Naringin extraction in the citrus by-products was reported using ultrasonic-assisted extraction (UAE) with a shorter extraction time. Naringin was extracted using a 1/2″ microtip probe at 37% amplitude and a 2 s pulse for 15 min and was eluted at 8.5 min using a 20 min acidic buffer linear gradient. High-performance liquid chromatography (HPLC) with ultraviolet visible detector is the preferred choice for separating polyphenolic compounds due to its selectivity, sensitivity and accuracy over the other colorimetric methods. Major bioactives have a significant absorptivity in the ultraviolet–visible region, making them suitable for HPLC-UV-based separation. In this protocol, we describe a simple, rapid analytical protocol for the reverse phase separation and quantification of naringin using HPLC–UV assisted by ultrasonic extraction with shorter extraction and detection time.