Improved Solid Phase Extraction Procedure for Assay of Cephalosporins in Human Urine Samples
Abstract Solid phase extraction technique has been evaluated for the treatment of urine samples in the analysis of cephalosporins before injection into an HP-Hypersil ODS-C18 column. Cephalexin, cefotaxime, cefazolin, cefuroxime, and cefoxitin were tested with seven different reversed-phase extraction column cartridges and the obtained urine extracts were not clean. However, 3M Empore extraction disk cartridges packed with octadecyl (C18) bonded silica provided clean extracts with a single extraction. The recoveries of the five cephalosporins ranged from 56 to 60 % in the 1.25–500 g/mL concentration range. The assay was accurate, precise, and adequate for testing the drug content in urine s…
Simple and sensitive reversed-phase liquid chromatographic assay for analysis of chlorthalidone in urine
Abstract This study describes a rapid method for the determination of chlorthalidone in human urine by reversed-phase liquid chromatography and UV detection at 230 nm, after clean-up over a C8 solid-phase extraction column. Liquid chromatography was carried out on a C18-bonded phase using acetonitrile-phosphate buffer (pH=3) gradient elution. Triamterene was used as internal standard. The system has been applied to the determination of chlorthalidone in the 0.10–10.0 μg/mL concentration interval; the limit of detection was 6 ng/mL.
Analysis of Diuretics in Urine by Column-Switching Chromatography and Fluorescence Detection
The potential of column-switching chromatography and fluorescence detection for the analysis of diuretics in urine is evaluated. Sample cleanup and chromatographic parameters have been optimized to achieve maximum sensitivity for the detection and quantification of some relevant diuretics. On the basis of these studies, an on-line procedure for the simultaneous determination of amiloride, furosemide, bumetanide and triamterene is presented. The linearity, precision, accuracy and sensitivity of the method are discussed. The utility of the described approach has been tested by analysing urine samples obtained after administration of bumetanide.
Nylon-Supported Plasmonic Assay Based on the Aggregation of Silver Nanoparticles: In Situ Determination of Hydrogen Sulfide-like Compounds in Breath Samples as a Proof of Concept
A procedure for supporting silver nanoparticles (AgNPs) on nylon is proposed. Besides, the membrane has been developed as a solid-phase colorimetric plasmonic sensor for volatile sulfide compounds (VSCs) like H2S, CH3SH, and (CH3)2S. AgNP behavior in the membrane has been studied by UV-vis diffuse reflectance spectrometry, Raman spectrometry, High-resolution transmission electron microscopy (HR-TEM), and Scanning electron microscopy (SEM). The sensor responded by changing its color from yellow in absence of VSCs to several orange/brown colors in the function of VSC concentration as occurs in solution; an increase in the hydrodynamic diameter, estimated by both asymmetrical flow field-flow f…
Delivering Inorganic and Organic Reagents and Enzymes from Zein and Developing Optical Sensors.
