to each type of propolis, and by combining the methods, solutions with different propolis extracts were obtained. Firstly, the total content of flavonoids in the propolis extracts was determined for the assessment and prediction of bioactivity. The properties of the extracts relevant for the preparation of electrospinning solutions were also evaluated. Secondly, the most appropriate choice of PVA molecular weight was made in order not to subject the propolis to too high temperatures (to save resources and not reduce the bioactivity of propolis) during the solution preparation process and to obtain fibers with the smallest possible diameter (for larger surface-to-volume ratios of nanofibers and high porosity). Third, electrospinning solutions were evaluated (viscosity, pH, conductivity and density, shelf life) before and after the addition of propolis to predict the maximum propolis content in the fibers and spinning stability. Each solution combination was spun using a cylindrical type electrode (suitable for industrial production) and tested for a stable electrospinning process. Using adapted solution-mixing sequences, all the obtained solutions were spun stably, and homogeneous fibers were obtained without major defects.     

毛细管区带电泳法测定3种香豆素的离解常数

采用毛细管区带电泳法(CZE)测定瑞香苷(DN)、瑞香素(DP)和7-羟基香豆素(7-HC)的离解常数(p Ka).通过测试不同酸度条件下各香豆素的迁移时间,以香豆素在电泳中有效淌度和运行液中OH-浓度的关系,推导了毛细管区带电泳-紫外分光光度(CZE-UV)法测定3种香豆素离解常数(p Ka)的线性模型,并建立了这3种香豆素的p Ka测定方法.测得3种香豆素的p Ka值分别是7.43(7-HC)、6.91(DN)和6.86(DP).同时也采用传统方法 -紫外分光光度法对这些物质的p Ka进行测定以验证CZE的可靠性,两种测定结果相一致,说明CZE-UV法用于3种香豆素的离解常数的测定,快速、简便、结果可靠.     

高效毛细管电泳对克伦特罗对映体的分离及定量检测

以羟丙基-β-环糊精(HP-β-CD)作为手性添加剂,采用毛细管区带电泳(CZE)成功分离了克伦特罗(Clenbuterol,Cle)对映体.研究了β-CD种类与浓度,缓冲液pH值及浓度,操作温度等对分离的影响.结果表明,以30 mmol/L的HP-β-CD为手性添加剂,50 mmol/L的磷酸盐缓冲溶液(pH 2.5)为缓中液,分离电压24 kV,操作温度20℃,可使Cle对映体实现基线分离,其分离度为6.78.对拆分机理也进行了探讨,测定了HP-β-CD与两对映体的结合常数及热力学参数;对CZE定量能力(线性,精度)进行了考察,R和S型的线性相关系数都大于0.998,两者的相对标准偏差(RSD)均低于2.2%.     

高效毛细管电泳,激光干涉检测快速分离二糖研究

首次报道一种高效毛细管电泳、激光干涉折射检测快速分离二糖的新方法。详细研究了四硼酸钠溶液中，ｐＨ值、四硼酸钠浓度及有机添加剂对二糖－硼酸络离子迁移行为的影响，发现在ｐＨ为９．７的０．１ｍｏｌ／Ｌ四硼酸钠溶液中，１０ｍｉｎ内４种二糖得到基线分离。采用自制激光干涉型折射指数检测器，不需对二糖衍生化处理，可直接进行在柱检测。     

砷酸根和硒酸根的毛细管电泳分离

在铬酸盐和硼酸盐溶液中,以十六烷基三甲基溴化铵(CTAB)为电渗流改性剂,通过添加有机添加剂甲醇,成功地分离了砷酸根、硒酸根及常见酸根离子。优化分离条件发现,采用210 nm直接检测砷酸根比在260 nm处间接检测其检出限更低,而且可以减少大量C l-的干扰。迁移时间重复性的相对标准偏差(RSD)不大于3.0%,当信噪比(S/N)为2时,在260 nm处,HAsO42-与SeO42-的检出限分别为0.019 mg/L和0.078mg/L;在210 nm处,HAsO42-检出限可达0.004 3 mg/L。加标回收率均在95%~108%之间,满足定量要求。但NO3-会干扰HAsO42-的定量分析。     

Capillary Zone Electrophoresis of some organic acids in milk whey

This paper describes a method for analysing some acids of milk whey by Capillary Zone Electrophoresis. After eliminating the whey proteins by ultrafiltration, the whey underwent electrophoretic separation in the presence of anodic electroosmotic flow. The following analytes were detected: citric, orotic, uric, and hippuric acids. A procedure is described for sample preparation and the operating conditions for electrophoretic capillary separation established. Finally, orotic acid is quantitatively determined.     

