目的 建立HPLC-MS/MS法測定NB4細(xì)胞培養(yǎng)液中全反式維甲酸的濃度。方法 采用ACQUITY UPLCTM BEH C₁₈（50 mm×2.1 mm，1.7 μm）色譜柱，以布洛芬為內(nèi)標(biāo)，流動相為乙腈-水（含0.1%乙酸），體積流量為0.2 mL/min，梯度洗脫方式分離全反式維甲酸，同時采用ESI源負(fù)離子檢測方式，定量分析時的離子反應(yīng)分別為m/z 299.2→m/z 255.2（全反式維甲酸）和m/z 205.4→m/z 161.3（內(nèi)標(biāo)布洛芬）。結(jié)果 全反式維甲酸在2.55~255 ng/mL線性關(guān)系良好，定量限為2.55 ng/mL，日內(nèi)精密度RSD ≤ 10.3%，日間精密度RSD ≤ 12.9%。結(jié)論 建立的HPLC-MS/MS法可用于測定NB4細(xì)胞含維甲酸的培養(yǎng)液中全反式維甲酸的濃度，以及在加入維甲酸代謝阻斷劑時全反式維甲酸濃度的變化。

Objective To develop an HPLC-MS/MS method for determination of all-trans retinoic acid (ATRA) in NB4 cell medium. Methods Following liquid-liquid extraction with anhydrous ether, the analyte (ATRA) and internal standard (ibuprofen) were separated from medium using an gradient elution with acetonitrile and aqueous phase, containing 0.1% acetic acid, on an ACQUITY UPLCTM BEH C₁₈ (50 mm×2.1 mm, 1.7 μm) column at the flow rate of 0.2 mL/min. The detection was carried out by means of electrospray ionization mass spectrometry in negative ion mode. Multiple reaction monitoring (MRM) mode with the transitions of m/z 299.2→m/z 255.2 and m/z 205.4→m/z 161.3 was used to quantify ATRA and internal standard, respectively. Results The calibration curves were linear in the range of 2.55-255 ng/mL. The limit of quantification was 2.55 ng/mL. The intra-and inter-day precisions (RSD) were less than 10.3% and 12.9%. Conclusion It can be used to determinate all-trans retinoic acid (ATRA)in NB4 cell medium, and the variety of ATRA with metabolic inhibitor of ATRA.