Quercetin, a key flavonoid in Anoectochilus roxburghii (Wall.) Lindl., plays an important role in determining the pharmacological value of this medicinal herb. However, traditional methods for quercetin quantification are destructive and time-consuming, limiting their application in real-time quality monitoring. This study investigates the hyperspectral response characteristics of quercetin using near-infrared hyperspectral imaging and establishes a feature-based model to explore its detectability in A. roxburghii leaves. We scanned standard quercetin solutions of known concentration under the same imaging conditions as the leaves to produce a dilution series. Feature-selection methods used included the successive projections algorithm (SPA), Pearson correlation, and competitive adaptive reweighted sampling (CARS). A 1D convolutional neural network (1D-CNN) trained on SPA-selected wavelengths yielded the best prediction performance. These key wavelengths—particularly the 923 nm band—showed strong theoretical and statistical relevance to quercetin’s molecular absorption. When applied to plant leaf spectra, the standard-trained model produced continuous predicted quercetin values that effectively distinguished cultivars with varying flavonoid contents. PCA visualization and ROC-based classification confirmed spectral transferability and potential for functional evaluation. This study demonstrates a non-destructive, spatially resolved, and biochemically interpretable strategy for identifying bioactive markers in plant tissues, offering a methodological basis for future hyperspectral inversion studies and intelligent quality assessment in herbal medicine.