Tehran University of Medical Sciences Traditional and Integrative Medicine 2476-5104 1 3 2016 10 04 96 100 EN Seyede Nargess Sadati Lamardi Department of Traditional Pharmacy, School of Persian Medicine, Tehran University of Medical Sciences, Tehran, Iran Somayeh Jahantab Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran Parisa Ziyarati Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran Mahnaz Khanavi Department of Pharmacognosy and Persian Medicine AND Pharmacy Research Center, School of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran Mohammad Reza Shams-Ardekani Department of Pharmacognosy and Persian Medicine AND Pharmacy Research Center, School of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran 2016 10 02 The genus Ajuga is used for the treatment of joint pain, gout, jaundice, and wound healing in Iranian traditional medicine. Ajuga chamaecistus ssp. tomentella is an exclusive subspecies of Ajuga chamaecistus in the flora of Iran. Plants belong to Ajuga species are advantageous sources of phytoecdysteroids. 20-hydroxyecdysone (20E) is an important phytoecdysteroid with anabolic property. This study aimed to determine and quantify 20E in methanolic extract of aerial parts of Ajuga chamaecistus ssp. tomentella. The standard reference of 20E was isolated from n-butanolic fraction of aerial parts of Ajuga chamaecistus ssp. tomentella using high-performance liquid chromatography (HPLC). The analysis was carried out on an ODSA (C18) column with isocratic elution using water–acetonitrile (75:25). The standard calibration curve represented good linearity (r2 = 0.9997). The limit of quantification (S/N = 10) and detection (S/N = 3) were determined as 7.93  and  2.38  µg/ml,  respectively.  The  content  of  20E  in  methanolic  extract  of  Ajuga chamaecistus ssp. tomentella was determined to be 2.58% (w/w) (0.46% in dry plant). The quantitative proportion of the 20E found in Ajuga chamaecistus ssp. tomentella extract proposes the possible uses of this plant in commercial formulations or as a source of 20E. https://jtim.tums.ac.ir/index.php/jtim/article/view/41 https://jtim.tums.ac.ir/index.php/jtim/article/download/41/55