Rapid determination of the synthetic pyrethroid insecticide, deltamethrin, in rat plasma and tissues by HPLC

Deltamethrin (DLM), [(S)-alpha-cyano-d-phenoxybenzyl-(1R,3R)-e-(2,2 dibromovinyl)-2,2-dimethylcyclo-propane-1-carboxylate], is a pyrethroid insecticide widely used in agriculture and households. There are several methods for analysis of DLM in biological fluids and tissues, but these methods are time consuming. They generally involve the extraction of DLM with lipid-soluble solvents such as n-pentane, n-hexane, diethylether or acetone, and subsequent evaporation of the solvent. A more rapid and sensitive high-performance liquid chromatography (HPLC) method to analyze DLM in plasma and tissues (liver, kidney, and brain) was developed and validated according to U.S. Food and Drug Administration (U.S. FDA) and International Conference on Harmonization (ICH) of Technical Requirements for Registration of Pharmaceuticals for Human Use guidelines. The limit of detection (S/N of 3/1) for DLM was 0.01 microg/ml for plasma, liver, kidney and brain. The method performances were shown to be selective for DLM and linear over the concentration range 0.01-20.0 microg/ml. For five replications of samples at 0.05, 0.1, 0.2, 1.5 and 4.0 microg/ml, intraday precision and accuracy values were in the range of 0.7-13.1% relative standard deviation (%R.S.D.) and 1.8-14.1%Error, respectively. Interday (n = 15) precision and accuracy values at 0.05, 0.1, 0.2, 1.5, and 4.0 microg/ml were in the range of 3.2-15.2% (%R.S.D.) and 3.7-14.8%Error, respectively. The absolute recoveries of DLM ranged from 93 to 103% for plasma, 95 to 114% for liver, 97 to 108% for kidney, and 95 to 108% for brain. This method can be quite useful for DLM pharmacokinetic and tissue distribution studies, for which multiple plasma and tissue samples have to be analyzed quickly with high reproducibility.     

Exposure risks to pesticide applicators posed by the use of electric backpack sprayers and stretcher-mounted sprayers in orchards

Abstract

The introduction of the pesticide registration system in pesticide risk assessments has promoted the scientific and safe use of pesticides, and the assessment of occupational exposure risk. In the present study, we performed an experiment in a citrus orchard subject to normal orchard management practices. By measuring the exposure of applicators’ (farmers and technicians) body parts to 45% malathion emulsifiable concentrate during its application using electric backpack sprayers (0.25?KPa) and stretcher-mounted sprayers (1.5?KPa), the unit exposure (UE) was determined. The risks of exposure for pesticide applicators who adopted five different protective measures (A: no personal protective equipment (PPE), i.e., no clothes, no gloves, no caps, and no socks; B: short-sleeved top and shorts; C: short-sleeved top, shorts, and a single pair of gloves; D: short-sleeved top, shorts, a single pair of gloves, and a cap; E: long-sleeved top, long pants, a single pair of gloves, and a cap) were also assessed. The results were as follows: 1) The total levels of exposure for pesticide applicators using electric backpack sprayers and stretcher-mounted sprayers were 3613.63?µg and 5654.28?µg, respectively. When electric backpack sprayers were used, the body parts that had the highest exposure were the head (13.8%), hands (19.9%) and back (14.0%), and when stretcher-mounted sprayers were used, the hands (32.5%) and lower legs (21.1%) had the highest level of exposure; 2) In the absence of PPE, the UE values for farmers who used electric backpack sprayers and farmers who used stretcher-mounted sprayers were significantly different. However, when PPE was used, the difference in UE values between the farmers using the two different types of sprayers was not significant; 3) When protective measure A was adopted, the risk quotient (RQ) values of the farmers and technicians who used electric backpack sprayers for the application of malathion were 1.44 and 0.54, respectively; the corresponding RQ values when protective measure B was adopted were 0.97 and 0.28, respectively. When stretcher-mounted sprayers were used for the application of chlorpyrifos, the RQ value of the farmers who adopted protective measure E was 0.43 while other types of PPE use resulted in RQ values greater than 1. In contrast, the RQ value for technicians was 1.62 when protective measure A was used and 1.02 when protective measure B was adopted, whereas other types of PPE use resulted in RQ values less than 1. Therefore, besides increasing the awareness of personal protection among pesticide applicators, improvement in the management of pesticide use and the enhancement of standard operations are of practical significance for controlling occupational exposure to pesticides.     

