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Scopoletin has previously been reported as a biomarker for the standardization of <i>Paederia foetida</i> twigs. This study is the first report on the determination and quantification of scopoletin using quantitative nuclear magnetic resonance (qNMR) in the different extracts of <i>Paederia foetida</i> twigs. The validated qNMR method showed a good linearity (<i>r</i>² = 0.9999), limit of detection (LOD) (0.009 mg/mL), and quantification (LOQ) (0.029 mg/mL), together with high stability (relative standard deviation (RSD) = 0.022%), high precision (RSD < 1%), and good recovery (94.08–108.45%). The quantification results of scopoletin concentration in chloroform extract using qNMR and microplate ultraviolet-visible (UV-vis) spectrophotometer was almost comparable. Therefore, the qNMR method is deemed accurate and reliable for quality control of <i>Paederia foetida</i> and other medicinal plants without extensive sample preparation.