Abstract: AIM To isolate and determine the structure of flavonol glycoside from glandless cotton seeds. METHODS The flavonol glycosides were extracted from glandless cotton seeds with 60% ethanol and purified by polyaromatic resin, polyamide, silica gel and Sephadex LH-20 on column chromatography. UV,¹H and ¹³CNMR ,¹³C-¹H COSY, HMBC, ¹H-¹H COSY and TOCSY spectra were used for the structural identification. RESULTS Five flavonol glycosides (I-V) were obtained. They were identified as kaempferol 3-O-β-D-apiosyl(1→2)-α-L-rhamnosyl(1→6)-β-D-glucoside (I), quercetin-3-O-β-D-apiosyl(1→2)-α-L-rhamnosyl(1→6)-β-D-glucoside (II), quercetin-3-O-β-D-apiosyl(1→2)-α-D-glucoside (III), rutin (IV) and quercetin-3-O-β-D-glucoside (V). CONCLUSION Compound I is a new natural product. Compounds II and III were isolated from this genus for the first time.