Brega A, Villa P, Quadrini G, Quadri A, Lucarelli C
Laboratorio Biomed., Concesio, Brescia, Italy.
J Chromatogr. 1991 Aug 16;553(1-2):249-54. doi: 10.1016/s0021-9673(01)88495-8.
A method for the determination of acetone in plasma or urine by high-performance liquid chromatography (HPLC) was developed. Plasma specimens are deproteinized with acetonitrile (1:1, v/v) 2,4-dinitrophenylhydrazine (DNPH) is added to the supernatant or to filtered urine samples, similarly treated with acetonitrile (2:1, v/v) to prevent crystallization of the synthesized phenylhydrazone. An aliquot (20 microliters) of the reaction mixture was subjected to HPLC at ambient temperature using a reversed-phase Pecosphere 3 x 3 C18 column with acetonitrile-water (45:55, v/v) as eluent at a flow-rate of 1 ml/min and detection at 365 nm. Hydroxyacetone and acetoacetate phenylhydrazone derivatives do not interfere. The identification of acetone by its retention time was confirmed by comparison with a laboratory-synthesized acetone DNPH derivative. The concentration of acetone, eluted within 3 min, was determined by the peak-height method. The detection limit was 0.034 mmol/l; the relative standard deviations were less than 5% within run (n = 20) and less than 10% between run (n = 20).
开发了一种通过高效液相色谱法（HPLC）测定血浆或尿液中丙酮的方法。血浆标本用乙腈（1:1，v/v）进行脱蛋白处理，向上清液或过滤后的尿液样本中加入2,4-二硝基苯肼（DNPH），同样用乙腈（2:1，v/v）处理以防止合成苯腙结晶。取20微升反应混合物的等分试样，在室温下使用反相Pecosphere 3×3 C18柱进行HPLC分析，以乙腈-水（45:55，v/v）作为洗脱剂，流速为1毫升/分钟，在365纳米处进行检测。羟基丙酮和乙酰乙酸苯腙衍生物不产生干扰。通过与实验室合成的丙酮DNPH衍生物比较，以保留时间确认丙酮的鉴定。在3分钟内洗脱的丙酮浓度通过峰高法测定。检测限为0.034毫摩尔/升；批内相对标准偏差小于5%（n = 20），批间相对标准偏差小于10%（n = 20）。
