Evaluation of sources of variability in a nitric oxide screening assay for engineered nanomaterials

Published

November 18, 2025

Author(s)

Tariq Fahmi, Robert Gutierrez, Ana Barrios, Tae Joon Cho, John Elliott, Sanghamitra Majumdar, Bryant Nelson, Aaron Johnston-Peck, Alessandro Tona, Anil Patri, Elijah Petersen

Abstract

Assays to detect potential biocompatibility issues can play a key role in supporting the development of new technologies such as medical products containing engineered nanomaterials (ENMs). In this paper, we describe an evaluation of sources of uncertainty in a nitric oxide screening assay to monitor nitric oxide production in RAW 264.7 cells when exposed to ENMs. Protocol testing was conducted during draft standard development and post-publication to better understand these sources of variability such as the impact of insufficient removal of the ENM, the number of cells seeded, the selection of positive control compounds, and the culture techniques of the cells prior to the experiments. Several in-process control measurements were used to monitor the performance of intermediate steps in the assay procedure. Two gold nanoparticles with different surface coatings and nano-sized polystyrene particles were used to demonstrate the applicability of some of the control measurements.

Citation

Nanomaterials

Pub Type

Journals

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Citation

Fahmi, T. , Gutierrez, R. , Barrios, A. , Cho, T. , Elliott, J. , Majumdar, S. , Nelson, B. , Johnston-Peck, A. , Tona, A. , Patri, A. and Petersen, E. (2025), Evaluation of sources of variability in a nitric oxide screening assay for engineered nanomaterials, Nanomaterials, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=959244 (Accessed December 4, 2025)