Determination of phosphate/arsenate by a modified molybdenum blue method and reduction of arsenate by S2O4

A modified molybdenum blue method for fast and accurate measurement of arsenate and phosphate in aqueous solution at concentrations below 10 μmol l⁻¹ is reported. The modification consists of optimizing the composition of the molybdenum-containing solution (potassium antimony tartrate, ammonium molybdate, sulfuric acid).Selective reduction of arsenate by sodium dithionite is used to determine phosphate concentrations, and for the speciation of arsenate and arsenite, in an aqueous mixture according to the scheme developed by Johnston and Pilson. Sodium dithionite is used for the first time to achieve complete, fast (<10 min), and selective reduction of arsenate in neutral solution.These two significant improvements afforded a colorimetric limit for As detection near 1 ppb, which easily meets the requirements imposed by the revised EPA threshold levels for As in drinking water.     

Dual fluorescence/contactless conductivity detection for microfluidic chip

A new dual detection system for microchip is reported. Both fluorescence detector (FD) and contactless conductivity detector (CCD) were combined together and integrated on a microfluidic chip. They shared a common detection position and responded simultaneously. A blue light-emitting diode was used as excitation source and a small planar photodiode was used to collect the emitted fluorescence in fluorescence detection, which made the device more compact and portable. The coupling of the fluorescence and contactless conductivity modes at the same position of a single separation channel enhanced the detection characterization of sample and offered simultaneous detection information of both fluorescent and charged specimen. The detection conditions of the system were optimized. K⁺, Na⁺, fluorescein sodium, fluorescein isothiocyanate (FITC) and FITC-labeled amino acids were used to evaluate the performance of the dual detection system. The limits of detection (LOD) of FD for fluorescein Na⁺, FITC, FITC-labeled arginine (Arg), glycine (Gly) and phenylalanine (Phe) were 0.02 μmol L⁻¹, 0.05 μmol L⁻¹, 0.16 μmol L⁻¹, 0.15 μmol L⁻¹, 0.12 μmol L⁻¹ respectively, and the limits of detection (LOD) of CCD achieved 0.58 μmol L⁻¹ and 0.39 μmol L⁻¹ for K⁺ and Na⁺ respectively.     

Sequential injection determination of orthophosphate as ion associate of 12-molybdophosphate with Astra Phloxine

A simple and fast reaction between 12-molybdophosphate heteropoly anion and the polymethine dye Astra Phloxine was used for the development of a new SIA method for the determination of orthophosphate. The suggested method has higher sensitivity and a broader calibration range than existing SIA methods. It can be used to detect phosphate in the range from 0.2 to 10 μmol L⁻¹ with a detection limit of 0.1 μmol L⁻¹ and an acceptable throughput of 20 samples h⁻¹. The comparably low molybdate (5.6 mmol L⁻¹) and dye (0.1 mmol L⁻¹) concentrations led to an improvement in the stability of the base line. Inter-day reproducibility was better than 5%, while the intra-day RSD was in the range 0.8-3.5%. The method was used for the analysis of natural water samples and phosphorus-containing chemicals having a low content of orthophosphate.     

A portable and low cost equipment for flow injection chemiluminescence measurements

A compact, reliable and low cost flow injection chemiluminescence system is described. The flow system consists of a set of solenoid micro-pumps that can dispense reproductive micro-volumes of solutions. The luminometer was based on a coiled cell constructed from polyethylene tubing that was sandwiched between two large area photodiodes. The whole equipment costs about US$ 750 and weights ca. 3 kg. Equipment performance was evaluated by measuring low concentrations of hydrogen peroxide by oxidation of luminol and for the determination of ammonium, based on its inhibition of the luminescence provided by the reaction of luminol and sodium hypochlorite. Linear responses were achieved within 1.0-80 μmol L⁻¹ H₂O₂ and 0.6-60 μmol L⁻¹ NH₄⁺ with detection limits estimated as 400 nmol L⁻¹ H₂O₂ and 60 nmol L⁻¹ NH₄⁺ at the 99.7% confidence level. Coefficients of variation were 1.0 and 1.8%, estimated for 20 μmol L⁻¹ H₂O₂ and 15 μmol L⁻¹ NH₄⁺ (n = 20), respectively. Reagent consumption of 55 μg luminol, effluent volume of 950 μL per determination and sampling rate of 120 samples per hour were also achieved.     