高效液相色谱法测定雷帕霉素

建立了雷帕霉素高效液相色谱测定方法。其色谱条件：色谱柱Zorbax Eclipse XDB-C8（4.6 mm×150 mm, 3.5 μm）反相柱,以乙腈和水溶液为流动相,紫外检测波长278 nm,流速1.0 mL/min,柱温55 ℃。结果表明：此方法标准曲线范围是0.2～20 μg,线性良好,加样回收率为99.4%～100.79%,日间相对标准偏差（RSD）（n=4）为1.25%～2.25%。本方法操作简便、准确,可有效应用于雷帕霉素提取纯化过程中的检测。     

Biological monitoring of eutrophication in rivers

There is an increasing awareness of the need to assess the impact of nutrient enrichment on river ecosystems separately from the impacts of organic effluents. A range of methods have been proposed and some have moved from the development stage to practical use by water management organizations. The methods can be applied to broad surveys or provide baseline information to assess possible future change. In the latter case it is recommended that several different methods are used, especially where it is important to get reliable information on the long-term impact of improvements in effluent quality. Estimates of biomass measured as chlorophyll a have often been used for phytoplankton and sometimes also for benthic communities. However, a lot of care is needed in applying this method, because of the range of factors besides nutrient concentration which can influence values. Approaches based on the whole community have been developed by a number of research groups, usually involving semiquantitative estimates of abundance. There has also been a rapid increase in the use of indices based on the relative proportions of epilithic diatom species. The methodologies used by a number of research groups in Europe are broadly similar, making it possible to compare results between different regions. The development of indices based on macrophyte floristic composition in relation to river nutrient status is also under development, especially in France and UK. However, interpretation of the results is complicated where long-term changes are taking place in nutrient concentrations in the water, because of the varying contributions of sediment and water to different species of rooted plant. Bioassays can be especially helpful where it is desired to establish whether either N or P is limiting for a population of community.     

Biological monitoring by citizens using Web-based photographic databases of fishes

The Internet has changed the way biologists communicate; this includes the collection of information on fish biology. This technological change may allow the possibility for biological monitoring by general citizens via the Internet. The Japanese Internet atlas of fishes, WEB sakana-zukan, has been in operation since 2002. It provides an opportunity to communicate and accumulate information on fish biology by amateur users, who consist mainly of sports fishing fans. This website has functioned not only as an entertainment and educational tool for users with an environmental and ichthyological interest. To date, more than 35,000 photographs of fishes have been registered on the website by more than 1,000 unique users, and more than 37,500 photographs of fishes have been posted in its bulletin board system (linking approximately 28,000 photographs to the website) by more than 2,000 unique users. As these photographs almost always include time and locality data, it is possible to provide distribution data by publishing the information as scientific papers or registering them as collections at the Kanagawa Prefectural Museum of Natural History (the Image Database of Fishes (KPM-NR) contributing to the Global Biodiversity Information Facility). If the communication method is developed and improved, various scientists will be able to collect much more biodiversity data from general citizens via the Internet.     

Biological monitoring among benzene-exposed workers in Bangalore city, India.

