Quick analysis of baicalin in Scutellariae Radix by enzyme-linked immunosorbent assay using a monoclonal antibody

To establish an immunoassay for baicalin (BA), a hybridoma cell line (9D6) secreting a monoclonal antibody (MAb) against BA was prepared by cell fusion with splenocytes derived from a mouse immunized with BA-bovine serum albumin (BSA) conjugate and a myeloma cell line, SP2/0-Ag14. MAb 9D6 shows specific reactivity against BA and its aglycone, baicalein, but not against other natural products. We developed an enzyme-linked immunosorbent assay (ELISA) using MAb 9D6 in a competitive manner, ranging from 200 ng/mL to 2 μg/mL. After validating the developed ELISA on the basis of intra- and inter-assays and a recovery experiment, it was found that the ELISA was not only simple, but also sufficiently reliable and accurate for quality control of Scutellariae Radix. It allowed determination of BA in complex and mixed materials, such as Kampo medicines.     

Development and validation of an immunochromatographic assay for rapid multi-residues detection of cephems in milk

A one-step immunochromatographic assay (ICA) was developed for the detection of seven kinds of cephems in milk. Polyclonal antibodies (PcAb) with group-specific to cephems were raised in rabbits after immunization with cephalexin-keyhole limpet hemocyanin (KLH) conjugate. The specificity of anti-sera was determined by indirect competitive enzyme-linked immunosorbent assay (icELISA), and the 50% inhibitions (IC₅₀) of cephalexin and cefadroxil were obtained at 1.5 ng mL⁻¹; IC₅₀ of cefatiofur, cefapirin, cefazolin, cefalothin and cefotaxine were 4, 3.7, 3.2, 4.5 and 5 ng mL⁻¹, respectively. The PcAb against cephems were conjugated to colloidal gold particles as the detection reagent for ICA strips to test for cephems. This method achieved semi-quantitative detection of cephems in <5 min, with high sensitivity to cephalexin and cefadroxil (both 0.5 ng mL⁻¹). At the same time, cefatiofur, cefapirin, cefazolin, cefalothin and cefotaxine were detected at <100 ng mL⁻¹ in spiked processed-milk samples. This method was compared with an enzyme-linked immunosorbent assay by testing 40 milk samples, and the positive samples were validated by a high-performance liquid chromatographic method, with an agreement rate of 100% for both comparisons. In conclusion, the method was rapid and accurate for the multi-residue detection of cephems in milk.     

Highly sensitive multianalyte immunochromatographic test strip for rapid chemiluminescent detection of ractopamine and salbutamol

A novel immunochromatographic assay (ICA) was proposed for rapid and multiple assay of β₂-agonists, by utilizing ractopamine (RAC) and salbutamol (SAL) as the models. Owing to the introduction of chemiluminescent (CL) approach, the proposed protocol shows much higher sensitivity. In this work, the described ICA was based on a competitive format, and horseradish peroxidase-tagged antibodies were used as highly sensitive CL probes. Quantitative analysis of β₂-agonists was achieved by recording the CL signals of the probes captured on the two test zones of the nitrocellulose membrane. Under the optimum conditions, RAC and SAL could be detected within the linear ranges of 0.50–40 and 0.10–50 ng mL⁻¹, with the detection limits of 0.20 and 0.040 ng mL⁻¹ (S/N = 3), respectively. The whole process for multianalyte immunoassay of RAC and SAL can be completed within 20 min. Furthermore, the test strip was validated with spiked swine urine samples and the results showed that this method was reliable in measuring β₂-agonists in swine urine. This CL-based multianalyte test strip shows a series of advantages such as high sensitivity, ideal selectivity, simple manipulation, high assay efficiency and low cost. Thus, it opens up new pathway for rapid screening and field analysis, and shows a promising prospect in food safety.     

