检验医学
檢驗醫學
검험의학
LABORATORY MEDICINE
2015年
8期
821-824
,共4页
25-羟基维生素D2%25-羟基维生素D3%液相色谱串联质谱%内标法%血清
25-羥基維生素D2%25-羥基維生素D3%液相色譜串聯質譜%內標法%血清
25-간기유생소D2%25-간기유생소D3%액상색보천련질보%내표법%혈청
25-hydroxyvitamin D2%25-hydroxyvitamin D3%Liquid chromatography tandem-mass spectrometry%Internal standard method%Serum
目的:建立液相色谱串联质谱法(LC-MS/MS)测定血清25-羟基维生素D2[25(OH)D2]及25-羟基维生素D3[25(OH)D3]含量的方法。方法采用乙醇沉淀蛋白,正己烷液液萃取目标组分,乙腈复溶。采用LC-MS/MS[正离子电喷雾离子化(ESI+)的多反应监测模式(MRM)]氘代同位素内标法检测血清25(OH)D2及25(OH)D3含量并进行相关方法学验证。结果 LC-MS/MS检测血清25(OH)D2及25(OH)D3的批内精密度为0.88%~7.69%,批间精密度为1.56%~9.90%;25(OH)D2在0.5~10.0 ng/mL、25(OH)D3在5~100 ng/mL范围内线性良好,线性相关系数分别为0.9991、0.9999,校准品测试结果正确度为95.5%~101.2%。281名志愿者男、女之间25(OH)D2含量差异无统计学意义(P>0.05),25(OH)D3含量差异有统计学意义(P<0.05)。结论 LC-MS/MS检测血清25(OH)D2及25(OH)D3的敏感性高,结果准确、稳定,可应用于临床分析。
目的:建立液相色譜串聯質譜法(LC-MS/MS)測定血清25-羥基維生素D2[25(OH)D2]及25-羥基維生素D3[25(OH)D3]含量的方法。方法採用乙醇沉澱蛋白,正己烷液液萃取目標組分,乙腈複溶。採用LC-MS/MS[正離子電噴霧離子化(ESI+)的多反應鑑測模式(MRM)]氘代同位素內標法檢測血清25(OH)D2及25(OH)D3含量併進行相關方法學驗證。結果 LC-MS/MS檢測血清25(OH)D2及25(OH)D3的批內精密度為0.88%~7.69%,批間精密度為1.56%~9.90%;25(OH)D2在0.5~10.0 ng/mL、25(OH)D3在5~100 ng/mL範圍內線性良好,線性相關繫數分彆為0.9991、0.9999,校準品測試結果正確度為95.5%~101.2%。281名誌願者男、女之間25(OH)D2含量差異無統計學意義(P>0.05),25(OH)D3含量差異有統計學意義(P<0.05)。結論 LC-MS/MS檢測血清25(OH)D2及25(OH)D3的敏感性高,結果準確、穩定,可應用于臨床分析。
목적:건립액상색보천련질보법(LC-MS/MS)측정혈청25-간기유생소D2[25(OH)D2]급25-간기유생소D3[25(OH)D3]함량적방법。방법채용을순침정단백,정기완액액췌취목표조분,을정복용。채용LC-MS/MS[정리자전분무리자화(ESI+)적다반응감측모식(MRM)]도대동위소내표법검측혈청25(OH)D2급25(OH)D3함량병진행상관방법학험증。결과 LC-MS/MS검측혈청25(OH)D2급25(OH)D3적비내정밀도위0.88%~7.69%,비간정밀도위1.56%~9.90%;25(OH)D2재0.5~10.0 ng/mL、25(OH)D3재5~100 ng/mL범위내선성량호,선성상관계수분별위0.9991、0.9999,교준품측시결과정학도위95.5%~101.2%。281명지원자남、녀지간25(OH)D2함량차이무통계학의의(P>0.05),25(OH)D3함량차이유통계학의의(P<0.05)。결론 LC-MS/MS검측혈청25(OH)D2급25(OH)D3적민감성고,결과준학、은정,가응용우림상분석。
Objective To establish a method for the determinations of 25-hydroxyvitamin D2[25(OH)D2]and 25-hydroxyvitamin D3[25(OH)D3] contents in serum by liquid chromatography tandem-mass spectrometry (LC-MS/MS) .Methods After proteins in serum were precipitated by ethanol, liquid-liquid extraction by n-Hexane, then the residuals were re-dissolved in acetonitrile and analyzed by LC-MS/MS in the positive electrospray ionization ( ESI+) mode and multiple reaction monitor(MRM) mode.The quantitative analysis for 25(OH)D2 and 25(OH)D3 contents was carried out by deuterium isotope as internal standard, and the methodology validation was performed.Results The within-run precision of 25 ( OH) D2 and 25 ( OH) D3 was 0.88%-7.69%, and between-run precision was 1.56%-9.90%.The good correlation coefficients were 0.999 1 and 0.999 9 at the concentration of 0.5-10.0 ng/mL 25(OH) D2 and 5-100 ng/mL 25(OH)D3, respectively.The accuracy of quality control materials of 25(OH)D2 and 25(OH) D3 was 95.5%-101.2%.25(OH)D2 content did not have statistical significance(P>0.05), and 25(OH)D3 content had statistical significance(P <0.05) between males and females of 281 volunteers.Conclusions This method is sensitive, accurate and stable, satisfying the quantification requirements of 25(OH)D2 and 25(OH)D3 in serum, and it can be used in clinical analysis.