A Simple and Validated LC Method for the Simultaneous Determination of Three Compounds in Mikania laevigata Extracts

The Mikania genus is widely known as guaco and is used to treat fever, rheumatism, influenza and respiratory diseases. This article deals with the simultaneous quantification of three commercially available phenolic markers (o-coumaric acid, coumarin and syringaldehyde) in M. laevigata extracts, through LC-PDA. The validation data show that the method is specific, accurate, precise and robust, and also indicative of the stability of guaco extract. The method was linear, over a range of 1.25–20.0 μg mL⁻¹ for o-coumaric acid, 2.5–40.0 μg mL⁻¹ for coumarin, and 0.25–4.0 μg mL⁻¹ for syringaldehyde. The range of recovery was 94.3–96.4% for all the components, at a level of 100%.

     

Isocratic Reversed-Phase HPLC for Simultaneous Separation and Determination of Seven Antiepileptic Drugs and Two of their Active Metabolites in Human Plasma

A simple reversed-phase high-performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of the antiepileptic drugs (AEDs) zonisamide (ZNS), primidone (PRI), lamotrigine (LTG), phenobarbital (PB), phenytoin (PHT), oxcarbazepine (OXC), and carbamazepine (CBZ) and two of their active metabolites, monohydroxycarbamazepine (MHD) and carbamazepine 10,11-epoxide (CBZE) in human plasma. Plasma (100 μL) was pretreated by deproteinization with 300 μL methanol containing 20 μg mL⁻¹ propranolol hydrochloride as internal standard. HPLC was performed on a C₈ column (4.6 mm × 250 mm; particle size 5 μm) with methanol–acetonitrile–0.1% trifluoroacetic acid, 235:120:645 (v/v), as mobile phase at a flow rate of 1.5 mL min⁻¹. ZNS, OXC, and CBZ were monitored by UV detection at 235 nm, and PRI, LTG, MHD, PB, PHT, and CBZE by UV detection at 215 nm. Relationships between response and concentration were linear over the concentration ranges 1–80 μg mL⁻¹ for ZNS, 5–50 μg mL⁻¹ for PRI, 1–25 μg mL⁻¹ for LTG, 1–50 μg mL⁻¹ for MHD, 5–100 μg mL⁻¹ for PB, 1–10 μg mL⁻¹ for CBZE, 0.5–25 μg mL⁻¹ for OXC, 1–50 μg mL⁻¹ for PHT, and 1–25 μg mL⁻¹ for CBZ. Intra-day and inter-day reproducibility were adequate (coefficients of variation were ≤11.6%) and absolute recovery ranged from 95.2 ± 6.13 to 107.7 ± 7.76% for all the analytes; for the IS recovery was 98.69 ± 1.12%. The method was proved to be accurate, reproducible, convenient, and suitable for therapeutic monitoring of the nine analytes.     

Simultaneous RP-LC Determination of Losartan Potassium, Ramipril, and Hydrochlorothiazide in Pharmaceutical Preparations

A simple, rapid, and precise reversed-phase high-performance liquid chromatographic method has been developed for simultaneous determination of losartan potassium, ramipril, and hydrochlorothiazide. The three drugs were separated on a 150 mm × 4.6 mm i.d., 5 μm particle, Cosmosil C₁₈ column. The mobile phase was 0.025 m sodium perchlorate–acetonitrile, 62:38 (v/v), containing 0.1% heptanesulphonic acid, pH adjusted to 2.85 with orthophosphoric acid, at a flow rate of 1.0 mL min⁻¹. UV detection was performed at 215 nm. The method was validated for linearity, accuracy, precision, and limit of quantitation. Linearity, accuracy, and precision were acceptable in the ranges 35–65 μg mL⁻¹ for losartan, 1.75–3.25 μg mL⁻¹ for ramipril, and 8.75–16.25 μg mL⁻¹ for hydrochlorothiazide.     

