Application of flavonoids – quercetin and rutin – as new matrices for matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometric analysis of Pt(II) and Pd(II) complexes

Attempts are being made to overcome the resistance of tumour cells to platinum (Pt) drugs by the synthesis of new generations of Pt complexes, and it is important to find appropriate and simple methods for the characterization of those novel complexes. The additional applicability of such a method for the analysis of the interactions of metal complexes with biomolecules would be advantageous. Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS) seems to possess the capability to become this method of choice, since it could be applied to low‐mass complexes as well as for the analysis of large biomolecules. In this work the applicability of flavonoids – quercetin and rutin – as matrices for MALDI‐TOFMS analysis of dichlorido(ethylendiamine)platinum(II) ([PtCl₂(en)]), dichlorido(diaminocyclohexane)platinum(II) ([PtCl₂(dach)]) and chloride (diethylenetriamine) palladium(II) chloride ([PdCl(dien)]Cl) complexes is demonstrated. Spectra of Pt(II) and Pd(II) complexes recorded in the presence of quercetin and rutin are rather simple: Pt(II) complexes generate [M+Na]⁺ or [M+K]⁺ions, whereas the investigated Pd(II) complex gives ions generated by the loss of one Cl^? or HCl. Flavonoids give a relatively small number of well‐defined ions in the low‐mass region (at m/z 303.3 for quercetin and m/z 633.5 for rutin). Quercetin and rutin can be applied in much lower concentrations than other common MALDI matrices and require rather low laser intensity. We speculate that flavonoids stabilize the structures of the metal complexes and that they may be useful for the analysis of other biologically active metal complexes, thus implying their broader applicability. Copyright © 2009 John Wiley & Sons, Ltd.     

Cyclic chronopotentiometric determination of sugars at Au and Pt microelectrodes in flowing solutions

The main advantage of the application of cyclic chronopotentiometry (CCP) in end-column CE detection arises from the fact that the detection parameters and the magnitude of the analytical signal are (in contrast with other electrochemical detection methods) independent of the ohmic polarization of the solution caused by the separation current at the detection end of the capillary. CCP was used to determine sugars on platinum and gold microelectrodes after separation by CE. The results obtained with a gold microelectrode were better. Subsequently this detection method was used for quantitative determination of sugars in honeys and for their authentication.     

Dependence of the solubility of natural flavonoids in water on the concentration of miramistin,polyvinylpyrrolidone, and human serum albumin

In organized media of the cationic surfactant miramistin and the polymers polyvinylpyrrolidone and human serum albumin, the solubility of natural flavonoids quercetin and rutin increased by one or two orders of magnitude. The increase was more significant for hydrophobic quercetin than for hydrophilic rutin. The solubility also depended on the structure and self-organization of molecules in organized media and the site of flavonoids in them. The calculated binding constants increased in the series polyvinylpyrrolidone < miramistin < human serum albumin.     

Capillary Electrophoresis Chemiluminescence for the Analysis of Flavonoids in Pharmaceuticals and Human Plasma

A capillary electrophoresis (CE)-chemiluminescence (CL) method has been developed for the determination of the pharmacologically active flavonoids, including rutin and quercetin in pharmaceuticals and human plasma containing Ginkgo biloba leaves extract (EGb). The separation was conducted in borate buffer and luminol. The post-column CL reagent was K₃Fe(CN)₆ in NaOH medium. Rutin and quercetin were baseline separated within 10 min with detection limits of 1.0 and 5.0 nM, respectively. The maximum intra- and inter-day relative standard deviations (RSD) of migration time of analytes were <3.5 %. Moreover, the high selectivity of the CL detection and the high-separation efficiency of CE render the method the potential of quick analyzing the pharmacologically active substances in complex matrix with satisfactory results.

     

Capillary Electrophoresis Chemiluminescence for the Analysis of Flavonoids in Pharmaceuticals and Human Plasma

A capillary electrophoresis (CE)-chemiluminescence (CL) method has been developed for the determination of the pharmacologically active flavonoids, including rutin and quercetin in pharmaceuticals and human plasma containing Ginkgo biloba leaves extract (EGb). The separation was conducted in borate buffer and luminol. The post-column CL reagent was K₃Fe(CN)₆ in NaOH medium. Rutin and quercetin were baseline separated within 10 min with detection limits of 1.0 and 5.0 nM, respectively. The maximum intra- and inter-day relative standard deviations (RSD) of migration time of analytes were <3.5 %. Moreover, the high selectivity of the CL detection and the high-separation efficiency of CE render the method the potential of quick analyzing the pharmacologically active substances in complex matrix with satisfactory results.     