Biomimetic sensor based on MnMn complex as manganese peroxidase mimetic for determination of rutin

A novel biomimetic sensor for rutin determination based on a dinuclear complex [Mn^IIIMn^II(Ldtb)(μ-OAc)₂]BPh₄ containing an unsymmetrical dinucleating ligand, 2-[N,N-bis(2-pyridylmethyl)-aminomethyl]-6-[N-(3,5-di-tert-butyl-2-oxidoben-zyl)-N-(2-pyridylamino)aminomethyl]-4-methylphenol (H₂Ldtb), as a manganese peroxidase mimetic was developed. Several parameters were investigated to evaluate the performance of the biomimetic sensor obtained after the incorporation of the dinuclear complex in a carbon paste. The best performance was obtained in 75:15:10% (w/w/w) of the graphite powder:Nujol:Mn^IIIMn^II complex, 0.1 mol L⁻¹ phosphate buffer solution (pH 6.0) and 4.0 × 10⁻⁵ mol L⁻¹ hydrogen peroxide. The response of the sensor towards rutin concentration was linear using square wave voltammetry in the range of 9.99 × 10⁻⁷ to 6.54 × 10⁻⁵ mol L⁻¹ (r = 0.9998) with a detection limit of 1.75 × 10⁻⁷ mol L⁻¹. The recovery study performed with pharmaceuticals ranged from 96.6% to 103.2% and the relative standard deviation was 1.85% for a solution containing 1.0 × 10⁻³ mol L⁻¹ rutin (n = 6). The lifetime of this biomimetic sensor was 200 days (at least 750 determinations). The results obtained for rutin in pharmaceuticals using the biomimetic sensor and those obtained with the official method are in agreement at the 95% confidence level.     

Simultaneous spectrophotometric determination of phosphate and silicate ions in river water by using ion-exclusion chromatographic separation and post-column derivatization

The simultaneous spectrophotometric determination of phosphate and silicate ions in river water was examined by using ion-exclusion chromatography and post-column derivatization. Phosphate and silicate ions were separated by the ion-exclusion column packed with a polymethacrylate-based weakly acidic cation-exchange resin in the H⁺-form (TSKgel Super IC-A/C) by using ultra pure water as an eluent. After the post-column derivatization with molybdate and ascorbic acid, so-called molybdenum-blue, both ions were determined simultaneously by spectrophotometry. The effects of sulfuric acid, sodium molybdate and ascorbic acid concentrations and reaction coil length, which have relation to form the reduced complexes of molybdate and ions, on the detector response for phosphate and silicate ions were investigated. Under the optimized conditions (color-forming reactant, 50 mM sulfuric acid-10 mM sodium molybdate; reducing agent, 50 mM ascorbic acid; reaction coil length, 6 m), the calibration curves of phosphate and silicate ions were linear in the range of 50-2000 μg L⁻¹ as P and 250-10,000 μg L⁻¹ as Si. This method was successfully applied to water quality monitoring of Kurose-river watershed and it suggested that the effluent from a biological sewage treatment plant was significant source of phosphate ion in Kurose-river water.     

