Values Of LOD Research Articles

Stability-Indicating Method of Entacapone-Related Substances Using UPLC in Finished Dosage Form

A rapid, specific, sensitive and robust method for entacapone-related substances in carbidopa, entacapone and levodopa film-coated tablet was developed using ACQUITY UPLC with BEH C18 column. Mobile phase (A), i.e. 0.1% orthophosphoric acid and mobile phase (B), i.e. acetonitrile with water in the ratio 75 : 25 (v/v) with gradient method were employed. The method was evaluated for identification of process impurities and unknown impurities. It has been validated according to ICH (Q2) R1 guidelines. The values of LOD and LOQ for impurity and entacapone were found to be 0.01% and 0.03% respectively. Degradation studies were carried out in peroxide, acid, alkali conditions and contribution to mass balance was established. It was stable under heat, light-exposed and humid conditions.

A Novel HPLC Method Validation Based on Analytical Techniques of Metoclopramide Benzamide Derivative (Metoclopramide Base) and its Determination from Solid Dispersion by Solvent Evaporation Method

The crucial endeavor in this method is focused on the study analysing recovery and column efficiency (theoretical plate) after extraction of metoclopramide from PVP K30-Metoclopramide (3:1 w/w) solid dispersion (SD). The proposed work also presents a reversed-phase high performance liquid chromatography method for the detection of very small amount of metoclopramide base followed by different analytical techniques. In this context, it may be noted that the values of LOD and LOQ for the metoclopramide base are obtained as, 0.052 µg/mL and 0.159 µg/mL respectively. The linearity is performed in the range of 2-20 µg/mL; performance of column remained satisfactory throughout the analysis, N= 4927-2434. The percentage recovery of pure and extracted metoclopramide is ranging from 103%-105% and 86%-120% approximately. The results collectively demonstrate that the proposed method is selective. Clear isolation of metoclopramide from SD is achieved without any solid state interference.

LISINOPRIL AN OVERVIEW

Lisinopril is an oral long-acting angiotensin converting enzyme inhibitor. It is a lysinederivative of Enalprilate and has structural similarity with its substrate. It differs from captopril by lacking the sulfhydryl group [1,2]. Lisinopril is chemically described as (S)-1-[N2-(1carboxy-3-phenylpropyl)-L-lysyl]-Lproline dihydrate. Its empirical formula is C21H31N3O5.2H2O.The present study was aimed at developing a simple, sensitive, precise and accurate HPLC method for the estimation of Lisinopril from bulk samples and tablet dosage forms. A non-polar C8 analytical chromatographic column was chosen as the stationary phase for the separation and determination of Lisinopril. Mixtures of commonly used solvents like water, methanol and acetonitrile with or without buffers in different combinations were tested as mobile phases. The choice of the optimum composition is based on the chromatographic response factor, a good peak shape with minimum tailing. A mixture of buffer and methanol in the ratio of 35:65 v/v was proved to be the most suitable of all the combinations since the chromatographic peak obtained was well defined, better resolved and almost free from tailing. The retention time of Lisinopril was found to be 2.29 min. The linearity was found satisfactory for the drug in the range 20.0-60.0 μg/mL (Table 1.4). Precision of the method was studied by repeated injection of tablet solution and results showed lower % RSD values (Table 1.5-1.7). This reveals that the method is quite precise. The percent recoveries of the drug solutions were studied at three different concentration levels. The percent individual recovery and the % RSD at each level were within the acceptable limit (Table 1.8). This indicates that the method is accurate. The absence of additional peaks in the chromatogram indicates non-interference of the commonly used excipients in the tablets and hence the method is specific. The deliberate changes in the method have not much affected the peak tailing, theoretical plates and the percent assay. This indicates that the present method is robust (Table 1.9). The system suitability studies were carried out to check various parameters such as theoretical plates and tailing factor (Table 1.10). The lowest values of LOD and LOQ as obtained by the proposed method indicate that the method is sensitive (Table 1.11). The solution stability studies indicate that the drug was stable up to 24 hours (Table 1.12). The forced degradation studies indicate that the drug was stable in stability studies (Table 1.13).