Colloidal gold nanoparticle probe-based immunochromatographic assay for the rapid detection of chromium ions in water and serum samples

An immunochromatographic assay (ICA) using gold nanoparticles coated with monoclonal antibody (McAb) for the detection of chromium ions (Cr) in water and serum samples was developed, optimized and validated. Gold nanoparticles coated with affinity-purified monoclonal antibodies against isothiocyanobenzyl-EDTA (iEDTA)-chelated Cr³⁺ were used as the detecting reagent in this completive immunoassay-based one-step test strip. The ICA was investigated to measure chromium speciation (Cr³⁺ and Cr⁶⁺ ions) in water samples. Chromium standard samples of 0–80 ng mL⁻¹ in water were determined by the test strips. The results showed that the visual lowest detection limit (LDL) of the test strip was 50.0 ng mL⁻¹. A portable colorimetric lateral flow reader was used for the quantification of Cr. The results indicated that the linear range of the ICA with colorimetric detection was 5–80 ng mL⁻¹. The ICA was also validated for the detection of chromium ions in serum samples. The test trips showed high stability in that they could be stored at 37 °C for at least 12 weeks without significant loss of activity. The test strip also showed good selectivity for Cr detection with negligible interference from other heavy metals. Because of its low cost and short testing time (within 5 min), the test strip is especially suitable for on-site large-scale screening of Cr-polluted water samples, biomonitoring of Cr exposure, and many other field applications.     

Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for the analysis of glycyrrhizic acid

Glycyrrhizic acid (GL) is a major active compound of licorice. The specific monoclonal antibody (MAb) (designated as 8F₈A₈H₄2H₇) against GL was produced with the immunogen GL–BSA conjugate. The dissociation constant (K _d) value of the MAb was approximately 9.96×10⁻¹⁰ M. The cross reactivity of the MAb with glycyrrhetic acid was approximately 2.6%. The conventional indirect competitive enzyme-linked immunosorbent assay (icELISA) and simplified icELISA adapted with a modified procedure were established using the MAb. The IC₅₀ value and the detect range by the conventional icELISA were 1.1 ng mL⁻¹ and 0.2–5.1 ng mL⁻¹, respectively. The IC₅₀ value and the detect range by the simplified icELISA were 5.3 ng mL⁻¹ and 1.2–23.8 ng mL⁻¹, respectively. The two icELISA formats were used to analyze GL contents in the roots of wild licorice and different parts of cultivated licorice (Glycyrrhiza uralensis Fisch). The results obtained with the two icELISAs agreed well with those of the HPLC analysis. The correlation coefficient was more than 0.98 between HPLC and the two icELISAs. The two icELISAs were shown to be appropriate, simple, and effective for the quality control of raw licorice root materials.     

Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B1 detection in cereal

A rapid and sensitive one-step competitive enzyme immunoassay for the detection of FB₁ was developed. The anti-idiotypic nanobody–alkaline phosphatase (Ab2β−Nb−AP) was validated by the AP enzyme activity and the properties of bounding to anti-FB1-mAb (3F11) through colorimetric and chemiluminescence analyses. The 50% inhibitory concentration and the detection limit (LOD) of colorimetric enzyme-linked immunosorbent assay (ELISA) for FB₁ were 2.69 and 0.35 ng mL⁻¹, respectively, with a linear range of 0.93–7.73 ng mL⁻¹. The LOD of the chemiluminescence ELISA (CLIA) was 0.12 ng mL⁻¹, and the IC₅₀ was 0.89 ± 0.09 ng mL⁻¹ with a linear range of 0.29–2.68 ng mL⁻¹. Compared with LC-MS/MS, the results of this assay indicated the reliability of the Ab2β−Nb−AP fusion protein based one-step competitive immunoassay for monitoring FB₁ contamination in cereals. The Ab2β−Nb−AP fusion proteins have the potential to replace chemically-coupled probes in competitive enzyme immunoassay systems.     

Chemiluminescence determination of barbituric acid using Ru(phen)(3)(2+)-Ce(IV) system

A chemiluminescence (CL) method for the determination of barbituric acid (BA) was proposed, which is based on the enhancement of BA to the CL intensity of Tris-(1,10-phenanthroline)ruthenium(II) (Ru(phen)₃²⁺)-cerium(IV) (Ce(IV)) system. The concentration of BA is proportional to the CL intensity in the range of 5.0×10⁻³-2.0 μg ml⁻¹. The detection limit is 6.9×10⁻⁴ μg ml⁻¹. The relative standard deviation (R.S.D.) of determining 11 samples containing 0.20 μg ml⁻¹ BA is 3.2%. This CL method has been successfully applied to the determination of BA in the synthetic samples. The mechanism of CL reaction was studied.     