Cobalt phthalocyanine as a novel molecular recognition reagent for batch and flow injection potentiometric and spectrophotometric determination of anionic surfactants

Cobalt(II) phthalocyanine [Co(II)Pc] is used as both an ionophore and chromogen for batch and flow injection potentiometric and spectrophotometric determination of anionic surfactants (SDS), respectively. The potentiometric technique involves preparation of a polymeric membrane sensor by dispersing [Co(II)Pc] in a plasticized PVC membrane. Under batch mode of operation, the sensor displays a near-Nernstian slope of −56.5 mV decade⁻¹, wide response linear range of 7.8 × 10⁻⁴ to 8.0 × 10⁻⁷ mol L⁻¹, lower detection limit of 2.5 × 10⁻⁷ mol L⁻¹ and exhibits high selectivity for anionic surfactants in the presence of many common ions. Under hydrodynamic mode of operation (FIA), the slope of the calibration plot, limit of detection, and working linear range are −51.1 mV decade⁻¹, 5.6 × 10⁻⁷ and 1.0 × 10⁻³ to 1.0 × 10⁻⁶ mol L⁻¹, respectively. The spectrophotometric method is based on the use of [Co(II)Pc] solution in dimethylsulfoxide (DMSO) as a chromogenic reagent. The maximum absorption of the reagent at 658 nm linearly decreases with the increase of anionic surfactant over the concentration range 2-30 μg mL⁻¹. The lower limit of detection is 1 μg mL⁻¹ and high concentrations of many interfering ions are tolerated. Flow injection spectrophotometric measurements are carried out by injection of the surfactant test solution in a stream of the reagent in DMSO. The sample throughput, working range and lower detection limit are 25-30 samples h⁻¹, 4-60 and 2 μg mL⁻¹, respectively. The potentiometric and spectrophotometric techniques are applied to the batch and flow injection measurements of anionic surfactants in some commercial detergent products. The results agree fairly well with data obtained using the standard methylene blue spectrophotometric method.     

Microtiterplate phosphate assay based on luminescence quenching of a terbium complex amenable to decay time detection

We describe a terbium-ligand complex (TbL) for a microtiterplate assay for phosphate (P) in the 0.3-100 μmol L⁻¹ range based on luminescence quenching. As the pH optimum is at neutral pH (7.4) the probe is quenched by both, primary (H₂PO₄⁻) and secondary phosphate (HPO₄²⁻). The LOD is 110 nmol L⁻¹. A Stern-Volmer study revealed that quenching is mostly static. Due to the ms-decay time of TbL, the first luminescence lifetime assay for phosphate could also be developed. The lifetime-based calibration plot is linear between 0.5 and 5 μmol L⁻¹ of P. The effect of various surfactants on assay performance and a study on interferents are presented. The probe was successfully applied to determination of P in commercial plant fertilizers and validated against the molybdenum blue test. The probe is the most sensitive lanthanide-based probe for phosphate.     

Simultaneous determination of Mn(II), Cu(II) and Fe(III) as 2-(5′-bromo-2′-pyridylazo)-5-diethylaminophenol complexes by adsorptive cathodic stripping voltammetry at a carbon paste electrode

A simple and precise square-wave adsorptive cathodic stripping voltammetry (SW-AdCSV) method has been described for simultaneous determination of Mn(II), Cu(II) and Fe(III) in water samples using a carbon paste electrode. In 0.1 mol L⁻¹ acetate buffer (pH 5) containing 50 μmol L⁻¹ of 2-(5′-bromo-2′-pyridylazo)-5-diethylaminophenol (5-Br-PADAP), Mn(II), Cu(II) and Fe(III) were simultaneously determined as metal-complexes with 5-Br-PADAP following preconcentration onto the carbon paste electrode by adsorptive accumulation at +1.0 V (vs. Ag/AgCl/3 M KCl). Insignificant interference from various cations (K⁺, Na⁺, Mg²⁺, Ca²⁺, Al³⁺, Bi³⁺, Sb³⁺, Se⁴⁺, Zn²⁺, Ni²⁺, Co²⁺, Cd²⁺, Pb²⁺, V⁵⁺, Ti⁴⁺ and NH₄⁺), anions (HCO₃⁻, Cl⁻, NO³⁻, SO₄²⁻ and PO₄³⁻) and ascorbic acid was noticed. Limits of detection of 0.066, 0.108 and 0.093 μg L⁻¹ and limits of quantitation of 0.22, 0.36 and 0.31 μg L⁻¹ Mn(II), Cu(II) and Fe(III), respectively, were achieved by the described method. The described stripping voltammetry method was successfully applied for simultaneous determination of Mn(II), Cu(II) and Fe(III) in ground, tap and bottled natural water samples.     

