Yang Yang, Zhang Feng, Gao Shouhong, Wang Zhipeng, Li Mingming, Wei Hua, Zhong Renqian, Chen Wansheng
Department of Pharmacy, Changzheng Hospital, The Second Military Medical University of CPLA, Shanghai 200003, China.
Department of Pharmacy, The 71st Group Army Hospital of CPLA Army, The Affiliated Huaihai Hospital of Xuzhou Medical University, Xuzhou 221004, China.
J Anal Methods Chem. 2020 Aug 1;2020:4641709. doi: 10.1155/2020/4641709. eCollection 2020.
A targeted ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established and validated for the simultaneous determination of 34 amino acids in tissue samples from colorectal cancer (CRC) patients. The chromatographic separation was achieved on an Agilent ZORBAX SB-C column (3.0 × 150 mm, 5 m) with a binary gradient elution system (A, 0.02% heptafluorobutyric acid and 0.2% formic acid in water, v/v; B, methanol). The run time was 10 min. The multiple reaction monitoring mode was chosen with an electrospray ionization source operating in the positive ionization mode for data acquisition. The linear correlation coefficients were >0.99 for all the analytes in their corresponding calibration ranges. The sample was pretreated based on tissue homogenate and protein precipitation with a 100 mg aliquot sample. The average recovery and matrix effect for 34 amino acids and 3 internal standards were 39.00%∼146.95% and 49.45%∼173.63%, respectively. The intra- and interday accuracy for all the analytes ranged from -13.52% to 14.21% (RSD ≤8.57%) and from -14.52% to 12.59% (RSD ≤10.31%), respectively. Deviations of stability under different conditions were within ±15% for all the analytes. This method was applied to simultaneous quantification of 34 amino acids in tissue samples from 94 CRC patients.
建立并验证了一种靶向超高效液相色谱-串联质谱（UHPLC-MS/MS）方法，用于同时测定结直肠癌（CRC）患者组织样本中的34种氨基酸。采用安捷伦ZORBAX SB-C柱（3.0×150 mm，5μm），二元梯度洗脱系统（A，0.02%七氟丁酸和0.2%甲酸的水溶液，v/v；B，甲醇）进行色谱分离。运行时间为10分钟。选择多反应监测模式，采用电喷雾电离源在正离子模式下运行以采集数据。所有分析物在其相应校准范围内的线性相关系数均>0.99。基于组织匀浆和用100 mg等分试样进行蛋白质沉淀对样品进行预处理。34种氨基酸和3种内标的平均回收率和基质效应分别为39.00%∼146.95%和49.45%∼173.63%。所有分析物的日内和日间准确度分别为-13.52%至14.21%（RSD≤8.57%）和-14.52%至12.59%（RSD≤10.31%）。所有分析物在不同条件下的稳定性偏差均在±15%以内。该方法应用于对94例CRC患者组织样本中的34种氨基酸进行同时定量分析。