Nowadays, interest in using environmentally friendly materials is increasing in many fields. However, the rational design of sensors with biodegradable materials is a challenge. The main aim of this work is to show the possibility of using zein, a protein from corn, as a biodegradable and low-cost material for immobilizing, stabilizing, and delivering different kind of reagents for developing optical sensors. Enzymes, metallic salts, and aromatic and small organic compounds were tested. In addition, different techniques of immobilization, entrapment and adsorption, were used, and different formats, such as solid devices and also multiwell platforms, were proposed. The capacity of zein for i…
Color study of historic silks
The chemical characterization of silk textiles of historic value may be necessary to achieve a better understanding of the production processes applied, to evaluate their preservation, to detect manipulations or forgeries, and to value the combination of tradition and innovation in contemporary manufacturing techniques. The main objective of this work was to study four historical silks from the Garín collection, all of them from the 20th century, although with 19th century designs. To this end, non-invasive spectroscopic and microscopy techniques were applied in order to obtain information on the dyes used during their production and their link with those used in the silk industry in previo…
On-Line Derivatization into Precolumns for the Determination of Drugs by Liquid Chromatography and Column Switching: Determination of Amphetamines in Urine
A chromatographic system for the on-line derivatization of drugs using column switching is described. The system uses a 20 mm × 2.1 mm i.d. precolumn packed with a unmodified ODS stationary phase. This column is used for sample cleanup and enrichment of the analytes. Next, the trapped analytes are derivatized by injection of the derivatization reagent into the precolumn. Finally, the derivatives are transferred to the analytical column for their separation under reversed-phase conditions. The influence of several parameters such as the reaction time, the amount of derivatization reagent, or the system design has been studied using some amphetamines as model compounds and three derivatizatio…
Elimination of the unknown irrelevant matrix absorbance by using the H-Point Standard Additions Method (HPSAM)
It is demonstrated how the H-Point Standard Additions Method (HPSAM) using DeltaA as analytical signal (from data at three previously selected wavelengths) is only related with analyte concentration when unknown irrelevant matrix absorbance is present. The method is compared with the most common previously reported methods, such as derivative spectroscopy or the compensation method. The obtained results show that the proposed HPSAM leads to the same found concentration of analyte as the other reported methods, except detection limits and standard deviation for six replicates which are lower, because of the use of absorbance data (instead of first derivative data as usual). In addition, the …
Determination of acetazolamide in human urine samples by reversed-phase high-performance liquid chromatography in the presence of xanthines.
A simple, rapid and selective high-performance liquid chromatographic assay for the determination of acetazolamide in urine samples is described. After extraction with ethyl acetate, the drug is chromatographed on an HP-Hypersil ODS-C18 column with a mobile phase of acetonitrile-phosphate buffer (pH 3) and ultraviolet detection at 275 nm. The efficiency of the extraction, the linearity and the reproducibility of the method permit the evaluation of acetazolamide urinary excretion a long time after its administration.
A new derivatization procedure for the determination of cephalexin with 1,2-naphthoquinone 4-sulphonate in pharmaceutical and urine samples using solid-phase extraction cartridges and UV–visible detection
Abstract The present report shows how to derivatize cephalexin with 1,2-naphthoquinone-4-sulphonate (NQS) into solid-phase extraction cartridges (C18) using UV–visible detection. Optimum conditions for this new procedure are: hydrogen carbonate/carbonate buffer pH=10.5, 5 min reaction time at 25°C and NQS concentration of 7.1×10−3 mol l−1. The accuracy and the precision of the method were tested. The procedure was used to measure cephalexin in pharmaceutical and urine samples. The results obtained were contrasted with those reported by UV-spectrophotometric and HPLC methods for pharmaceutical samples and with HPLC method for urine samples. The H-point standard additions method was used to m…
Solid phase extraction of amines
Abstract The objective of this paper is to provide information about solid phase extraction (SPE) as an alternative to liquid–liquid extraction of amines from several matrices. Different sorbents ranging from non-polar phases, such as C 18 silica to more polar such as cyanopropylsilica (CN) have been tested for analysis of aliphatic amines as monoamines, diamines and polyamines. Phenylalkylamines such as amphetamine or methamphetamine and heterocyclic amines such as histamine or cephalosporins (which also contain a carboxylic group), have also been studied. The different steps involved in the extraction procedure have been tested (conditioning, retention, pre-concentration, washing and elut…
New Reusable Solid Biosensor with Covalent Immobilization of the Horseradish Peroxidase Enzyme: In Situ Liberation Studies of Hydrogen Peroxide by Portable Chemiluminescent Determination.
