Use of cerium oxide (CeO(2)) as a packing material for the chromatographic separation of C(60) and C(70) fullerenes

Cerium oxide (CeO₂) was tested as a packing material in liquid chromatography for the separation of C₆₀ and C₇₀ fullerenes. The separation of C₆₀ and C₇₀ fullerenes could be achieved within 20 min by using pure n-hexane as the mobile phase. Furthermore, some higher fullerenes could also be separated in less than 40 min. The peak area was reproducible to a large extent. The separation of fullerenes by liquid chromatography on CeO₂ is shown to be an effective method for their isolation in large amounts. The column efficiency of the CeO₂ column was compared with commercial silica gel and ODS columns. The main advantage of the CeO₂ column is its ability to separate large amounts of fullerenes (C₆₀ and C₇₀) in toluene.     

Retention of C60 and C70 fullerenes on reversed-phase high-performance liquid chromatographic stationary phases.

The separation of C60 and C70 fullerenes on four different polysiloxane stationary phases was examined. It was determined that polar solvents can be used as mobile phases effectively for the separation of fullerene molecules. Unlike previously published work, a polymeric octadecyl siloxane (ODS) stationary phase provided higher separation factors for C70/C60 than did monomeric ODS stationary phases or phenyl substituted stationary phases. For example, for a methanol-diethyl ether (50:50, v/v) mobile phase and C60, k' approximately 5.0 separation factors, alpha = 3.3, were achieved with polymeric ODS compared to alpha = 2.2, with a monomeric ODS stationary phase. A linear solvation energy relationship (LSER) was used to model the importance of solvent interactions and stationary phase interaction to solute retention.     

Separation of C60 and C70 fullerenes on silica modified by polyaromatic and π-acid aromatic compounds

Separation of C₆₀ and C₇₀ fullerenes by HPLC was studied using sorbents synthesized by reaction of perylenedicarboxylic anhydride, dimethoxyviolanthrene, the tetramer of chromotropic acid with formaldehyde (TCA), trinitrobenzoyl chloride, or chlorotrinitrobenzene with γ-aminopropyl silica. These sorbents possess satisfactory chromatographic properties. The sorbent based on TCA is effective for separation of preparative amounts of fullerenes. Translated fromIzvestiya Akademii Nauk. Seriya Khimicheskaya, No. 8, pp. 1544–1546, August, 1997.     

Optimisation of data handling and detection conditions for automated chromatographic assay software

Summary The principle of automated chromatographic peak detection and analysis software is summarized, and critical steps are systematically studied. As the only parameter to be entered is the acquisition frequency, evaluation of its effect on software performance is discussed. In the case of relatively noisy chromatograms, it is shown experimentally that numerous points per peak have to be taken, leading to quite fast computer acquisition procedures. The use of discrete Fourier transform filtration techniques can modify peak shapes and a comparative study evaluates the relative errors induced in the shapes and characteristics of the chromatographic profiles. Optimisation of filtering conditions is achieved and it is shown that for a filter position only 2% of the Nyquist frequency no deformation occurs in the chromatographic profile. Detection of the start and finish of chromatographic peaks is optimized according to a simple four step iterative procedure. In the case of simulations, the difference between the values used to simulate peaks and those calculated by the software are less than 1%.     

Rapid and sensitive gas chromatographic method for detection of barbital and pentobarbital in blood using flash-heater methylation and nitrogen-specific detection

Summary A sensitive and simplified method is described for the determination of barbital (B) and pentobarbital (PB) in blood using gas chromatography with a nitrogen-specific flame ionization detector. After a simple one-step extraction, B and PB were reconstituted in trimethylanilinium hydroxide and introduced directly into the gas chromatograph. Lower limits of detection was 0.14 μg/ml for B and PB in either water or blood solutions. The recoveries for B from water and blood were 90 and 88%, respectively; for PB they were essentially complete. The average between-run and within-run coefficients of variation for B from water and blood were lower than 7.8%, whereas those of PB were lower than 3.0%. The scale and sensitivity of detection for B and PB were found to be suitable for use in pharmacokinetic studies in the rat.     

Analysis and chromatographic separation of some steroid hormones on NH2 layers

Summary The previously described analytical method for carbohydrates, catecholamines, uric acid, creatine and creatinine using thin-layer chromatography on aminomodified HPTLC plates and subsequent thermal activation of the chromatogram zones is expanded to include several steroid hormones. Specifically, they are the pharmacologically relevant compounds cortisone and hydrocortisone, estradiol and estradiol benzoate, estriol, estrone, methyltestosterone, testosterone and testosterone propionate, prednisolone, pregnandiol and triol, progesterone and Reichstein's S.     