Use of molybdate as novel complex-forming selector in the analysis of polyhydric phenols by capillary zone electrophoresis

Molybdate was examined as a complex-forming additive to the CE background electrolytes (BGE) to affect the selectivity of separation of polyhydric phenols such as flavonoids (apigenin, hyperoside, luteolin, quercetin and rutin) and hydroxyphenylcarboxylic acids (ferulic, caffeic, p-coumaric and chlorogenic acid). Effects of the buffer concentrations and pH and the influence of molybdate concentration on the migration times of the analytes were investigated. In contrast to borate (which is a buffering and complex-forming agent generally used in CE at pH ≥9) molybdate forms more stable complexes with aromatic o-dihydroxy compounds and hence the complex-formation effect is observed at considerably lower pH. Model mixtures of cinnamic acid, ferulic acid, caffeic acid and 3-hydroxycinnamic acid were separated with 25 mM morpholinoethanesulfonic acid of pH 5.4 (adjusted with Tris) containing 0.15 mM sodium molybdate as the BGE (25 kV, silica capillary effective length 45 cm × 0.1 mm I.D., UV-vis detection at 280 nm). With 25 mM 2-hydroxy-3-[4-(2-hydroxyethyl)-1-piperazinyl]propanesulphonic acid/Tris of pH^* 7.4 containing 2 mM sodium molybdate in aqueous 25% (v/v) methanol as the BGE mixtures of all the above mentioned flavonoids, p-coumaric acid and chlorogenic acid could be separated (the same capillary as above, UV-vis detection at 263 nm). The calibration curves (analyte peak area versus concentration) were rectilinear (r > 0.998) for ≈8-35 μg/ml of an analyte (with 1-nitroso-2-naphthol as internal standard). The limit of quantification values ranged between 1.1 mg l⁻¹ for p-coumaric acid and 2.8 mg l⁻¹ for quercetin. The CE method was employed for the assay of flavonoids in medicinal plant extracts. The R.S.D. values ranged between 0.9 and 4.7% (n = 3) when determining luteolin (0.08%) and apigenin (0.92%) in dry Matricaria recutita flowers and rutin (1.03%) and hyperoside (0.82%) in dry Hypericum perforatum haulm. The recoveries were >96%.     

Carbon nanotube/polystyrene composite electrode for microchip electrophoretic determination of rutin and quercetin in Flos Sophorae Immaturus

In this report, carbon nanotube/polystyrene (CNT/PS) composite electrodes have been fabricated as sensitive amperometric detectors of microchip capillary electrophoresis (CE) for the determination of rutin and quercetin in Flos Sophorae Immaturus. The composite electrode was fabricated on the basis of the in situ polymerization of a mixture of CNT and styrene in the microchannel of a piece of fused silica capillary under heat. The surface morphologies of the composite in the electrodes were observed by using a scanning electron microscope. The performance of this unique system has been demonstrated by separating and detecting rutin and quercetin. The new CNT-based CE detector offered significantly lower detection potentials, yielded substantially enhanced signal-to-noise characteristics, and exhibited resistance to surface fouling and hence enhanced stability. It demonstrated long-term stability and reproducibility with a relative standard deviation of less than 5% for the peak current (n = 20) and should also find a wide range of applications in conventional CE, flowing injection analysis, and other microfluidic analysis systems.     

Quantitative HPLC determination of main flavonoid content of <Emphasis Type="Italic">Rhododendron adamsii</Emphasis> leaves and stems

The content of main flavonoids from Rhododendron adamsii R. leaves and stems was determined quantitatively using HPLC. It was found that myricetin and quercetin dominated the identified compounds (myricetin, quercetin, dihydroquercetin, rutin) in leaves; dihydroquercetin, in stems (1.1, 1.0, and 2.5 mass% of raw material, respectively). Dihydroquercetin and rutin were found for the first time in R. adamsii. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 26–29, January–February, 2009.     

