Physical therapists employ several exercises to alleviate low back pain (LBP). Electromyography (EMG) examination of exercises can monitor muscle activation to help clinicians determine the exercise's effect on stabilisation, endurance, or strength. This study evaluated surface EMG activity comparison for Flying Squirrel Exercise (FSE) and the novel 3-Point Quadripod Exercise (3-PQE) to find the most effective exercise for stimulating the lumbar multifidus (LM) muscle. The study recruited 64 healthy young females (19-24 years). Raw data were normalized and are expressed as the percentage of maximum voluntary isometric contraction (%MVIC). The test-retest reliability of the EMG recordings was estimated using intraclass correlation coefficient (ICC3,1). One-way ANOVA was used to statistically analyse and compare the EMG amplitudes during the two exercises. The ICCs for 3-PQE and FSE were 0.94 (SEM, 21.7% MVIC) and 0.87 (SEM, 19.05% MVIC), respectively. The 3-PQE (69 ± 26% MVIC) demonstrated significantly higher activity than did FSE (30 ± 18% MVIC) (F = 15.573, p = 0.001). Thus, 3-PQE might be a feasible strategy for the prevention and rehabilitation of LBP in females.