Simultaneous separation and determination of Cu(II), Fe(III), and Pb(II) by reversed-phase ion-pair chromatography withN,N,N′, N′-ethylene-diaminetetrakis(methylenephosphonic Acid)

Summary A reversed-phase ion-pair chromatographic (RPIPC) method withN,N,N′, N′-ethylenediaminetetrakis(methylenephosphonic acid) (EDTMP) as coordinating agent has been developed for simultaneous separation and detection of Cu(II), Fe(III), and Pb(II) ions. Response is linearly dependent on amount of sample over the range 9.52–50.8 μg mL⁻¹ for Cu(II), 8.31–41.8 μg mL⁻¹ for Fe(III), and 37.3–51.8 μg mL⁻¹ for Pb(II). The method has been applied successfully to an artificial mixed-ore sample.     

Resonance Rayleigh scattering spectra of tetracycline antibiotic–Cu(II)–titan yellow systems and their applications in analytical chemistry

Tetracycline antibiotics (TCs) such as doxycycline (DOTC), chlortetracycline (CTC), oxytetracycline (OTC), and tetracycline (TC) react with Cu(II) in pH 3.5 BR buffer medium to form 1:1 cationic chelates, which further react with titan yellow to form 2:1 ion association complexes. These result in great enhancement of resonance Rayleigh scattering (RRS) and the appearance of new RRS spectra. The ion association complexes of DOTC, CTC, OTC, and TC have similar spectral characteristics and their maximum RRS wavelengths are all located at 464 nm. The quantitative determination ranges and the detection limits (3σ) of the four TCs are 0.037–4.8 μg mL⁻¹ and 11.2 ng mL⁻¹ for DOTC, 0.041–5.2 μg mL⁻¹ and 12.4 ng mL⁻¹ for CTC, 0.050–4.8 μg mL⁻¹ and 15.1 ng mL⁻¹ for TC, and 0.088–5.0 μg mL⁻¹ and 26.3 ng mL⁻¹ for OTC, respectively. The optimum reaction conditions, the effects of foreign substances, the structure of ternary complexes, and the reaction mechanism are discussed. A sensitive, rapid, and simple RRS method for the determination of DOTC has been developed.     

Simultaneous Determination of Baicalin, Baicalein, Wogonin, Oxysophocarpine, Oxymatrine and Matrine in the Chinese Herbal Preparation of Sanwu-Huangqin-Tang by Ion-Paired HPLC

A sensitive and reliable ion-paired high-performance liquid chromatographic method has been established for the simultaneous quantification of six major active ingredients, namely baicalin, baicalein, wogonin, oxysophocarpine, oxymatrine and matrine in the Chinese herbal preparation, Sanwu-Huangqin-Tang. HPLC analyses were performed on a Phenomenex luna C₁₈ column with mobile phase of methanol–acetonitrile–aqueous phosphoric acid at a flow rate of 0.9 mL min⁻¹. The complete separation was achieved within 35 min for the six target constituents. A good linear regression relationship between peak-areas and concentrations was obtained over the range of 12.10–242.0 μg*mL⁻¹ for baicalin, 5.05–101.0 μg*mL⁻¹ for baicalein, 0.95–19.0 μg*mL⁻¹ for wogonin, 2.75–55.0 μg*mL⁻¹ for oxysophocarpin, 2.75–55.0 μg*mL⁻¹ for oxymatrine and 4.90–98.0 μg*mL⁻¹ for matrine, respectively. The repeatability was evaluated by intra- and inter-day assays with relative standard deviation (RSD) being less than 5.1%. The recoveries, measured at three concentration levels, varied from 93.8 to 102.1%. The assay was successfully applied for determination of six bioactive compounds in Sanwu-Huangqin-Tang. The interaction of chemical constituents was observed when the herbs were used in compatibility. The results indicated that the developed assay method was rapid, accurate and could be readily utilized as a quality control method for Sanwu-Huangqin-Tang.     