Multicenter evaluation of the new immunoassay method for TSH measurement using the automated DxI platform

Aim of the studyRecently, Beckman Coulter Diagnostics set up a new TSH immunoassay for the automated DxI platform. The aim of this study was to evaluate and compare the analytical performance and clinical results of this method with those of previous method. Material and methodsA multicenter study (named TSH ELAS Study) was organized using 593 serum samples, collected from healthy subjects and patients with thyroid disorders, and 13 control samples, circulated in an External Quality Assessment (EQA) scheme. ResultsThe values of LoB and LoD, and LoQ at 20% CV were 0.0004mIU/L, 0.001mIU/L and 0.0023mIU/L, respectively. Moreover, TSH concentrations >0.01mIU/L actually show imprecision values lower than 5% CV. This new TSH assay showed a systematic underestimation (on average of 6.25%) compared to old method, which is mainly due to larger differences between methods for samples with low TSH concentrations, related to the better analytical sensitivity of new compared to old method. In a reference population, including 279 apparently healthy adult subjects, Caucasian volunteers (mean age 43.6years, age 20–63years, 138 women and 141 males) the distribution of TSH concentrations was: mean (CI 95%) 1.694mIU/L (1.588–1.779), median 1.495mIU/L (1.412–1.588mIU/L), 97.5th percentile 3.707mIU/L. ConclusionsThe new TSH immunoassay for DxI platform shows some relevant improvements compared to the previous one: use of the most recent WHO 3rd IRP 81/563 standard and monoclonal antibodies (instead of polyclonal antibodies of the old method), and better analytical sensitivities and reproducibility.

Substitution influence of halo polyurethane foam on the removal of bismuth, cobalt, iron and molybdenum ions from environmental samples

Abstract Polyurethane foam was modified by the addition of halogen atoms and amino groups into the foam matrix in order to increase its polarity for the sorption of ionic species. The halo-modified sorbents (X-PUFs, X = Cl, Br or I) were characterized using FTIR, UV/Vis and Raman spectroscopy, EA, XRD and SEM. The average values of pHZCP, maximum value of ∆pH (pH 9) and bulk DC conductivity for X-PUFs are 3.8, −1.9 and 2.6 × 10−5 Ω−1 cm−1, respectively. The activity of the surface functional groups of X-PUFs were evaluated by the extraction of Bi(III), Co(II), Fe(III) and Mo(III) ions and extraction conditions including pH, shaking time, flow rate, temperature and initial metal concentration were optimized. Maximum extraction of those ions was achieved as their thiocyanate complexes in acidic medium (pH 1–3, 5–6 and 1.5–2 mol L−1 H2SO4) within 1–30 min. A perfect isotherm curve with almost zero intercept (0.008), good correlation coefficient R2 (0.973) was obtained. The values of LOD, LOQ and RSD (n = 6) for the determination of Bi(III), Co(II), Fe(III) and Mo(III) ions are 2 ng L−1, 7 ng L−1 and 0.75%, respectively. The sorption capacities of the tested metal ions onto Cl-PUF, Br-PUF and I-PUF are 0.16, 0.14 and 0.14 mmol g−1, respectively. The accuracy of the procedure was verified by the analysis of food, ground water and pharmaceutical samples with recovery and average RSD values of 99–100% and 2.39%, respectively.

Assessment of the effect of three-dimensional mantle density heterogeneity on Earth rotation in tidal frequencies

In this paper, we report the assessment of the effect of the three-dimensional (3D) density heterogeneity in the mantle on Earth orientation parameters (EOP) (i.e., the polar motion, or PM, and the length of day, or LOD) in the tidal frequencies. The 3D mantle density model is estimated based upon a global S-wave velocity tomography model (S16U6L8) and the mineralogical knowledge derived from laboratory experiment. The lateral density variation is referenced against the preliminary reference earth model (PREM). Using this approach the effects of the heterogeneous mantle density variation in all three tidal frequencies (zonal long periods, tesseral diurnal, and sectorial semidiurnal) are estimated in both PM and LOD. When compared with mass or density perturbations originated on the Earth's surface such as the oceanic and barometric changes, the heterogeneous mantle contributes less than 10% of the total variation in PM and LOD in tidal frequencies. However, this is the gap that has not been explained to close the gap of the observation and modeling in PM and LOD. By computing the PM and LOD caused by 3D heterogeneity of the mantle during the period of continuous space geodetic measurement campaigns (e.g., CONT94) and the contribution from ocean tides as predicted by tide models derived from satellite altimetry observations (e.g., TOPEX/Poseidon) in the same period, we got the lump-sum values of PM and LOD. The computed total effects and the observed PM and LOD are generally agree with each other. In another word, the difference of the observed PM and LOD and the model only considering ocean tides, at all tidal frequencies (long periods, diurnals, and semidiurnals) contains the contributions of the lateral density heterogeneity of the mantle. Study of the effect of mantle density heterogeneity effect on torque-free Earth rotation may provide useful constraints to construct the reference earth model (REM), which is the next major objective in global geophysics research beyond PREM.

