黑龙江医药
黑龍江醫藥
흑룡강의약
HEILONGJIANG MEDICAL JOURNAL
2015年
3期
457-459
,共3页
UPLC-MS/MS( ESI)%皮质醇%内标%溯源%标准化
UPLC-MS/MS( ESI)%皮質醇%內標%溯源%標準化
UPLC-MS/MS( ESI)%피질순%내표%소원%표준화
UPLC MS/MS (ESI)%cortisol%internal standard%traceability%standardization
目的：建立一种同位素稀释-超高效液相色谱串联质谱法检测血浆中的皮质醇。方法：使用C18固相抽提小柱纯化、富集人体血浆中的总皮质醇。采用LC/MS/MS-ESI法分析血浆中的皮质醇含量。梯度流动相为0 min,50%甲醇；3 min,90%甲醇；3.5 min 90%甲醇；3.6min 50%甲醇；5 min 50%甲醇。水相含0.2%甲酸。MRM检测模式选择性监测cortisol(m/z363.2/121.4)、内标cortisol-d4(m/z367.4/121.4)。结果：血浆中加入梯度标准品的加标回收率分别为102.5%，101.1%，103.3%。检测冻存血浆中皮质醇的实验里，组内变异为0.54%～2.47%，组间变异为0.96%～1.69%，线性相关系数为R2≥0.999。LC/MS/MS信噪比为10时检出限为12pg。结论：该方法具有良好的特异性、灵敏度、精密度，能够准确的检测血浆中总皮质醇含量，为实现皮质醇溯源打下基础。
目的：建立一種同位素稀釋-超高效液相色譜串聯質譜法檢測血漿中的皮質醇。方法：使用C18固相抽提小柱純化、富集人體血漿中的總皮質醇。採用LC/MS/MS-ESI法分析血漿中的皮質醇含量。梯度流動相為0 min,50%甲醇；3 min,90%甲醇；3.5 min 90%甲醇；3.6min 50%甲醇；5 min 50%甲醇。水相含0.2%甲痠。MRM檢測模式選擇性鑑測cortisol(m/z363.2/121.4)、內標cortisol-d4(m/z367.4/121.4)。結果：血漿中加入梯度標準品的加標迴收率分彆為102.5%，101.1%，103.3%。檢測凍存血漿中皮質醇的實驗裏，組內變異為0.54%～2.47%，組間變異為0.96%～1.69%，線性相關繫數為R2≥0.999。LC/MS/MS信譟比為10時檢齣限為12pg。結論：該方法具有良好的特異性、靈敏度、精密度，能夠準確的檢測血漿中總皮質醇含量，為實現皮質醇溯源打下基礎。
목적：건립일충동위소희석-초고효액상색보천련질보법검측혈장중적피질순。방법：사용C18고상추제소주순화、부집인체혈장중적총피질순。채용LC/MS/MS-ESI법분석혈장중적피질순함량。제도류동상위0 min,50%갑순；3 min,90%갑순；3.5 min 90%갑순；3.6min 50%갑순；5 min 50%갑순。수상함0.2%갑산。MRM검측모식선택성감측cortisol(m/z363.2/121.4)、내표cortisol-d4(m/z367.4/121.4)。결과：혈장중가입제도표준품적가표회수솔분별위102.5%，101.1%，103.3%。검측동존혈장중피질순적실험리，조내변이위0.54%～2.47%，조간변이위0.96%～1.69%，선성상관계수위R2≥0.999。LC/MS/MS신조비위10시검출한위12pg。결론：해방법구유량호적특이성、령민도、정밀도，능구준학적검측혈장중총피질순함량，위실현피질순소원타하기출。
Objective:To establish a stable isotope dilution-ultra performance liquid chromatography tandem mass spectrometry method for the detection of plasma cortisol. Methods:Total cortisol concentration in human plasma and purification, using C18 solid phase extraction column. Cortisol concentrations in plasma were analyzed by using LC/MS/MS-ESI method. A gradient mobile phase was 0 min, 3 min, 50%methanol, 90%methanol;methanol;3.5 min 90%3.6min 50%;5 min 50%of methanol;methanol. The aqueous phase containing 0.2% formic acid. MRM detection mode selective monitoring of cortisol (m/z363.2/121.4) cortisol-d4 (m/z367.4/121.4), the internal standard. Results:The recovery of added standard gradient in plasma was respectively 102.5%, 101.1%, 103.3%. Detection of plasma cortisol in freezing experiment, within group variation is 0.54%~2.47%, 0.96%~1.69%between group variation, linear correlation coefficient is R2=0.999. LC/MS/MS signal to noise ratio is 10, the detection limit is 12pg. Conclusion:The method has good specificity, sensitivity, precision, detection of plasma total cortisol levels can be accurate, lay the foundation for the realiza-tion of cortisol.