Simple and rapid determination of piperacillin and ceftazidime in human plasma samples by HPLC

Summary A simple and rapid liquid chromatographic method has been developed for the determination of therapeutic levels of piperacillin (I) and ceftazidime (II) in human plasma. Plasma and p-propionamidophenol (internal standard) were precipitated with methanol (I) or 20% trichloroacetic acid (II). The supernatant was analysed on a 5 μm Spherisorb ODS C₁₈ column with acetonitrile-0.05 M phosphate buffer pH 3.8 as mobile phase and ultraviolet detection at 254 nm. The calibration graph was linear from 10 to 250 μg mL⁻¹, for (I), and from 5 to 200 μg mL⁻¹ for (II). Intra and inter-day CV did no exceed 2.29% for (I), and were 10.76–11.13%–2.00–5.62 for (II) at concentrations of 10 μg mL⁻¹ and 250 μg mL⁻¹.     

Resolution of an intense sweetener mixture by use of a flow injection sensor with on-line solid-phase extraction

An integrated solid-phase spectrophotometry–FIA method is proposed for simultaneous determination of the mixture of saccharin (1,2-benzisothiazol-3(2H)-one-1,1-dioxide; E-954) (SA) and aspartame (N-l-α-aspartyl-l-phenylalanine-1-methyl ester; E-951) (AS). The procedure is based on on-line preconcentration of AS on a C₁₈ silica gel minicolumn and separation from SA, followed by measurement, at λ=210 nm, of the absorbance of SA which is transiently retained on the adsorbent Sephadex G-25 placed in the flow-through cell of a monochannel FIA setup using pH 3.0 orthophosphoric acid–dihydrogen phosphate buffer, 3.75×10^–3 mol L⁻¹, as carrier. Subsequent desorption of AS with methanol enables its determination at λ=205 nm. With a sampling frequency of 10 h⁻¹, the applicable concentration range, the detection limit, and the relative standard deviation were from 1.0 to 200.0 μg mL⁻¹, 0.30 μg mL⁻¹, and 1.0% (80 μg mL⁻¹, n=10), respectively, for SA and from 10.0 to 200.0 μg mL⁻¹, 1.4 μg mL⁻¹, and 1.6% (100 μg mL⁻¹, n=10) for AS. The method was used to determine the amounts of aspartame and saccharin in sweets and drinks. Recovery was always between 99 and 101%. The method enabled satisfactory determination of blends of SA and AS in low-calorie and dietary products and the results were compared with those from an HPLC reference method.     

Detection of proteases using an immunochemical method with haptenylated–gelatin as a solid-phase substrate

A simplified method for the measurement of proteases utilising solid-phase substrates incorporating an ELISA end-point detection step is described. Gelatin–hapten conjugates adsorbed onto polystyrene surfaces were found to be efficient substrates for proteases. Digestion of the solid-phase protein–hapten complexes resulted in proportional desorption of the attached conjugates and decrease in the detectable hapten species. Gelatin–cholic acid conjugates, affinity-purified sheep anti-cholic acid antibody–HRP and a chromogenic substrate were incorporated into a convenient and highly sensitive solid-phase immunochemical method. The detectable signal is inversely proportional to enzyme activity. Bacterial proteases (alpha-chymotrypsin Type II, Type IX from Bacillus polymyxa, Type XIV from Streptomyces griseus, Type XXIV from Bacillus licheniformens) were assayed. Dose–response curves for enzyme activities were measured within ranges of 0–550 μunits mL⁻¹ for chymotrypsin, 0–12 μunits mL⁻¹ for type IX, 0–35 μunits mL⁻¹ for type XIV and 0–100 μunits mL⁻¹ for type XXIV. The detection limits of the proteases studied were 89 μunits mL⁻¹ for chymotrypsin, 0.26 μunits mL⁻¹ for type IX, 5.8 μunits mL⁻¹ for type XIV and 6.5 μunits mL⁻¹ for type XXIV. It was demonstrated that the two-step immunochemical method combines the simplicity and sensitivity of solid-phase enzyme immunoassays, the broad specificity of gelatin as a protease substrate and the flexibility of the solid-phase format.     