Spectral and fluorescent kinetics features of Nd3+ ion in Nb2O5, Ta2O5 and La2O3 mixed lithium zirconium silicate glasses

A sensitive competitive flow injection chemiluminescence (CL-FIA) immunoassay for immunoglobulin G (IgG) was developed using gold nanoparticle as CL label. In the configuration, anti-IgG antibody was immobilized on a glass capillary column surface by 3-(aminopropyl)-triethoxysilane and glutaraldehyde to form immunoaffinity column. Analyte IgG and gold nanoparticle labeled IgG were passed through the immunoaffinity column mounted in a flow system and competed for the surface-confined anti-IgG antibody. CL emission was generated from the reaction between luminol and hydrogen peroxide in the presence of Au (III), generated from chemically oxidative dissolution of gold nanoparticle by an injection of 0.10 mol L⁻¹ HCl–0.10 mol L⁻¹ NaCl solution containing 0.10 mmol L⁻¹ Br₂. The concentration of analyte IgG was inversely related to the amount of bound gold nanoparticle labeled IgG and the CL intensity was linear with the concentration of analyte IgG from 1.0 ng mL⁻¹ to 40 ng mL⁻¹ with a detection limit of 5.2 × 10⁻¹⁰ g mL⁻¹. The whole assay time including the injections and washing steps was only 30 min for one sample, which was competitive with CL immunoassays based on a gold nanoparticle label and magnetic separation. This work demonstrates that the CL immunoassay incorporation of nanoparticle label and flow injection is promising for clinical assay with sensitivity and high-speed.     

A liposome immune lysis assay for enrofloxacin in carp and chicken muscle

A homogenous complement-mediated liposome immune lysis assay (LILA) was developed for the determination of enrofloxacin (ENRO) in carp and chicken muscle. ENRO was covalently coupled to DPPE, and then immobilized onto the surface of liposomes by reverse-phase evaporation method. The performed liposome would be specifically lysed by the sequential additions of anti-ENRO monoclonal antibody (MAb) and guinea pig complement. Through a competitive assay format, the performed liposome can be used to detect ENRO in a range of 5.0-20 ng mL⁻¹ in assay buffer. The limit of detection of ENRO in carp and chicken muscle was 1 ng g⁻¹ and the limit of quantification was 2 ng g⁻¹. Recoveries ranged from 58.3% to 65.2% for carp and 55.6-63.8% for chicken muscle at spiked levels of 2-8 ng g⁻¹, with intra-assay and inter-assay variations 5.6-12.3% and 7.1-19.2%, respectively.     

A bare-eye based one-step signal amplified semiquantitative immunochromatographic assay for the detection of imidacloprid in Chinese cabbage samples

A novel bare-eye based one-step signal amplified semi-quantitative immunochromatographic assay (SAS-ICA) was developed for detection of the pesticide imidacloprid. This method was based on competitive immunoreactions. Signal amplification was achieved by dual labeling of the test lines (TLs) on the strip using high affinity nanogold-biotinylated anti-imidacloprid mAb (BAb) and nanogold-streptavidin (Sa) probes. The relative color intensities of three TLs (TL-I, TL-II and TL-III) on a nitrocellulose (NC) membrane were used for direct visual analysis of the SAS-ICA strips, and could be used for semi-quantitation of analyte concentrations by observing what TLs disappeared in the amplification zone. Under optimized conditions, the following imidacloprid concentration ranges would be detected by visual examination of the SAS-ICA strip: 0–5 ng mL⁻¹ (negative samples), and 5–25 ng mL⁻¹, 25–250 ng mL⁻¹, 250–1000 ng mL⁻¹ and >1000 ng mL⁻¹ (positive samples). The sensitivity (the visual detection limit (VDL) of TL-III) and semi-quantitative analytical capacity (when TL-III disappeared completely) of the SAS-ICA strip were 10-fold and 160-fold higher than those of traditional ICA, respectively. The developed SAS-ICA strip was applied to the analysis of spiked and authentic contaminated Chinese cabbage samples in the laboratory and under field conditions, and the results were validated by high-performance liquid chromatography (HPLC). This process could be adopted as a potential generous technique for all ICA-based detection methods.     