Cloud point extraction for ultra-trace Cd determination in microwave-digested biological samples by ETAAS

Cloud point extraction (CPE) has been used for the preconcentration of cadmium, after the formation of a complex with 2-(5-bromo-2-pyridylazo)-5-(diethylamino)-phenol (5-Br-PADAP), and further determination by graphite furnace atomic absorption spectrometry (ETAAS) using polyethyleneglicolmono-p-nonyphenylether (PONPE 7.5) as surfactant. The chemical variables that affect the cloud point extraction were optimized. The separation of the two phases was easily accomplished by cooling the mixture in order to make more viscous the surfactant-rich phase. In order to establish the optimum conditions for the determination of Cd by ETAAS, Pd + Mg, Pt, Ir, Rh and Ru were studied as chemical modifiers. The best thermal stabilization was obtained with Pd + Mg, with a maximum pyrolysis temperature of 1100 °C. Under the optimum conditions i.e., pH 9.0, [5-Br-PADAP] = 2.0 × 10⁻⁵ mol L⁻¹, [PONPE 7.5] = 0.02% (w/v), an enhancement factor of 22-fold was reached. The lower limit of detection (LOD) obtained under the optimal conditions was 0.008 μg L⁻¹. The precision for 10 replicate determinations at 0.2 μg L⁻¹ Cd was 3.5% relative standard deviation (R.S.D.). The calibration graph using the preconcentration method was linear with a correlation coefficient of 0.9984 at levels close to the detection limit up to at least 1.0 μg L⁻¹. The method was successfully applied to the determination of cadmium in urine samples and in a water standard reference material.     

Speciation analysis of mercury in sediments, zoobenthos and river water samples by high-performance liquid chromatography hyphenated to atomic fluorescence spectrometry following preconcentration by solid phase extraction

A high-pressure microwave digestion was applied for microwave-assisted extraction (MAE) of mercury species from sediments and zoobenthos samples. A mixture containing 3 mol L⁻¹ HCl, 50% aqueous methanol and 0.2 mol L⁻¹ citric acid (for masking co-extracted Fe³⁺) was selected as the most suitable extraction agent. The efficiency of proposed extraction method was better than 95% with R.S.D. below 6%. A preconcentration method utilizing a “homemade” C18 solid phase extraction (SPE) microcolumns was developed to enhance sensitivity of the mercury species determination using on-column complex formation of mercury-2-mercaptophenol complexes. Methanol was chosen for counter-current elution of the retained mercury complexes achieving a preconcentration factor as much as 1000. The preconcentration method was applied for the speciation analysis of mercury in river water samples. The high-performance liquid chromatography-cold vapour atomic fluorescence spectrometric (HPLC/CV-AFS) method was used for the speciation analysis of mercury. The complete separation of four mercury species was achieved by an isocratic elution of aqueous methanol (65%/35%) on a Zorbax SB-C18 column (4.6 mm × 150 mm, 5 μm) using the same complexation reagent (2-mercaptophenol). The limits of detection were 4.3 μg L⁻¹ for methylmercury (MeHg⁺), 1.4 μg L⁻¹ for ethylmercury (EtHg⁺), 0.8 μg L⁻¹ for inorganic mercury (Hg²⁺), 0.8 μg L⁻¹ for phenylmercury (PhHg⁺).     