Environmental and biological monitoring was carried out in the winter season of 2004 for 30 gasoline station workers (study subjects) and 30 office workers (controls) of Bangalore city, India. Personal air sampling was carried out in the breathing zone of workers using an Anasorb CSC sorbent tube (SKC 226-01) fitted to the low-flow personal samplers (PCXR4 and pocket pump Model No. 210-1002) at a flow rate of 200 ml min(-1) during the shift work of 8 h. The benzene content adsorbed in the sorbent tube (SKC 226-01) was desorbed with 1 ml of benzene-free carbon disulfide on a developing vibrator and later analysed by Trace GC fitted with MXT-624 column and flame ionization detector. The mean time weighted average benzene concentration found among study and controls was 1.10+/-1.08 and 0.070+/-0.035 mg m(-3), respectively. Biological monitoring for benzene exposure was performed by measuring trans,trans muconic acid (t,t-MA) in the end shift urine samples using HPLC-UV technique. End-shift urine samples (1 ml) were adjusted to pH 7-9 with phosphate buffer pH 7.4 passed through the preconditioned Q-SAX anion-exchange cartridge and the (t,t-MA) is extracted with 10% acetic acid and later analysed by HPLC-UV detection The mean t,t-MA found among study and controls were 563.16+/-281.81 and 266.88+/-110.65 microg g(-1) creatinine. About 50% of the study subjects (15) have higher t,t-MA values than the biological exposure index of the American Conference of Government Industrial Hygienist (ACGIH). Correlation is significant at 5% level (p<0.05) between personal air benzene concentration and urinary t,t-MA in the study group. Based on these findings, the t,t-MA can be used as a biomarker for benzene exposure.     

Determination of aflatoxins in medicinal herbs by HPLC. An efficient method for routine analysis

A fast and easy to perform method for the routine determination of aflatoxins in medicinal herbs was developed. The described method involves a single-step extraction with a non-chlorinated solvent, an immunoaffinity clean-up and HPLC with fluorescence detection. Whilst assays with naturally contaminated and with spiked samples of several herbs showed that the recoveries were somewhat low and dependent on the kind of sample and the degree of grinding, the intra-batch reproducibility was good, allowing a reliable quantitation by the standard-addition method. Good linearity, repeatability and accuracy were demonstrated in assays involving several medicinal herbs. The limit of quantitation was of the order of 0.05-0.1 ng/g, being dependent of the species analysed, and the method required no tedious concentration or back-extraction steps.     

Frequencies of chromosomal aberrations in pesticide sprayers working in plastic green houses.

The frequency of chromosome aberrations was studied in peripheral blood lymphocytes of 29 workers occupationally exposed to a mixture of pesticides and in 14 age- and sex-matched healthy controls. There was a significant increase in chromosome aberrations in sprayers when compared to unexposed persons (2.39% compared to 0.54%). No positive correlation between the frequency of chromosome aberrations and the duration of exposure was observed. No significant difference between smokers and non-smokers was found.     

Biological monitoring of n-hexane exposure in shoe workers

To analyse working conditions and to provide information about the degree to which shoe workers are exposed to n-hexane, the urinary excretion of total 2,5-hexanedione (2,5-HD) was determined in 81 employees in 12 shoe factories. Twenty-five individuals who had experienced no exposure to solvents were used as controls. 2,5-HD was measured in spot urine samples collected from workers at the end of shift. In the urine of shoe workers, the 2,5-HD presented a mean value of 2.33 mg g^-1 creatinine, a median of 1.96 mg g^-1 creatinine. The mean 2,5-HD concentration in the urine samples from non-exposed subjects was 0.28 mg g^-1 creatinine, the median value was 0.18 mg g^-1 creatinine. The mean time-weighted average (TWA) concentration of n-hexane in 12 shoe workshops was 126.1 ppm, ranging from 23 to 215 ppm. We found a significant, but low, correlation (r = 0.40; p < 0.001) between TWA intensity of environmental exposure to n-hexane and the concentration of 2,5-HD in urine. The probable effect of toluene on the concentration of 2,5-HD was also discussed in the present study.     

Study of aminoglycoside-nucleic acid interactions by an HPLC method

The interactions of a number of aminoglycoside antibiotics with tRNA and DNA were studied by an HPLC method. based on tRNA and DNA peak size exclusion. Among the compounds studied (deoxystreptamine, neamine, neomycin B, kanamycin A, gentamicin A, netilmicin, streptomycin, and the synthetic neamine analogue BKN3), neomycin B and the synthetic analogue of neamine were proved to be the most potent binders.     