New detection modes for the determination of cephalosporins and their decomposition products

Summary The feasibility of using fluorescence and electrochemical detection after high performance liquid chromatography separation is investigated for a sensitive and selective determination of cephalosporins and their decomposition products. The sensitivity and detectability of fluorescence and electrochemical detection are compared to those of UV absorption detection. The methods have been validated for the determination of cephalosporins in biological fluids and for stability studiesin vitro.     

分离富勒烯的新型高效液相色谱固定相

从分子结构与分子间作用力的关系出发，以富电子性化合物甲苯、α－二甲苯、α－甲基萘修饰的聚苯乙烯－二乙烯苯（ＰＳ－ＤＶＢ）树脂作为固定相分离Ｃ６０、Ｃ７０混合物，获得很好的结果，其中以α－甲基萘修饰ＰＳ－ＤＶＢ和为固定相分离效果最理想。     

Electroactivity of nitriles on platinum electrodes and its use in chromatographic detection

Acetonitrile yields two oxidative peaks, first at ca. +0.30 and second at ca. +1.15 V vs. Ag/AgCl in cyclic voltammetry with platinum electrodes in 0.10 M methanesulfonic acid (MSA) containing 0.05–5 mM concentrations of acetonitrile. This electroactivity of the nitrile group was used for a direct detection of nitriles after their chromatographic separation. Three organic nitriles (acetonitrile, propionitrile and butanenitrile) were separated with an IonPac ICE-AS 1 column, eluted with 0.10 M MSA and detected on a platinum electrode via pulsed amperometric detection. Analytical performance was evaluated with a three potential waveform (+0.30 V, +1.15 V, −0.30 V vs. Ag/AgCl, current integration at +1.15 V). Numerical values of detection limits, linearity of calibration and reproducibility are reported for all three organic nitriles.     

Simple and sensitive liquid chromatographic method with fluorimetric detection for the analysis of gamma-amino-n-butyric acid in human urine

A simple and sensitive liquid chromatographic method is described for the analysis of γ-amino-n-butyric acid (GABA) in human urine. GABA is increased in the urine of cancer patients and could be used as a biomarker in the diagnosis and treatment of related patients. The method is based on derivatizing GABA with a fluorescent reagent (naproxen acyl chloride) for transforming the non-chromophoric GABA to a derivative with chromophoric and fluorophoric properties. The resulting derivative is highly responsive to a fluorimetric detector (λ_ex = 230 nm, λ_em = 350 nm). The lower quantitation of the method is attainable at 100 nM GABA with a detection limit about 10 nM (S/N = 3 with 20 μL injected). Application of the method to the analysis of GABA in the urine of patients with ovarian and uterine cancer was demonstrated.     

Elaboration and use of a double channel detection ion chromatographic method for determination of ions in aqueous samples

Summary In this paper we report our research in developing a simple and reliable method for determination of various anions in aqueous samples. Among the several methods existing for determining ions in aqueous samples, ion chromatography is becoming more and more important, due to its reliability and ability to determine the concentration of more ions from one sample. Although several detection methods are available, in this work we used conductivity detection, and indirect UV photometric detection. Because in some cases it can be important to gather more detailed information on the composition of a sample, we present a way to meet this demand by using double channel detection. Presented at Balaton Symposium on High Performance Separation Methods, Siófok, Hungary, September, 1–3, 1999     

A highly sensitive method for the determination of adriamycin and adriamycinol in human plasma using HPLC with fluorimetric detection

Summary A simple and very sensitive HPLC method, for the simultaneous determination in human plasma of adriamycin and its metabolite adriamycinol, is described. Plasmas from patients were stored frozen. Thawed samples were extracted by absorption of anthracyclin onto a small C18 column. After evaporation of the eluate and reconstitution of the residue with methanol (100L), 30 to 40L of the mixture were injected into the chromatograph. Separation was obtained using an RP 8 column with a mobile phase of formate buffermethanol-acetonitrile (502327, v/v). A spectrofluorimeter was used as detector. The limit of sensitivity of the assay was 50 pcg/ml of plasma.     