Herein, we reported a chemiluminescent biosensor based on the covalent immobilization of the horseradish peroxidase (HRP) enzyme on a polydimethylsiloxane (PDMS) support to quantify in situ hydrogen peroxide (H2O2). The chemiluminescent reaction based on the use of luminol as an oxidizable substrate, with HRP as the catalyst, has been used in order to quantify H2O2 as the oxidizing agent. The performance of the proposed biosensor has been demonstrated to determine H2O2 liberated by cells in a culture medium and for evaluating the delivery of H2O2 from denture cleaner tablets, as examples of application. For both analyses, the results indicated that the biosensor is cost-effective, sensitive…
Study of the Stability of Citrate Capped AgNPs in Several Environmental Water Matrices by Asymmetrical Flow Field Flow Fractionation
Asymmetrical flow field-flow fractionation (AF4) coupled to UV-Vis and dynamic light scattering (DLS) detectors in series, was tested for stability studies of dispersions of citrate-capped silver nanoparticles (AgNPs) in several water matrices. The main goal is to provide knowledge to understand their possible behavior in the environment for short times since mixturing (up to 180 min). Ultrapure (UPW), bottled (BW1, BW2), tap (TW), transitional (TrW) and sea water (SW) matrices were assayed. Observations were compatible with the aggregation of AgNPs, a change in the plasmon band and a size growth with time were done. Fractograms showed different evolution fingerprints in the function of the…
Derivatization of amines in solid-phase extraction supports with 9-fluorenylmethyl chloroformate for liquid chromatography
Abstract A new method based on reaction in solid-phase extraction cartridges with 9-fluorenylmethyl chloroformate is described for improved derivatization of amines prior to liquid chromatography. The effects of the reaction conditions, the kinetics of the reaction and the kind of packing material on the derivatization have been evaluated. The reliability of the described approach has been tested by derivatizing propylamine, diethylamine, diethanolamine, glycine, glutamic acid, p-aminobenzoic acid and β-phenylethylamine. Reaction yields compared with those of the analogous solution derivatizations were in the 44–169% range, and satisfactory linearity was achieved at concentrations of the am…
Strategies for the enantiomeric determination of amphetamine and related compounds by liquid chromatography.
This paper summarizes recent research on the stereospecific analysis of amphetamine, its analogs and metabolites, by liquid chromatography. The different methods proposed have been evaluated and compared in terms of resolution power, time of analysis, sensitivity, or potential for automation. Chiral derivatization, followed by separation of the diastereomers formed in achiral chromatographic systems, is still the method preferred for the analysis of amphetamines at trace levels, as derivatization also improves analyte detectability. This is the method of choice for the enantiomeric analysis of amphetamines at the low concentrations typically encountered in biological samples. In recent year…
Towards sarcosine determination in urine for prostatic carcinoma detection
Abstract Sarcosine, a potential biomarker for prostate cancer, can be detected in a solid state enzyme based biosensor using sarcosine oxidase, with particle immobilised reagents. A novel fusion protein of the fluorescent protein, mCherry, sarcosine oxidase (SOx), and the polypeptide R5 (R52-mCherry-SOx-R5-6 H), was explored, which allowed self-immobilization on silica microparticles and long-term (90 days +) retention of activity, even at room temperature. In contrast, commercial wildtype SOx lost activity in a few days. A silica-R52-mCherry-SOx-R5-6H microparticle sensor for determination of sarcosine in urine, linked the SOx coproduct, H2O2, to a measurement catalysed by horseradish pero…
Column-switching techniques for high-performance liquid chromatography of drugs in biological samples.
In recent years, an increasing number of publications have demonstrated the potential of column-switching techniques for the chromatographic separation, determination and preparative isolation of analytes from biological matrices. Column-switching systems greatly facilitate drug analysis, by on-line sample clean-up and trace enrichment, or by improving the analytical separative process. In this paper, the main applications of column-switching techniques to drug analysis in biological samples, are reviewed.
