How to further improve the throughput of capillary electrophoresis (CE) is a fascinating question. Herein an idea to substantially increase the throughput of CE has been proposed together with theory and experimental demonstration. The key is to introduce samples for CE, one after another, by a short suspension of voltage application, which was hence termed separation-interrupted sequential injections (Sisi). The idea was demonstrated to be feasible on a laboratory-built CE instrument coupled with tandem C4D (contactless capacitively-coupled conductivity) detectors. At least 50 injections of a testing sample (mixture of NH4+, K+, Ca2+, Na+ and Mg2+) were successfully separated in only a single run. The separation took 145 min in total, equivalent to 2.9 min per analysis which is only 21% of that of normal CE. Quantification of the separated ions was performed, with a limit of detection of 1.1-2.6 μM, a limit of quantification of 3.2-8.9 μM, and a linear range up to 1000 μM (R2 > 0.99). The recovery was between 88% and 112% measured by spiking standards into samples at low, middle and high levels. The real applicability of Sisi-CE was evaluated by direct injection and analysis of 45 mineral water samples also in a single run. Its clinical application potential was demonstrated by high throughput assay of the calcium and zinc gluconate oral solution formula, and the blood potassium of hyperkalemia and hypokalemia from patients with renal failure disease. This method can be extended to other applications such as omics studies through the use of more suitable detectors. The theory proposed may also be applicable to other high throughput methods.
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http://dx.doi.org/10.1039/d1ay00223f | DOI Listing |
J Am Soc Mass Spectrom
January 2025
Department of Chemistry, Washington State University, Pullman, Washington 99164, United States.
Phased structures for lossless ion manipulation offer significant improvements over the scanning second gate method for coupling with ion trap mass analyzers. With an experimental run time of under 1 min for select conditions and an average run time of less than 4 min, this approach significantly reduces experimental time while enhancing the temporal duty cycle. The outlined SLIM system connects to an ion trap mass analyzer via a PCB stacked ring ion guide, which replaces the commercial ion optics and capillary inlet.
View Article and Find Full Text PDFAnal Chem
December 2024
Department of Chemistry and Chemical Biology, McMaster University, Hamilton, Ontario L8S 4M1, Canada.
Mass spectrometry (MS)-based metabolomics often rely on separation techniques when analyzing complex biological specimens to improve method resolution, metabolome coverage, quantitative performance, and/or unknown identification. However, low sample throughput and complicated data preprocessing procedures remain major barriers to affordable metabolomic studies that are scalable to large populations. Herein, we introduce PeakMeister as a new software tool in the R statistical environment to enable standardized processing of serum metabolomic data acquired by multisegment injection-capillary electrophoresis-mass spectrometry (MSI-CE-MS), a high-throughput separation platform (<4 min/sample) which takes advantage of a serial injection format of 13 samples within a single analytical run.
View Article and Find Full Text PDFBiotechniques
December 2024
Department of Human and Animal Cell Lines, Leibniz-Institute DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany.
The strict suppression of telomerase activity (TA) in terminally differentiated human cells causes a shortening of the chromosome ends after each cell division. This tumor suppression surveillance mechanism is associated with a limited number of cell divisions known as Hayflick limit. Here we present an optimized protocol for measuring TA that combines a fluorescently labeled bait primer and polymerase chain reaction (PCR) amplification with analytical capillary electrophoresis (CE) to achieve a detection limit of one telomerase-positive cell per ten thousand negative cells.
View Article and Find Full Text PDFBrief Bioinform
November 2024
Beijing Institute of Genomics, Chinese Academy of Sciences and China National Center for Bioinformation, Beijing 100101, China.
Short tandem repeats (STRs) represent one of the most polymorphic variations in the human genome, finding extensive applications in forensics, population genetics and medical genetics. In contrast to the traditional capillary electrophoresis (CE) method, genotyping STRs using massive parallel sequencing technology offers enhanced sensitivity and accuracy. However, current methods are mainly designed for target sequencing with higher coverage for a specific STR locus, thereby constraining the utility of STRs in low- and medium-coverage whole genome sequencing (WGS) data.
View Article and Find Full Text PDFJ Chromatogr A
January 2025
School of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, China; Key Laboratory of Advanced Mass spectrometry and Molecular Analysis of Zhejiang Province, Ningbo University, Ningbo, 315211, China.
B-complex vitamins are essential micronutrients that maintaining health, and provide (individually/simultaneously) many important biological actions in organism. Therefore, sensitive, reliable analytical method to determine B-complex vitamins simultaneously in actual samples is significant. Conventional analytical methods for vitamins analysis are usually labor-intensive, time-consuming and mostly do not allow the simultaneous determination.
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