Fast batch injection analysis of H(2)O(2) using an array of Pt-modified gold microelectrodes obtained from split electronic chips

A fast and robust analytical method for amperometric determination of hydrogen peroxide (H(2)O(2)) based on batch injection analysis (BIA) on an array of gold microelectrodes modified with platinum is proposed. The gold microelectrode array (n=14) was obtained from electronic chips developed for surface mounted device technology (SMD), whose size offers advantages to adapt them in batch cells. The effect of the dispensing rate, volume injected, distance between the platinum microelectrodes and the pipette tip, as well as the volume of solution in the cell on the analytical response were evaluated. The method allows the H(2)O(2) amperometric determination in the concentration range from 0.8 μmolL(-1) to 100 μmolL(-1). The analytical frequency can attain 300 determinations per hour and the detection limit was estimated in 0.34 μmolL(-1) (3σ). The anodic current peaks obtained after a series of 23 successive injections of 50 μL of 25 μmolL(-1) H(2)O(2) showed an RSD<0.9%. To ensure the good selectivity to detect H(2)O(2), its determination was performed in a differential mode, with selective destruction of the H(2)O(2) with catalase in 10 mmolL(-1) phosphate buffer solution. Practical application of the analytical procedure involved H(2)O(2) determination in rainwater of S?o Paulo City. A comparison of the results obtained by the proposed amperometric method with another one which combines flow injection analysis (FIA) with spectrophotometric detection showed good agreement.     

高效液相色谱法测定不同产地枇杷叶中的3种黄酮类成分

建立了高效液相色谱同时测定枇杷叶中3种黄酮类成分的分析方法。该方法分析了不同产地枇杷叶中芦丁、槲皮素和山柰酚的含量差异。枇杷叶粉末用甲醇超声提取后,加盐酸回流,制备样品测试液。采用Diamonsil C18色谱柱（250 mm×4.6 mm,5 μm）,以0.4%（v/v）磷酸水溶液-乙腈为流动相,梯度洗脱。分别对7个不同产地的枇杷叶样品中的芦丁、槲皮素和山柰酚进行测定。结果表明,芦丁、槲皮素、山柰酚在各自的质量浓度范围内线性关系良好（r>0.99）,加标回收率分别为96.33%、95.81%和95.80%,RSD分别为6.48%、0.90%和3.02%。该方法操作简单、分离度好、重复性高。不同产地枇杷叶中3种黄酮类成分的含量存在差异,其中芦丁的差异最大,而山柰酚的含量最稳定且在不同产地样品中均可检出,或可用作枇杷叶药材质量控制的标志成分。     

液相色谱法同时测定维药蜀葵花中芦丁、槲皮素和山柰酚

维药是祖国医药学不可分割的组成部分。维药现代化,即利用现代技术研究维药的有效成分,是维药科学化、标准化、规范化、商品化和产业化的必经之路。本文建立了维药蜀葵花中有效成分芦丁、槲皮素和山柰酚的选择性提取方法,优化了高效液相色谱法(HPLC)同时测定这3种有效成分的分析条件。采用HC-C₁₈色谱柱(250 mm×4.6 mm, 5 μm)和甲醇-0.4%磷酸(50:50, v/v)流动相,在柱温30 ℃和流速1.00 mL/min的条件下实现了3种物质之间以及和干扰物之间的基线分离。维药蜀葵花中芦丁、槲皮素及山柰酚的线性范围分别为12.5~150 μg/mL (r=0.9998), 12.5~125 μg/mL (r=0.9999)及12.5~125 μg/mL (r=0.9988),加标回收率(n=5)分别为100.3%(RSD=1.1%)、97.60%(RSD=0.47%)、97.75%(RSD=0.71%)。该方法实现了同时测定维药蜀葵花中芦丁、槲皮素及山柰酚,为其他黄酮类物质的开发应用提供了科学依据,同时也可为其他维药分析提供借鉴。     

