Journal of Applied Botany and Food Quality 91, 126 - 134 (2018), DOI:10.5073/JABFQ.2018.091.018 1 University of Belgrade, Faculty of Biology, Institute of Botany and Botanical Garden “Jevremovac”, Belgrade, Serbia 2 Institute for Medicinal Plant Research “Dr. Josif Pančić”, Belgrade, Serbia Antineurodegenerative, antioxidant and antibacterial activities and phenolic components of Origanum majorana L. (Lamiaceae) extracts Sonja Duletić-Laušević 1 , Ana Alimpić Aradski 1* , Stoimir Kolarević 1 , Branka Vuković-Gačić 1 , Mariana Oalđe 1 , Jelena Živković 2 , Katarina Šavikin 2 , Petar D. Marin 1 (Submitted: July 4, 2017; Accepted: March 7, 2018) * Corresponding author Summary The aim of this study was to examine chemical composition, as well as antineurodegenerative, antioxidant and antibacterial activity of aqueous and ethanolic extracts of Origanum majorana L. (Lamia- ceae) originating from Serbia, Greece, Egypt and Libya. Total phe- nolics and lavonoids, antioxidant activities, and acetylcholinesterase and tyrosinase inhibitory activities were measured spectrophotome- trically. Determination of phenolic compounds in extracts was done using HPLC-DAD technique. Antibacterial activity included deter- mination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using the microdilution method. The highest phenolic and lavonoid contents were recorded in the ethanolic extract of the Egyptian sample and in aqueous extract of Serbian sample. The HPLC analysis showed high content of rosma- rinic acid, with the highest amount found in the ethanolic extract of the plants from Egypt. Water extracts showed prevalently better antioxidant and antineurodegenerative activity in applied tests than the ethanolic extracts. Gram-positive bacterial strains showed higher sensitivity to tested extracts. According to the obtained results, sweet marjoram samples from Serbia and Egypt can be marked as more promising, due to the highest content of total phenolics and lavo- noids and the best antioxidant, antibacterial and tyrosinase inhibitory activity. Keywords: Origanum majorana, extracts, chemical composition, antioxidant activity, antibacterial activity, antineurodegenerative ac- tivity List of abbreviations AAE - ascorbic acid equivalent; ABTS - 2,2-azino-bis(3-ethyl- benzothiazoline-6-sulfonic acid) diammonium salt; AChE - acetyl- cholinesterase; AD - Alzheimer disease; BHA - 2(3)-t-butyl-4-hy- droxyanisole; BHI - Brain-Heart infusion; BHT - 3,5-di-tert-butyl- 4-hydroxytoluene; DMSO - dimethylsulfoxide; DPPH - 2,2-dyphe- nyl-1-pikrylhydrazyl (DPPH); DTNB - 5,5’-dithiobis(2-nitrobenzoic acid); FRAP - ferric reducing ability of plasma; GAE - gallic acid equivalent; L-DOPA - 3,4-dihydroxy-L-phenylalanine; MBC - minimal bactericidal concentration; MHB - Mueller-Hinton broth; MIC - minimal inhibitory concentration; QE - quercetin equivalent; TFC - total lavonoid content; TPC - total phenolic content; TPTZ - 2,4,6-tripyridil-s-triazin; TYR - tyrosinase; β-CB assay - β-Carotene Bleaching assay Introduction The mint family (Lamiaceae) includes aromatic plants widely used for culinary, medicinal, cosmetic and ornamental purposes, such as basil, rosemary, sage, oregano, lavender, thyme and mint (Raja, 2012). The genus Origanum is composed of 42 species and 18 hy- brids widely distributed in Eurasia and North Africa (IetswaaRt , 1980), being native to the mountainous areas of Mediterranean and Asia (ChIshtI et al., 2013). Species belonging to the genus Origa- num are used since the ancient times as spices, medicinal, aromatic and ornamental plants (MeyeRs, 2005). In vitro pharmacological in- vestigations showed their antibacterial, antifungal, antioxidant, anti- spasmodic, antimutagenic, antitumoral, analgesic, antithrombin and antihyperglycaemic activities (ChIshtI et al., 2013). Origanum majorana L. (sweet marjoram) is herbaceous perennial shrub, inhabiting dry slopes and rocky places, native to Cyprus and Eastern Mediterranean area (IetswaaRt , 1980). It is used as a medi- cinal plant, against cold, as a spasmolytic, antirheumatic, diuretic, and antiasthmatic drug, and as a culinary herb, for lavouring sau- sages, sauces, soups and condiments (ChIshtI et al., 2013). The essential oil of marjoram of different origin was previously analyzed for the composition and biological activities (teIxeIRa et al., 2013; hajlaouI et al., 2016). Various extracts were stud- ied for antioxidant (jun et al., 2001; V ágI et al., 2005a; el-MaatI et al., 2012; Roby et al., 2013; ayaRI et al., 2013; benChIkha et al., 2013; V asudeVa et al., 2014; afIfI et al., 2014; eRenleR et al., 2015) and antimicrobial activities (V ágI et al., 2005b; leeja and thoppIl, 2007; abdel-MassIh et al., 2010). In the presented manuscript for the irst time the marjoram extracts were tested for the antineurodegenerative effects, which is of grow- ing scientiic and public interest. The marjoram spices used in the analyses were produced by respectable companies for culinary herbs and spices, while Libyan marjoram was purchased at the local mar- ket. In several papers the commercially purchased marjoram mate- rial was subjected to various analyses (Mossa et al., 2015; wahby et al., 2015; feRnandes et al., 2016), because of importance of testing the commercial, widely used spices in daily diet for possible beneits for consumers health. Considering the insuficiency of data on medicinal properties of O. majorana extracts, the goals of this study were chemical analysis as well as investigation of antibacterial, antioxidant and antineurodegenerative activity of marjoram aqueous and ethanolic extracts obtained from plants cultivated in Serbia and three Mediterranean countries (Greece, Egypt, Libya). Material and methods Chemicals and reagents Methanol, ethanol, distilled water, glacial acetic acid, hydrochloric acid and chloroform were purchased from Zorka Pharma, Šabac (Serbia). Gallic acid, quercetin, ascorbic acid, 2(3)-t-butyl-4-hydroxy- anisole (BHA), 3,5-di-tert-butyl-4-hydroxytoluene (BHT), 2,2-dy- phenyl-1-pikrylhydrazyl (DPPH), 2,2’-azino-bis(3-ethylbenzothia- zoline-6-sulfonic acid) diammonium salt (ABTS), 2,4,6-tripyridil-s- triazin (TPTZ), potassium acetate (C 2 H 3 KO 2 ), potassium-persulfate (K 2 S 2 O 8 ), dimethylsulfoxide (DMSO), sodium carbonate anhydrous (Na 2 CO 3 ), aluminium nitrate nonahydrate (Al(NO 3 ) 3 9H 2 O), sodium acetate (C 2 H 3 NaO 2 ), iron(III) chloride (FeCl 3 ), iron(II)-sulfate hep- tahydrate (FeSO 4 ∙ 7H 2 O), β-carotene, Folin-Ciocalteu phenol rea- gent, sodium phosphate monobasic (NaH 2 PO 4 ), sodium phosphate