Non-ionic detergents are indispensable tools for investigating cell membranes. Herein, we address the question of how molecular- and supramolecular properties of detergents can be leveraged to improve membrane protein purification and drug amplification. We introduce a modular synthesis for linear triglycerol detergents and observe an inverse correlation between detergents that are good for protein purification and antibiotic amplification. Non-ionic detergents that form worm-like micelles are linked with poor protein purification and antibiotic amplification. The utility of micelle-forming detergents depends on polarity. To enable a better finetuning of micellar polarity, we develop a liquid chromatography method to refine structural contributions of chemical linkers in detergents to polar heads and nonpolar tails. Interrogations on protein purification and antibiotic amplification outcomes led us to propose how the structural integrity of outer membranes in Gram-negative bacteria determines solubilization by non-ionic detergents. Our findings facilitate the assessment of detergents for protein purification and antibiotic amplification.