Recovery and determination of palladium from its alloys using iminodiacetic polyurethane/carbon nanofibers sorbent

ABSTRACTNew sorbent was synthesized by coupling iminodiacetic polyurethane foam with carbon nanofibers to increase its surface area and sorption capacity. FTIR, UV/Vis, Raman spectroscopy, elemental analysis, and scanning electron microscopy were used to characterize the IDAPUF/CNFs sorbent. By using these techniques, it was found that the sorbent contains 0.58, 0.62, and 2.23 mmol g−1 of amino, carboxylic, and phenolic groups, respectively. The maximum sorption (99–100%) of palladium(II) ions onto IDAPUF/CNFs was achieved within 10–15 min at pH 5.0. A perfect isotherm curve with a zero intercept (0.0003) and good correlation (R2 = 1) was obtained. The capacity of the IDAPUF/CNFs sorbent preloaded onto a glass column was calculated to be 0.58 mmol g−1. The values of LOD, LOQ, and RSD% (n = 6) are 0.004 ng mL−1, 0.013 ng mL−1, and 1.17%, respectively. The accuracy of the procedure was verified by the recovery of Pd(II) ions (100%) from some palladium alloys and road dust samples (RSD% = 0.36).

Development and validation of a stability-indicating RP-LC method for the simultaneous determination of otilonium bromide and its expected degradation product in bulk drug and pharmaceutical preparation

A simple, precise, accurate and rapid stability indicating liquid chromatography method had been developed for the simultaneous determination of otilonium bromide (OB) and its expected degradation product; p -aminobenzoic acid (PABA) in bulk powder and pharmaceutical preparation. Chromatographic separation was carried out by isocratic elution on Waters Atlantis C18 column (4.6 × 150 mm, 5 µm) using 2 mM ammonium acetate buffer (pH = 2.35) containing 0.05% TFA: acetonitrile (30:70, v : v ) as the mobile phase. The flow rate was 0.8 mL/min with UV detection at 290 nm. Linearity was obtained over a concentration range of 0.5-100 μg/mL with regression coefficient of 1 for OB, and over concentration range of 1-50 μg/mL with regression coefficient of 0.9998 for PABA. The values of LOD and LOQ were found to be 0.0665 and 0.2018 μg/mL for OB and 0.1974 and 0.598 μg/mL for PABA, respectively. The method was validated as per ICH guidelines. The method was successfully applied to the determination of the drug in bulk powder and pharmaceutical preparation.

UV Spectrophotometric Method for the Estimation of Roflumilast in Human Serum

Ultra violet spectrophotometric method for the estimation roflumilast in spiked human serium. The present study was spectrophotometric estimation of roflumilast in spiked human serum is based on measurement at the maximum wavelength 248 nm using 0.2M Hcl as a solvent. The roflumilast stock solution was prepared with distilled water to produse calibration curve. The standard solution of roflumilast shows absorption maxima at 248 nm. The roflumilast obeyed beer lamberts law in the concentration range of 40-88μg/ml with regression 0.9987 at 248nm. The overall percentage recovery of roflumilast was found to be 99.52% which reflects that the method was free from the interference. The % RSD values of intra day and inter day precision was found to be 0.031 and 0. 046% respectively, which is less than 2 and hence this method is precise. The values of LOD and LOQ were found to be 0.783 and 2.375 μg/ml, respectively and it indicates this method was sensitive.

Spectroscopic Studies and Applications of the Reactions of Some Anti-Diabetic and Anti-Hypertensive Drugs with Rose Bengal

Simple, rapid and sensitive spectrophotometric methods were developed and validated for the microdetermination of terazosin HCl, doxazosin mesylate and pioglitazone HCl drugs in pure and pharmaceutical dosage forms. These methods are based on ion-pair formation reaction between these drugs and a chromogenic reagent Rose Bengal (RBeng). These reactions were studied under various conditions and the optimum parameters were selected. The spectrophotometric microdeterminations have been done at λmax=570 nm for terazosin HCl and pioglitazone HCl and at λmax=575 nm for doxazosin mesylate. Under proper conditions the suggested procedures were successfully applied for microdetermination of these drugs in pure and in pharmaceutical dosage forms. The values of SD, RSD, recovery %, LOD, LOQ and Sandell sensitivity refer to the high accuracy and precession of the applied procedures. The results obtained were compared with the data obtained by the official methods, referring to confidence and agreement with rose bengal procedure results. The solid drugs-reagent ion-pairs were prepared, separated and their structures were investigated using elemental analysis, FT-IR, 1H-NMR and thermal analyses and the results confirm the structures proposed by the stoichoimetric ratio in the solution work. The biological activities of the drugs and their solid ion-pairs against some types of (G-) and (G+) bacteria and fungai were studied and compared with each other. It is found that terazosin-RBeng and pioglitazone-RBeng reaction products have antibacterial effect higher than the parent drugs, but doxazosin-RBeng reaction product has almost the same antibacterial effect of the parent drug.

