目的 采用頂空進(jìn)樣氣相色譜三重四級(jí)桿質(zhì)譜聯(lián)用（HS-GC-MS/MS）法測(cè)定富馬酸丙酚替諾福韋中微量的基因毒性雜質(zhì)N-亞硝基二甲胺（NDMA）。方法 采用三重四極桿GC-MS/MS，Agilent VF-WAX ms（30 m×0.25 mm，1 μm）色譜柱，載氣：氦氣；恒流模式1.0 mL·min^-1；程序升溫，進(jìn)樣口溫度230℃，頂空溫度130℃；質(zhì)譜采用電子轟擊電離源（EI），電離能量為70 eV，離子源溫度230℃，多反應(yīng)監(jiān)測(cè)（MRM）模式進(jìn)行檢測(cè)，溶劑為N-甲基吡咯烷酮（NMP）。進(jìn)行專屬性、系統(tǒng)適用性、檢測(cè)限與定量限、線性與范圍、準(zhǔn)確度、精密度、溶液穩(wěn)定性、耐用性考察。結(jié)果 NDMA與相鄰色譜峰之間分離效果良好；NDMA在7.0～105.0 ng·mL^-1線性關(guān)系良好，檢測(cè)限為3.5 ng·mL^-1，定量限為7.0 ng·mL^-1；NDMA低、中、高質(zhì)量濃度（56、70、84 ng·mL^-1）回收率為95.6%～109.3%，RSD為4.0%（n=9）；重復(fù)性試驗(yàn)NDMA質(zhì)量濃度RSD為6.5%，中間精密度RSD為6.1%；對(duì)照品溶液室溫放置24 h穩(wěn)定，供試品溶液室溫放置90 h內(nèi)溶液穩(wěn)定；保持其他條件不變，分別改變進(jìn)樣口溫度（230、225、235℃）、離子源溫度（230、225、235℃）、載氣體積流量（1.0、0.9、1.1 mL·min^-1）、頂空溫度（130、128、132℃），方法耐用性良好。結(jié)論 所建立的方法準(zhǔn)確度好、靈敏度高、簡(jiǎn)便可靠，對(duì)儀器污染小，可用于富馬酸丙酚替諾福韋中NDMA的質(zhì)量控制。

Objective To establish a HS-GC-MS/MS method for the microdetermination of N-nitrosodimethylamine (NDMA) in tenofovir alafenamide fumarate. Methods GC-MS/MS was used, and the separation was performed on a Agilent VF-WAX ms column (30 m×0.25 mm, 1 μm) with temperature programmed. Carrier gas:helium, Constant current mode of 1.0 mL·min^-1. The inlet temperature was 230℃ and flow rate was 1.0 mL·min^-1, headspace temperature was 130℃. Mass spectrometry was used with EI source, the voltage was 70 eV, ion source temperature was 230℃, and detected with selected ion monitoring (MRM) data acquisition mode. The samples were dissolved in NMP. Specificity, system applicability, limit of detection and limit of quantification, linearity and range, accuracy, precision, solution stability and durability were investigated. Results The separation between NDMA and adjacent autobiographic peaks was effective, NDMA was well correlated with the linearity range between 7.0-105.0 ng·mL^-1. The limit of detection of NDMA was 3.5 ng·mL^-1 and the limit of quantitation was 7.0 ng·mL^-1. The average recoveries of NDMA were between 95.6% and 109.3% at low, middle and high concentration, with the RSD (n=9) of 4.0%. RSD of reproducibility test NDMA was 6.5%, and RSD of intermediate precision was 6.1%. The control solution was stable at room temperature for 24 h, and the test solution was stable at room temperature for 90 h. The inlet temperature (230, 225, 235℃), ion source temperature (230, 225, 235℃), carrier gas volume flow rate (1.0, 0.9, 1.1 mL·min^-1), and headspace temperature (130, 128, 132℃) were changed with other conditions unchanged. The durability of the method was good. Conclusion The method is accurate, highly sensitivite, simple and reliable with less pollution to the instrument, and it can be used for the quality control of the genotoxic impurity NDMA in tenofovir alafenamide fumarate.

R991

中國(guó)醫(yī)學(xué)科學(xué)院醫(yī)學(xué)與健康科技創(chuàng)新工程項(xiàng)目資助（2019-I2M-5-020）；天津市科技計(jì)劃項(xiàng)目（18YFZCSY00090）