Radio gas chromatography of steroid metabolites by collection of radioactive fractions on thin layer chromatography plates

A simple and inexpensive technique for the monitoring of radioactive compounds in gas chromatographic effluent is described. Use has been made of the adsorption properties of thin layer chromatography plates to trap eluting radioactive peaks. Performance of the technique is demonstrated by analysis of radioactive steroid mixtures. The efficiency and sensitivity of this technique equal that of more complicated techniques.     

Solubilized substrates for the on-line measurement of lipases by flow injection analysis during chromatographic enzyme purification.

A flow injection analysis (FIA) system for the on-line measurement of lipases in chromatographic processes has been developed. The photometrically detectable substrates para-nitrophenylpalmitate, S,O,O'-tripropyryl-1-thioglycerol, and 1,2-O-dilauryl-rac-glycero-3-glutaric-resorufinester were investigated. Different detergents and qualities of assay emulsions were tested for optimal results in FIA applications. Emphasis was placed on increasing the stability of the assay emulsion. Lipases of different origin and specificity were detected. The linear detection range was adapted to the requirements of the chromatographic purification procedures. The connection of the FIA with a fast protein liquid chromatography system permitted the automatization of lipase purification by monitoring protein content, salinity, and enzyme activity of the effluent from column chromatography.     

An evaluation of systems for paper chromatography of C21 steroids.

A group of moderately polar C21 steroids (3-5 oxygen functions) has been chromatographed in 9 solvent systems. Using the concept that standard deviation of the mean RF is an index of chromatographic resolution, and that coefficients of correlation between sets of RF data can be used to quantify the similarities of chromatographic systems, we have evaluated the resolving properties of the systems when used individually, and also when used in combinations of two, three and four. The discriminating powers of some of the most effective individual systems, and some of the sequences of systems which are most efficient, are shown graphically as chromatography trees. The relationship between the total effective standard deviation of a group of systems which are used in sequence and the probability that a pair of compounds will be separated by more than 0.10 RF is discussed.     

The use of a pre-column for the direct high-performance liquid chromatographic determination of the anti-depressants clovoxamine and fluvoxamine in plasma

A high-performance liquid chromatographic system for the direct determination of the antidepressants clovoxamine and fluvoxamine in plasma is described. The primary amines are derivatized with the fluorogenic reagent fluorescamine in order to increase the sensitivity and selectivity, but also to decrease the polarity of the amines. The band broadening of some combinations of pre-column and analytical column is compared for the fluorescamine derivative of fluvoxamine. The combination of a pre-column containing RP-2 and an analytical column containing RP-8 has been successfully used for the determination of the two antidepressants in plasma. At relatively low concentrations of the drugs, a simple step-gradient elution is required for the removal of a large proportion of the more polar components of the samples. Concentrations in the range of 10—1000 ng clovoxamine per ml plasma were determined by means of external standardization and show a good correlation with the data of a more laborious gas chromatographic method. The detection limits for clovoxamine and fluvoxamine are approximately 3 ng/ml in plasma.     

草酸脱羧酶及其应用

草酸脱羧酶是一种含锰的酶,在白腐菌中广泛存在,少数低等真菌和细菌中也能产生。目前,至少10多种草酸脱羧酶得到了分离和纯化。该酶是一种氨基酸残基在379个左右的单体酶,一般都为酸性糖蛋白,含有2个锰离子,形成2个活性区域;表面一些氨基酸被不同程度地糖基化。晶体结构和其它一些波谱学研究解释了其空间结构和可能的电子传递机制。运用PCR技术和cDNA文库技术,越来越多的草酸脱羧酶基因被克隆。已研究的该酶基因中都含有17个左右的内含子,这些内含子在活性域位置上有比较高的保守性。一些特殊氨基酸序列的存在决定了该酶的表达形式为诱导型,菌株的基因调控序列中含有一段受草酸化合物作用的序列。该酶在一些酵母和植物等异源表达系统中有成功表达的报道。该酶的应用主要集中在以下几方面：造纸废水中的草酸盐降解;食品中的草酸降解;草酸生物检测（如,临床诊断）等。     

Volatile Flavor of Black Tea

The first effluent of essential oil of black tea in the gas chromatographic analysis is important for characterizing black tea flavor. The isolation and identification of main unknown components were conducted by means of gas chromatography, and also by infrared spectra and ultraviolet spectra. Ethylacetate, ethylalcohol and β-myrcene were identified as the increasing compounds during fermentation. cis-β-Ocimene and trans-β-ocimene were also identified only in completely manufactured black tea. These three terpenoid hydrocarbons are newly found constituents of essential oil of black tea and are supposed to contribute considerably to the black tea flavor.     

