Selective Plate-Based Assay for Trace EDTA Analysis via Boron Trifluoride-methanol Derivatization UHPLC-QqQ-MS/MS Enabling Biologic and Vaccine Processes

Abstract

The employment of ethylenediaminetetraacetic acid (EDTA) across several fields in chemistry and biology has required the creation of a high number of quantitative assays. Nonetheless, the determination of trace EDTA, especially in biologics and vaccines, remains challenging. Herein, we introduce an automated high-throughput approach based on EDTA esterification in 96-well plates using boron trifluoride-methanol combined with rapid analysis by ultra-high-performance liquid chromatography–triple quadrupole tandem mass spectrometry (UHPLC-QqQ-MS/MS). Derivatization of EDTA to its methyl ester (Me-EDTA) serves to significantly improve chromatographic performance (retention, peak shape, and selectivity), while also delivering a tremendous enhancement of sensitivity in the positive ion mode electrospray ionization (ESI+). This procedure, in contrast to previous EDTA methods based on complexation with metal ions, is not affected by high concentration of other metals, buffers, and related salts abundantly present in biopharmaceutical processes (e.g., iron, copper, citrate, etc.). Validation of this assay for the determination of ng·mL–1 level EDTA in monoclonal antibody and vaccine products demonstrated excellent performance (repeatability, precision, and linear range) with high recovery from small sample volumes while also providing an advantageous automation-friendly workflow for high-throughput analysis.

Digitally Enabled Generic Analytical Framework Accelerating the Pace of Liquid Chromatography Method Development for Vaccine Adjuvant Formulations

“…4a). The components in this adjuvant mixture are cholesterol ( 4), DOPC ( 6), 3D-PHAD ® (11), and MPLA (12), which are commercially available and used in similar ASO1b and ALFQ formulations. 17,20,51 This final method uses a mobile phase A of 1% acetic acid with 0.08% TEA and a mobile phase B blend of ACN and IPA at a solvent ratio of 90:10 (ACN: IPA).…”

“…2). This is in contrast to C18, the most commonly used stationary phase for lipids and adjuvants 23,43 , that required a longer gradient slope at high organic composition to elute highly retained analytes (9)(10)(11)(12). An overlay of chromatograms obtained using C18 and Phenyl hexyl is shown in Fig.…”

“…[8][9][10][11] However, analytical assays that quantify small molecules present in vaccine products are needed throughout initial development to commercialization. [12][13][14] Among these, adjuvants are crucial components that enhance the immune response triggered by vaccines. Adjuvants boost vaccine effectiveness, enable the use of smaller vaccine doses, and potentially expand availability.…”

Upaya Meningkatkan Skill Komunikasi Siswa Kelas Ix-E Mts Negeri 2 Demak Melalui Layanan Group Conseling Dengan Strategi Simulasi Pada Tahun Pelajaran 2022/2023

“…Ketrampilan komunikasi antar pribadi dapat dilatih melalui beberapa cara antara lain: wawancara, permainan, bimbingan, diskusi, berpidato, menulis (Bennett et al, 2021). menjelaskan bahwa "group prosedur yang lebih intensif dan lebih mendalam adalah group therapy".…”

Extrapolation performance improvement by quantum chemical calculations for machine-learning-based predictions of flow-synthesized binary copolymers

“…Therefore, a high-throughput reactor is one of the critical elements for more efficient studies using ML. Material exploration [15][16][17] and analysis [18][19][20] using high-throughput instruments, whether in organic or inorganic chemistry, are already being actively studied in combination with ML. Coley et al, for example, used ML to predict the synthetic pathways for organic compounds, created a platform for their synthesis using a robotic ow device and demonstrated the platform's efficacy for 15 drugs or drug analogues.…”