Development of an immunoassay for rapid screening of vardenafil and its potential analogues in herbal products based on a group specific monoclonal antibody

Vardenafil is a phosphodiesterase-5 (PDE-5) inhibitor for the treatment of erectile dysfunction (ED). Undeclared vardenafil and related analogues adulterated in herbal products are a threat to public health. To screen vardenafil and its analogues in herbal matrix rapidly, an immunoassay based on a group specific monoclonal antibody (McAb) was developed.Glutaraldehyde was used to link vardenafil to immunogen and coating-antigen, respectively. Through the assessment of the structural specificity of eight anti-vardenafil McAbs, the McAb of 4B9 was characterized as being specific to the common structure of vardenafil and its analogues. An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was established based on this McAb, the limit of detection of vardenafil was 5.0 ng mL⁻¹, the calibration curve was linear from 5.0 to 40 ng mL⁻¹ (R² = 0.952) with an IC₅₀ value of 18.2 ng mL⁻¹. In the extracts of 20 Chinese traditional drugs, the detection capability (CCβ) of vardenafil was 0.08 mg g⁻¹, the recoveries were 76-116% and the coefficients of variation (CV%) were 9.7%-16.2%. The ic-ELISA was in good agreement with LC-UV when detected herbal products containing vardenafil and its analogue.The method is a suitable tool for screening vardenafil and its analogues as illegal additives in herbal products.     

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.     

Determination of citrate in tablets and of oxytetracycline in serum using europium (III) luminescence

A direct method for the determination of citrate and oxytetracycline in samples containing complex matrices like tablets or serum has been developed using the luminescence of the ternary complex formed with Eu(III) ions. The triplet-state energy level of oxytetracycline (OxTc), the excitation maximum (412 nm) and the luminescence lifetime of Eu-OxTc (58 μs) were determined. A 17-fold luminescence enhancement at 615 nm occurs upon addition of citrate within a short 5-min incubation time at neutral pH. This is accompanied by a threefold increase of the luminescence decay time. The optimal conditions for determination of OxTc are equal concentrations of Eu (III) and citrate (C = 1 · 10^− 4 mol L^− 1) and pH 7.2. For determination of citrate, the optimal concentrations of Eu(III) and OxTc are 1 : 0.5 (C_Eu = 1 · 10^− 4 mol L^− 1, C_OxTc = 5 · 10^− 5 mol L^− 1) at pH 7.2. The linear range for determination of OxTc in serum is 0.25-250 μg mL^− 1, and for citrate in tablets from 0.5 to 10.0 μg mL^− 1 (2.3 · 10^− 6- 4 · 10^− 5 mol L^− 1). The detection limit was 0.1 μg mL^− 1 for OxTc and 0.2 μg mL^− 1 (1 · 10^-6 mol L^− 1) for citrate, respectively. A comparison of the new method with other methods for determination of citrate is given.     

An indirect competitive enzyme-linked immunosorbent assay for determination of norfloxacin in waters using a specific polyclonal antibody

A specific polyclonal anti-norfloxacin antibody was obtained, and a sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed for determining trace amounts of norfloxacin in various waters. Good linearity was achieved in the range from 0.1 to 10 μg L⁻¹. The average IC₅₀ value was determined to be 2.2 μg L⁻¹ and the limit of detection was 0.016 μg L⁻¹ at a signal-to-noise ratio of 3 in phosphate-buffered saline buffer. Recoveries of norfloxacin at various spiking levels ranged from 74 to 105% in groundwater, surface water, treated and untreated wastewater samples, with relative standard deviations of 3–5%. The assay was applied for determining norfloxacin in municipal wastewater, surface water, and groundwater collected in a metropolis of China. Raw wastewater samples were only submitted to filtration and pH adjustment while the other water samples were pre-concentrated by solid phase extraction prior to the icELISA assay. Good agreement of the results obtained by the icELISA and liquid chromatography tandem mass spectrometry further confirmed the reliability and accuracy of the icELISA for rapid detection of norfloxacin in waters.     