聚乙烯醇接枝微粒对黄酮类物质的吸附特性

以硅胶表面接枝有聚乙烯醇(PVA)的接枝微粒PVA/SiO₂为固体吸附剂, 对槲皮素和芦丁2种黄酮类化合物进行了吸附研究, 结果表明, 由于接枝微粒PVA/SiO₂表面含有高密度的羟基, 使得该接枝微粒与黄酮类化合物分子之间可形成多位点的常规氢键和π型氢键, 溶剂的竞争吸附对黄酮化合物的吸附容量产生很大的影响. 以弱极性的1,2-二氯乙烷(DCE)为溶剂时, 几乎不存在溶剂的竞争吸附, 槲皮素和芦丁具有最高的吸附容量, 分别为0.32 mmol/g(96 mg/g)和0.23 mmol/g(140 mg/g); 而在质子溶剂乙醇中, 强烈的溶剂竞争吸附使两者的吸附容量降至0.22 mmol/g(65 mg/g)和0.14 mmol/g(87 mg/g). 升高温度会减弱氢键作用, 甚至使氢键断裂, 导致吸附容量减小. 质子溶剂中电解质的存在, 对吸附作用产生负性影响. 功能微粒PVA/SiO₂对黄酮类化合物的吸附为放热过程, 且为焓驱动的吸附过程.     

Microsequential injection analysis: Determination of rutin and quercetin in food supplements and pharmaceutical products

Conditions for the oxidative coupling of rutin and quercetin with 3-methylbenztiazolinon-2-hydrazone have been determined under the conditions of microsequential flow analysis (‘lab-on-a-valve’). To provide the high sensitivity of the determination, methods aimed at improving the overlapping of the zones in the flow have been utilized; these are stopped flow and injection of a ‘spacer’ zone. Good results have been obtained using a new approach, the injection of a ‘spacer’ zone with a rate gradient. A program of operations for the automatic spectrophotometric determination of rutin and quercetin has been composed. The analytical range is (6–50) × 10⁻⁵ M for the determination of rutin and (5–50) × 10⁻⁵ M for the determination of quercetin, the productivity of analysis being 59 samples per hour. The procedure has been applied to the analysis of plant extracts and pharmaceutical product Askorutin.     

Voltammetric examinations of ferrocene on microelectrodes and microarrayelectrodes

Voltammetric investigations of ferrocene in organic media with planar, disk and band shaped microelectrodes (ME) and microarrayelectrodes (MAE) were carried out in stationary solutions and under flow-through conditions with flow-injection analysis (FIA). The electrodes were embedded in cylindrical electrode bodies made of epoxy resin. During the FIA experiments miniaturized flow-through cells with a three electrode configuration (wall-jet, thin-layer principle) were used. As detectors microelectrodes and microarrayelectrodes consisting of platinum wires (radius: 10 μm), gold wires (radius: 12.5 μm), carbon fibres (radius: 3.5 μm) and gold foils (foil thickness: 30 μm) were constructed using fine precision manufacturing methods by sealing the electrode materials in insulating resins.     

Flow injection measurements of <Emphasis Type="Italic">S</Emphasis>-nitrosothiols species in biological samples using amperometric nitric oxide sensor and soluble organoselenium catalyst reagent

A novel flow injection analysis (FIA) system suitable for measurement of S-nitrosothiols (RSNOs) in blood plasma is described. In the proposed (FIA) system, samples and standards containing RSNO species are injected into a buffer carrier stream that is mixed with the reagent stream containing 3,3′-dipropionicdiselenide (SeDPA) and glutathione (GSH). SeDPA has been shown previously to catalytically decompose RSNOs in the presence of a reducing agent, such as GSH, to produce nitric oxide (NO). The liberated NO is then detected downstream by an amperometric NO sensor. This sensor is prepared using an electropolymerized m-phenylenediamine (m-PD)/resorcinol and Nafion composite films at the surface of a platinum electrode. Using optimized flow rates and reagent concentrations, detection of various RSNOs at levels in the range of 0.25–20 μM is possible. For plasma samples, detection of background sensor interference levels within the samples must first be carried out using an identical FIA arrangement, but without the added SeDPA and GSH reagents. Subtraction of this background sensor current response allows good analytical recovery of RSNOs spiked into animal plasma samples, with recoveries in the range of 90.4–101.0%.     