Three-Component EC-SPR Biosensor Based on Graphene Oxide, SiO2 and Gold Nanoparticles in NADH Determination

The main goal of this paper was to develop a biosensor containing three types of structures: graphene oxide (GO), silica nanoparticles (SNs) and gold nanoparticles (GNPs) for effective electrochemical and optical determination of dihydronicotinamide adenine dinucleotide dehydrogenase (NADH). The subsequent steps in biosensor formation, the manner of structure organization and the layer thickness were verified by different methods. Electrochemical oxidation of NADH was performed with the usage of differential pulse voltammetry method (DPV). Results show that obtained biosensor catalyzes the process of NADH electrochemical oxidation. The shift of oxidation potential toward less positive values and the increment of oxidation current were clearly noted. Additionally, the oxidized NAD form was found to exhibit an electrostatic affinity to the top layer of sensor giving a possibility to SPR detection. DPV and SPR signals registered for different amount of NADH were correlated with its concentration into linear relationship: y = 8.6388x − 0.0109; R2 = 0.9997 and y = 1305.8x − 74.098; R2 = 0.9913, respectively. Enhancement of the electrochemical sensitivity toward NADH, as a result of electrode modification, was proved by lower values of LOD (0.0236 mM) and LOQ (0.0707 mM) in comparison with corresponding parameters calculated for non-modified gold electrode.

Investigation of electrochemical behavior of anisomycin on gold electrode followed by HPLC–MS/MS analysis

Anisomycin is an immunosuppressant in low doses (< 0.1 μM) with possible application in treatment of some autoimmune diseases and in inhibiting transplantation rejection. Anisomycin suppresses malignant tumor cell growth and affects memory. For the first time it was the subject of the electrochemical investigations by cyclic voltammetry and square wave voltammetry on gold electrode in 0.05 M NaHCO3 using its electrochemical activity. The cyclic voltammetry experiments at different sweep rates show that electrochemical process is irreversible and diffusion controlled. Based on square wave voltammetry measurements, the calculated values of LOD and LOQ were 1 and 4 nM (in the absence of biological fluid), as well as 2 and 6 nM (in the presence of spiked urine) indicating the high sensitivity of the proposed electroanalytical method. High performance liquid chromatography–tandem mass spectrometry was a reference method for quantification of anisomycin and served for structural identification of its hydrolysis product (deacetylanisomycin).

Determination of ketosteroid hormones in meat by liquid chromatography tandem mass spectrometry and derivatization chemistry.

A method for the determination and quantification of ketosteroid hormones in meat by mass spectrometry, based on the derivatization of the carbonyl moiety of steroids by O-methylhydroxylamine, is presented. The quantitative assay is performed by means of multiple-reaction-monitoring (MRM) scan mode and using the corresponding labelled species, obtained by reaction with d 3-methoxylamine, as internal standard. The accuracy of the method was established by evaluating artificially spiked samples, obtaining values in the range 90-110%. Recovery tests were performed on blank matrix samples spiked with non-natural steroids including trenbolone and melengestrol acetate. The latter experiment revealed that the yield of the extraction processes was approximately 60%. Good values of LOQ and LOD were achieved, making this method competitive with current hormone assay methods.

Development and validation of an RP-HPLC method for quantification of trans-resveratrol in the plant extracts

New, simple, cost effective, accurate and reproducible RP-HPLC method was developed and validated for the quantification of trans-resveratrol in the extracts of grape exocarp and seeds. The method has proved to be simpler and faster than available methods. Methanol was used as a mobile phase with a flow rate of 1.0 cm3 min-1, while the quantification was effected at 306 nm. The separation was performed at 35?C using a C18 column. The results showed that the peak area response was linear in the concentration range of 1-40 ?g cm-3. The values of LOD and LOQ were found to be 0.125 and 0.413 ?g cm-3, respectively. The antioxidant activity of the extracts was determined using DPPH assay. The ability of DPPH radicals inhibition decreases in the following order: the extract of grape exocarp &gt; trans-resveratrol standard &gt; the extract of grape seeds.

