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目的观察不同大小及不同放置时间的滤纸洗脱尿半乳糖醇气相色谱/质谱(GC/MS)检测结果与原尿液半乳糖醇GC/MS检测结果之间差异,探讨滤纸尿送检筛查希特林蛋白缺陷所致新生儿肝内胆汁淤积症(NICCD)的可行性。方法选择既往GC/MS分析提示含有半乳糖醇的原尿液10份,每份制成3种大小不同的滤纸吸附,晾干,在室温下放置1~7 d,然后洗脱。取原尿液和洗脱尿100μL,用生化分析仪测定尿肌酐水平。取原尿液和洗脱尿100μL,原尿液先采用尿素酶处理后进行尿液代谢产物GC/MS检测。用质谱仪检索NIST图谱库定性检测到的尿液代谢产物。用正十七烷酸作为内标物质,应用内标法以内生肌酐定量代谢产物。结果同一份含有半乳糖醇的尿液,制成3种大小不同的滤纸尿,放置第1、3、5、7天的大小不同滤纸洗脱尿与其原尿液均经GC/MS检测,并进行比较,发现不同时间/不同大小的滤纸洗脱尿与其原尿液半乳糖醇的出峰时间及水平,均无统计学差异,不同时间/不同大小的滤纸洗脱尿半乳糖醇的出峰时间及水平均无统计学差异。结论在常温下,7 d内当洗涤滤纸的液体与滤纸吸收的尿液量一致时,滤纸尿可代替-80℃储存的原尿。建议使用的滤纸片规格是5 cm×8 cm,可吸收1.0 mL尿液。
Objective To observe the difference between GC-MS and urinary galactitol GC / MS in different size and time of filter paper, Feasibility of neonatal intrahepatic cholestasis (NICCD) due to Hittingin deficiency. Methods Select past 10 GC / MS analysis tips of raw urine containing galactitol, each made of three different sizes of filter paper adsorption, dried, at room temperature for 1 ~ 7 d, and then eluted. Take the original urine and urine eluted 100μL, with biochemical analyzer urine creatinine levels. Take the original urine and urine eluted 100μL, the original urine after urease treatment of urine metabolites by GC / MS detection. Mass spectrometry was used to retrieve urine metabolites that were qualitatively detected by the NIST Mapping Library. Using n-heptadecanoic acid as internal standard substance, endogenous creatinine was quantified using internal standard method. Results The same urine containing galactitol made of 3 different sizes of filter paper urine, placed on the 1st, 3rd, 5th and 7th days of different sizes of different types of filter paper urine and urine were detected by GC / MS, and The results showed that there was no significant difference in the time and level of the peak of urinary excretion of galactitol from urine of different times and different sizes of filter paper, and the peak of urinary galactitol elution from different time and different sizes of filter paper There was no significant difference in time and level. Conclusions Filter paper urine can replace original urine stored at -80 ° C when the amount of urine absorbed by filter paper is equal within 7 days at room temperature. The recommended filter paper size is 5 cm × 8 cm and absorbs 1.0 mL of urine.