Biomarker mass spectrometry assays are in high demand, and analysis of pro-gastrin releasing peptide (ProGRP) as a small cell lung cancer marker has been recently investigated by mass spectrometry after immunoextraction. In this article, we introduce an assay based on molecularly imprinted polymers (MIPs) targeting the proteotypic peptide of ProGRP as a possible alternative to current immuno-based assay. The MIPs were prepared by surface-initiated reversible addition-fragmentation chain transfer polymerization and were introduced as sorbents for the cleanup and enrichment of a ProGRP signature peptide from tryptically treated serum samples. The use of an appropriate solid-phase extraction protocol allowed specific extraction of the target peptide while depleting other peptides that arose from the sample digestion, hence resulting in reduced background. The selective extraction of a ProGRP signature peptide, after digestion of serum samples, translates into a time- and cost-effective method suited for bottom-up analysis wherever targeted peptide extraction from complex matrices is required.
Affinity-based solid-phase extraction (SPE) is an attractive low-cost sample preparation strategy for biomarker analysis. Molecularly imprinted polymers (MIPs) as affinity sorbents offer unique opportunities for affinity SPE, due to their low manufacturing cost and high robustness. A limitation is the prediction of their affinity; therefore, screening of analyte recovery and specificity within a large range of SPE conditions is important in order to ensure high-sensitivity detection and assay reproducibility. Here, a µ-SPE method for screening of the MIP-SPE materials using a commercial 384-well filter plate is presented. The method allows for rapid and automated screening using 10-30 µL of packed SPE sorbent per well and sample volumes in the range of 10-70 µL. This enables screening of many different SPE sorbents while simultaneously identifying optimal SPE conditions. In addition, the 384-well format also facilitates detection with a multitude of analytical platforms. Performance of the µ-MIP-SPE method was investigated using a series of MIPs designed to capture pro-gastrin-releasing peptide (ProGRP). Fractions coming from sample load, cartridge wash, and elution were collected and analyzed using mass spectrometry (MS). The top-performing MIPs were identified, together with proper SPE conditions.
A mathematical model that expresses Total trihalomethane (TTHM) concentration in terms of initial chlorine concentration, total organic carbon, bromide ion concentration, contact time, and pH is developed for Zai water treatment plant which supplies water to Jabal Amman. The developed mathematical model is for constant temperature of 20 • C. To adjust model calculated TTHM concentrations for temperatures other than 20 • C, another mathematical model that expresses TTHM growth rate as function of temperature is also developed. To test the ability of the two developed models in predicting TTHM concentrations throughout water supplies, a sampling program that aimed at measuring TTHM concentrations in addition to the predictors in the two developed mathematical models namely; chlorine concentration, bromide ion concentration, total organic carbon, temperature and pH throughout Jabal Amman water supply was conducted. The two developed mathematical models and WaterCad, which was used to determine water age, were used to predict TTHM concentrations throughout Jabal Amman water supply. Predicted TTHM concentrations were compared to actual TTHM concentrations measured during the sampling program. Results showed that there is good agreement between measured and, calculated TTHM concentrations, which means that the method presented in this paper, can be used to obtain good estimates of TTHM concentrations throughout networks.
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