Deng Chunhui, Zhang Wei, Zhang Jie, Zhang Xiangmin
Department of Chemistry, Fudan University, Shanghai 200433, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Jun 15;805(2):235-40. doi: 10.1016/j.jchromb.2004.03.001.
Acetone is an important volatile disease marker. Due to its nature of activity and volatility, it is a difficult task to measure the concentration of acetone in biological samples with accuracy. In this paper, we developed a novel method for determination of trace amount acetone in human plasma by solid-phase microextraction technique with on-fiber derivatization. In this method, the poly(dimethylsiloxane)/divinylbenzene (PDMS/DVB) fiber was used and O-2,3,4,5,6-(pentafluorobenzyl) hydroxylamine hydrochloride (PFBHA) was first loaded on the fiber. Acetone in plasma sample was agitated into headspace and extracted by solid-phase microextraction (SPME) fiber and subsequently derivatized with PFBHA on the fiber. Acetone oxime was analyzed by gas chromatography-mass spectrometry (GC-MS). Quantitative analysis of acetone in plasma was carried out by using external standard method. The SPME conditions (extraction temperature and time) and the method validation were studied. The present method was tested by determination of acetone in diabetes plasma and normal plasma. Acetone concentration in diabetes plasma was found to be higher than 1.8mM, while in normal plasma was lower than 0.017 mM. The results show that the present method is a potential tool for diagnosis of diabetes.
丙酮是一种重要的挥发性疾病标志物。由于其活性和挥发性的特性,准确测量生物样品中丙酮的浓度是一项艰巨的任务。在本文中,我们开发了一种通过固相微萃取技术结合纤维上衍生化来测定人血浆中痕量丙酮的新方法。在该方法中,使用了聚(二甲基硅氧烷)/二乙烯基苯(PDMS/DVB)纤维,并且首先将O-2,3,4,5,6-(五氟苄基)盐酸羟胺(PFBHA)负载到纤维上。血浆样品中的丙酮被搅动到顶空中,通过固相微萃取(SPME)纤维进行萃取,随后在纤维上与PFBHA进行衍生化。丙酮肟通过气相色谱-质谱联用(GC-MS)进行分析。血浆中丙酮的定量分析采用外标法进行。研究了SPME条件(萃取温度和时间)以及方法验证。通过测定糖尿病患者血浆和正常血浆中的丙酮对本方法进行了测试。发现糖尿病患者血浆中的丙酮浓度高于1.8mM,而正常血浆中的丙酮浓度低于0.017mM。结果表明,本方法是一种潜在的糖尿病诊断工具。