Strategy For High-throughput Analysis Research Articles

Direct analysis of urinary 1-hydroxypyrene using extractive electrospray ionization ion trap tandem mass spectrometry

Fast detection (0.5 min) of 1-hydroxypyrene (1-OHP) in urine and hydrolyzed urine without pre-treatment has been successfully achieved by using extractive electrospray ionization ion trap tandem mass spectrometry (EESI-MS/MS) under optimized EESI and MS/MS conditions. Experimental results indicated that for MS/MS analysis the operating parameter of activation Q (AQ) was critical for 1-OHP fragmentation in collision induced dissociation (CID) experiments, and in the EESI process the primary ESI solvent was a key factor for extractive ionization of urinary 1-OHP. The limit of detection (LOD) and limit of quantification (LOQ) were 0.75 and 2.25 μM for both urine and hydrolyzed urine samples. A five-point working curve ranging from 2.29 to 22.91 μM for 1-OHP in urine or hydrolyzed urine was obtained (R2 = 0.9941 for urine and R2 = 0.9983 for hydrolyzed urine), and the relative standard deviations (RSD, n = 6) were 2.6–9.7% and 1.5–6.4%, respectively. The developed EESI-MS/MS method was validated by detecting 1-OHP in both urine and hydrolyzed urine samples. Recoveries were determined to >50% and 0.5 min was taken for each measurement, indicating that the proposed method is a promising strategy for high throughput analysis of urinary 1-OHP required for health risk assessment of exposure to polycyclic aromatic hydrocarbons (PAHs).

Fluorescence-based tools to improve biopharmaceutical process development

Background Process optimization and control are essential to match biopharmaceutical market demands. Early-steps in the development of new processes require screening hundreds of transfected clones in order to select those with the best expression characteristics, and identifying optimal environmental conditions for these clones to grow and express the protein of interest. While some culture systems include in situ analysis of cell density, most often by optical density or capacitance measurements, the majority of them still rely on off-line, laborious and time-consuming protocols for recombinant protein analysis. Spectroscopic methods have been proposed in literature to support bioprocesses development, because they are non-invasive and provide data on multiple components present in the culture bulks, thereby increasing information on process performance [1]. Here, we developed a fluorescence-based method for real-time monitoring of viable cells and antibody titers in bioreactor cultures, and extended this strategy for high throughput analysis of cellular productivity in 96well plates.

Chromatography in Industry

This review focuses on the chromatography research that has been carried out within industry or in close cooperation with industry and that has been reported in the scientific literature between 2006 and mid-2008. Companies in the health care sector, such as pharmaceutical and biotechnology companies, are the largest contributors. Industrial research seems to take place in an open environment in cooperation with academia, peer companies, and institutions. Industry appears ready to embrace new technologies as they emerge, but they focus strongly on making chromatography work robustly, reliably, rapidly, and automatically. "Hyphenated" systems that incorporate on-line sample-preparation techniques and mass-spectrometric detection are the rule rather than the exception. Various multidimensional separation methods are finding numerous applications. Strategies aimed at speeding up the development of new chromatographic methods remain the focus of attention. Also, there is a clear trend toward exploring chromatographic methods for parallel processing along with other strategies for high-throughput analysis.