Development and validation of a liquid chromatographic method for Casiopeina IIgly in rat plasma

A sensitive and specific liquid chromatographic method using solid-phase extraction with Sep-pak cartridges has been developed for the determination of Casiopeina IIgly and validated over the linear range 2.5-50 microg/ml in rat plasma. The analysis was performed on a Symetry C(18) (5 microm) column with a Phenomenex C(18) precolumn. The mobile phase was methanol-water (58:42, v/v). The column effluent was monitored at 273 nm. The results showed that the assay is sensitive at 2.5 microg/ml. Maximum intra-day coefficient of variation was 11.47%. The recovery based upon addition of internal standard to rat plasma was 80.98%. The method was used to perform preclinical pharmacokinetic studies in rat plasma and was found to be satisfactory.     

Advances in techniques for ecdysteroid analysis

Summary

Recent improvements in chromatographic methods for ecdysteroids are reviewed. Minor improvements in liquid chromatography have increased the separation and range of compounds which can be accomodated, some useful techniques have been introduced in thin layer chromatography, and with new radioactivity scanners and direct mass spectrometry from TLC, this technique retains an important place among the tools of the ecdysonist. Supercritical fluid chromatography has been suggested as a rapid and more sensitive alternative to liquid chromatography. In mass spectrometry, fast atom bombardment has proved most useful, though coupling HPLC through thermospray and ptasmaspray methods has been attempted, but as yet, these techniques lack sensitivity. New solid phase immunoassay methods using colorimetric rather than radioactivity measurements promise to give a better means of analysis of small amounts of ecdysteroids, capable of use in the presence of radiolabeled compounds. A solid phase separation of C-20, 22 diol ecdysteroids with immobilized phenylboronic acid has been reported.     

Chromatographic screening techniques in systematic toxicological analysis

A review of techniques used to screen biological specimens for the presence of drugs was conducted with particular reference to systematic toxicological analysis. Extraction systems of both the liquid–liquid and solid-phase type show little apparent difference in their relative ability to extract a range of drugs according to their physio-chemical properties, although mixed-phase SPE extraction is a preferred technique for GC-based applications, and liquid–liquid were preferred for HPLC-based applications. No one chromatographic system has been shown to be capable of detecting a full range of common drugs of abuse, and common ethical drugs, hence two or more assays are required for laboratories wishing to cover a reasonably comprehensive range of drugs of toxicological significance. While immunoassays are invariably used to screen for drugs of abuse, chromatographic systems relying on derivatization and capable of extracting both acidic and basic drugs would be capable of screening a limited range of targeted drugs. Drugs most difficult to detect in systematic toxicological analysis include LSD, psilocin, THC and its metabolites, fentanyl and its designer derivatives, some potent opiates, potent benzodiazepines and some potent neuroleptics, many of the newer anti-convulsants, alkaloids colchicine, amantins, aflatoxins, antineoplastics, coumarin-based anti-coagulants, and a number of cardiovascular drugs. The widespread use of LC–MS and LC–MS–MS for specific drug detection and the emergence of capillary electrophoresis linked to MS and MS–MS provide an exciting possibility for the future to increase the range of drugs detected in any one chromatographic screening system.     

Ganglioside alterations in stimulated murine macrophages

A two-dimensional thin-layer chromatographic technique has been used to separate and display gangliosides from murine peritoneal macrophages in different functional states. Resident macrophages have a relatively simple ganglioside pattern with about 15 resorcinol-positive spots. Gangliosides from resident cells contained mostly (90%) N-glycolylneuraminic acid. Thioglycolate-elicited and Corynebacterium parvum-activated macrophages have much more complex patterns with about 40 resorcinol-positive spots. Although ganglioside sialic acid content of stimulated macrophages was only slightly higher than that of resident cells, it consisted of nearly equal amounts of N-acetyl- and N-glycolylneuraminic acid. The shift in the ganglioside sialic acid type and the expression of different gangliosides in macrophages upon stimulation may help explain some of the differences in function and responsiveness noted in these macrophage populations.     

Issues in the extinction of specific stimulus-outcome associations in Pavlovian conditioning

This paper reviews a variety of studies designed to examine the effects of extinction upon control by specific stimulus-outcome (S-O) associations in Pavlovian conditioning. Studies conducted with rats in a magazine approach conditioning paradigm have shown that control by specific S-O associations is normally unaffected by extinction treatments, although other aspects of conditioned responding seem affected in a more enduring way. However, recent work suggests that extinction can undermine control by such associations if it is administered after the conditioned stimulus is weakly encoded. The results from these studies suggest that it may be important to consider multiple response systems in assessing the impact of extinction. Studies conducted with the flavor preference learning paradigm in rats also show that specific S-O associations can be undermined by procedures that involve presenting a flavor cue in the absence of its associated nutrient. These findings provide no support for the view that flavor preference learning necessarily entails some unique learning process that differs from more conventional processes. As in other situations, some of these effects likely involve a masking process, but the extent to which masking or true associative weakening occurs in extinction more generally is a topic that is not well understood. Finally, we present some data to suggest that extinction also involves conditional "occasion-setting" control by contextual cues. Special procedures are recommended in assessing such learning when the goal is to distinguish this form of learning from other more conventional mechanisms of extinction.     