LC Determination of Four Isoflavone Aglycones in Red Clover (Trifolium pratense L.)

Red clover (Trifolium pratense L.) is an important forage plant that contains the isoflavones daidzein, genistein, formononetin, and biochanin A. These compounds have been studied lately due to their human health benefits. The aim of this study was to develop and validate an HPLC method with simplified sample preparation to quantify daidzein, genistein, formononetin and biochanin A simultaneously in red clover leaves. The validation showed that the method is specific, accurate, precise and robust, not to mention that the sample preparation is easier and faster than those described earlier. The response was linear over a range of 0.01–0.2 μg mL⁻¹ for daidzein, 0.05–0.5 μg mL⁻¹ for genistein, 4–40 μg mL⁻¹ for formononetin and 2–20 μg mL⁻¹ for biochanin A. The range of recoveries was 85.6–101.0%. The RSD for intra- and inter-day precision were <2.54 and <7.22%, respectively. Five populations of red clover, from the National Plant Germplasm System-USDA were analyzed and the content of daidzein, genistein, formononetin and biochanin A ranged from 7.87–91.31, 51.60–131.30, 6568.33–23461.82, to 2499.55–10337.33 μg g⁻¹ of dried material, respectively.     

A Validated HPLC Method for Simultaneous Determination of 2-Hydroxy-4-methoxybenzaldehyde and 2-Hydroxy-4-methoxybenzoic Acid in Root Organs of Hemidesmus indicus

A reverse phase HPLC method was developed and validated for the simultaneous determination of 2-hydroxy-4-methoxybenzaldehyde and 2-hydroxy-4-methoxybenzoic acid in the root extracts of Hemidesmus indicus. A comprehensive validation of the method including sensitivity, linearity, reproducibility, accuracy, limit of detection (LOD) and limit of quantification (LOQ) was conducted using the optimized chromatographic conditions. The method was found to be linear (r > 0.998) in the range of 5–350 μg mL⁻¹ for 2-hydroxy-4-methoxybenzaldehyde and for 2-hydroxy-4-methoxybenzoic acid (r > 0.999) in the range 10–300 μg mL⁻¹. The method was found to be precise with inter-day precision values (% RSD) in the ranges of 0.61–1.76% for 2-hydroxy-4-methoxybenzaldehyde and 1.3–2.8% for 2-hydroxy-4-ethoxybenzoic acid while intra-day precisions (% RSD) of two analytes were in the range of 0.41–1.07 and 0.95–2.5%. The limits of detection (LODs) for 2-hydroxy-4-methoxybenzaldehyde and 2-hydroxy-4-methoxybenzoic acid were 0.84 and 2.34 μg mL⁻¹. The described method was fast, sensitive and reproducible, and thus well suited for routine analysis of these two compounds from root extracts of H. indicus and other plants.     

HPLC Determination of DCJW in Rat Plasma and Its Application to Pharmacokinetics Studies

A high-performance liquid chromatography–UV method for determining DCJW concentration in rat plasma was developed. The method described was applied to a pharmacokinetics study of intramuscular injection in rats. The plasma samples were deproteinized with acetonitrile in a one-step extraction. The HPLC assay was carried out using a VP-ODS column and the mobile phase consisting of acetonitrile–water (80:20, v/v) was used at a flow rate of 1.0 mL min⁻¹ for the effective eluting DCJW. The detection of the analyte peak area was achieved by setting a UV detector at 314 nm with no interfering plasma peak. The method was fully validated with the following validation parameters: linearity range 0.06–10 μg mL⁻¹ (r > 0.999); absolute recoveries of DCJW were 97.44–103.46% from rat plasma; limit of quantification, 0.06 μg mL⁻¹ and limit of detection, 0.02 μg mL⁻¹. The method was further used to determine the concentration–time profiles of DCJW in the rat plasma following intramuscular injection of DCJW solution at a dose of 1.2 mg kg⁻¹. Maximum plasma concentration (C _max) and area under the plasma concentration–time curve (AUC) for DCJW were 140.20 ng mL⁻¹ and 2405.28 ng h mL⁻¹.     