Second-order calibration for simultaneous determination of pharmaceuticals in water samples by solid-phase extraction and fast high-performance liquid chromatography with diode array detector

A fast high-performance liquid chromatography–diode array detection (HPLC–DAD) approach combined to solid phase extraction (SPE) as a pre-concentration step is developed for simultaneous determination of five selected pharmaceuticals (carbamazepine, naproxen, diclofenac, gemfibrozil and mefenamic acid) in water samples. The effective factors on the efficiency of SPE procedure are optimized using faced-centered central composite design (FCCD). In addition, multi-response optimization by using Derringer's desirability function is used to find the optimum experimental conditions for extraction of analytes from well and river waters. Due to the complexity of water matrices and the presence of different chromatographic issues, new combination of multivariate curve resolution–correlation optimized warping to parallel factor analysis (MCR–COW–PARAFAC) and multivariate curve resolution-alternating least squares with trilinearity constraint (MCR–COW–MCRtril) is proposed for simultaneous determination of five target pharmaceuticals in these samples. This strategy allowed us to overcome matrix effects and to exploit second-order advantage. Recoveries ranging from 80.12% to 102.71% and relative standard deviations below 8.0% for all pharmaceuticals proved the accuracy and precision of the proposed method. In addition, the values of relative error in calibration curves (RE, %) were below 11.24% and 12.19% for MCR–COW–PARAFAC and MCR–COW–MCR, respectively. Furthermore, the values of detection limits (LODs) and quantification limits (LOQs) were between 0.02 to 0.32ngmL−1 and 0.07 to 1.07ngmL−1, respectively. In addition, analytical figures of merit using multivariate approaches were calculated for both methods. In this regard, the values of sensitivity, LOD and LOQ showed an improvement compared to univariate techniques. Inspection of the results showed that due to the efficient correction of elution time shifts using MCR–COW method and preserving trilinear data structure, both MCR–COW–PARAFAC and MCR–COW–MCRtril give similar results. These results confirmed that coupling optimized SPE–HPLC–DAD method with second-order calibration algorithms can be considered as an efficient method for fast, simple and cost effective quantification of pharmaceuticals in highly contaminated samples, such as river and well waters.

Analysis and Evaluation of Fatty Acid in Rattan Tea by Highly Selective and Sensitive HPLC- FLD-MS Method Coupled with Pre-Column Fluorescent Labeling

Rattan tea regarded as one of the most promising dietary supplements for the prevention and treatment of many chronic diseases has gained extensive attention. Free fatty acids play an important role in human body as essential nutrients. Contents of 31 free fatty acids in Rattan Tea were firstly investigated using toluene-4-sulfonic acid 2-(naphthalene-1-yl-phenyl-amino)-ethyl ester as pre-column fluorescence derivatization reagent by RE-HPLC coupled with an atmospheric chemical ionization-mass spectrometry source (APCI/ MS). The results indicated that lower values of LOD (0.52-1.53 ng mL(-1) ) provided the guarantee for trace analysis and evaluation of free fatty acids. The experimental recoveries were in the range of 88.9-103.6 %. Results of the present study indicated the highest free fatty acids content of Rattan Tea was in April. Thirteen free fatty acids were found, including C18:2 omega 6, C18:3 omega 3, omega 6 and C20:4 omega 6 essential to life. The developed method exhibited powerful potential for the systematic detection of bioactive components of medicinal plants.

Development and validation of a new RP-HPLC method for determination of quercetin in green tea

Green tea (Camellia sinensis) contains quercetin as a bioactive compound. Quercetin has anti-inflammatory and anticancer effects. The aim of this paper was to develop and validate an RP-HPLC method for determination of quercetin in green tea simpler and faster than other available methods. RP-HPLC analysis was performed by isocratic elution with a flow rate of 1.0 mL/min. Pure methanol was used as a mobile phase, while the quantification was effected at 370 nm. The separation was performed at 35°C using a C18 column (4.6 × 250 mm, 5 μm). The results showed that the peak area response is linear within the concentration range of 10–70 μg/mL (r = 0.9986). The values of LOD and LOQ were 1.2 and 4 μg/mL, respectively. For the intra-day and intra-instrument reproducibility, RSD were in the range of 0.05–0.84% and 0.89–1.55%, respectively. The results of accuracy for the different concentrations of quercetin (40, 50 and 60 μg/mL) were 101.3, 98.4, and 98.2%. The developed and validated method was successfully applied to the determination of quercetin in green tea extract.

