Dorrani Masoumeh, Zhao Jifang, Bekhti Nihel, Trimigno Alessia, Min Sangil, Ha Jongwon, Han Ahram, O'Day Elizabeth, Kamphorst Jurre J
Olaris, Inc., 175 Crossing Boulevard Suite 410, Framingham, MA 01702, USA.
Seoul National University Hospital, 101, Daehak-ro, Jongno-gu, Seoul 03080, Republic of Korea.
Metabolites. 2024 May 11;14(5):280. doi: 10.3390/metabo14050280.
Mass spectrometry (MS)-based clinical metabolomics is very promising for the discovery of new biomarkers and diagnostics. However, poor data accuracy and reproducibility limit its true potential, especially when performing data analysis across multiple sample sets. While high-resolution mass spectrometry has gained considerable popularity for discovery metabolomics, triple quadrupole (QqQ) instruments offer several benefits for the measurement of known metabolites in clinical samples. These benefits include high sensitivity and a wide dynamic range. Here, we present the Olaris Global Panel (OGP), a HILIC LC-QqQ MS method for the comprehensive analysis of ~250 metabolites from all major metabolic pathways in clinical samples. For the development of this method, multiple HILIC columns and mobile phase conditions were compared, the robustness of the leading LC method assessed, and MS acquisition settings optimized for optimal data quality. Next, the effect of U-C metabolite yeast extract spike-ins was assessed based on data accuracy and precision. The use of these U-C-metabolites as internal standards improved the goodness of fit to a linear calibration curve from r < 0.75 for raw data to >0.90 for most metabolites across the entire clinical concentration range of urine samples. Median within-batch CVs for all metabolite ratios to internal standards were consistently lower than 7% and less than 10% across batches that were acquired over a six-month period. Finally, the robustness of the OGP method, and its ability to identify biomarkers, was confirmed using a large sample set.
基于质谱（MS）的临床代谢组学在发现新的生物标志物和诊断方法方面极具前景。然而，数据准确性和可重复性较差限制了其真正潜力，尤其是在对多个样本集进行数据分析时。虽然高分辨率质谱在发现代谢组学中已颇受欢迎，但三重四极杆（QqQ）仪器在临床样本中已知代谢物的测量方面具有诸多优势。这些优势包括高灵敏度和宽动态范围。在此，我们介绍了Olaris全球检测板（OGP），这是一种亲水作用色谱（HILIC）液相色谱 - 三重四极杆质谱方法，用于全面分析临床样本中所有主要代谢途径的约250种代谢物。为开发此方法，比较了多种HILIC色谱柱和流动相条件，评估了领先液相色谱方法的稳健性，并优化了质谱采集设置以获得最佳数据质量。接下来，基于数据准确性和精密度评估了U - C代谢物酵母提取物加标的效果。使用这些U - C代谢物作为内标，改善了线性校准曲线的拟合优度，从原始数据的r < 0.75提高到尿液样本整个临床浓度范围内大多数代谢物的r > 0.90。所有代谢物与内标物比率的批内CV中位数始终低于7%，在六个月期间采集的批次间CV小于10%。最后，使用大量样本集证实了OGP方法的稳健性及其识别生物标志物的能力。
