Wang Qihui, Gao Pan, Cheng Fei, Wang Xiaoyi, Duan Yixiang
Research Center of Analytical Instrumentation, Analytical & Testing Centre, Sichuan University, Chengdu 610064, PR China.
Department of Head and Neck Oncology, West China College of Stomatology, Sichuan University, Chengdu 610064, PR China.
Talanta. 2014 Feb;119:299-305. doi: 10.1016/j.talanta.2013.11.008. Epub 2013 Nov 21.
This study aimed to set-up an ultra performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS) method for the determination of salivary L-phenylalanine and L-leucine for early diagnosis of oral squamous cell carcinoma (OSCC). In addition, the diagnostic accuracy for both biomarkers was established by using receiver operating characteristic (ROC) analysis. Mean recoveries of l-phenylalanine and L-leucine ranged from 88.9 to 108.6% were obtained. Intra- and inter-day precision for both amino acids was less than 7%, with acceptable accuracy. Linear regression coefficients of both biomarkers were greater than 0.99. The diagnostic accuracy for both biomarkers was established by analyzing 60 samples from apparently healthy individuals and 30 samples from OSCC patients. Both potential biomarkers demonstrated significant differences in concentrations in distinguishing OSCC from control (P<0.05). As a single biomarker, L-leucine might have better predictive power in OSCC with T1-2 (early stage of OSCC including stage I and II), and L-phenylalanine might be used for screening and diagnosis of OSCC with T3-4 (advanced stage of OSCC including stage III and IV). The combination of L-phenylalanine and L-leucine will improve the sensitivity (92.3%) and specificity (91.7%) for early diagnosis of OSCC. The possibility of salivary metabolite biomarkers for OSCC diagnosis is successfully demonstrated in this study. This developed method shows advantages with non-invasive, simple, reliable, and also provides lower detection limits and excellent precision and accuracy. These non-invasive salivary biomarkers may lead to a simple clinical tool for the early diagnosis of OSCC.
本研究旨在建立一种超高效液相色谱 - 电喷雾电离 - 质谱（UPLC - ESI - MS）方法，用于测定唾液中的L - 苯丙氨酸和L - 亮氨酸，以早期诊断口腔鳞状细胞癌（OSCC）。此外，通过使用受试者工作特征（ROC）分析确定了这两种生物标志物的诊断准确性。L - 苯丙氨酸和L - 亮氨酸的平均回收率在88.9%至108.6%之间。两种氨基酸的日内和日间精密度均小于7%，准确性可接受。两种生物标志物的线性回归系数均大于0.99。通过分析60份来自明显健康个体的样本和30份来自OSCC患者的样本，确定了这两种生物标志物的诊断准确性。两种潜在生物标志物在区分OSCC与对照组时，其浓度显示出显著差异（P<0.05）。作为单一生物标志物，L - 亮氨酸在T1 - 2期（OSCC早期，包括I期和II期）的OSCC中可能具有更好的预测能力，而L - 苯丙氨酸可用于T3 - 4期（OSCC晚期，包括III期和IV期）的OSCC的筛查和诊断。L - 苯丙氨酸和L - 亮氨酸的联合使用将提高OSCC早期诊断的灵敏度（92.3%）和特异性（91.7%）。本研究成功证明了唾液代谢物生物标志物用于OSCC诊断的可能性。所开发的方法具有无创、简单、可靠的优点，并且还具有较低的检测限以及出色的精密度和准确性。这些非侵入性唾液生物标志物可能会带来一种用于OSCC早期诊断的简单临床工具。