Characterization of the Interaction between Bovine Serum Albumin and Lomefloxacin by Capillary Zone Electrophoresis

Three capillary zone electrophoresis (CZE) methods of the frontal analysis (FA), vacancypeak (VP) and simplified Hummel-Dreyer (SHD) were applied to investigate interaction betweenbovine serum albumin (BSA) and lomefloxacin, the experimental condition was established after alarge number of tests. Based on the site-binding model, the binding parameters were measuredaccording to the site model by Scatchard.     

毛细管区带电泳法测定卷烟中的生物碱

在磷酸缓冲体系中采用毛细管区带电泳法测定卷烟中的生物碱时,检测灵敏度低,分离度差。考察了卷烟中生物碱的 提取条件,分离缓冲溶液的类型、pH值和浓度,卷烟中生物碱测定方法的线性范围、检出限、重现性和回收率。结果发 现,当采用410 mmol/L的酒石酸溶液（pH 2.8）为缓冲体系时,卷烟中生物碱的检测灵敏度和分离度均有明显改善,烟碱 的线性范围为0.06~0.80 mg/L（其他生物碱为0.006~0.10 mg/L）,检出限为0.002~0.01 mg/L,相对标准偏差为2.2%~10%,回收率为87.6%~102%。     

Prediction of Migration Times of Organic Ions in Capillary Zone Electrophoresis

The electrophoretic mobility ratio (R value) of any two ions is constant and independent of the capillary type and electrophoretic conditions if their electrical charges and hydration radii are constant. The use of strong acid salts and quaternary ammonium salts is therefore proposed for the determination of R values. Such analytes are called markers. The following determinations can be carried out: (i) the determination of the migration time corresponding to the electroosmotic flow (EOF) in any capillary under any electrophoretic condition by measuring the migration times of two markers in the condition studied (useful when the EOF is weak); (ii) the determination of the migration time of an analyte in any capillary by knowing the migration time of the markers in the capillary studied. If the pH is changed and the ionization of the analyte is pH dependent, the resulting migration time for the analyte can be calculated. The constancy of the mobility ratios of seven markers was checked experimentally at eight different pH values (between pH 3 and 10), at three temperatures, and for two buffer concentrations. The predicted and experimental migration times were also compared in two different types of capillaries.     

Potential Use of Propolis in Phytocosmetic as Phytotherapeutic Constituent

Phytocosmetic is an important aspect of traditional medicine in several cultures. Researchers are now focusing to find new and effective ingredients of natural origin. Propolis is a natural beehive product extensively used in traditional medicine. We aimed in the present study to investigate the potential use of propolis as an aesthetic and phytotherapeutic constituent in phytocosmetics. Propolis was extracted using 80% ethanol. Total phenolic and flavonoid contents were determined calorimetrically. Free radical scavenging ability and reducing capacity were evaluated using four assays and expressed as IC₅₀ values. Antibacterial activity was evaluated by the determination of minimum inhibitory concentration (MIC) on 11 Gram-positive and Gram-negative bacteria. The wound healing activity of 30% ethanolic extract and propolis ointment was studied using excision wounds in the anterio-dorsal side of the rats. The phenolic acid composition of the tested propolis was investigated using UFLC/MS-MS analysis. The tested propolis was rich in phenolic and flavonoid content and demonstrated an interesting antibacterial and antioxidant activity. Wounds treated with propolis appear to display a lesser degree of inflammation. Chemical analysis led to the identification of 11 phenolics. Among them, five are considered as main compounds: Chlorogenic acid (48.79 ± 5.01 ng/mL), Gallic acid (44.25 ± 6.40 ng/mL), Rutin (21.12 ± 3.57 ng/mL), Caffeic acid (28.19 ± 4.95 ng/mL), and trans-cinnamic acid (20.10 ± 6.51 ng/mL). Our results indicated that propolis can not only be used as a cosmetic ingredient but also be used as a preventative and curative constituent, which might be used as a barrier when applied externally on infected and non-infected skin.     