Review: total doses in fractionated radiotherapy--implications of new radiobiological data

The total dose in radiotherapy has been adjusted in the past for different fractionation schedules by the use of empirical formulae such as NSD, TDF and CRE. It is now appropriate to consider fractionation factors which include more biological insight in their formulation than was possible earlier. It has become clear, from both clinical and experimental animal data, that the total dose in multi-fraction irradiations depends more critically on size of dose-per-fraction for late than for early damage to normal tissues. This difference has been interpreted as due to different shapes of the underlying dose-response curves. The late reactions respond with more curvature in the dose-response curve, i.e. with more repair capability at very low doses per fraction, than the early tissue reactions. A linear-quadratic relationship for the dose-response curves has been found to fit experimental data well, with few exceptions. This paper reviews this interpretation and explores some of its implications for radiotherapy and for radiobiology applied to therapy. Of many repair factors that have been suggested, the ratio alpha/beta (of the linear to the quadratic coefficients) is one that should be independent of the level of damage assayed. Values of alpha/beta of about 10 Gy have been reported for a number of early tissue responses but a range of values from about 1 to 5 or 6 Gy for late responses. It is a current challenge to radiobiology to explain why this difference occurs. Once such values are known for different tissues--and the dangers of premature assumptions are emphasized--calculations are possible which might be useful in radiotherapy as an alternative to NSD, TDF, CRE etc. Some data are presented on the magnitude of differences from these previously used empirical formulae, with a discussion about how easily detected the discrepancies might be in clinical practice. Applications to hypofractionation, hyperfractionation and accelerated fractionation are illustrated.     

红葡萄酒中“马厩味”不良风味物质的检测及其抑制方法的研究进展

葡萄酒是大众喜爱的酒种之一,已在人们的日常消费中占有越来越高的比重。但许多葡萄酒在生产,窖藏及贮运环节,可能会遭受微生物的污染。有些微生物在酒体中会产生不良风味。酒香酵母属酵母菌就是这样一种微生物。它在酒体中所产生的马厩味不良风味物质4-乙基酚或4-乙基愈创木酚对红酒会造成风味上的巨大影响。本文综述了马厩味风味物质产生的机理,以及国外马厩味不良风味物质的检测及抑制方法等,以期对国内红葡萄酒产业相关不良风味物质的检测及其抑制方法的开发有所借鉴。     

Quantifying process tradeoffs in the operation of chromatographic sequences

A method for the rapid representation of key process tradeoffs that need to be made during the analysis of chromatographic sequences has been proposed. It involves the construction of fractionation and maximum purification factor versus yield diagrams, which can be completed easily on the basis of chromatographic data. The output of the framework developed reflects the degree of tradeoff between levels of yield and purity and provides a fast and precise prediction of the sample fraction collection strategy needed to meet a desired process specification. The usefulness of this approach for the purposes of product purification and contaminant removal in a single chromatographic step has been successfully demonstrated in an earlier paper and it is now extended by application to a chromatographic sequence: the separation of a hypothetical three-component protein system by hydrophobic interaction chromatography (HIC) followed by size exclusion chromatography (SEC). The HIC operation has a strong impact upon the subsequent SEC step. The studies show how the analysis of performance in such a chromatographic sequence can be carried out easily and in a straightforward fashion using the fractionation diagram approach. The methodology proposed serves as a useful tool for identifying the process tradeoffs that must be made during operation of a sequence of chromatographic steps and indicates the impact on further processing of the cut-point decisions that are made.     

Quantitative Microdialysis: Analysis of Transients and Application to Pharmacokinetics in Brain

