Microfluidic capillary zone electrophoresis mass spectrometry analysis of alkaloids in Lobelia cardinalis transgenic and mutant plant cell cultures

Abstract

Application of a microfluidic CE* device for CZE‐MS allows for fast, rapid, and in‐depth analysis of large sample sets. This microfluidic CZE‐MS device, the 908 Devices ZipChip, involves minimal sample preparation and is ideal for small cation analytes, such as alkaloids. Here, we evaluated the microfluidic device for the analysis of alkaloids from Lobelia cardinalis hairy root cultures. Extracts from wild‐type, transgenic, and selected mutant plant cultures were analyzed and data batch processed using the mass spectral processing software MZmine2 and the statistical software Prism 8. In total 139 features were detected as baseline resolved peaks via the MZmine2 software optimized for the electrophoretic separations. Statistically significant differences in the relative abundance of the primary alkaloid lobinaline (C27H34N2), along with several putative “lobinaline‐like” molecules were observed utilizing this approach. Additionally, a method for performing both targeted and untargeted MS/MS experiments using the microfluidic device was developed and evaluated. Coupling data‐processing software with CZE‐MS data acquisition has enabled comprehensive metabolomic profiles from plant cell cultures to be constructed within a single working day.

Bladder cancer hunting: A microfluidic paper‐based analytical device

“…Relying on the appearance of the wax-printing printer, the fabrication procedure of μPAD can be greatly simplified. The application of colloidal gold-labeling technology makes the test results visible to the naked eye without any equipment, and only about 3-10 min are needed to complete the experiment, which is better than that of any other methods such as clinical cystoscopy [37], cytology [2,38], detection of urinary extracellular vesicles [39], or CTCs [40,41] with the microfluidic chip and any biomarker [42]. More importantly, although simple, the chip can simultaneously find two specific biomarkers, and double the accuracy of bladder cancer detection results, making the detection efficiency of the μPAD better than the existing detection methods.…”

“…Bladder cancer is a urinary system tumor disease, which is common in men with slow healing and high recurrence rate [1]. Cystoscopy is the best standard method for detecting and monitoring the recurrence of bladder cancer in clinics [2]. However, this method is painful and less sensitive to early diagnosis.…”

Pharmacokinetic and metabolic analysis of an Alzheimer's disease therapeutic in rat serum via microfluidic CZE–MS

“…The microfluidic CZE–MS device utilized in this work, commercially known as the 908 Devices ZipChip, consists of a small, etched glass chip, enclosed in a polymer casing, that can perform fast and efficient electrophoretic separations at lower flow rates (150 nl/min) than most nano/microflow LC systems (Kelley et al, 2019; Mellors et al, 2008; 908 Devices, 2016). The device mounts directly to the inlet of many modern mass spectrometers in a specially designed housing unit that supplies both the pressure and voltage required for its function.…”

“…Additionally, the device couples both CZE separations and ESI into one system, eliminating the need to integrate the separation capillary into the instrument’s ionization source. A variety of applications for this separation device have been reported in the literature, including intact antibody analysis, protein analysis and comprehensive metabolomic profiling (Beri et al, 2018; Chen et al, 2018; Fussl et al, 2020; Kelley et al, 2019; Khatri et al, 2017; Wang et al, 2017; Zhang et al, 2018). The fast and efficient attributes of this device can be observed in its ability to chromatographically resolve all 20 amino acids in less than 2 min with more complicated mixtures displaying similar resolution in 3–6 min (Beri et al, 2018; 908 Devices, 2016; Kelley et al, 2019).…”

“…A variety of applications for this separation device have been reported in the literature, including intact antibody analysis, protein analysis and comprehensive metabolomic profiling (Beri et al, 2018; Chen et al, 2018; Fussl et al, 2020; Kelley et al, 2019; Khatri et al, 2017; Wang et al, 2017; Zhang et al, 2018). The fast and efficient attributes of this device can be observed in its ability to chromatographically resolve all 20 amino acids in less than 2 min with more complicated mixtures displaying similar resolution in 3–6 min (Beri et al, 2018; 908 Devices, 2016; Kelley et al, 2019). Additionally, very low sample volumes are required for analyses with the standard injection volume being only 5 nl.…”