The petentiometric determination of peroxide hydrogen and glucose on the glassy electrode modified by the calix[4]arene

A new petentiometric method to determine peroxide hydrogen and glucose had been studied. This method had been applied on the petentiometric determination of peroxide hydrogen and glucose in the total ionic strength adjustment buffer (TISAB) (pH 7.5) solution with the glassy electrode modified by the calix[4]arene. The glassy carbon electrode covered with the calix[4]arene depended on the H₂O₂ concentration in the range of log[H₂O₂] from −3.3 to −1.2 in the solution of TISAB (pH 7.5) with nearly Nernstian slope of about 65.6 ± 3 mV and the detection limit of peroxide hydrogen was 4.0 × 10⁻⁵ mol L⁻¹. The glassy carbon electrode covered with the calix[4]arene depended on the glucose concentration in the range of log[glucose] from −3.6 to −2.8 in the solution of TISAB (pH 7.5) with nearly Nernstian slope of about 50.2 ± 2 mV and the detection limit of glucose was 2.0 × 10⁻⁵ mol L⁻¹. The electrode had the good selectivity, sensitivity, stability and repeatability.     

Flow injection analysis of ethyl xanthate by in-line dialysis and UV spectrophotometric detection

A simple and sensitive spectrophotometric flow method for determination of low concentrations of the flotation collector O-ethyldithiocarbonate (ethyl xanthate, CH₃CH₂-O-CS₂⁻) in solutions is described. The method is based on ethyl xanthate detection at 301 nm in medium of NaOH 50 mmol L⁻¹. By injection of 200 μL of sample, the analytical method shows linear response for the ethyl xanthate concentration from 0.5 up to 500 μmol L⁻¹. Successive injections of 4 μmol L⁻¹ ethyl xanthate (n = 23) show a coefficient of variation lower than 0.6%, denoting high repeatability. The detection limit is 0.3 μmol L⁻¹. At a flow rate of 2.0 mL min⁻¹, a frequency of 120 injections/h of ethyl xanthate can be attained. By introduction of a tangential dialysis cell in the FIA system, the manual sample filtration step with 0.22 μm filter was eliminated and the residual interference of suspended material, was completely overcome even for unfiltered sludge suspension samples, an important advantage that compensates for the frequency reduction to 25 injections/h elevation and detection limit elevation to 2 μmol L⁻¹, still outreaching for many applications. Potential applications of the method embrace the at line determination of ethyl xanthate in the ore processing industry, control of the concentration at its optimal level during the flotation process, as well as monitoring of residues in the effluents.     

Linear polyamines as carriers in thiocyanate-selective membrane electrodes

The polyamines, octyl-[2-(2-octylamino-ethylamino)-ethyl]-amine (L¹) and octyl-{2-[2-(2-octylamino-ethylamino)-ethylamino]-ethyl}-amine (L²), have been used as anion ionophores in PVC-based membrane ion-selective electrodes. Different electrodes were prepared containing L¹, or L², and o-nitrophenyl octyl ether (NPOE) or bis(2-ethylhexyl)sebacate (DOS) as plasticizers. The response of the electrodes was tested in two different buffers, HEPES-KOH (pH 7) and MES-KOH (pH 5.6). Electrodes containing L¹ and L² with NPOE (E1 and E2, respectively) showed a Nernstian response for thiocyanate with a good response time. The detection limit, linear range and slope for electrode E1 were 3.8 × 10⁻⁶ mol dm⁻³, 1 × 10⁻⁵ to 1 × 10⁻¹ mol dm⁻³ and −57.2 mV decade⁻¹ at pH 5.6 and 4.47 × 10⁻⁶ mol dm⁻³, 1.95 × 10⁻⁵ to 1 × 10⁻¹ mol dm⁻³ and −58.1 mV decade⁻¹ at pH 7.0. For electrode E2 the detection limit, linear range and slope found were 2.63 × 10⁻⁶ mol dm⁻³, 7.94 × 10⁻⁶ to 1 × 10⁻¹ mol dm⁻³ and −58.5 mV decade⁻¹ at pH 5.6 and 1.23 × 10⁻⁵ mol dm⁻³, 7.95 × 10⁻⁵ to 1 × 10⁻¹ mol dm⁻³ and −46.0 mV decade⁻¹ at pH 7. In contrast, electrodes containing DOS as plasticizers gave only response at pH 5.6 (detection limit, linear range and slope at pH 5.6 were 3.16 × 10⁻⁵ mol dm⁻³, 1 × 10⁻⁴ to 1 × 10⁻¹ mol dm⁻³ and −52.6 mV decade⁻¹). Selectivity coefficients for different anions with respect to thiocyanate were calculated. The electrode E2 at pH 5.6 was also used for the determination of SCN⁻ by potentiometric titrations with Ag⁺ ions with good results. The electrode E2 was also used to determine concentrations of thiocyanate in biological samples.     

