High-performance size exclusion chromatography of biological products

Summary High-performance size exclusion chromatography has been applied to the analysis and separation of a range of biological products — substances and preparations used in human medicine that cannot be completely defined by chemical and physical means. A variety of proteins and polysaccharides have been examined and results are described for a number of blood products, and polysaccharides of bacterial origin. This method is superior to conventional size esclusion for characterization of such substances and permits their isolation for further examination, without work-up, by biological and immunological techniques. The selection of appropriate column supports is considered. The limitations are considered of this procedure for studying the interaction between antithrombin III and heparin as the basis of a non-biological assay method.Presented at the 14th International Symposium on Chromatography London, September, 1982     

Rapid and sensitive gas chromatographic method for detection of barbital and pentobarbital in blood using flash-heater methylation and nitrogen-specific detection

Summary A sensitive and simplified method is described for the determination of barbital (B) and pentobarbital (PB) in blood using gas chromatography with a nitrogen-specific flame ionization detector. After a simple one-step extraction, B and PB were reconstituted in trimethylanilinium hydroxide and introduced directly into the gas chromatograph. Lower limits of detection was 0.14 μg/ml for B and PB in either water or blood solutions. The recoveries for B from water and blood were 90 and 88%, respectively; for PB they were essentially complete. The average between-run and within-run coefficients of variation for B from water and blood were lower than 7.8%, whereas those of PB were lower than 3.0%. The scale and sensitivity of detection for B and PB were found to be suitable for use in pharmacokinetic studies in the rat.     

层层自组装技术在生物医用材料领域中的应用研究进展

基于聚电解质阴阳离子交替组装的层层自组装技术由于可在温和的条件下实现多种生物大分子在材料表面的固定,并通过对组装条件的控制实现多种生物功能,已成为生物医用材料表面设计的重要手段。本文对层层自组装技术在构建血液相容性界面、组织工程表面、药物控释涂层等生物医用材料领域的应用研究进行了比较系统的阐述。     

骨髓增生异常综合症患者血液红细胞^31P—NMR谱的特征

本文介绍了用３１Ｐ－ＮＭＲ谱研究骨髓增生异常综合症（ＭＤＳ）患者血液红细胞的方法。分析了患者和正常人的血液红细胞在３１Ｐ－ＮＭＲ谱上的差异，探讨这些差异在临床医学上对ＭＤＳ患者进行诊断和疗效跟踪检测的应用途径。     

固相萃取—紫外导数光谱法测定血中茚满二酮类抗凝血杀鼠剂

报道了血中敌鼠、氯敌鼠、杀鼠酮的题示测定方法。该法操作简便快速、回收率高，灵敏度较高，萃取使用国产高分子多孔微球ＧＤＸ３０１，价廉易得。三种杀鼠剂以１０ｍｇ／Ｌ量分别加于血中，回收率依次为１００．５％±２．９％、１０１．５％±１．３％及９９．３％±１．５％；血中三种杀鼠剂检出限分别为０．４、０．６及０．４ｍｇ／Ｌ。     

人体血清中钾、钠、钙、镁测定的新方法——OCEP-1型离子分析仪的应用

本文报道用毛细管等速电泳法分离并同时测定人体血清中的四种阳离子(K、Na、Ca、Mg)的新方法。以0.01mol/L HC1溶液(含7‰18-冠醚)为前导电解质溶液,以0.01mol/L Tris溶液为尾随电解质溶液,可得到区带间界面鲜明的谱图。具有试液用量少、操作方便、分辨力高等特点。用于血清样品的测定,结果与常规分析一致。方法回收率为98.0～102.5％,可推广应用。     

Development and Validation of a Sensitive LC-MS/MS Method for the Simultaneous Analysis of Three-Tropane Alkaloids in Blood and Urine

A simple and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous analysis of three tropane alkaloids in blood and urine. After 1 mL of a blood or urine sample was extracted using a liquid–liquid extraction method with ethyl acetate at pH 8 and homatropine as the internal standard, the tropane alkaloids were separated. An Allure PFP propyl column (50 mm × 2.1 mm, 5 µm) separated the tropane alkaloids using an acetonitrile-buffer solution (20 mmol/L ammonium acetate and 0.1% formic acid, pH 4) (70:30) as the mobile phase at a flow-rate of 0.2 mL/min in isocratic mode, with the LC-MS/MS in the positive ionization mode. For each compound, detection was related to two daughter ions (scopolamine: m/z 304.4 → 138.1 and 155.9; atropine: m/z 290.3 → 124.0 and 93.1; anisodamine: m/z 306.3 → 140.1 and 91.1; and homatropine: m/z 276.3 → 124.3 and 142.1). The tropane alkaloids exhibited excellent linearity in the range of 0.05–100 ng/mL in blood and 0.2–100 ng/mL in urine, with a limit of detection range from 0.02 to 0.05 ng/mL for biological materials. The extraction recoveries of atropine, scopolamine, and anisodamine were more than 53% in the blood and urine; the interday and intraday RSDs were less than 10%; the within-day and between-day accuracy were between ?9.8% and +8.8%. The present method is simple and rapid, as shown by its application to a clinical case. This method is useful for routine analysis of tropane alkaloids in cases of suspected tropane alkaloid poisoning.     

Electro-kinetically driven route for highly sensitive blood pathology on a paper-based device

Enhancing the sensitivity of colorimetric detection in paper-devices is a quintessential step in achieving frugal diagnosis. Here, we demonstrate an effective way of improving the detection sensitivity of paper-based devices, as mediated by electro-kinetic mechanisms. By directly employing blood plasma, we investigate the electro-kinetic clustering of glucose, a neutral molecule in paper devices. Under the influence of uniform electric field, dispersed glucose gets accumulated in the paper strips. Due to the combination of EOF and electrophoretic migration, we achieve twofold increase in the colour intensity for both normal and diabetic samples. This approach is robust and possesses better sensitivity than conventional colorimetric assays and can be easily extended to other body fluid based diagnosis. These results may turn out to be of profound importance in improving the quality of pathological diagnosis in low-cost paper-based point-of-care devices deployed in resource-limited settings.     

Magnetic characterization of human blood in the atherosclerotic process in coronary arteries

In the last decades there has been an increasing interest in biomagnetism—a field of biophysics concerned with the magnetic properties of living organisms. Biomagnetism focuses on the measurement of magnetic properties of biological samples in the clinical environment. Progress in this field can provide new data for the understanding of the pathomechanism of atherosclerosis and support the diagnostic options for the evaluation and treatment of atherothrombotic complications. Lyophilized human blood samples from patients with atherosclerotic lesions (calcium scoring (CS) CS>0) and without atherosclerotic lesions (CS=0) were magnetically investigated. Magnetic measurements (performed in room and low temperature) indicated significant magnetic differences between these two groups of patients. Atherosclerotic blood samples are characterized by higher concentration of ferrimagnetic particles (magnetite and/or maghemite) and significant changes in the superparamagnetic behaviour. This research presents that magnetometry, in combination with medical research can lead to a better understanding of iron physiology in the atherosclerotic process.