Investigation of Chemiluminescence Behavior of Flavonoids with Cerium (IV)‐Rhodamine B System

Abstract

The chemiluminescence (CL) behavior of five major flavonoid types in cerium (IV)‐rhodamine B system was investigated by flow‐injection. Strong CL was observed when cerium (IV) reacted with rhodamine B in sulfuric acid medium in the presence of flavonoids. This reaction system has been established as a simple, rapid, and highly sensitive flow injection CL analysis for quercetin and kaempferol, and their detection limit (3σ) was 2.7 and 0.22 nmol/L, respectively. The relative standard deviation (n=8) was 1.2% for 1.0 µmol/L quercetin and 1.9% for 0.5 µmol/L kaempferol. This method was successfully applied to the determination of quercetin in the hydrolysate of rutin and compared well with the high performance liquid chromatography (HPLC) method. From a comparison of several related flavonoids, it was concluded that only flavonoids that contain a free 3‐hydroxyl and 2, 3‐double bond in conjugation with 4‐oxo function could produce a relatively strong CL emission.     

Fast and simultaneous detection of prominent natural antioxidants using analytical microsystems for capillary electrophoresis with a glassy carbon electrode: a new gateway to food environments

This paper examines for the first time the analytical possibilities of fast and simultaneous detection of prominent natural antioxidants including examples of flavonoids and vitamins using a CE microchip with electrochemical detection (ED). Unpinched injection conditions, zone electrophoretic separation and amperometric detection were carefully assayed and optimised. Analysis involved the zone electrophoretic separation of arbutin, (+)-catechin and ascorbic acid in less than 4 min using a borate buffer (pH 9.0, 50 mM), employing 2 kV as the separation voltage and +1.0 V as the detection potential. In addition, the separation of different 'couples' of natural antioxidants of food significance including (+)-catechin and ascorbic acid, (+)-catechin and rutin, as well as arbutin and phlorizdin is proposed. To demonstrate the potential and future role of CE microsystems, analytical possibilities and a new route in the raw sample analysis are presented. The preliminary results obtained allow the proposal of CE-ED microchips as a real gateway to microanalysis in foods.     

类黄酮化合物对G-四链体及双链DNA选择性的电喷雾质谱研究

利用电喷雾质谱(ESI-MS)研究了4种常见的类黄酮化合物芦丁、 槲皮素、 葛根素和柚皮苷与2种不同形态结构的G-四链体DNA和3种双链DNA的非共价相互作用, 比较了这些小分子化合物与不同形态结构DNA结合的强弱及形成复合物的化学计量. 结果表明, 芦丁和槲皮素对G-四链体DNA具有一定的选择性, 同时它们对双链DNA的选择性也较高; 而葛根素和柚皮苷对G-四链体DNA仅显示了较低的选择性.     

微波辅助提取刺五加中黄酮类化合物过程中的化学变化研究

利用HPLC-UV和HPLC-ESI-MSn技术系统地研究了传统中药刺五加中黄酮类化合物在微波辅助常压和高压提取过程中的化学变化, 并考察了提取压力和提取时间对其化学变化规律的影响, 结果表明, 在提取压力超过300 kPa时芦丁开始失去一个芸香糖转化为槲皮素; 随着微波照射时间的延长, 金丝桃苷、芦丁、槲皮苷和槲皮素提取产率先增加, 而后下降. 提取压力越大, 提取速率越快, 分解的速度也越快,达到最高提取产率的时间越短.     

Disentangling diffusion information of individual components in a mixture with a 3D COMPACT‐IDOSY NMR experiment

A new 3D diffusion‐ordered heteronuclear NMR experiment COMPACT‐IDOSY (cross‐polarization optimized multisite polarized accelerated time internally encoded diffusion ordered spectroscopy) has been designed and experimentally implemented on a mixture of flavonoids rutin and quercetin. The pulse sequence uses a cross‐polarization mixing period and diffusion encoding gradients internally incorporated into the coherence transfer interval of a long‐range heteronuclear correlation experiment. Substantial reduction in experimental time, good sensitivity and excellent resolution of signal overlap lead to the accurate determination of translational diffusion coefficients of individual components in the mixture. Copyright © 2012 John Wiley & Sons, Ltd.