NEW GENERATION HALO COLUMN FOR FAST ANALYSES OF ASPIRIN AND ATORVASTATIN IN PHARMACEUTICAL PREPARATION

A super-fast (within 4 min only) HPLC method is described for the analyses of aspirin and atorvastatin simultaneously in single dosage formulation. The mobile phase used was water (pH 2.0, adjusted by o-phosphoric acid)-acetonitrile (50:50, v/v) at 1.0 mL/min flow rate with 210 nm detection. The column used was new generation Halo C18(100 × 46 mm; shell particles; 2.7 µm). The capacity factors of aspirin and atorvastatin were 1.20 and 2.01, while the separation and resolution factors were 1.68 and 5.63, respectively. The values of LOD and LOQ for aspirin were 0.10 and 0.50 µg/mL, while these values for atorvastatin were 0.50 and 2.5 µg/mL, respectively. The linearity was observed in the ranges of 0.1 to 5.0 mg/mL for both drugs. The reported method can be used for the quality control of aspirin and atorvastatin in pharmaceutical preparations with good economy. Additionally, this method can also be used for the analyses of aspirin and atorvastatin drugs in biological and environmental samples.

Residue depletion of sulfachlorpyrazine in edible tissues of broiler chickens

The residue depletion profile of sulfachlorpyrazine was studied in healthy broilers after oral administration of Sulfatyf®, given at a dose of 50 mg sulfachlorpyrazine (SCP) per kg body weight once daily for 3 consecutive days. Twenty-five medicated broilers were slaughtered on the 5th, 10th, 14th and 16th day of post-medication, and muscle, fat with skin, liver and kidney tissues were sampled and analysed using an high-performance liquid chromatography (HPLC) method with satisfactory recovery (74.5% ± 2.9%) and specificity. The estimated values of LOD and LOQ were 18.40 and 55.70 ng ml−1, respectively. On the 5th and 10th day of post-medication, the concentrations of SCP residue in all samples examined were higher than the corresponding MRLs established by the European Union and the highest SCP concentrations were measured in muscle. A maximum withdrawal time of 14 days was calculated to ensure consumer safety.

Highly sensitive and specific determination of mercury(II) ion in water, food and cosmetic samples with an ELISA based on a novel monoclonal antibody

Mercury is one of the most toxic heavy metals present in the environment. In this study, a highly sensitive and specific monoclonal antibody (mAb)-based indirect competitive enzyme-linked immunosorbent assay (ELISA) for the determination of Hg(2+) was developed. A new bifunctional ligand, 6-mercaptonicotinic acid (MNA), which contains a pyridine ring bearing a carboxylic group and a mercapto group, was selected for the preparation of immunogen. After immunization of mice and performing the hybridoma technique, the obtained mAb was characterized for its binding affinity and selectivity for Hg(2+). Based on this novel mAb, an ELISA was established. At optimal experimental conditions, the standard curve of the ELISA for Hg(2+) was constructed in concentration range of 0.1-100 ng mL(-1). The values of IC(50) and LOD of the assay were found to be 1.12 and 0.08 ng mL(-1). The cross-reactivity was lower than 2% with MNA, CH(3)Hg, and CH(3)Hg-MNA and was 11.5% and 4.4% for Hg(+) and Au(3+), respectively. No cross-reactivity was found with other metal ions such as Cu(2+), Sn(2+), Ni(2+), Mn(2+), Pb(2+), Zn(2+), Cd(2+), Fe(2+), Co(2+), Mg(2+), Ca(2+), and anions such as Cl(-), NO(3)(-), NO(2)(-), HCO(3)(-), F(-), and SO(4)(2-), indicating that the assay displays not only high sensitivity but also high selectivity. Different kinds of samples including water, milk, green vegetable, kelp, facial cleanser, and night cream were spiked with Hg(2+) and the extracts were analyzed by ELISA. Acceptable recovery rates of 80.0-113.0% and coefficients of variation of 1.9-18.6% were obtained. A linear relationship between ELISA and cold-vapor atomic fluorescence spectroscopy (CV-AFS) as indicated by a correlation coefficient of 0.97 for liquid samples (water samples) and 0.98 for other samples was obtained. The proposed mAb-based ELISA provides a feasible analytical method for highly sensitive and specific, fast, simple, and accurate determination of uncomplexed trace Hg(2+) in environmental and food samples.