Development of direct competitive enzyme-linked aptamer assay for determination of dopamine in serum

A rapid and cost-effective screening method based on a competitive enzyme-linked aptamer assay (ELAA) for dopamine (DA) in serum has been optimized and validated. In this paper, we report advantageous sensitivity and specificity of aptamer assays as compared to the existing antibody based-immunoassays. The RNA aptamer (67 mer) was immobilized via site-directed immobilization with biotin both at the 3′-end on aptamer and at neutravidin plate. Various factors such as incubation temperature, divalent ion – Mg²⁺ ion and treatment of serum solution were evaluated for the performance of ELAA. The aptamer was incubated for 1 h at 4 °C in the assay buffer containing 5 mM Mg²⁺ ion, and serum was diluted (1:9, serum:assay buffer) and filtrated through a 3 kDa dialysis membrane to extract the proteins present in the serum. Assay was performed with 0.01 μg mL⁻¹ of aptamer and 1.205 × 10⁻⁷ M DA-HRP conjugate using the optimized method. A dose–response curve was constructed, and the limit of detection and a dynamic range for the DA were determined as 1.0 × 10⁻¹² M and four orders (1.0 × 10⁻⁷ M to 5.0 × 10⁻¹¹ M) of magnitude, respectively. The correlation diagram of the absorbance obtained both in buffer and in serum has shown a good agreement with the correlation coefficient (R² = 0.9872): Abs. (in serum) = 0.9612 × Abs. (in buffer) − 0.0556. The cross-reactivity evaluation demonstrated that norepinephrine showed some cross-reactivity (3.68%) whereas 3-methoxytyramine, epinephrine, homovanillic acid and 3,4-dihydroxyphenylacetic acid showed almost no cross-reactivity (<1%). Percent recoveries of DA in serum were quite satisfactory (∼95%). This paper describes usefulness of the aptamer assay in monitoring DA in human serum.     

Determination of N-methylcarbamate insecticides in water samples using dispersive liquid–liquid microextraction and HPLC with the aid of experimental design and desirability function

A rapid and simple method for the extraction and preconcentration of N-methylcarbamates (NMCs) (carbofuran, carbaryl and promecarb) in water samples using dispersive liquid–liquid microextraction (DLLME) using chemometrics was developed. Influence variables such as volume of extracting (CHCl₃) and dispersing solvents (ACN), pH and ionic strength, extraction time and centrifugation time and speed were screened in a 2^7–4 Plackett–Burman design was investigated. The significant variables were optimized by using a central composite design (CCD) combined with desirability function (DF). At optimum conditions values of variables set as 126 μL chloroform, 1.5 mL acetonitrile, 1 min extraction time, 10 min centrifugation at 4000 rpm min⁻¹, natural pH, 4.7% (w/v) NaCl, the separation was reached in less than 14 min using a C₁₈ column and an isocratic binary mobile phase (acetonitrile: water (50:50, v/v)) with flow rate of 1.0 mL min⁻¹. At optimum conditions method has linear response over 0.001–10 μg mL⁻¹ with detection limit between 0.0001 and 0.0005 μg mL⁻¹ with relative standard deviations (RSDs) in the range 2.18–5.06% (n = 6).     

Simultaneous determination of salbutamol sulphate, bromhexine hydrochloride and etofylline in pharmaceutical formulations with the use of four rapid derivative spectrophotometric methods

Four simple, rapid, accurate, precise, reliable and economical spectrophotometric methods have been proposed for simultaneous determination of salbutamol sulphate (SS), bromhexine hydrochloride (BH) and etofylline (ET) in pure and commercial formulations without any prior separation or purification. They were first derivative zero crossing spectrophotometry (method 1), simultaneous equation method (method 2), derivative ratio spectra zero crossing method (method 3) and double divisor ratio spectra derivative method (method 4). The ranges for SS, BH and ET were found to be 1-35 μg mL⁻¹, 4-40 μg mL⁻¹ and 5-80 μg mL⁻¹. For methods 1 and 2, the values of limit of detection (LOD) were 0.2314 μg mL⁻¹, 0.4865 μg mL⁻¹ and 0.2766 μg mL⁻¹ and the values of limit of quantitation (LOQ) were 0.7712 μg mL⁻¹, 1.6217 μg mL⁻¹ and 0.9221 μg mL⁻¹ for SS, BH and ET, respectively. For method 3, LOD values were 0.3297 μg mL⁻¹, 0.2784 μg mL⁻¹ and 0.7906 μg mL⁻¹ and LOQ values were 0.9325 μg mL⁻¹, 0.9282 μg mL⁻¹ and 2.6352 μg mL⁻¹ for SS, BH and ET, respectively. For method 4, LOD values were 0.3161 μg mL⁻¹, 0.2495 μg mL⁻¹ and 0.2064 μg mL⁻¹ and LOQ values were 0.9869 μg mL⁻¹, 0.8317 μg mL⁻¹ and 0.6879 μg mL⁻¹ for SS, BH and ET. The precision values were less then 2% R.S.D. for all four methods. The common excipients and additives did not interfere in their determinations. The results obtained by the proposed methods have been statistically compared by means of Student t-test and by the variance ratio F-test.     

