联苯菊酯酶联免疫吸附分析方法研究

建立了定量测定联苯菊酯的间接竞争酶联免疫吸附分析方法(ic-ELISA)。利用联苯菊酯的代谢物联苯醇合成了联苯菊酯的半抗原LBc(2-甲基-3-苯基苄基氧基羰基丙酸)和LBy(2-甲基-3-苯基苯甲酸)。通过碳二亚胺法将LBc交联于牛血清蛋白(BSA)作为免疫抗原(LBc-BSA),通过活泼酯法将LBc和LBy分别交联于卵清蛋白(OVA)作为包被抗原(LBc-OVA和LBy-OVA),LBc-BSA为免疫原制备了联苯菊酯的兔抗血清,间接非竞争酶联免疫吸附分析方法测得其效价达5.12×104。通过同源异源分析,发现异源分析的灵敏度较高,对pH值、离子强度、甲醇含量等影响因素进行了研究,0.3mol/L钠离子强度的磷酸缓冲液(pH7.5)和30%的甲醇确定为联苯菊酯间接竞争酶联免疫吸附分析方法的最佳工作条件,该方法的IC50为2.16±0.32mg/L,检出限(LDL)为0.016±0.002mg/L。对大部分拟除虫菊酯,如三氟氯氰菊酯、溴氰菊酯、氯氰菊酯、氰戊菊酯、甲氰菊酯和拟除虫菊酯的代谢物3-苯氧基苯甲酸没有明显的交叉反应。     

4-烃基-3-氰基-双环笼状磷酸酯类化合物的合成及生物活性研究

以2-烃基-2-羟甲基-1,3-丙二醇为原料, 经保护两个羟基, PCC氧化, 引入氰基, 脱保护后与POCl3或PSCl3环合, 成功合成了14个4-烃基-3-氰基双环笼状磷酸酯类化合物, 目标化合物的结构通过1H NMR, MS和元素分析进行了表征. 采用放射性配体受体结合实验, 测定了14个化合物抑制[3H]EBOB [4’-ethynyl-4-n-propylbicycloorthobenzoate]与家蝇及大鼠GABA受体的结合活性. 结果表明部分化合物在10－5 mol/L具有较好的抑制活性, 且在大鼠与家蝇GABA受体之间具有一定选择性.     

Effect of hapten structures on specific and sensitive enzyme-linked immunosorbent assays for N-methylcarbamate insecticide metolcarb

Five different haptens of the N-methylcarbamate insecticide metolcarb were designed and synthesized. All of the haptens were conjugated with ovalbumin (OVA) for the coating antigen, and one hapten containing all of the structure of metolcarb was conjugated with bovine serum albumin (BSA) for the immunogen. Two polyclonal antisera were raised against the BSA conjugate, and ten antibody/coating conjugate combinations were selected for studies of assay sensitivity and specificity for metolcarb. A class-specific combination was found, with the I₅₀ of the assay ranged from 0.64 to 20.98 μg mL⁻¹ for seven tested N-methylcarbamate insecticides except for pirimicarb. Considering titer, I₅₀ and cross-reactivity of all combinations of antibody/coating conjugate, a competitive indirect enzyme-linked immunosorbent assay (ELISA) in a homologous system, whose limit of detection (LoD) reached 1.4 ng mL⁻¹, was presented. The results of competitive ELISAs indicated that coating hapten structure can significantly affect not only assay sensitivity but also its specificity.     

Influence of high pressure treatment on rheological and other properties of egg white

Abstract

The paper deals with the influence of high pressure treatment of fresh egg white on its properties and protein composition (individual amino-acids predicted as a function of pressure and time levels). The rheological properties are changed by high pressure from Newtonian to non-Newtonian behaviour, with increasing apparent viscosity as the pressure and time increased. The pH, whipping ability, foam stability, gel strength of heat induced gels after treatment and the whole protein content, were also predicted.

The results showed that the foam stability is increased with increasing pressure and time of processing. The foam volume is also increased with pressure. The pH did not change with pressure or time of processing. Composition of proteins as indicated by individual amino-acids did not exhibit statistically important changes. Gel strength of heat induced gels prepared from previously pressured liquid whites showed no important change of values with pressure or time of treatment. The modulus of elasticity showed a decrease for samples pressured to 400 MPa for 5 up to 15 minutes.     

On The Fluorometric Method for the Measurement of β-Cyanoalanine Synthase Activity

Abstract

A modification of a fluorescence assay method for β-cyanoalanine synthase has been described which involves the direct coupling of the product of cyanide and cysteine to resazurin, to produce the fluorescent resosurfin, The fluorescence produced was found to be directly proportional to the rate of the reaction.     

