Ophiopogonis Radix, also called Maidong in Chinese, is the dried root of Ophiopogon japonicus (L.f) Ker-Gawl. Saponins and homoisoflavonoids are the two most important groups of active components in Ophiopogon japonicus, but their content is very low. In this work, a fast and efficient microwave-assisted extraction method was developed to extract saponins and homoisoflavonoids from Ophiopogon japonicus. Three main factors were selected using univariate approach, and then Box-Behnken design was employed for the optimization of extraction conditions. The optimal conditions were as follows: solid-solvent ratio of 1:24, microwave power of 630W, and extraction time of 300s. Ultra-high performance liquid chromatography – charged aerosol detector (UHPLC–CAD) method was developed to quantitatively analyze 10 saponins and homoisoflavonoids in 49 samples with different growing years and/or producing location together with its analogue Liriope spicata. The developed method is rapid, accurate and specific, which can be used for the quality evaluation of Ophiopogon japonicus. This study was supported by the National Natural Science Foundation of China (File no. 82104354), Guangdong Basic and Applied Basic Research Foundation (File no. 2022A1515012154), the funding grants from University of Macau (File no. MYRG2022-00198-ICMS), and the Science and Technology Development Fund, Macau SAR (File no. 0070/2021/AGJ).