Background: Supplementation with dietary palmitoleate was reported to lower serum high-sensitivity C-reactive protein (hsCRP) for those with elevated hsCRP. The objective of the study was to evaluate subjectively and objectively measured outcomes of an omega-7 fatty acid (n7FA) dietary supplement on people with non-specific chronic musculoskeletal discomfort. Methods: A double-blind, placebo-controlled, 1:1 randomized single crossover trial was composed of Group “A” which was given placebo first and verum second while group “B” was given verum first and placebo second. Volunteers with uncomplicated chronic musculoskeletal discomfort or a baseline serum hsCRP >1.0 mg/L were recruited from 2017 – 2019. Verum capsules contained palmitoleate and the placebo contained medium chain fatty acids (MCT). Participants took two capsules per day from the first bottle for three weeks followed by two capsules per day from the second bottle without washout for an additional three weeks. Subjects were instructed to maintain their current diet and lifestyle.Results: From 211 volunteers consented and screened, 56 subjects were randomized. Six subjects dropped out, and fifty subjects contributed to the statistical analyses. At baseline, none of the investigated biomarkers were significantly correlated to subjectively assessed musculoskeletal discomfort levels, and Group B showed higher HDL and lower triglyceride than Group A. Main Outcomes of crossover analysis found no statistical significance in all measurements. The mean difference of subjectively measured PROMIS® scores between the first and second visits showed improvement for both placebo and verum groups. Conclusions: Supplementation for three weeks with 688mg per day of a palmiteolate dietary supplement did not reduce serum inflammatory biomarkers nor did it improve subjectively measured quality of life (QoL) for those with chronic non-specific musculoskeletal discomfort. Future studies should explore time, dosing and verification of cis-palmiteolate versus trans-palmiteolate. Future studies should also consider selection of patients with chronic inflammation, for example individuals with higher body-mass index (BMI).Trial registration: NCT03669575. Registered 13 September 2018 – Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT03669575.