■慢性肾病的筛查依赖于准确和精确的肌酐测量。传统上,使用高通量化学分析仪测量血清或血浆中的肌酐。然而,干血斑(DBS)也可以用来改善测试访问。
■样品是从6毫米DBS冲头获得的,在经历乙腈崩溃之前在水中重建。在注射前使用80:20乙腈:水稀释所得上清液。使用等度梯度鉴定肌酐,并使用API4000三重四极杆质量分析仪进行检测。定量依赖于矩阵匹配的校准物,值与罗氏Cobas酶促测定法一致。评估方法性能的验证研究包括精度,线性度准确度,方法比较,稳定性,干扰,和矩阵效应。
LC-MSMS测定从0.3至20mg/dL是线性的(y=1.02x-0.11;R2=0.996)。使用跨越分析测量范围的矩阵匹配对照(n=4),精度范围为5.2%至8.1%。LC-MSMS结果对应于酶测定(Roche),拟合线方程为y=0.956x-0.07(R2=0.995;n=173)。与BeckmanJaffe测定相比,Siemens和Roche酶促测定在与DBS肌酐浓度(n=40成对的静脉/DBS集合)相关方面具有更高的准确性(-2.5%和-0.8%对-6.3%和-4.1%,分别)或iSTAT(-28.4%和-27.1%,分别)。准确性不受血细胞比容的影响,血斑体积,过量的IgG或IgA,或高甘油三酯血症。没有观察到基质效应,提取和加工效率都很稳健。环境稳定性延长到至少10天,暴露于极端温度并不影响肌酐结果.
■我们成功开发了一种准确,精确的LC-MSMS方法,用于定量DBS中的肌酐。
UNASSIGNED: Screening for chronic kidney disease relies on accurate and precise creatinine measurements. Traditionally, creatinine is measured in serum or plasma using high-throughput chemistry analyzers. However, dried blood spots (DBS) can also be utilized to improve testing access.
UNASSIGNED: Samples were obtained from a 6 mm DBS punch, which was reconstituted in water before undergoing an acetonitrile crash. The resulting supernatant was diluted using an 80:20 acetonitrile: water before injection. Creatinine was identified using an isocratic gradient, and detected using an API 4000 triple quadrupole mass analyzer. Quantification relied on matrix-matched calibrators, with values harmonized to the Roche Cobas enzymatic assay. Validation studies assessing method performance included precision, linearity, accuracy, method comparison, stability, interference, and matrix effects.
UNASSIGNED: The LC-MSMS assay was linear from 0.3 to 20 mg/dL (y = 1.02x-0.11; R2 = 0.996). Precision ranged from 5.2 to 8.1 % using matrix-matched controls (n = 4) that spanned the analytical measurement range. LC-MSMS results corresponded to the enzymatic assay (Roche) with a fitted line equation of y = 0.956x-0.07 (R2 = 0.995; n = 173). The Siemens and Roche enzymatic assays demonstrated higher accuracy in correlating to the DBS creatinine concentration (n = 40 paired venous/DBS collections) compared to the Beckman Jaffe assay (-2.5 % and -0.8 % versus -6.3 % and -4.1 %, respectively) or the iSTAT (-28.4 % and -27.1 %, respectively). Accuracy was unaffected by hematocrit, blood spot volume, excess IgG or IgA, or hypertriglyceridemia. No matrix effects were observed, and both extraction and processing efficiency were robust.Ambient stability extended to at least 10 days, and exposure to extreme temperature did not affect the creatinine results.
UNASSIGNED: We successfully developed an accurate and precise LC-MSMS method for quantifying creatinine in DBS.