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-C₁₈ column (3.0 × 150 mm, 5 μm) 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.

中文翻译：

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基于靶向UHPLC-MS / MS方法同时测定大肠癌患者肿瘤组织中34种氨基酸。

建立了靶向超高效液相色谱-串联质谱（UHPLC-MS / MS）方法，并验证了该方法可同时测定大肠癌（CRC）患者组织样品中的34个氨基酸。色谱分离在Agilent ZORBAX SB-C达到₁₈柱（3.0×150毫米，5  μm）具有二元梯度洗脱系统（A，0.02％的七氟丁酸和0.2％的甲酸水溶液，v / v; B，甲醇）。运行时间为10分钟。选择多反应监测模式，其中电喷雾电离源以正电离模式运行以获取数据。所有分析物在其相应的校准范围内的线性相关系数均> 0.99。基于组织匀浆和蛋白质沉淀，使用100 mg等分样品对样品进行预处理。34种氨基酸和3种内标物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％之内。该方法用于同时定量来自94名CRC患者的组织样本中的34种氨基酸。