Propolis Efficacy: The Quest for Eco-Friendly Solvents

Propolis, a natural product made by bees with resins and balsams, is known for its complex chemical composition and remarkable bioactivities. In this study, propolis extraction was studied seeking extracts with strong bioactivities using less orthodox solvents, with some derived from apiary products. For that, a propolis sample collected from Gerês apiary in 2018 (G18) was extracted by maceration with six different solvents: absolute ethanol, ethanol/water (7:3), honey brandy, mead, propylene glycol and water. The solvent influence on the chemical composition and antioxidant and antimicrobial activities of the extracts was investigated. Antioxidant potential was assessed by the DPPH free-radical-scavenging assay and the antimicrobial activity by the agar dilution method. Chemical composition of the extracts was determined in vitro by three colorimetric assays: total ortho-diphenols, phenolics and flavonoids contents and the LC-MS technique. To our knowledge, this is the first time that solvents such as honey brandy and mead have been studied. Honey brandy showed considerable potential to extract propolis active compounds able to inhibit the growth of bacteria such as the methicillin-sensitive Staphylococcus aureus and Propionibacterium acnes (MIC values of 100 and 200 µg/mL, respectively) and the fungi Candida albicans and Saccharomyces cerevisiae (MIC = 500 µg/mL, for both). Mead extracts displayed high antioxidant capacity (EC₅₀ = 1.63 ± 0.27 µg/mL) and great activity against resistant bacteria such as the methicillin-resistant Staphylococcus aureus and Escherichia coli (MIC = 750 µg/mL, for both). The production of such solvents made from beehive products further promotes a diversification of apiary products and the exploration of new applications using eco-friendly solutions.     

Analysis of Flavonoids and Phenolic Acid in Propolis by Capillary Electrophoresis

A high-performance capillary electrophoresis (CE) is developed for the determination of six ingredients – rutin, ferulic acid, apigenin, luteolin, quercetin and caffeic acid – in Propolis and its extract capsules in this work. The effects of several factors such as the acidity and concentration of the running buffer, the separation voltage, the injection time and the wavelength of UV detector were investigated to find the optimum conditions. Under the optimum conditions, the analytes were separated within 14 min in a borax buffer (pH 9.2). Notably, excellent linearity was obtained over two orders of magnitude with detection limits (S/N=3) ranged from 3.7 × 10^–7 to 1.8 × 10^–6 g mL^–1 for all six analytes. The relative standard deviations of the migration times of six constituents ranged between 1.06% and 1.53%. The recoveries of six constituents ranged from 94.9% to 108.4%. This method was successfully used in the analysis of Propolis and its extract capsules with a relatively simple extraction procedure, and the assay results were satisfactory.     

A Computer Method for Predicting the Electrophoretic Mobility of Peptides

A computer model is presented for the prediction of the electrophoretic mobilities of peptides from physical constants derived from their amino acid sequences. The model assumes that the electrophoretic mobility can be represented by a product of four functions according to the relation:  l_comp= l(L)w(W)q(Q)c(CC), where L (a length parameter) is represented by the number of amino acid residues of the peptide, W (a width parameter) is represented by the average residue mass, Q = the charge of the peptide, and CC = the position of the center of charge relative to the center of mass. The model was used to calculate the electrophoretic mobilities of peptides in a 50 mM phosphate buffer at pH 2.5. Sixty-four test peptides ranging in size from 2 to 39 amino acid residues were used for this study. The calculated mobilities show excellent correlation with experimental measurements with a correlation coefficient greater than 0.98.     

Activity of Compounds from Temperate Propolis against Trypanosoma brucei and Leishmania mexicana

Ethanolic extracts of samples of temperate zone propolis, four from the UK and one from Poland, were tested against three Trypanosoma brucei strains and displayed EC₅₀ values < 20 µg/mL. The extracts were fractionated, from which 12 compounds and one two-component mixture were isolated, and characterized by NMR and high-resolution mass spectrometry, as 3-acetoxypinobanksin, tectochrysin, kaempferol, pinocembrin, 4′-methoxykaempferol, galangin, chrysin, apigenin, pinostrobin, cinnamic acid, coumaric acid, cinnamyl ester/coumaric acid benzyl ester (mixture), 4′,7-dimethoxykaempferol, and naringenin 4′,7-dimethyl ether. The isolated compounds were tested against drug-sensitive and drug-resistant strains of T. brucei and Leishmania mexicana, with the highest activities ≤ 15 µM. The most active compounds against T. brucei were naringenin 4′,7 dimethyl ether and 4′methoxy kaempferol with activity of 15–20 µM against the three T. brucei strains. The most active compounds against L. mexicana were 4′,7-dimethoxykaempferol and the coumaric acid ester mixture, with EC₅₀ values of 12.9 ± 3.7 µM and 13.1 ± 1.0 µM. No loss of activity was found with the diamidine- and arsenical-resistant or phenanthridine-resistant T. brucei strains, or the miltefosine-resistant L. mexicana strain; no clear structure activity relationship was observed for the isolated compounds. Temperate propolis yields multiple compounds with anti-kinetoplastid activity.     

