气质联用技术检测尿琥珀酰丙酮及其在酪氨酸血症诊断中的应用  

The urine succinylacetone determination by gas chromatography-mass spectrometry and its application in diagnosis for tyrosinemia

作　　者：孙卫华[1] 曹迪[1] 王艺[1] 杨毅[1] 

机构地区：[1]复旦大学附属儿科医院儿科研究所,上海201102

出　　处：《检验医学》2009年第5期373-377,共5页Laboratory Medicine

摘　　要：目的建立气相色谱质谱用技术（GC-MS）测定尿琥珀酰丙酮（SA）的方法,用于临床上鉴别诊断酪氨酸血症Ⅰ型患者。方法尿SA先用盐酸羟胺进行污化反应,然后用乙酰乙酸2次萃取,双三甲基硅烷-三氟乙酰胺（BSTFA-TMCS）衍生化,衍生化后的SA用气相色谱质谱仪进行测定分析。质谱分析模式采用选择离子监测（SIM）模式进行检测,SA选择212/182 m/z,内标丙二酸选择233/248 m/z。分析该法的精确性、稳定性、样本回收率、残留分析和实测浓度与加入浓度之间的相关性。用该法检测肝肿大伴酪氨酸升高的13例患者。结果SA批内变异系数（CV）为6.8%;批间CV为7.8%;回收率为94.0%-102.0%;前处理CV为12.4%;残留分析〈1%。在5-120 mg/mL之间,实测浓度与加入浓度之间的相关系数（r）为0.997。内标丙二酸回收率为95.0%-103.0%。确诊酪氨酸血症Ⅰ型患儿2例,尿SA浓度分别为26、54 mmol/mol肌酐。结论GC-MS测定尿SA具有较高的回收率、精确性和准确性,为临床上鉴别诊断酪氨酸血症Ⅰ型提供了新的方法。Objective To establish the urine succinylacetone （SA） determination by gas chromatography-mass spectrometry （GC-MS） for the differential diagnosis of tyrosinemia I patients. Methods The SA in urine was oximated by hydroxylamine hydrochloride, then extracted with ethyl acetate twice, and was derivatized with nitrogen, oxygen-bis （trimethylsilyl） trifluoroacetamide and trimethylchlorosilane （BSTFA-TMCS） and assayed by GC-MS using selected ion monitoring （SIM）. The SA was determinated with 212/182 m/z and malonic acid（ internal standard）with 233/248 m/ z. The precision, accuracy, recoveries of the spiked samples, carry-over, linearity and the correlation of practical determination level and addition level was analyzed. Urine samples of 13 patients with hepatomegaly and tyrosinemia were determined. Resets The intra and inter assay coefficient of variation （CV） were 6.8% and 7.8% ; Inter-assay CV of sequential preparations of the same sample was 12.4%. The recovery of the spiked samples were 94.0% -102.0%. The carry-over analysis was less than 1%. The correlation coefficient of linearity was 0.997. The recovery of malonic acid was 95.0% -103.0%. Two patients with tyrosinemia I were diagnosed,and the levels of SA were 26 and 54 mmol/mol creatinine respectively. Conclusions The urine SA determination by GC-MS shows high recovery, precision and accuracy. It can be used in the differential diagnosis of tyro-sinemia Ⅰ.

关 键 词：琥珀酰丙酮 气相色谱 质谱 酪氨酸血症Ⅰ型 

分 类 号：R446.1[医药卫生—诊断学]