Orlova N V, Proskurnin M A, Samburova V A, Dryagleva I D, Brusnichkin A V
Chemistry Department, M.V. Lomonosov Moscow State University, Vorob'evy Hills, 119992, GSP-2, Moscow, Russia.
Anal Bioanal Chem. 2003 Apr;375(8):1038-44. doi: 10.1007/s00216-003-1753-0. Epub 2003 Feb 28.
Based on the optimized spectrophotometric determination of pyrogens (of various classes ( p-aminophenol and endotoxins), thermal lensing was applied to the determination of these substances at the submicrogram level. The limit of detection of p-aminophenol, a pyrogenic impurity in pharmaceutical formulations of paracetamol, by reaction with resorcinol in alkaline solutions is 100 ng mL(-1). Phloroglucinol was considered as an analog of resorcinol as a reagent in this reaction. The conditions of spectrophotometric determination of pyrogenic lipopolysaccharides (endotoxins) by ion-pair formation with methylene blue (the limit of detection is 100 ng mL(-1)), by ion-pair formation with Stains-All (1-ethyl-2-[3-(1-ethylnaphtho[1,2-d]thiazolin-2-ylidene)-2-methylpropenyl]naphtho[1,2-d]thiazolium bromide) (the limit of detection is 500 ng mL(-1)), and by reaction of 2-keto-3-deoxyoctonic acid with thiobarbituric acid (the limit of detection is 800 ng mL(-1)) were proposed. The optimized procedure for 2-keto-3-deoxyoctonic acid was applied for thermal lensing that provided a decrease in the limit of detection to 70 ng mL(-1) and was also used for lipopolysaccharide determination in the endotoxin standard from E. coli.
基于对热原（各类热原，如对氨基苯酚和内毒素）的优化分光光度法测定，采用热透镜法在亚微克水平测定这些物质。对乙酰氨基酚药物制剂中的致热杂质对氨基苯酚，在碱性溶液中与间苯二酚反应的检测限为100 ng mL⁻¹。在该反应中，间苯三酚被视为间苯二酚的类似试剂。提出了通过与亚甲蓝形成离子对（检测限为100 ng mL⁻¹）、与全染剂（1-乙基-2-[3-(1-乙基萘并[1,2-d]噻唑啉-2-亚基)-2-甲基丙烯基]萘并[1,2-d]噻唑溴化物）形成离子对（检测限为500 ng mL⁻¹）以及通过2-酮-3-脱氧辛酸与硫代巴比妥酸反应（检测限为800 ng mL⁻¹）分光光度法测定致热脂多糖（内毒素）的条件。将优化后的2-酮-3-脱氧辛酸测定方法应用于热透镜法，使检测限降至70 ng mL⁻¹，该方法还用于测定大肠杆菌内毒素标准品中的脂多糖。
