Quantification of 20-hydroxyecdysone, a Major Phytoecdysteroid, in Ajuga Chamaecistus ssp. Tomentella Using High-Performance Liquid Chromatography
AbstractThe 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.
Aghili Alavi Khorasani MH. Makhzan Al-Advieh.Tehran University of Medical Sciences Publications. Tehran, Iran 2009. [In Persian].
Naghibi F, Mosaddegh M, Mohammadi Motamed M, Ghorbani A. Labiatae family in folk medicine in Iran: from ethnobotany to pharmacology. Iran J Pharm Res 2005; 4: 63-79.
Mozafarian V. A dictionary of Iranian plant names.Farhangmoaser Publications. Tehran, Iran 2007;pp. 21-22. [In Persian].
Ono Y, Fukaya Y, Imai S, Yamakuni T. Beneficial effects of Ajuga decumbens on osteoporosis and arthritis. Biol Pharm Bull 2008; 31: 1199-1204.
Gautam R, Jachak SM, Saklani A. Anti- inflammatory effect of Ajuga bracteosa Wall Ex Benth. Mediated through cyclooxygenase (COX) inhibition. J Ethnopharmacol 2011; 133: 928-930.
El-Hilaly J, Lyoussi B. Hypoglycaemic effect of the lyophilised aqueous extract of Ajuga iva in normal and streptozotocin diabetic rats. Journal of Ethnopharmacology 2002; 80: 109-113.
Kuria KA, de Coster S, Muriuki G, Masengo W, Kibwage I, Hoogmartens J, et al. Antimalarial activity of Ajuga remota Benth (Labiatae) and Caesalpinia volkensii Harms (Caesalpiniaceae): in vitro confirmation of ethnopharmacological use. J Ethnopharmacol 2001; 74: 141-148.
Mamadalieva NZ, El-Read MZ, Ovidi E, Ashou ML, Hamoud R, Sagdullaev SS, et al. Antiproliferative, antimicrobial and antioxidant activities of the chemical constituents of Ajuga turkestanica. Phytopharmacology 2013; 4: 1-18.
Khanavi M, Davoodipoor AM, Sadati SN, Ardekani MR, Sharifzadeh M. Antinociceptive effect of some extracts from Ajuga chamaecistus Ging. ssp. tomentella (Boiss.) Rech. f. aerial parts. Daru 2014; 22: 56.
Sadati N, Ostad S, Karimian Z, Shams Ardekani MR, Akbarzadeh T, Hadjiakhoondi A. Phytochemical study and in vitro cytotoxic effect of Ajuga chamaecistus ssp. tomentella. Asian Journal of Chemistry 2012; 24: 2871-2874.
Sadati N, Jenett-Siems K, Siems K, Ardekani MR, Hadjiakhoondi A, Akbarzadeh T, et al. Major constituents and cytotoxic effects of Ajuga chamaecistus ssp. tomentella. Z Naturforsch C 2012; 67: 275-281.
Ramazanov N Sh. Phytoecdysteroids and other biologically active compounds from plants of the genus Ajuga. Chemistry of Natural Compounds 2005; 41: 361-369.
Gorelick-Feldman J, Cohick W, Raskin I.Ecdysteroids elicit a rapid Ca2+ flux leading to Akt activation and increased protein synthesis in skeletal muscle cells. Steroids 2010; 75: 632-7.
Hamden K, Ayadi F, Jamoussi K, Masmoudi H, Elfeki A. Therapeutic effect of phytoecdysteroids rich extract from Ajuga iva on alloxan induced diabetic rats liver, kidney and pancreas. Biofactors 2008; 33: 165-175.
Kutepova TA, Syrov VN, Khushbaktova ZA, Saatov Z. Hypoglycemic activity of the total Ecdysteroid extract from Ajuga turkestanica. Pharmaceutical Chemistry Journal 2001; 35:608-609.
Dinan L. The Karlson lecture. Phytoecdysteroids:what use are they? Arch Insect Biochem Physiol 2009; 72: 126-141.
Cai YJ, Dai JQ, Fang JG, Ma LP, Hou LF, Yang L, et al. Antioxidative and free radical scavenging effects of ecdysteroids from Serratula strangulata. Can J Physiol Pharmacol 2002; 80: 1187-1194.
Ghosh D, Laddha KS. Extraction and monitoring of phytoecdysteroids through HPLC. J Chromatogr Sci 2006; 44: 22-26.
Wu JJ, Cheng KW, Wang H, Ye WC, Li ET, Wang M. Simultaneous determination of three phytoecdysteroids in the roots of four medicinal plants from the genus Asparagus by HPLC. Phytochem Anal 2009; 20: 58-63.
Gallo MB, Beltrame FL, Vieira PC, Cass QB, Fernandes JB, da Silva MF. Quantitative determination of 20-hydroxyecdysone in methanolic extract of twigs from Vitex polygama Cham. J Chromatogr B Analyt Technol Biomed Life Sci 2006; 832: 36-40.