Langmuir-Blodgett film of p-tert-butylthiacalix[4]arene modified glassy carbon electrode as voltammetric sensor for the determination of Hg(II)

The π-A isotherms and UV-vis spectra of the transferred films suggested that the monolayer of p-tert-butylthiacalix[4]arene can coordinate with Hg²⁺ at the air-water surface. From these observations, a glassy carbon electrode coated with Langmuir-Blodgett film of p-tert-butylthiacalix[4] arene as a new voltammetric sensor is designed for the determination of trace amounts of Hg²⁺. Compared with bare glassy carbon electrode and modified glassy carbon electrode using direct coating method, the Langmuir-Blodgett film-modified electrode can greatly improve the measuring sensitivity of Hg²⁺. Under the selected conditions, the Langmuir-Blodgett film-modified electrode in 0.1 mol L⁻¹ H₂SO₄ + 0.01 mol L⁻¹ KCl solution shows a linear voltammetric response for Hg²⁺ in the range of 5.0 × 10⁻¹⁰ to 1.5 × 10⁻⁷ mol L⁻¹, with a detection limit of 2.0 × 10⁻¹⁰ mol L⁻¹. The proposed method was also applied to determine Hg²⁺ in water samples (tap, lake and river water). In addition, the fabricated electrode exhibited a distinct advantage of simple preparation, non-toxicity, good reproducibility and good stability.     

The use of a gold disc microelectrode for the determination of copper in human sweat

A novel approach of using a gold disc microelectrode to analyze sweat samples for copper ions by anodic square wave stripping voltammetry (SW stripping voltammetry) is described. Sweat was collected from the lower back of four subjects after physical exercise and the sample volume required for the determinations was 100 μL. Under the optimized conditions, the calibration plot was linear over the range 1-100 μmol L⁻¹ Cu(II) with a limit of detection of 0.25 μmol L⁻¹. The precision was evaluated by carrying out five replicate measurements in a 1 μmol L⁻¹ Cu(II) solution and the standard deviation was found to be 1.5%. Measurements were performed by inserting the microelectrode into sweat drops and Cu(II) concentrations in the analyzed samples ranged from 0.9 to 28 μmol L⁻¹. Values obtained by the proposed voltammetric method agreed well with those found using graphite furnace atomic absorption spectroscopy (GFAAS).     

Synchronous fluorescence determination of DNA based on the interaction between methylene blue and DNA

The fluorescence intensity of methylene blue (MB) quenched by DNA in the pH range of 6.5-8.0 was studied with synchronous fluorescence technology. A novel method for detecting single-stranded and double-stranded DNA was developed. The decreased fluorescence intensity at 664 nm is in proportion to the concentration of DNA in the range of 0.28-11.0 μmol L⁻¹ for ctDNA, 0.14-8.25 μmol L⁻¹ for thermally denatured ctDNA and 0.28-8.25 μmol L⁻¹ for hsDNA. The detection limits (S/N = 3) are 0.11, 0.04 and 0.04 μmol L⁻¹, respectively. The method is rapid, selective, and the reagents are lower toxic. It has been used for the determination of DNA in synthetic samples with good satisfaction. In addition, the interaction modes between MB and ctDNA and the mechanism of the fluorescence quenching were also discussed in detail. The experimental results from absorption spectra and fluorescence polarization indicate that the possible interaction modes between MB and DNA are the electrostatic binding and the intercalation binding.     

