检验医学
檢驗醫學
검험의학
LABORATORY MEDICINE
2015年
5期
427-432
,共6页
类固醇激素%血清%稳定同位素稀释%液相色谱%串联质谱
類固醇激素%血清%穩定同位素稀釋%液相色譜%串聯質譜
류고순격소%혈청%은정동위소희석%액상색보%천련질보
Steroid hormone%Serum%Stable isotope dilution%Liquid Chromatography%Tandem mass spectrometry
目的：建立一种能同时测定血清中多种类固醇激素[包括脱氢表雄酮（ DHEA）、17α－羟孕酮（17α－OHP4）、雄烯二酮（AD）、雌酮（E1）、皮质酮（CORT）、双氢睾酮（DHT）、孕烯醇酮（P5）和17－羟孕烯醇酮（17－OHP5）]的快速、灵敏的稳定同位素稀释高效液相色谱串联质谱（ID－HPLC－MS／MS）方法。方法血清样品经甲基叔丁基醚（MTBE）提取，上清液吹干后加入羟胺进行柱前衍生化反应后检测。采用 Phenomenex C18反相色谱柱和 Phenomenex C18预保护柱。流动相 A 为含0．1％甲酸的水，流动相 B 为含0．1％甲酸的甲醇。质谱采用电喷雾电离（ESI）模式进行离子化，正离子多反应监测模式（MRM）扫描，内标法定量分析。结果血清中8种类固醇激素最低定量限[LLOQ，以信噪比（S／N）≥10为标准]为0．03～0．625 ng／mL；在0．05～50．00 ng／mL范围内线性良好，相关系数（r）均≥0．9980；批内精密度为1．93％～13．81％，批间精密度为4．90％～16．18％，加标回收率为80．0％～130．6％。结论建立了同时测定8种类固醇激素的 ID－HPLC－MS／MS。该法灵敏度高、特异性强、准确且高效，可在10 min 内完成分离和检测，适用于人体血清中类固醇激素的定量分析。
目的：建立一種能同時測定血清中多種類固醇激素[包括脫氫錶雄酮（ DHEA）、17α－羥孕酮（17α－OHP4）、雄烯二酮（AD）、雌酮（E1）、皮質酮（CORT）、雙氫睪酮（DHT）、孕烯醇酮（P5）和17－羥孕烯醇酮（17－OHP5）]的快速、靈敏的穩定同位素稀釋高效液相色譜串聯質譜（ID－HPLC－MS／MS）方法。方法血清樣品經甲基叔丁基醚（MTBE）提取，上清液吹榦後加入羥胺進行柱前衍生化反應後檢測。採用 Phenomenex C18反相色譜柱和 Phenomenex C18預保護柱。流動相 A 為含0．1％甲痠的水，流動相 B 為含0．1％甲痠的甲醇。質譜採用電噴霧電離（ESI）模式進行離子化，正離子多反應鑑測模式（MRM）掃描，內標法定量分析。結果血清中8種類固醇激素最低定量限[LLOQ，以信譟比（S／N）≥10為標準]為0．03～0．625 ng／mL；在0．05～50．00 ng／mL範圍內線性良好，相關繫數（r）均≥0．9980；批內精密度為1．93％～13．81％，批間精密度為4．90％～16．18％，加標迴收率為80．0％～130．6％。結論建立瞭同時測定8種類固醇激素的 ID－HPLC－MS／MS。該法靈敏度高、特異性彊、準確且高效，可在10 min 內完成分離和檢測，適用于人體血清中類固醇激素的定量分析。
목적：건립일충능동시측정혈청중다충류고순격소[포괄탈경표웅동（ DHEA）、17α－간잉동（17α－OHP4）、웅희이동（AD）、자동（E1）、피질동（CORT）、쌍경고동（DHT）、잉희순동（P5）화17－간잉희순동（17－OHP5）]적쾌속、령민적은정동위소희석고효액상색보천련질보（ID－HPLC－MS／MS）방법。방법혈청양품경갑기숙정기미（MTBE）제취，상청액취간후가입간알진행주전연생화반응후검측。채용 Phenomenex C18반상색보주화 Phenomenex C18예보호주。류동상 A 위함0．1％갑산적수，류동상 B 위함0．1％갑산적갑순。질보채용전분무전리（ESI）모식진행리자화，정리자다반응감측모식（MRM）소묘，내표법정량분석。결과혈청중8충류고순격소최저정량한[LLOQ，이신조비（S／N）≥10위표준]위0．03～0．625 ng／mL；재0．05～50．00 ng／mL범위내선성량호，상관계수（r）균≥0．9980；비내정밀도위1．93％～13．81％，비간정밀도위4．90％～16．18％，가표회수솔위80．0％～130．6％。결론건립료동시측정8충류고순격소적 ID－HPLC－MS／MS。해법령민도고、특이성강、준학차고효，가재10 min 내완성분리화검측，괄용우인체혈청중류고순격소적정량분석。
Objective To establish a rapid, sensitive and stable isotope dilution high performance liquid chromatography-tandem mass spectrometry ( ID-HPLC-MS/MS ) for simultaneous quantitative determination of dehydroepiandrosterone(DHEA), 17 alpha-hydroxyprogesterone(17α-OHP4), androstenedione (AD), estrone (E1), corticosterone(CORT), dihydrotestosterone ( DHT), pregnenolone ( P5) and 17-hydroxylpregnenolone (17-OHP5). Methods The steroid hormones were extracted from human serum by methyl tert-butyl ether ( MTBE), and the supernatant was treated with hydroxylamine to produce post-column derivatives before detection.Steroid hormones were separated by Phenomenex reversed-phase C18 with precolumn.Water containing 0.1% formic acid and methanol containing 0.1% formic acid were used as mobile phase A and B,respectively.Multiple reaction monitoring(MRM)with the positive ion detection mode was applied to selectively detect these 8 steroid hormones ionized by electrospray ionization (ESI) interface, and the quantitative analysis for them was carried out by using deuterium isotope as internal standard.Results The lower limits of quantitation(LLOQ) of these 8 hormones can reach 0.03-0.625 ng/mL on the basis of signal-noise ratio ( S/N) ≥10.The correlation coefficients (r) were ≥0.998 0 with a good linear at the concentration of 0.05-50.00 ng/mL.The within-run precision was 1.93%-13.81%, and the between-run precision was 4.90%-16.18%.The recoveries were 80.0%-130.6%.Conclusions The ID-HPLC-MS/MS is established for simultaneous quantitative determination of 8 kinds of steroid hormones.It has the advantages of sensitivity, specificity and accuracy, which is suitable for the separation and determination of steroid hormones for human serum samples within 10 min.
