**Background:** Kiwano (Cucumis metuliferus) is an exotic fruit with increasing global consumption due to its nutritional properties and potential health benefits. Despite growing production and trade, no studies had previously assessed the chemical safety of kiwano regarding multiple organic contaminants. Pesticides, polychlorinated biphenyls (PCBs), and brominated flame retardants (BFRs) are persistent environmental pollutants that can accumulate in food and pose health risks including cancer, endocrine disruption, and neurotoxicity. The European Green Deal and Farm to Fork Strategy aim for a 50% reduction in pesticide use by 2030, underscoring the need for sensitive monitoring methods.
**Methods:** Ten kiwano fruits were collected in February 2019 from a farm in Cinfães, Douro, Portugal. The pulp was separated, homogenized, and stored at −18°C. A QuEChERS-based method was optimized by testing two extraction buffers (AOAC vs. EN) and two d-SPE clean-up compositions (CL1: 150 mg MgSO₄, 50 mg PSA, 50 mg GCB; CL2: 150 mg MgSO₄, 50 mg PSA, 25 mg GCB). The optimal protocol used 5 g pulp, 8 mL acetonitrile, 2 mL ultrapure water, EN QuEChERS salts (4 g MgSO₄, 1 g NaCl, 1 g NaCitrate, 0.5 g disodium citrate sesquihydrate), and CL2 clean-up. Extracts were dried under nitrogen, redissolved in n-hexane, and analyzed by GC-ECD (for OCPs, PCBs, BFRs) and GC-FPD (for OPPs), with GC/MS confirmation. Method validation followed Eurachem guidelines and SANTE/11312/2021, assessing linearity (1.5–18.7 µg/kg matrix-matched calibration), recovery at three spiking levels (7.5, 11.2, 14.9 µg/kg, n=5), intra- and inter-day precision, LOD, LOQ, and matrix effects.
**Key Results:** The QuEChERS EN + CL2 combination provided the best extraction efficiency, with recoveries for most compounds within the acceptable 70–120% range. The two-way ANOVA showed significantly different recoveries between CL1 and CL2 for OCP and BFR groups with AOAC, and for all chemical groups with EN. Matrix effects showed signal enhancement for all target compounds, most pronounced with lower GCB amounts. Validation results: recoveries ranged from 90% to 122% (mean 99%), RSD <15% for precision, LODs 0.2–2.2 µg/kg, LOQs 0.6–7.4 µg/kg, and linearity R² ≥0.991. Among 10 kiwano samples, PCB 101 was detected and confirmed by GC/MS (three m/z ions: 326 > 324 > 286) at 5.1 µg/kg in one sample. No other contaminants (12 OCPs, 6 OPPs, 7 BFRs, 4 other PCBs) were found above LOQ.
**Clinical Implications:** This is the first reported method for simultaneous determination of 30 organic contaminants in kiwano fruit. The detection of PCB 101, a persistent organic pollutant associated with anthropogenic sources, in one sample demonstrates that exotic fruits can accumulate environmental contaminants beyond pesticides. Although the detected level (5.1 µg/kg) is low, the findings underscore the importance of including PCBs, BFRs, and other legacy pollutants in routine food safety monitoring programs. The validated method provides a sensitive, reliable tool for regulatory surveillance and supports the European Green Deal objectives by enabling comprehensive chemical safety assessment of emerging fruit crops. Larger-scale monitoring across different production regions is warranted.