發(fā)布時間：2018-05-14 05:24

  本文選題：液相色譜-串聯(lián)質(zhì)譜 + 自動移液平臺　； 參考：《色譜》2016年06期

【摘要】：建立了一種高通量液相色譜-串聯(lián)質(zhì)譜技術檢測干血點(DBS)中同型半胱氨酸(homocycteine,Hcy)的方法。以DBS為樣本,homocystine-D8為同位素內(nèi)標,二硫蘇糖醇(DTT)為蛋白結(jié)合態(tài)Hcy的還原劑,使用含0.1%(v/v)甲酸、0.05%(v/v)三氟乙酸的乙腈溶液萃取。整個前處理過程使用自動移液平臺及96孔板實現(xiàn)高通量自動化操作。處理后的樣本經(jīng)過Phenomenex CN柱分離,使用多反應監(jiān)測模式進行LC-MS/MS分析。結(jié)果表明:Hcy的檢出限為0.12μmol/L(S/N=3),定量限為0.46μmol/L(S/N=10)。Hcy在1.16~148.00μmol/L范圍內(nèi)線性關系良好,R~2=0.994。Hcy的平均回收率為(103.0±4.97)%~(112.0±2.13)%,日內(nèi)相對標準偏差(RSD)為1.9%~4.6%,日間RSD為1.5%~7.1%。DBS樣本在不同溫度(-4、-20、22和37℃)下儲存不同時間(0、1、2、3、4、5、6、14天)后的穩(wěn)定性試驗顯示樣本總體RSD15%,經(jīng)前處理后的樣本在48 h內(nèi)的穩(wěn)定性試驗顯示樣本總體RSD5%。該方法與傳統(tǒng)生化分析方法的相關性好(R~2=0.9818,n=47)。
[Abstract]:A high throughput liquid chromatography-tandem mass spectrometry (HPLC-MS) method was developed for the determination of homocysteine homocyte-homocycteine (Hcyine) in DBS. Homocystine-D8 was used as the internal standard of DBS, DTT was used as the reductant of protein-bound Hcy, and acetonitrile solution containing 0.1% v / v) formic acid was used to extract the acetonitrile solution containing 0.05% V / v) trifluoroacetic acid. The whole preprocessing process uses automatic fluid transfer platform and 96 hole plate to realize high-throughput automatic operation. The treated samples were separated by Phenomenex CN column and analyzed by LC-MS/MS using multi-reaction monitoring model. The results showed that the detection limit was 0.12 渭 mol 路L ~ (-1) 路L ~ (-1) S / N ~ (3) and the quantitative limit was 0.46 渭 mol/L(S/N=10).Hcy. The linear range of 0.46 渭 mol/L(S/N=10).Hcy was 1.16 ~ 148.00 渭 mol/L. The average recovery of 0.994.Hcy was 103.0 鹵4.97 ~ 0.994.Hcy was 112.0 鹵2.133.The relative standard deviation (RSD) was 1.9 鹵4.6. the daily RSD for 1.5%~7.1%.DBS samples was stored at different temperatures (-4-2022 and 37 鈩，

本文編號：1886546