Subsequently, the optimized approach was transferred to 96-well plate formats and rigorously tested for analytical performance and throughput capacity. Finally, the potential for NTA was explored in standard reference materials (SRMs) for urine and plasma. The results indicate the feasibility of the approach for scaling and large-scale exposomewide association studies. Experimental Section Biological samples. Human pooled plasma was purchased from Innovative Research (IPLAWBLIH-31982) (Novi, MI, USA). Pooled urine was obtained from a female volunteer who abstained from consuming food and beverages stored in plastic containers, foods rich in phytoestrogens, and cosmetics containing parabens for two days before sample collection . Standard reference materials, including urine (SRM 3672) and plasma (SRM 1950) were obtained from the National Institute of Standards and Technology (NIST) (Gaithersburg, USA) for NTA. Detailed information on standards, including selection and spiking levels can be found in Tables . Method development and optimization. The sample pretreatment workflow was developed and optimized in several steps. Initially, two universal SPE sorbents, commercial hydrophilic-lipophilic balanced sorbent (HLB) from Waters and in-house prepared mixedmode SPE were compared. The mixed sorbent was made of two SPE sorbents, primary second amine (PSA) and C18 (PSA+C18), by mixing them in equal weights before packing. Details of the preparation are described in the Supplementary Information (SI). The SPE cartridges packed with the suitable sorbent (HLB or PSA+C18) were then used to optimize the processes of sample loading, washing, and elution. The trapping efficiency, or analyte loss, during loading, and washing was investigated by analyzing the effluents collected during the loading and washing steps. Analyte loss was estimated as the fraction of non-retained analyte by the SPE sorbent. An analyte loss of 0% indicates optimal analyte trapping during the SPE process, and a loss of 100% indicates insufficient interaction between the sorbent and the analyte. Additionally, extraction recovery (RE) was used to assess extraction efficiency, i.e., the combined efficiency of analyte trapping and analyte elution from the SPE. A recovery of 100% indicates perfect trapping and elution during the entire process, a RE of 0% either indicates insufficient analyte trapping, or insufficient analyte elution, whereas RE >100% can be used as an indication of contaminations introduced during the sample preparation process. Additionally, matrix effects in terms of signal suppression and signal enhancement (SSE) were monitored to assess the capability of the individual sample preparation workflows to remove interfering matrix components. Thus, an SSE of 100% means no influence of the sample matrix on the detector response and analyte quantification. Peak areas of pre-extraction spiked and post-extraction spiked samples with single concentrations were used to calculate the RE and SSE values. Performance was evaluated and classified into three categories: "good," "acceptable," and "poor". "Good" performance was defined as analyte loss <10% during loading and washing, and RE or signal SSE between 60%-140%. "Acceptable" performance was defined by an analyte loss between 10-20% during loading and washing and RE or SSE within 20%-60% or 140%-180%. Any values outside these ranges were classified as "poor" method performance.