The behavior of a microdialysis probe in vivo is mathematically described. A diffusion-reaction model is developed that not only accounts for transport of substances through tissues and probe membranes but also accounts for transport across the microvasculature and metabolism. Time-dependent equations are presented both for the effluent microdialysate concentration and for concentration profiles about the probe. The analysis applies either to measuring the tissue pharmacokinetics of drugs administered systemically, or for sampling of endogenously produced substances from tissue. In addition, an expression is developed for the transient concentration about the probe when it is used as an infusion device. All mathematical expressions are found to be a sum of an algebraic and an integral term. Theoretical prediction of time-dependent probe behavior in brain has been compared with experimental data for acetaminophen administered at 15 mg/kg to rats by intravenous bolus. Plasma and whole striatal tissue samples were used to describe plasma kinetics and to estimate a capillary permeability-area product of 0.07 min-1. Theoretical prediction of transient effluent dialysate concentrations exhibited close agreement with experimental data over 60 min. Terminal decline of the dialysate effluent concentration was slightly overestimated but theoretical concentrations still lay within the 95% confidence interval of the experimental data at 112 min. Microvasculature transport and metabolism play major roles in determining microdialysate transient responses. Extraction fraction (recovery) has been shown to be a declining function in time for five probe operating conditions. High rates of metabolism and/or capillary transport affect the time required to approach steady-state extraction, shortening the time as the rates increase. Conversely, for substances characterized by low permeabilities and negligible metabolism, experimental situations exist that are predicted to have very slow approaches to microdialysis steady state.     

Whey proteins as a model system for chromatographic separation of proteins

Although chromatographic separation of whey proteins has been considered too expensive, whey may serve as an excellent model mixture to investigate and validate the use of simulation tools in the development and optimization of chromatographic separations and the outcome could easily be utilized since the model system has an intrinsic value. Besides, milk from transgenic animals could be an attractive source of pharmaceuticals which must be separated from the other proteins in the milk. Several whey proteins are of interest especially, alpha-lactalbumin, beta-lactoglobulins, immunoglobulins, lactoperoxidase, and lactoferrin. The scope of the project is to develop a consistent set of chromatographic data for whey proteins including isotherms, transport properties and scale-up studies and to develop the appropriate models for the anion exchangers Q-Sepharose XL, Source 30Q, Ceramic Q-HyperD F, and Merck Fractogel EMD TMAE 650 (S). In this work we have determined and correlated gradient and isocratic retention volumes in the linear range of the isotherm for alpha-lactalbumin, beta-lactoglobulin A and B, and bovine serum albumin at a pH from 6 to 9 at various NaCl concentrations.     

Quantitative high-performance liquid chromatography of nucleosides and bases in human plasma

A reversed-phase high-performance liquid chromatographic (HPLC) method has been developed for the estimation of purines, pyrimidines and their congeners in biological fluids. An initial HPLC separation allowed the collection of a number of effluent fractions, each of which contained a single component of interest. The re-application of these fractions to a second HPLC separation permitted the resolution and quantification of nanogram amounts of these components. Isocratic elution with volatile buffers renders the samples amenable to automatic sampling procedures or lyophilisation. Data are presented on the application of the method to the analysis of nucleosides and bases in human plasma.     

Determination of phenelzine in human plasma with gas chromatography—mass spectrometry using an isotope labeled internal standard

A quantitative gas chromatographic—mass spectrometric assay was developed for the determination of phenelzine in human plasma. Phenelzine, in aqueous solution or in plasma reacts at room temperature with pentafluorobenzaldehyde to form quantitatively a hydrazone derivative. The derivative has good gas chromatographic characteristics. The assay utilizes selected ion monitoring in a gas chromatographic effluent, the molecular ion generated by electron impact ionization of phenelzine derivative. Phenelzine-d₇ was synthesized and used as an internal standard. The assay can measure 2 ng/ml of the drug with about 10% precision.The method was used for the determination of steady state levels of phenelzine in the plasma of patients taking a therapeutic dose of the drug.     

紫藤凝集素的分离纯化及理化性质研究

用常规方法处理的ＤＥＡＥ离子交换纤维素柱,通过线性离子强度梯度洗脱,从紫藤种子的蛋白粗提液中得到一定纯度的紫藤凝集素。纯化的凝集素凝集兔红血球的比活提高４０倍,总活力回收率为１９．２％。紫藤凝集素的分子量经ＰＡＧＥ鉴定为２０５ｋｄ,是由两种亚基构成的四聚体,这两种亚基各有２个,分子量ＳＤＳ－ＰＡＧＥ鉴定分别为７７６００ｄ和２５１００ｄ。紫藤凝集素是一种糖蛋白,等电点约为４．６０。它可凝集人的各种血     

Computer system for equipment management

A computer system has been developed to meet the requirements of equipment management. The system was originally developed to run on personal computers, but has been upgraded to provide true multiuser facilities and more advanced program capabilities. This has been achieved using improved hardware and a relational database. The manner in which the sofware operates is described, with some examples examined in more detail. The system provides a wide range of information, including inventory data, repair costs and time as well as service records and worksheets. In addition it meets the other basic requirements of the department, including wordprocessing, budgetary control and stock control. The system also provides an immediate and repid overview of the repair state of all equipment whilst reducing administration time for many aspects of the service. Emphasis has been placed on the integrity of the data and ease of entry of additional data.