Stereospecific analysis of flurbiprofen and its major metabolites in plasma and urine by chiral-phase liquid chromatography

Summary Direct chiral-phase HPLC methods have been developed for the determination of flurbiprofen and its major metabolites, namely 4′-hydroxyflurbiprofen and 3′-hydroxy-4′-methoxyflurbiprofen, in biological fluids using a derivatized amylose chiral stationary phase (CSP; Chiral-pak AD). Quantification of all three analytes, both free and conjugated, in urine was carried out following liquid-liquid extraction using tandem ultraviolet (UV) and fluorescence detection. Determination of flurbiprofen and the 4′-hydroxy-metabolite in plasma utilized the same CSP but required modification in the mobile phase composition and sole use of fluorescence detection. The urine assay was linear (r>0.998) between 0.05–10 μg mL⁻¹, 0.1–20 μg mL⁻¹ and 0.01–2 μg mL⁻¹ for the enantiomers of flurbiprofen, 4′-hydroxyflurbiprofen and 3′-hydroxy-4′-methoxyflurbiprofen respectively. The plasma assay was linear (r>0.997) between 0.1–6 μg mL⁻¹ and 0.01–0.6 μg mL⁻¹ for the enantiomers of flurbiprofen and 4′-hydroxyflurbiprofen respectively. Both assays, typically yielded within- and between-day imprecision and accuracy values less than 10% for the enantiomers of the different analytes. Initial volunteer studies suggest that the disposition of flurbiprofen displays modest enantioselectivity in humans.     

Spectrofluorimetric determination of uric acid based on its activation of catalytic oxidation of pyronine Y

A novel kinetic spectrofluorimetric method for the determination of uric acid based on the activation effect of uric acid on the Cu(II) ion catalyzed oxidation of pyronine Y by hydrogen peroxide was developed. The influence of different buffer solutions was tested and the Britton-Robinson buffer solution with pH 2.2 was found to be the optimum. The detection limit and the linear range for uric acid are 0.09 μg mL⁻¹ and 0.3–3.0 μg mL⁻¹, respectively. The RSD for eleven determinations of 1.6 μg mL⁻¹ uric acid was 1.6 %. Satisfactory results were obtained when using this method of uric acid determination in human urine.     

Optimization of solid-phase microextraction procedures for the determination of tricyclic antidepressants and anticonvulsants in plasma samples by liquid chromatography

Simple, sensitive, and reproducible off-line solid-phase microextraction and liquid chromatography (SPME/LC) methods are described for the determination of seven anticonvulsants and tricyclic antidepressants in human plasma. Factorial design and simplex methodology were applied in the optimization of the SPME procedure for tricyclic antidepressants analyses. Important factors in the SPME efficiency are discussed, such as the fiber coatings (both lab-made and commercial), extraction time, pH, ionic strength, influence of plasma proteins, and desorption conditions. The development of the lab-made fiber coatings, namely, octadecylsilane, aminosilane, and polyurethane, are further described and applied to anticonvulsants analyses. The investigated plasmatic range for the evaluated anticonvulsants, using CW-TPR fiber, were the following: phenylethylmalonamide (3.00–40.0 μg mL⁻¹), phenobarbital (5.00–40.0 μg mL⁻¹), primidone (3.00–40.0 μg mL⁻¹), carbamazepine and carbamazepine-epoxide (2.00–24.0 μg mL⁻¹), phenytoin (2.00–40.0 μg mL⁻¹), and lamotrigine (0.50–12.0 μg mL⁻¹). The antidepressants’ linear plasmatic concentration ranged from 75.0 to 500 ng mL⁻¹ for imipramine, amitriptyline, and desipramine, and from 50.0 to 500 ng mL⁻¹ for nortriptyline, being in all cases, the limit of quantification represented by the lowest value. The precision (interassays) for all investigated drugs in plasma sample spiked with different concentrations of each analyte and submitted to the described procedures were lower than 15%. The off-line SPME/LC methodologies developed allow anticonvulsants and antidepressants analyses from therapeutic to toxic levels for therapeutic drug monitoring.     