Microflow injection chemiluminescence system with spiral microchannel for the determination of cisplatin in human serum

A new microflow injection chemiluminescence (μFI-CL) system was described for the determination of cisplatin in human serum. By using the microchip with double spiral channel configuration, the sensitivity was greatly enhanced due to more efficient mixing of the analyte and reagent solutions. Experimental results revealed that common ions in human serum, such as Mn²⁺, Co²⁺, Fe³⁺, Cu²⁺, Zn²⁺, Ni²⁺, Na⁺, K⁺, Ca²⁺, Cl⁻, NO₃⁻, Ac⁻, CO₃²⁻, PO₄³⁻, SO₄²⁻ did not cause interference with the detection of Pt(II) by using 1,10-phenanthroline as the masking agent. Under the optimized conditions, a linear calibration curve (R² = 0.998) over the range 2.0 × 10⁻⁸ to 2.0 × 10⁻⁶ mol L⁻¹ was obtained with the detection limit of 1.24 × 10⁻⁹ mol L⁻¹. The relative standard deviation was found to be 3.46% (n = 12) for 2.0 × 10⁻⁷ mol L⁻¹. The sample consumption was only 2 μL with the sample throughput of 72 h⁻¹. It had been used for trace platinum determination in cisplatin injection and human serum samples after the dosage of cisplatin. The recovery varied from 97.6 to 103.9%. The results proved that the proposed μFI-CL system had the advantages of high sensitivity and precision, low sample and reagents consumption, and high analytical throughput.     

Rosmarinic acid determination using biomimetic sensor based on purple acid phosphatase mimetic

A new heterodinuclear Fe(III)Zn(II) complex which mimics the active site of the hydrolytic enzyme red kidney bean purple acid phosphatase was synthesized and characterized by IR, CHN and X-ray crystallographic analyses. This complex, [Fe^IIIZn^II(μ-OH)bpbpmp-CH₃](ClO₄)₂, containing the ligand (H₂bpbpmp-CH₃ = {2-[bis(2-pyridylmethyl)aminomethyl]-6-[(2-hydroxy-5-methylbenzyl) (2-pyridyl-methyl) aminomethyl]-4-methyl-phenol}) was employed in the construction of a biomimetic sensor and used in the determination of rosmarinic acid in plant extract samples. The response parameters and optimization of the biomimetic sensor design were evaluated. The best performance of this sensor was obtained for 75:15:10% (w/w/w) of the graphite powder:nujol:Fe(III)Zn(II) complex, 0.1 mol L⁻¹ phosphate buffer solution (pH 7.5), 1.19 × 10⁻⁴ mol L⁻¹ hydrogen peroxide with frequency, pulse amplitude, and scan increment at 30 Hz, 100 mV, and 0.6 mV, respectively. The rosmarinic acid concentration was linear in the range of 2.98 × 10⁻⁵ to 3.83 × 10⁻⁴ mol L⁻¹ (r = 0.9991) with a detection limit of 2.30 × 10⁻⁶ mol L⁻¹. This biomimetic sensor demonstrated long-term stability (300 days; 900 determinations) and reproducibility, with a relative standard deviation of 12.0%. The recovery study of rosmarinic acid in plant extract samples gave values from 90.3 to 98.3% and the concentrations determined showed agreement when compared with those obtained using capillary electrophoresis at the 95% confidence level.