UCLA Living Biofoundry provides NEW resources for chemical separation and analysis

High-performance liquid chromatography meets mass spectrometry and fragmentation analysis at UCLA CNSI user facility

Numerous lines of investigation in the life sciences and chemistry depend on the ability to separate, measure, identify and quantify the composite molecules in complex mixtures. These capabilities are essential for analyzing metabolites and tracing series of biochemical or inorganic chemical reactions, to name a few applications.

This package of resources is available to UCLA researchers, other academic organizations and private companies at the California NanoSystems Institute at UCLA’s Living Biofoundry. Established with National Science Foundation funding through the BioPACIFIC Materials Innovation Platform, the high-throughput facility provides open access to a Vanquish Flex ultrahigh performance liquid chromatography system with a TSQ Altis Plus triple quadrupole mass spectrometer.

This instrumentation offers chromatographic separation, mass spectrometry and fragmentation analysis with unprecedented sensitivity, even in complex matrices, enabling measurements close to the physical limit — and thus the quantification of compounds at very low concentrations. In addition to affording extraordinary reliability and reproducibility, the system can reveal hidden details that may be crucial to users’ findings.

“Analyzing a single chromatography peak and a single precursor mass by fragmentation, we found that an improved chromatography method allowed us to separate the peak into two,” said Michael Lake, technical director of the Living Biofoundry. “One fragment was uniquely associated with the first chromatography peak, but three other fragments were identified uniquely with the second peak. This allowed us to conclude that we had two chiral isomers, something that would have been overlooked without fragmentation analysis.”

The system also offers high selectivity and superior acquisition speed — monitoring of 600 selected reactions per second and screening of thousands of compounds per method. The capacity of loading 400 samples and remote access to monitor the run, in combination with automated sample preparation, will open the possibility for new types of analyses and inquiry.

A concrete example in synthetic biology is using selected reaction monitoring to look at precursors, intermediates and even divergent final products from a biosynthetic pathway, where fragment analysis can ensure that even highly similar intermediates and products can be fully separated and accurately quantitated. Another example would be peptide-based drugs or nanobodies, which can be precisely quantitated, even from complex solutions such as plasma.

The Vanquish chromatographer has a 1024-pixel diode array detector with a broad, customizable spectral range for peak detection. For ionization, the system can switch between two modes: heated electrospray ionization for analyzing targeted metabolites, chemicals and natural products; and atmospheric pressure ionization for examining peptides, proteins and polymers.

With intelligent sample precompression, users benefit from increased retention-time precision. Compact and easy to operate, the system features Chromeleon 7 software that integrates control and analysis of chromatography and mass spectrometry measurements. Its simple and intuitive user interface has options for automatic startup and calibration as well as real-time troubleshooting.

Those interested in gaining general information, project consultation and proof-of-concept, or training and assistance should reach out to technical director Michael Lake at biopacificmip@cnsi.ucla.edu.

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