LC–MS–MS determination of ibuprofen, 2-hydroxyibuprofen enantiomers,and carboxyibuprofen stereoisomers for application in biotransformation studies employing endophytic fungi

The purpose of this study was the development and validation of an LC–MS–MS method for simultaneous analysis of ibuprofen (IBP), 2-hydroxyibuprofen (2-OH-IBP) enantiomers, and carboxyibuprofen (COOH-IBP) stereoisomers in fungi culture medium, to investigate the ability of some endophytic fungi to biotransform the chiral drug IBP into its metabolites. Resolution of IBP and the stereoisomers of its main metabolites was achieved by use of a Chiralpak AS-H column (150 × 4.6 mm, 5 μm particle size), column temperature 8 °C, and the mobile phase hexane–isopropanol–trifluoroacetic acid (95: 5: 0.1, v/v) at a flow rate of 1.2 mL min⁻¹. Post-column infusion with 10 mmol L⁻¹ ammonium acetate in methanol at a flow rate of 0.3 mL min⁻¹ was performed to enhance MS detection (positive electrospray ionization). Liquid–liquid extraction was used for sample preparation with hexane–ethyl acetate (1:1, v/v) as extraction solvent. Linearity was obtained in the range 0.1–20 μg mL⁻¹ for IBP, 0.05–7.5 μg mL⁻¹ for each 2-OH-IBP enantiomer, and 0.025–5.0 μg mL⁻¹ for each COOH-IBP stereoisomer (r ≥ 0.99). The coefficients of variation and relative errors obtained in precision and accuracy studies (within-day and between-day) were below 15%. The stability studies showed that the samples were stable (p > 0.05) during freeze and thaw cycles, short-term exposure to room temperature, storage at −20 °C, and biotransformation conditions. Among the six fungi studied, only the strains Nigrospora sphaerica (SS67) and Chaetomium globosum (VR10) biotransformed IBP enantioselectively, with greater formation of the metabolite (+)-(S)-2-OH-IBP. Formation of the COOH-IBP stereoisomers, which involves hydroxylation at C3 and further oxidation to form the carboxyl group, was not observed.     

Reversed Capillary Electrochromatography: Investigation of Peak Symmetry for Eluting Curves of Solutes

Summary A clean method without use of organic solvents has been developed for isolation and high-performance liquid chromatographic (HPLC) determination of oxytetracycline (OTC) and sulphadimidine (SDD) in cow's milk. Isolation is rapid and simple—homogenization with an inorganic acid solution by means of a handy ultrasonic homogenizer, which is easy-to-use and portable, followed by centrifugation. Reversed-phase HPLC was performed on a C₄ column, with 1.25 mmol L⁻¹ succinic acid solution as mobile phase, and identification was by means of a photodiode-array detector. Separation of the analytes was achieved in less than 8 min. Significant linearity was established over the concentration range of 0.1–1.0 μg mL⁻¹ for both target compounds (r>0.99,P<0.01). Average recoveries of OTC and SDD (each spiked at 0.1–1.0 μg mL⁻¹) were ≥88.8, and inter- and intra-assay variability was ≤2.8%. The total time required for analysis of one sample was <20 min. The limits of quantitation of the method (μg mL⁻¹ in milk) were 0.044 for OTC and 0.023 for SDD. No organic solvent was used at any stage of the analysis.     

