CAJ | 학술논문

利用阳离子交换固相萃取柱( Waters Oasis MCX)富集净化 DNA样品,建立了液相色谱串联质谱(LC-MS/MS)同时检测DNA中3-甲基腺嘌呤(N3-MeA)和3-乙基腺嘌呤(N3-EtA)的方法。采用氘代-3-甲基腺嘌呤( d3-N3-MeA)和氘代-3-乙基腺嘌呤( d5-N3-EtA)为内标；进样量3μL,分析时间为13 min；亲水相互作用色谱柱(Waters XBridge HILIC)进行液相分离,流动相为10 mmol/L甲酸铵-乙腈溶液(5：95, V/V, pH=4.0),流速250μL/min；质谱条件：电喷雾离子源,多反应监测正离子扫描方式；电喷雾电压：5500 V,雾化气：369 Pa,气帘气：185 Pa,电离温度：400℃,驻留时间：40 ms。本方法对N3-MeA和N3-EtA的检出限分别为0.043和0.007μg/L,方法回收率为87.8%~103.0%。采用本方法检测了卷烟烟气粒相物暴露的DNA中N3-MeA和N3-EtA含量。结果表明,卷烟烟气粒相物暴露后的小牛胸腺DNA中3-甲基腺嘌呤和3-乙基腺嘌呤可被本方法定量检出。
이용양리자교환고상췌취주( Waters Oasis MCX)부집정화 DNA양품,건립료액상색보천련질보(LC-MS/MS)동시검측DNA중3-갑기선표령(N3-MeA)화3-을기선표령(N3-EtA)적방법。채용도대-3-갑기선표령( d3-N3-MeA)화도대-3-을기선표령( d5-N3-EtA)위내표；진양량3μL,분석시간위13 min；친수상호작용색보주(Waters XBridge HILIC)진행액상분리,류동상위10 mmol/L갑산안-을정용액(5：95, V/V, pH=4.0),류속250μL/min；질보조건：전분무리자원,다반응감측정리자소묘방식；전분무전압：5500 V,무화기：369 Pa,기렴기：185 Pa,전리온도：400℃,주류시간：40 ms。본방법대N3-MeA화N3-EtA적검출한분별위0.043화0.007μg/L,방법회수솔위87.8%~103.0%。채용본방법검측료권연연기립상물폭로적DNA중N3-MeA화N3-EtA함량。결과표명,권연연기립상물폭로후적소우흉선DNA중3-갑기선표령화3-을기선표령가피본방법정량검출。
A liquid chromatography-tandem mass spectrometry ( LC-MSMS ) method has been developed for the simultaneous determination of N3-methyladenine ( N3-MeA ) and N3-ethyladenine ( N3-EtA ) in calf thymus DNA. The DNA samples has been purified and enriched by cation exchange cartridge ( Waters Oasis MCX) . d3-N3-MeA and d5-N3-EtA were used as isotope internal standard. The DNA samples were injected with autosampler. The injected volume was 3 μL and analysis time was 13 min. The sample separation was carried out on hydrophilic interaction chromatograph ( Waters XBridge HILIC ) with 10 mmol/L ammonium formate-acetonitrile (5:92, V/V, pH=4. 0) as mobile phase. The flow rate was set at 250 μL/min. Mass spectrometry was performed by electrospray ionization ( ESI ) with multi-reactions monitoring ( MRM ) . The optimized operation conditions of MS were as follows: nebulizer gas 369 Pa; curtain gas 185 Pa, turbo ionspray temperature 400 ℃, ionspray voltage 5500 V, dwell time 40 ms. The limits of detection were 0. 043 and 0. 007 μg/L for N3-MeA and N3-EtA, respectively. The recoveries were between 87. 8% and 103. 0%for N3-MeA and N3-EtA. This method was successfully applied to the determination of N3-MeA and N3-EtA in calf thymus DNA by cigarette smoke condensate ( CSC) exposure. This method is appropriate for routine analysis and accurate quantification of N3-MeA and N3-EtA by CSC exposure.