An Enzyme-Linked Immunosorbent Assay for Determination of Dicyclanil in Animal Tissue

Dicyclanil is a toxic, pyrimidine-derived insect growth regulator used in veterinary medicine for the prevention of myiasis. A competitive indirect enzyme-linked immunosorbent assay was developed firstly for the determination of dicyclanil in animal tissue. The antigen was prepared using the glutaraldehyde method and polyclonal antibodies were obtained by immunizing rabbits with a dicyclanil-bovine serum albumin conjugate. The antibody obtained was specific for dicyclanil with an IC₅₀ of 9 ng/mL. Recoveries from sheep tissue and liquid milk were in the range of 52.5–72.4%, with coefficients of variation between 4.7% and 11.9%. The results were further confirmed by high performance liquid chromatography–tandem mass spectrometry.     

Characterization and Bioavailability of Vitamin C Nanoliposomes Prepared by Film Evaporation-Dynamic High Pressure Microfluidization

Vitamin C nanoliposomes were prepared by combining a conventional method (film evaporation) with dynamic high pressure microfluidization. Their physicochemical characterizations (antioxidant activity, particle size, entrapment efficiency, morphology, in vitro drug release, and storage stability) and skin permeation behavior were investigated. The results showed that vitamin C nanoliposomes, having equivalent DPPH (2, 2-diphenyl-1-picrylhydrazyl) free radical scavenging capacity of pure vitamin C solution without loss of their biological activity, exhibited better storage stability at 37°C for 24 hours and at 4°C for 60 days, a more excellent sustained drug release as well as higher skin penetration rate than vitamin C liposomes.     

The Effect of Temperature and Standings Can Cause Deviations in Prostate Specific Antigen (PSA) Assays

Abstract

Blood samples were assayed for PSA values immediately after sampled or by standing for an assigned period. Variation of the free‐ (f), total‐ (t) values, and the free to total (f/t) ratios were determined. Mathematical models were used to interpret the phenomena of deviation. Smaller values of PSA values changed randomly with time and temperature of standings, resulting in varying f/t ratios, while larger initial PSA values were relatively unaffected to any significant extent.

Model interpreted that the changes of PSA values might be caused by higher temperature and time of standings, and conformational participation was also possible.     

Degradation effects in the extraction of antioxidants from birch bark using water at elevated temperature and pressure

Experiments with birch bark samples have been carried to enable a distinction between extraction and degradation effects during pressurised hot water extraction. Two samples, E80 and E180, contained birch bark extracts obtained after extraction at 80 and 180 °C for up to 45 min, respectively. Two other samples, P80 and P180, were only extracted for 5 min at the two temperatures and were thereafter filtered and hydrothermally treated at 80 and 180 °C, respectively. During the latter treatment, samples were collected at different times to assess the stability of the extracted compounds. An offline DPPH (2,2-diphenyl-1-picrylhydrazyl) assay, as well as a high performance liquid chromatographic separation coupled to an electrochemical detector, were used to determine the antioxidant capacity of the processed samples. The results obtained with the different techniques were compared to assess the yield of the extraction and degradation processes. In addition, an online hyphenated system comprising high performance liquid chromatography coupled to diode-array; electrochemical; and tandem mass spectrometric detection (HPLC-DAD-ECD-MS/MS) was used to study the compositions of the extracts in more detail. The results for the samples processed at 80 °C showed that the extraction reached a steady-state already after 5 min, and that the extracted compounds were stable throughout the entire extraction process. Processing at 180 °C, on the other hand, gave rise to partly degraded extracts with a multitude of peaks in both the diode array and electrochemical detectors, and a higher antioxidant capacity compared to for the extracts obtained at 80 °C. It is concluded that HPLC-DAD-ECD is a more appropriate technique for the determination of antioxidants than the DPPH assay. The mass spectrometric results indicate that one of the extracted antioxidants, catechin, was isomerised to its diastereoisomers; (+)-catechin, (−)-catechin, (+)-epicatechin, and (−)-epicatechin.     

Computer Assisted Spectrophotometric Analysis of Three Component Mixtures (Caffeine Mixtures and Codeine Mixtures)

Abstract

Two mixtures (caffeine mixture (I) and codeine mixture (II)) of 3-components, containing acetaminophen and salicylamide as majors, have been assayed using different computer-assisted spectrophotometric methods. These methods are the Unique Absorbance Method (UAM), the Unique Orthogonal Function Method (UOFM), and Unique Fourier Function Method (UFFM), and also their correspondences under the least squares approach. The parameters have been optimized for the accurate determination of the minor component caffeine in mixture (I). In mixture (II), however. codeine has been extracted - in two steps-prior to the correction of interferences from the major components acetaminophen and salicylamide.