Improved liquid chromatographic determination of nine currently used (fluoro)quinolones with fluorescence and mass spectrometric detection for environmental samples

An HPLC method using C18-modified silica as stationary phase has been developed for environmental trace analysis of nine (fluoro)quinolones. Detection is done by fluorescence measurement or MS using the modes of SIM and selected reaction monitoring (SRM). Best separation is achieved with a gradient consisting of 50 mM formic acid and methanol, which is fully compatible with MS coupling. LOQs (S/N of 10) for fluorescence detection are between 10 and 60 microg/L, depending on the analyte. MS detection (SIM and SRM) yields LOQs that are better by a factor of at least an order of magnitude. Sample preconcentration and sample clean-up is accomplished by SPE (preconcentration factor of 1000), leading to LOQs in the low ng/L range. Recoveries of the preconcentration procedure are better than 80% for all analytes. The suitability for real samples has been demonstrated by analyzing surface waters, municipal waste waters, sewage treatment plant effluents, sewage sludge, and sediment taken from rivers and fish ponds. The method should also be useful for determination of residues of (fluoro)quinolones in food or other matrices. The degradation of the (fluoro)quinolones has been examined over 5 days in order to get information about the decomposition rate and the degradation products eventually occurring in the environment.     

An immunoassay based on lab-on-a-chip for simultaneous and sensitive detection of clenbuterol and ractopamine

Both clenbuterol (CLB) and ractopamine (RAC) are β-adrenergic agonists. After long-term excessive intake, there will be adverse reactions such as headache, chest tightness, limb numbness, and serious life-threatening. Simultaneous detection of CLB and RAC in related samples is of great importance for human health. In this work, we outline a microfluidics-based indirect competitive immunoassay (MICI) system that can sensitively detect residual CLB and RAC in pork, swine blood and swine urine. The rapid detection of multiple samples can be achieved in one chip, which greatly improves the detection efficiency. This method has good stability and reproducibility and the microfluidic chips are easy to manufacture. The linear ranges for CLB and RAC detection by MICI are 0.1–2.5 ng/mL and 0.1–5 ng/mL, and the limits of detection (LODs) are 0.094 ng/mL and 0.091 ng/mL, respectively. This straightforward and portable immunoassay system provides a good platform for rapid detection of harmful substances in food samples.     

Development and validation of a sensitive quantification method for hematoporphyrin monomethyl ether in plasma using high-performance liquid chromatography with fluorescence detection

A rapid, sensitive, precise and specific method for determination of hematoporphyrin monomethyl ether (HMME), a novel photodynamic therapy (PDT) drug, was developed and validated using high-performance liquid chromatography (HPLC) with fluorescence detection. HMME was isolated from the plasma by a single-step liquid-liquid extraction with ethyl acetate. The analyte and internal standard fluorescein were baseline separated on a Diamonsil C(18) analytical column (4.6 x 150 mm, 5 microm) and analyzed using a fluorescence detector with the excitation and emission wavelengths set at 395 and 613 nm, respectively. The method was linear in the concentration range 0.025-5 microg/mL with a lower limit of quantitation (LLOQ) of 10 ng/mL. The inter- and intra-day accuracies and precisions were all within 10% and the mean recoveries of HMME and fluorescein were 95 +/- 3.7 and 90 +/- 2.3%, respectively. The analyte was stable during all sample storage, preparation and analysis periods. This method was successfully applied to a pharmacokinetic study after a single-dose intravenous administration of HMME (5 mg/kg) to beagle dogs. This method was reproducible and sensitive enough for the pharmacokinetic study of HMME. Based on the results of the pharmacokinetic study, we suggest that a rather long light-avoiding time is essential for patients under HMME therapy.     

Development and validation of a pre-column reversed phase liquid chromatographic method with fluorescence detection for the determination of primary phenethylamines in dietary supplements and phytoextracts

A sensitive and selective reversed-phase liquid chromatographic (RP-LC) method was developed and validated to determine octopamine, tyramine and Tyrosine (Tyr) in complex matrices as formulations and phytoextracts (Citrus aurantium), after pre-column derivatization with o-phthaldialdehyde (OPA) reagent. The chromatographic separations were performed at room temperature on a Phenomenex Luna C18 column using methanol and sodium acetate buffer (pH 5.5) by varying composition gradient elution as mobile phase and detected flurometrically at λ(em)=455 nm with λ(ex)=340 nm. The results obtained by the proposed method were compared with those achieved by a validated direct RP-LC method with fluorescence detection at λ(em)=310 nm with λ(ex)=275 nm, as reference method, using a Phenomenex Gemini C18 column under isocratic elution conditions with acetonitrile and sodium 1-heptanesulphonate (pH 3), as mobile phase. The higher sensitivity of the derivatization method (detection limit about 0.06 pmol) allowed the sure determination of octopamine present in traces in the examined samples. The repeatability of method (RSD) was ≤1.90% and there was no significant difference between repeatability and intermediate precision data. Recovery studies showed good results 99.5-101.3% with RSD ranging from 0.8 to 1.2%. All analyses were performed by mild conditions in absence of preliminary difficult extraction methodologies or laborious step of sample pre-treatment.     