Enantiomeric separation of amphetamine and related compounds by liquid chromatography using precolumn derivatization witho-phthaldialdehyde
The utility of the derivatization reagento-phthaldialdehyde (OPA) in combination with the chiral thiol N-acetyl-L-cysteine (NAC) for the enantioselective analysis of primary amphetamines and structurally related substances has been investigated. The diastereomers obtained have been chromatographed on an achiral, reversed phase C18 column (125 mm×4 mm i.d., 5 μm), and detected fluorimetrically. The influence of parameters affecting the chromatographic separation as well as the derivatization reaction has been optimized, using amphetamine, norephedrine, norepinephrine and 3,4-methylenedioxyamphetamine (MDA) as model compounds. On the basis of the results obtained, possible conditions for the …
Chiral separation of ephedrines by liquid chromatography using β-cyclodextrins
β-cyclodextrins (β-CDs) have been evaluated as chiral recognition agents for the enantioseparation of ephedrines by liquid chromatography (LC). The compounds tested were norephedrine, ephedrine, pseudoephedrine, N-methylephedrine and N-methylpseuoephedrine. Two possibilities have been investigated, the employment of a chiral stationary phase consisting of immobilized β-cyclodextrin, and the addition of the chiral selector to the mobile phase. In the latter approach the following chiral agents have been assayed, native β-cyclodextrin (β-CD), methyl-β-cyclodextrin (M-β-CD), carboxyethyl-β-cyclodextrin (CE-β-CD), carboxymethyl-β-cyclodextrin (CM-β-CD) and hydroxypropyl-β-cyclodextrin (HP-β-CD)…
Reduction of Nitrates in Waste Water through the Valorization of Rice Straw: LIFE LIBERNITRATE Project
An improved and more sustainable waste management system is required for successful development of technologies based on renewable sources. Rice straw is submitted to controlled combustion reactions and the produced ashes are chemically treated to produce silica. After a chemical activation step, the activated silica shows potential as an adsorbent agent and will be used to remove the excess of nitrates in groundwater and wells in the area of Alginet (Valencia, Spain), selected as a vulnerable zone within the Nitrates Directive. The demonstration activity aims to have a local impact on municipalities of 200 inhabitants or fewer, decreasing from current nitrate concentrations close to 50 mg/…
Extractive-spectrophotometric determination of furosemide with sodium 1,2-naphthoquinone-4-sulphonate in pharmaceutical formulations
ABSTRACT An extractive-spectrophotometric study based on the reaction of furosemide with 1,2-naphthoquinone-4-sulphonate (NQS) is described. Optimal conditions are: NaH2PO4-Na2HPO4 pH 7.5, NQS 7.7×...
Spectrophotometric determination of phenols in water samples by the GHPSAM method
Abstract The generalized H-point standard-additions method (GHPSAM) is proposed in order to obtain the phenol concentration in water samples when the matrix is completely unknown. The procedure involves solid-phase extraction in BondElut PPL cartridges and data handling of the UV-visible spectrophotometry measurements. The spectral regions where the unknown interferent behaviour can be considered as linear are found and the analyte concentration free from bias error is estimated. The percentages of recovery of phenols in spiked samples were similar to those obtained by HPLC. Cresols or chlorophenols can be also determined in real samples by this method. The concentration range tested was 0.…
A new tool for evaluating and/or selecting analytical methods: Summarizing the information in a hexagon
Abstract A quantitative evaluation tool is proposed, which aims to assess optimal selection or/and testing of analytical methods. Objective criteria related to analytical performance, sustainability, environmental impact and economic cost are evaluated through the definition of penalty points divided into five different blocks, namely, figures of merit, toxicity and safety, residues, carbon footprint and economic cost. For each block, the overall qualification is scaled from 0 to 4 and it is depicted on a regular hexagonal pictogram that allows a user friendly comparison of analytical procedures. The present evaluation tool aims to be a guideline for evaluating and/or selecting analytical p…
Rapid evaluation of ammonium in different rain events minimizing needed volume by a cost-effective and sustainable PDMS supported solid sensor.
Abstract The presence of ammonium ion in rainwater is due to atmospheric processes which involve its scavenging from gas phase and particulate matter. The fractionated samplings of rainwater can provide information about these processes and their potential sources. However, only a low sample volume may be available, which constrained the analysis in general and more particularly in situ mode. For minimizing this limitation, this work proposes a polydimethylsiloxane (PDMS)-salicylate sensor that produces a color change. The embedding of solid reagents into PDMS was optimized. Good analytical characteristics (analysis time of 10 min, sample volume of 500 μL, limit of detection 0.03 μgmL−1) we…