Center for Translational Medicine, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, PR China.
Shanghai Key Laboratory of Diabetes Mellitus, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2022 Nov 15;1211:123485. doi: 10.1016/j.jchromb.2022.123485. Epub 2022 Oct 10.
Abnormal salivary amino acid (AA) levels may indicate dysfunction of the body. Being noninvasive, sampling easily and cost-effective of saliva, a rapid, precise and simple analysis method has become very important for quantitative salivary AA profiles. After one-step to precipitate protein, the resultant extraction was derived with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) within 10 min. Quantitation of AA profile was achieved within 6 min in a single run by ultra-high performance liquid chromatography coupled with a single quadrupole mass spectrometry (UHPLC-QDA detector). The method was validated with acceptable accuracy ranging from 80.33 % to 122.31 %, appropriate linearity with the coefficient (R) more than 0.991, good intra- and inter-day precision, repeatability and stability (RSD < 15 %). The recoveries at three different spiked concentrations ranged over 79.18 %-125.36 % while the matrix effect was from -19.86 % to 11.95 %. This simple, rapid and robust method was successfully applied to quantify human salivary 30 amino acids, in which the levels of taurine, γ-aminobutyric acid, methionine and tryptophan in healthy people were close to the LOQs. Besides, the levels of histidine and cystine were not able to be measured due to their relatively high LOQs, which were considered as the limitations of this developed method.
唾液中氨基酸（AA）水平异常可能表明身体功能障碍。唾液具有非侵入性、采样容易且成本效益高的特点，因此建立一种快速、精确和简单的分析方法来定量分析唾液 AA 谱非常重要。在一步沉淀蛋白后，用 6-氨基喹啉基-N-羟基琥珀酰亚胺基甲酸酯（AQC）在 10 分钟内提取。通过超高效液相色谱-单四极杆质谱联用（UHPLC-QDA 检测器），在单次运行中可在 6 分钟内实现 AA 谱的定量分析。该方法的验证结果表明，准确度在 80.33%至 122.31%之间，适当的线性相关系数（R）大于 0.991，具有良好的日内和日间精密度、重复性和稳定性（RSD<15%）。在三个不同浓度的加标回收率在 79.18%至 125.36%之间，而基质效应在-19.86%至 11.95%之间。该方法简单、快速、稳健，成功应用于定量分析人唾液中的 30 种氨基酸，其中健康人群中牛磺酸、γ-氨基丁酸、蛋氨酸和色氨酸的水平接近定量下限。此外，由于其较高的定量下限，组氨酸和胱氨酸的水平无法测量，这被认为是该方法的局限性。