基于杂交链式反应和共价有机框架的新型荧光传感器用于ctDNA的高灵敏和选择性检测

共价有机框架是近年来发展起来的一种高度结晶的多孔聚合物,由有机单元连接形成。由于其优异的孔隙率,模块性,结晶性和生物相容性,在传感领域显示出良好的应用前景。本文开发了一种基于杂交链式反应 (HCR) 和共价有机框架的新型荧光传感平台,用于高灵敏的检测循环肿瘤DNA (ctDNA)。ctDNA可以作为HCR的激活剂,通过触发发夹1 (H1) 和发夹2 (H2) 的自主交叉打开,激活HCR形成延伸的有缺口的双链DNA,导致荧光信号的变化,从而实现ctDNA的定量检测。HCR产物和荧光信号分别采用琼脂糖凝胶电泳和荧光光谱进行表征。ctDNA浓度在0-10 nM范围内,ctDNA的浓度与荧光信号呈良好的线性关系,检测限为1.3 nM。该方法对ctDNA具有满意的选择性,并在人类血清样品中显示出良好的性能。该荧光传感平台为ctDNA的检测提供了新思路,在癌症早期诊断中显示出潜在的应用前景。     

Chromatographic behavior of selected dyes on silica and cellulose micro-TLC plates: Potential application as target substances for extraction,chromatographic, and/or microfluidic systems

ABSTRACT

The main goal of this research communication focuses on screening of chromatographic behavior of 18 colorants including amaranth, black PN, bromophenol blue, bromocresol green, bromocresol purple, bromothymol blue, carmine, dimethyl yellow, erythrosine, fluorescein, methyl red, naphthalene black, patent blue, phenol red, Sudan II, Sudan III, Sudan IV, and p-xylenol blue. They are commonly used as food and industrial colorants or sensing molecules for analytical and biomedical applications. We studied retention (R_F), peak base width (W_b), and peak asymmetry factor (A_S) data collected under thermostatic conditions (303?K) using silica and cellulose-coated microplates (5?×?5?cm) and simple as possible mono/binary eluents. Presented retention database may help to select the proper conditions for, for example, chromatographic separation, extraction to solid phase (e.g. bar-type extraction devices) or application of such chemicals as the internal standard substances for planar/column chromatography and/or microfluidic devices (e.g., microfluidic paper-based analytical devices—μPADs). Furthermore, the microplates peak capacity (maximum number of resolvable spots, n) was calculated and discussed. As the second goal, we presented results of literature concerning dyes quantification in complex samples. We compared latest methodologies involving simple spectrophotometric analysis and more sophisticated protocols like high-throughput separation/detection procedures based on high-performance planar/liquid chromatography, capillary electrophoresis and microfluidic devices.     

Reversed-phase liquid chromatographic determination of uranium based on its ternary complex with fluoride and 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol. New chromatographic probe for the detection of fluoride on C18 stationary phase surface

Optimum conditions for the reversed-phase liquid chromatographic separation of uranium as U(VI)-F(-)-(5-Br-PADAP) [5-Br-PADAP is 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol] ternary complex on the end-capped C18 column were evaluated. The developed sensitive, selective and versatile method allows to determine uranium in the wide concentration range 0.2-120.0 mug ml(-1) (detection limit, DL, is 0.15 mug ml(-1) for 20 mul loop). Solvent mixture, acetonitrile+water (65+35, v/v) containing fluoride in concentration 3x10(-3) mol l(-1) (pH 5.5) was used as eluent. Double action of fluoride present in eluent - as stabilising agent of the ternary system and modifier of the stationary phase was evaluated. In order to prove appearance of the modifying effect of fluoride on the stationary phase the new chromatographic probe - system Zr(IV)-(5-Br-PADAP)/Zr(IV)-F(-)-(5-Br-PADAP) was introduced as a tool for the detection of F(-) presence on the surface.