Development of a bioanalytical method for quantification of methoxy-polyethylene glycol propionic acid polymer based on ultra-high performance liquid chromatography tandem mass spectrometry technique and micro-solid phase extraction to enhance sensitivity and greenness

Ning Zhang; Yingxia Guo; Yijin Wang; Luyao Yu; Chunpeng Feng; Jiye Tian; Lei Yin; Meiyun Shi

doi:10.1016/j.chroma.2025.466163

Development of a bioanalytical method for quantification of methoxy-polyethylene glycol propionic acid polymer based on ultra-high performance liquid chromatography tandem mass spectrometry technique and micro-solid phase extraction to enhance sensitivity and greenness

J Chromatogr A. 2025 Jun 23:1757:466163. doi: 10.1016/j.chroma.2025.466163. Online ahead of print.

Authors

Ning Zhang¹, Yingxia Guo², Yijin Wang², Luyao Yu², Chunpeng Feng², Jiye Tian², Lei Yin³, Meiyun Shi⁴

Affiliations

¹ Cancer Hospital of Dalian University of Technology, Dalian University of Technology, Shenyang, 110042, China; School of Chemical Engineering, Ocean and Life Sciences, Dalian University of Technology, Panjin, Liaoning, 124221, China.
² School of Chemical Engineering, Ocean and Life Sciences, Dalian University of Technology, Panjin, Liaoning, 124221, China.
³ Cancer Hospital of Dalian University of Technology, Dalian University of Technology, Shenyang, 110042, China; School of Chemical Engineering, Ocean and Life Sciences, Dalian University of Technology, Panjin, Liaoning, 124221, China. Electronic address: leiyin@dlut.edu.cn.
⁴ Cancer Hospital of Dalian University of Technology, Dalian University of Technology, Shenyang, 110042, China; School of Chemical Engineering, Ocean and Life Sciences, Dalian University of Technology, Panjin, Liaoning, 124221, China. Electronic address: shimy@dlut.edu.cn.

PMID: 40580558
DOI: 10.1016/j.chroma.2025.466163

Abstract

Revealing the potential of cellular cytotoxic and pharmacokinetics for methoxy‑polyethylene glycol propionic acid (M-PEG-PA) polymers is critical for evaluating the biocompatibility of PEG-based nanocarriers. M-PEG₆-PA was employed as a target compound in this study. Cellular uptake of M-PEG₆-PA polymers was first investigated by a green and selective ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) assay. After micro-solid phase extraction (μ-SPE), M-PEG₆-PA and internal standard (IS) M-PEG₅-PA were separated on an ACQUITY UPLC® BEH C₁₈ (2.1 × 50 mm I.D, 1.7 μm) chromatographic column. Multiple reaction monitoring (MRM) of characteristic transitions at mass-to-charge ratio (m/z) 367.2 [M-H]^- → 118.8 and m/z 323.2 [M-H]^- → 75.1 were selected for quantification of M-PEG₆-PA and M-PEG₅-PA polymers, respectively. The established analytical method for quantitative analysis of M-PEG₆-PA polymer demonstrated strong linear correlation within 0.01 to 1 μg/mL. This validated assay was subsequently applied to investigate cellular internalization of M-PEG₆-PA in MCF-7 cell samples. The experimental results revealed minimal cytotoxic effects of M-PEG₆-PA at exposure levels spanning 3 μg/mL to 3 mg/mL, with intracellular accumulation remaining below 0.03 % of administered doses in MCF-7 cell samples.

Keywords: Cytotoxicity; PEG; Pharmacokinetics; Quantification; UHPLC-MS/MS.