ABSTRACT A flow injection with chemiluminescence (CL) detection system applying tris-2,2′-bipyridylruthenium(II)–potassium permanganate ([Ru(bpy)3]2+–KMnO4) CL reaction was established to determine lambda-cyhalothrin (LCT) and clodinafop-propargyl (CP) pesticides in freshwater samples. Experimental parameters were optimised, and solid phase extraction was employed as phase separation techniques. Linearity of calibration curves for the standard solutions ranges of LCT and CP were of 0.001–1.0 mg/L (y = 1230.3x +23.895, R 2 = 0.9998 (n = 7)) and 0.001–2.0 mg/L (y = 680.12x +18.5, R 2 = 0.9998 (n = 9)), respectively. The respective limits of detection (LODs) and limits of quantification (LOQs) for LCT and CP were found as 3.4 × 10−4 and 3.5 × 10−4 mg/L (3σ blank) and 9.3 × 10−4 and 9.6 × 10−4 mg/L (10σ blank). The injection throughput was 115 h−1. As a consequence of applying tests of significance, the obtained results by the suggested method were not significantly different from those of the reported methods at a 95% confidence level. Recoveries for LCT and CP were achieved over the ranges of 93.8–108% (relative standard deviation (RSD) = 1.7–3.4%) and 93.4–108.2% (RSD = 2.2–3.5%), respectively. The most possible CL reaction mechanism was investigated and formulated.