胶束毛细管电泳法同时分离四环素与青霉素类药物的研究

采用胶束毛细管电泳(MEKC),建立了四环素(TCs)和青霉素(PENs)两类7种药物同时分离的方法。考察了MEKC中缓冲液类型、离子浓度和pH值,以及表面活性剂(SDS)浓度、分离电压、温度等参数的影响。利用L16(45)正交试验,确立了最佳的电泳条件:缓冲液为40 mmol/L磷酸二氢钾-20 mmol/L硼砂,添加65 mmol/L SDS,pH 7.9,分离电压28 kV,分离温度28℃,紫外检测波长分别为350 nm和200nm。结果表明:7种药物在25 min内得到完全分离。在1.56~50 mg/L范围内呈良好的线性关系,相关系数(r2)为0.997 9~0.999 9,峰面积的相对标准偏差(n=6)为4.1%~7.3%;迁移时间的相对标准偏差(n=6)为0.33%~0.67%。在2.0,5.0,10.0 mg/kg的加标水平下,7种药物的回收率为83.6%~93.3%,相对标准偏差(n=6)为4.7%~7.6%。该法快速、简便、准确,具有较高的灵敏度,已应用于合肥市及周边地区水塘和湖水中7种药物的快速分离检测。     

Application of High Temperature High Resolution Gas Chromatography to Crude Extracts of Propolis

The underivatized acetone and hexane fractions from propolis samples (predominant flora Citrus spp. and Vernonia polyanthes) were analyzed by HT-HRGC (high temperature high resolution gas chromatography) and HT-HRGC coupled to mass spectrometry (HT-HRGC-MS). Several compounds, including flavonoid aglycones, phenolic acids, and high molecular weight compounds were characterized in crude extracts by HT-HRGC-MS. HT-HRGC and HT-HRGC-MS were shown to be quick and informative tools for rapid analysis of crude extracts without need for prior derivatization and purification.     

Chemical Variability and Pharmacological Potential of Propolis as a Source for the Development of New Pharmaceutical Products

This review aims to analyze propolis as a potential raw material for the development and manufacture of new health-promoting products. Many scientific publications were retrieved from the Scopus, PubMed, and Google Scholar databases via searching the word "propolis". The different extraction procedures, key biologically active compounds, biological properties, and therapeutic potential of propolis were analyzed. It was concluded that propolis possesses a variety of biological properties because of a very complex chemical composition that mainly depends on the plant species visited by bees and species of bees. Numerous studies found versatile pharmacological activities of propolis: antimicrobial, antifungal, antiviral, antioxidant, anticancer, anti-inflammatory, immunomodulatory, etc. In this review, the composition and biological activities of propolis are presented from a point of view of the origin and standardization of propolis for the purpose of the development of new pharmaceutical products on its base. It was revealed that some types of propolis, especially European propolis, contain flavonoids and phenolic acids, which could be markers for the standardization and quality evaluation of propolis and its preparations. One more focus of this paper was the overview of microorganisms’ sensitivity to propolis for further development of antimicrobial and antioxidant products for the treatment of various infectious diseases with an emphasis on the illnesses of the oral cavity. It was established that the antimicrobial activity of different types of propolis is quite significant, especially to Gram-negative bacteria and lipophilic viruses. The present study could be also of interest to the pharmaceutical industry as a review for the appropriate design of standardized propolis preparations such as mouthwashes, toothpastes, oral drops, sprays, creams, ointments, suppositories, tablets, and capsules, etc. Moreover, propolis could be regarded as a source for the isolation of biologically active substances. Furthermore, this review can facilitate partially overcoming the problem of the standardization of propolis preparations, which is a principal obstacle to the broader use of propolis in the pharmaceutical industry. Finally, this study could be of interest in the area of the food industry for the development of nutritionally well-balanced products. The results of this review indicate that propolis deserves to be better studied for its promising therapeutic effects from the point of view of the connection of its chemical composition with the locality of its collection, vegetation, appropriate extraction methods, and standardization.