Elution-band relaxation method. A method to analyze isomerization kinetics by HPLC and application to protein denaturation-renaturation

We present a novel method, the 'elution-band relaxation method', to analyze quantitatively reversible isomerization kinetics by elution chromatography, taking advantage of the high resolution and speed of high-performance liquid chromatography (HPLC). The kinetic information is obtained by measuring the first temporal moments of chromatograms of molecules undergoing isomerization and analyzing their dependence on the column length or flow rate. The major advantage of this method is that it is applicable to reactions as fast as the time of elution in HPLC, a speed which has not been attained previously in analysis of isomerization reactions based on the chromatographic property of molecules. We describe the method and report an experimental application to the denaturation-renaturation kinetics of bovine pancreatic ribonuclease A as an example.     

Activation of Drosophila sodium channels promotes modification by deltamethrin. Reductions in affinity caused by knock-down resistance mutations

kdr and super-kdr are mutations in houseflies and other insects that confer 30- and 500-fold resistance to the pyrethroid deltamethrin. They correspond to single (L1014F) and double (L1014F+M918T) mutations in segment IIS6 and linker II(S4-S5) of Na channels. We expressed Drosophila para Na channels with and without these mutations and characterized their modification by deltamethrin. All wild-type channels can be modified by <10 nM deltamethrin, but high affinity binding requires channel opening: (a) modification is promoted more by trains of brief depolarizations than by a single long depolarization, (b) the voltage dependence of modification parallels that of channel opening, and (c) modification is promoted by toxin II from Anemonia sulcata, which slows inactivation. The mutations reduce channel opening by enhancing closed-state inactivation. In addition, these mutations reduce the affinity for open channels by 20- and 100-fold, respectively. Deltamethrin inhibits channel closing and the mutations reduce the time that channels remain open once drug has bound. The super-kdr mutations effectively reduce the number of deltamethrin binding sites per channel from two to one. Thus, the mutations reduce both the potency and efficacy of insecticide action.     

Biological monitoring of human exposure to metals

Biological monitoring of exposure to metals and metalloids involves not only determination of these elements in selected body fluids and tissues but, in some cases, also determination of a certain biochemical indicator which signalises the presence of the monitored element in the organism. Biological monitoring of occupational exposure to metals has a long tradition whereas biological monitoring of exposure in general environment has developed more intensively only in the past two decades. New information about the toxic effect of some metals and metalloids and about their kinetics of absorption, distribution and excretion in experimental animals, and particularly in man, is necessary for elaborating suitable biological exposure tests. Of current interest is also the movement of persistent noxae in the environment, seen from the ecologic view point. This report outlines the present state of the problem in Czechoslovakia.     

改进的HPLC法测定红豆杉植物和细胞培养物中紫杉醇的含量

紫杉醇 (Ｔａｘｏｌ)是一种新型的抗癌药物。由于紫杉醇在红豆杉属 (ＴａｘｕｓＬ .)植物中的含量很低 ,而且运用植物细胞培养技术生产紫杉醇仍处实验阶段 ,因此 ,对于紫杉醇含量甚微的样品 ,寻找一种精确、灵敏、快速的检测方法就显得尤为重要。本实验采用改进的ＨＰＬＣ法对南方红豆杉〔Ｔａｘｕｓｃｈｉｎｅｎｓｉｓｖａｒ.ｍａｉｒｅｉ (Ｌｅｍｅ啨ｅｔＬ啨ｖｌ.)ＣｈｅｎｇｅｔＬ .Ｋ .Ｆｕ〕枝和叶 ,以及中国红豆杉〔Ｔａｘｕｓｃｈｉｎｅｎｓｉｓ (Ｐｉｌｇｅｒ)Ｒｅｈｄ .〕细胞培养物中的紫杉醇含量进行了测定。1　实验方法1 1　实…