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A series of poly( N -acryloyl glycinamide) (pNAGA) polymers were synthesized and studied as capture agents for surface-enhanced Raman scattering (SERS) detection of aflatoxin B1 (AFB1), a highly carcinogenic food-borne toxin. Four molecular weights of pNAGA were synthesized by reversible addition-fragmentation chain-transfer (RAFT) polymerization to study the dependence of affinity agent efficacy on chain length for this AFB1 sensing platform. Isothermal titration calorimetry (ITC) was used to verify the sign and magnitude of the enthalpic effects involved in polymer–AFB1 interactions in solution and to understand the effects of pNAGA chain length on AFB1 noncovalent binding. pNAGA–AFB1 interactions were found to be exothermic, and longer pNAGA chains generally resulted in smaller enthalpy decreases per repeat unit. With pNAGA 22 being thermodynamically the strongest affinity agent, we hypothesize that AFB1 affinity is determined by a balance between the configurational restrictions in pNAGA chains and the enthalpic advantage of binding AFB1. SERS spectral changes observed following AFB1 exposure were used to evaluate the influence of polymer molecular weight (2.0–5.2 kDa), order of attachment (pre- vs. post- functionalization of the substrate) and attachment chemistry (thiol vs. trithiocarbonate) on the sensitivity of AFB1 detection. The method by which target, polymer affinity agent, and signal transduction mechanism are combined was found to have significant impacts on the achieved sensitivity. The most effective polymer chain length (pNAGA 22 ), anchoring chemistry (thiol), and polymer/toxin assembly scheme (in-solution) allowed detection of 10 ppb AFB1 in water (below the FDA regulatory limit of 20 ppb), a hundred-fold improvement over SERS sensing without the pNAGA affinity agent.
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Analysis of interactions between pharmaceuticals and humic acid: Characterization using entrapment and high-performance affinity microcolumns
The presence of pharmaceuticals as microcontaminants in the environment has become a particular concern given the growing increase in water reuse and recycling to promote global sustainability of this resource. Pharmaceuticals can often undergo reversible interactions with soluble dissolved organic material such as humic acid, which may be an important factor in determining the bioavailability and effects of these compounds in the environment. In this study, high-performance affinity microcolumns containing non-covalently entrapped and immobilized humic acid are used to examine the binding strength and interactions of this agent for tetracycline, carbamazepine, ciprofloxacin, and norfloxacin, all common pharmaceutical microcontaminants known to bind humic acid. The binding constants, as measured with Aldrich humic acid, have good agreement with values reported in the literature. In addition, the effects of temperature, ionic strength, and pH on these interactions are examined with the humic acid microcolumns. This technique made it possible to determine the relative importance of electrostatic interactions vs non-polar interactions or hydrogen bonding on these binding processes. This study illustrates how affinity microcolumns can be used to screen and uniformly quantify binding by pharmaceuticals with humic acid, as well as to study the mechanisms of these interactions, with this information often being acquired in minutes and with small amounts of binding agent (~0.3 mg per microcolumn, which could be used over 200-300 experiments). Use of entrapment and affinity microcolumns can support similar research for a wide range of other microcontaminants with humic acid or alternative binding agents found in water and the environment.
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- PAR ID:
- 10556572
- Publisher / Repository:
- Elsevier
- Date Published:
- Journal Name:
- Journal of Chromatography A
- Volume:
- 1737
- Issue:
- C
- ISSN:
- 0021-9673
- Page Range / eLocation ID:
- 465427
- Subject(s) / Keyword(s):
- Humic acid Pharmaceuticals Interaction studies Affinity microcolumn High-performance affinity chromatography
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.1016/j.chroma.2024.465427