Fluorescence enhancement of the protein–curcumin–sodium dodecyl benzene sulfonate system and protein determination

Protein can greatly enhance the fluorescence of curcumin (CU) in the presence of sodium dodecyl benzene sulfonate (SDBS). Experiments indicate that under the optimum conditions, the enhanced intensity of fluorescence is proportional to the concentration of proteins in the range of 0.0050–20.0 μg mL⁻¹ for bovine serum albumin (BSA), 0.080–20.0 μg mL⁻¹ for human serum albumin (HSA), and 0.040–28.0 μg mL⁻¹ for egg albumin (EA). Their detection limits (S/N=3) are 1.4 ng mL⁻¹, 20 ng mL⁻¹, and 16 ng mL⁻¹, respectively. The method has been satisfactorily used for the determination of proteins in actual samples. In comparison with most of fluorimetric methods, this method is quick and simple, has high sensitivity and good stability. The interaction mechanism is also studied.     

Utility of <Emphasis Type="Italic">π</Emphasis>-acceptor reagents for spectrophotometric determination of sulphonamide drugs via charge-transfer complex formation

A simple, sensitive and accurate spectrophotometric method for the determination of sulphonamides (sulphamethoxazole (SMZ), sulphaguanidine (SGD), sulphaquinoxaline sodium (SQX), sulphametrole (SMR), and sulphadimidine sodium (SDD)) has been developed. The charge-transfer reactions between sulphonamides as n-electron donors and 7,7,8,8-tetracyanoquinodimethane (TCNQ), 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), and 2,5-dichloro-3,6-dihydroxy-1,4-benzoquinone (chloranilic acid, p-CLA) as π-acceptors resulting in highly coloured complexes were studied. Experimental conditions for these CT reactions were carefully optimised. Beer’s law is valid over the concentration ranges from 4–280 μg mL⁻¹, 4–260 μg mL⁻¹, 4–200 μg mL⁻¹, and 4–200 μg mL⁻¹ of SMZ, SGD, SQX, and SDD using DDQ reagent, respectively. While the calibration curves are linear in the concentration ranges from 4–180 μg mL⁻¹, 4–80 μg mL⁻¹, 4–60 μg mL⁻¹, 4–180 μg mL⁻¹, and 4–60 μg mL⁻¹ of SMZ, SGD, SQX, SMR, and SDD, respectively, using TCNQ reagent and from 4–380 μg mL⁻¹ and 4–300 μg mL⁻¹ of SQX and SDD, respectively, using p-CLA reagent, respectively. Different analytical parameters, namely molar absorptivity (ε), standard deviation, relative standard deviation, correlation coefficient, limit of detection, and limit of quantification, were calculated. The results obtained by the proposed methods are in good agreement with those obtained by the official method as indicated by the percent recovery values.     

Simultaneous Determination of Lidocaine, Proline and Lomefloxacin in Human Urine by CE with Electrochemiluminescence Detection

A new method was developed for the simultaneous determination of lidocaine, proline and lomefloxacin in human urine by capillary electrophoresis-electrochemiluminescence detection with Ru(bpy)₃ ²⁺. Conditions of the separation and detection were investigated and optimized. It was proved that 20 mM phosphate buffer at pH 6.7 could achieve the most favorable resolution, and the high sensitivity of detection was obtained by using the detection potential at 1.15 V and 5 mM Ru(bpy)₃ ²⁺–60 mM phosphate buffer at pH 7.6 in the detection reservoir. The detection limits were 0.02 μg mL⁻¹ for lidocaine, 0.03 μg mL⁻¹ for proline and 0.06 μg mL⁻¹ for lomefloxacin. Relative standard deviations of the ECL intensity and the migration time were 3.5 and 1.1% for 6 μg mL⁻¹ lidocaine, 3.2 and 1.0% for 6 μg mL⁻¹ proline and 3.7 and 1.2% for 6 μg mL⁻¹ lomefloxacin, respectively. A baseline separation for lidocaine, proline and lomefloxacin was achieved within 360 s. The developed method was successfully applied to determine the amounts of lidocaine, proline and lomefloxacin in human urine. The recovery and RSD were in the range of 93.3–97.2 and 3.8–4.9%, respectively.