CAJ | 학술논문

目的建立一种使用同位素稀释液相色谱串联质谱(ID-LC/MS/MS)测定人血清尿酸的候选参考方法 .方法用[1,3]-~(15)N_2尿酸作内标,与一定量的血清充分混匀,加入等量乙腈沉淀血清蛋白并提取尿酸,氯仿纯化上清液后用液相色谱串联质谱(LC/MS/MS)分离测定.优化LC/MS/MS的色谱条件,评价该法的精密度、回收率和偏倚,并计算不确定度.结果对3个浓度水平的室间质量评价用质控血清测定3批,批内、批间和总变异系数(CV)的均值(范围)分别为0.54%(0.44%～0.69 %)、0.66%(0.45 %～0.79 %)和0.86%((0.66%～)1.02 %).加样回收试验回收率范围为98.38%～102.64%.美国国家标准与技术研究所(NIST)标准物质SRM 909b-Ⅰ、Ⅱ的测定均值与靶值的偏倚分别为0.04%和0.25%.3种质控血清的相对扩展不确定度分别为1.80%、2.19%和2.04%.结论建立的ID-LC/MS/MS测定血清尿酸的方法精密、准确,有望作为血清尿酸测定的参考方法 .
목적 건립일충사용동위소희석액상색보천련질보(ID-LC/MS/MS)측정인혈청뇨산적후선삼고방법 .방법 용[1,3]-~(15)N_2뇨산작내표,여일정량적혈청충분혼균,가입등량을정침정혈청단백병제취뇨산,록방순화상청액후용액상색보천련질보(LC/MS/MS)분리측정.우화LC/MS/MS적색보조건,평개해법적정밀도、회수솔화편의,병계산불학정도.결과 대3개농도수평적실간질량평개용질공혈청측정3비,비내、비간화총변이계수(CV)적균치(범위)분별위0.54%(0.44%～0.69 %)、0.66%(0.45 %～0.79 %)화0.86%((0.66%～)1.02 %).가양회수시험회수솔범위위98.38%～102.64%.미국국가표준여기술연구소(NIST)표준물질SRM 909b-Ⅰ、Ⅱ적측정균치여파치적편의분별위0.04%화0.25%.3충질공혈청적상대확전불학정도분별위1.80%、2.19%화2.04%.결론 건립적ID-LC/MS/MS측정혈청뇨산적방법 정밀、준학,유망작위혈청뇨산측정적삼고방법 .
Objective To develop a candidate reference method for the measurement of serum uric acid by isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS). Methods An isotope labeled internal standard(IS) [1,3]-~(15)N_2 uric acid was weighed and spiked to serum samples. Acetonitrile was applied to precipitate serum proteins and extract uric acid and IS. After being treated with chloroform for further clean-up, the samples were analyzed by LC/MS/MS. Serum uric acid was quantified by a bracketing calibration. Results The within-run, between-run and total coefficients of variation(CV)of 3 batches of measurements of 3 levels of lyophilized sera used in external quality assessment ranged from 0.44% to 0.69%, 0.45 % to 0.79% and 0.66% to 1.02%, and the averages were 0.54%, 0.66% and 0.86%,respectively. The analytical recoveries ranged from 98.38% to 102.64% with an average of 100.55%. The results of analyzing the certified reference material SRM909b-Ⅰand Ⅱ showed 0.04% and 0.25% proportional bias compared with certified values. Evaluated uncertainty of measurement for the determination of (3 levels) of lyophilized sera and values of the relative expanded uncertainty were 1.80%,2.19% and 2.04%. Conclusions An ID-LC/MS/MS method for measuring serum uric acid has been developed. The method is highly precise and accurate and may be used as a candidate reference method for serum uric acid measurement.