A simple and rapid competitive enzyme-linked immunosorbent assay (cELISA) for high-throughput measurement of secretory immunoglobulin A (sIgA) in saliva

A simple competitive enzyme-linked immunosorbent assay (cELISA) was established for rapid measurement of secretory immunoglobulin A (sIgA) in saliva. The method was based on competitive reaction between the immobilized IgA and free IgA in the solution for the limited amount of horseradish peroxidase-conjugated rabbit anti-human IgA. In comparison with the conventionally used Sandwich ELISA, the cELISA is simpler, low-cost, and shows better reproducibility since it is not affected by the variation of capture antibodies from different batches. The assay time was also significantly reduced from more than 5 h to less than 3 h. Different curve-fitting models were compared, among which the fully specified logit-log model gave the best results. The linear working range and limit of detection were found to be 0.1-100 μg mL⁻¹ and 0.05 μg mL⁻¹, respectively. Matrix effects of saliva samples were investigated and a reasonable range of dilution factors were proposed. The developed method offers a very practical approach for high-throughput measurement of sIgA in saliva samples.     

Determination of trace formaldehyde in blood plasma by resonance fluorescence technology

A novel method for the determination of trace formaldehyde in blood plasma has been established by using resonance fluorimetry technique. It was based on the fact that oxidation of pyronine Y by potassium bromate was catalyzed by formaldehyde in sulfuric acid. When the wavelength interval was at Δλ = 0 nm, it was found that the decreased intensity (ΔF) of resonance fluorescence at 574.6 nm was proportional to the concentration of formaldehyde in the range of 1.27 × 10⁻² to 2.28 μg mL⁻¹. The limit of detection and the average recovery for formaldehyde were 3.80 ng mL⁻¹ and 101.6% (n = 6), respectively. The present method had been applied to the determination of trace formaldehyde in blood plasma, and the obtained results were in good agreement with those obtained by the resonance light scattering method.     

A naked-eye based strategy for semiquantitative immunochromatographic assay

It is critical to develop a cost-effective quantitative/semiquantitative assay for rapid diagnosis and on-site detection of toxic or harmful substances. Here, a naked-eye based semiquantitative immunochromatographic strip (NSI-strip) was developed, on which three test lines (TLs, TL-I, TL-II and TL-III) were dispensed on a nitrocellulose membrane to form the test zone. Similar as the traditional strip assay for small molecule, the NSI-strip assay was also based on the competitive theory, difference was that the analyte competed three times with the capture reagent for the limited number of antibody binding sites. After the assay, the number of TLs developed in the test zone was inversely proportional to the analyte concentration, thus analyte content levels could be determined by observing the appeared number of TLs. Taking aflatoxin B₁ as the model analyte, visual detection limit of the NSI-strip was 0.06 ng mL⁻¹ and threshold concentrations for TL-I–III were 0.125, 0.5, and 2.0 ng mL⁻¹, respectively. Therefore, according to the appeared number of TLs, the following concentration ranges would be detectable by visual examination: 0–0.06 ng mL⁻¹ (negative samples), and 0.06–0.125 ng mL⁻¹, 0.125–0.5 ng mL⁻¹, 0.5–2.0 ng mL⁻¹ and >2.0 ng mL⁻¹ (positive samples). That was to say, compared to traditional strips the NSI-strip could offer more parameter information of the target analyte content. In this way, the NSI-strip improved the qualitative presence/absence detection of traditional strips by measuring the content (range) of target analytes semiquantitatively.