Citation: Saidi, A.; Hambaba, L.; Bensaad, M.S.; Kadi, I.E.; Bensouici, C.; Sami, R.; Alharthi, S.; Elhakem, A.; Alsharari, Z.D.; Baty, R.S.; et al. Phytochemical Analysis Using cLC-DAD, Nutritional Importance and Assessment of Antioxidant, Antidiabetic and Anticholinesterase Activities of Ruta tuberculata Forssk Organic Extracts. Sustainability 2022, 14, 10451. https://doi.org/10.3390/ su141610451 Academic Editors: Ivan Savic and Ivana Savic Gajic Received: 19 July 2022 Accepted: 13 August 2022 Published: 22 August 2022 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- iations. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). sustainability Article Phytochemical Analysis Using cLC-DAD, Nutritional Importance and Assessment of Antioxidant, Antidiabetic and Anticholinesterase Activities of Ruta tuberculata Forssk Organic Extracts Asma Saidi 1,2 , Leila Hambaba 1 , Mohamed Sabri Bensaad 1,3 , Imed Eddine Kadi 4 , Chawki Bensouici 4 , Rokayya Sami 5, * , Sarah Alharthi 6 , Abeer Elhakem 7 , Zayed D. Alsharari 8 , Roua S. Baty 9 , Raja Saad Alruwais 10 , Mahmood D. Aljabri 11 , Mohammed M. Rahman 12 , Ali A. Shati 13 and Fahmy G. Elsaid 13,14 1 Laboratory of Biotechnology of Bioactive Molecules and Cellular Physiopathology (LBMBPC), Department of Microbiology and Biochemistry, Faculty of Natural and Life Sciences, University Batna 2, Batna 05078, Algeria 2 Department of Natural Sciences and Life, Faculty of Exact Sciences and Natural Sciences and Life, Mohamed Khider University, Biskra 07000, Algeria 3 Laboratory of Cellular and Molecular Physio-Toxicology-Pathology and Biomolecules (LPTPCMB), Department of Biology of Organisms, Faculty of Natural and Life Sciences, University Batna 2, Batna 05078, Algeria 4 Biotechnology Research Center, Ali Mendjli UV 03, Constantine 25000, Algeria 5 Department of Food Science and Nutrition, College of Sciences, Taif University, P.O. Box 11099, Taif 21944, Saudi Arabia 6 Department of Chemistry, College of Science, Taif University, P.O. Box 11099, Taif 21944, Saudi Arabia 7 Department of Biology, College of Science and Humanities in Al-Kharj, Prince Sattam bin Abdulaziz University, P.O. Box 11942, Al-Kharj 16273, Saudi Arabia 8 Department of Clinical Nutrition, Faculty of Applied Medical Sciences, University of Tabuk, P.O. Box 4279, Tabuk 71491, Saudi Arabia 9 Department of Biotechnology, College of Science, Taif University, P.O. Box 11099, Taif 21944, Saudi Arabia 10 Chemistry Department, Faculty of Science and Humanities, Shaqra University, P.O. Box 33, Dawadmi 17452, Saudi Arabia 11 Department of Chemistry, University College in Al-Jamoum, Umm Al-Qura University, P.O. Box 715, Makkah 21955, Saudi Arabia 12 Department of Chemistry, Faculty of Science, King Abdulaziz University, P.O. Box 80200, Jeddah 21589, Saudi Arabia 13 Biology Department, Science College, King Khalid University, P.O. Box 960, Abha 61421, Saudi Arabia 14 Zoology Department, Faculty of Science, Mansoura University, P.O. Box 70, Mansoura 35516, Egypt * Correspondence: rokayya.d@tu.edu.sa Abstract: Among the Mediterranean aromatic plants, Ruta tuberculata Forssk. (Rutaceae) has been widely used as a traditional natural remedy against various disorders resulting from its divers’ pharmacological virtues. The aim of this study is to characterize for the first time the phenolic profile of its ethyl acetate (EtOAcE) and acetonic (AcE) extracts and to screen their in vitro antioxidant, antidiabetic, and neuroprotective activities. Phenolic content was determined using spectrophoto- metric and cLC-DAD analysis. Pharmacologically, in vitro antioxidant power was evaluated using six different antioxidant methods. Moreover, the antidiabetic and neuroprotective capacities were assessed in vitro by determining the α-amylase, α-glucosidase, and acetylcholinesterase inhibitory activities. Phytochemically, the highest flavonoid content was found in EtOAcE where the major identified compounds were myrecetin, rutin, sylimarin, naringenin, and quercetin. In presence of other phenolic acids, gallic acid was exclusively detected in AcE. Furthermore, both R. tuberculata extracts showed significantly remarkable antioxidant activities, especially the EtOAcE. Interestingly, AcE strongly inhibited the acetylcholinesterase and α-glucosidase, with the respective IC 50 values of 20.48 ± 0.2 and 104.5 ± 1.8 μg/mL. In this study, we also reported the nutritional quality associated with the identified phytocompounds. R. tuberculata organic extracts may offer exciting reserves to achieve new anti-diabetic and anti-Alzheimer drugs which have also antioxidant potential. Sustainability 2022, 14, 10451. https://doi.org/10.3390/su141610451 https://www.mdpi.com/journal/sustainability