Evaluation, prediction and optimization the ultrasound-assisted extraction method using response surface methodology: antioxidant and biological properties of Stachys parviflora L.

Document Type: Original Article


1 School of Medicine, Gonabad University of Medical Sciences, Gonabad, Iran

2 Ferdowsi University of Mashhad, International campus, Department of Food Science and Technology, Mashhad, Iran

3 Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran

4 School of Pharmacy; Biotechnology Research Center; Mashhad University of Medical Sciences, Mashhad, Iran

5 Department of Food Science and Technology, Sabzevar Branch, Islamic Azad University, Sabzevar, Iran

6 Department of Food and Drug Control, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

7 Medical Surgical Nursing, Social Determinant of Health Center, Gonabad University of Medical Sciences, Gonabad, Iran


Objective(s):To optimize the extraction method using response surface methodology, extract the phenolic compounds, and identify the antioxidant and biological properties of Stachys parviflora L.  extracts.
Materials and Methods: Maceration and ultrasound-assisted extraction (UAE) (4, 7, 10 min treatment time, 40, 70, 100 % high-intensity and 60, 80, 100 % (v v-1) methanol purity) were applied to obtain the extracts. SEM was conducted to provide the microstructure of the extracted plant. MICs (colorimetric assay), MFCs (colony diameter), total phenolic content, total flavonoid content, radical scavenging capacity and extraction efficiency were determined. HPLC analysis was applied to measure the existent phenolic compounds.
Results: A quadratic model (4 min treatment time, 74.5 % high-intensity and 74.2 % solvent purity) was suggested as the best (TPC: 20.89 mg GAE g-1 d.m., TFC: 6.22 mg QEs g-1 d.m., DPPH IC50: 21.86 µg ml-1 and EE: 113.65 mg g-1 d.m.) UAE extraction model. The optimized UAE extract was generally more effective against Gram-positive microorganisms (MIC: 10-20; MBC: 10-40 (mg ml-1)) than Gram-negative ones (MIC: 40; MBC: >40 (mg ml-1)). Moreover, it (MGI: 2.32-100 %) revealed more anti-mold activity than maceration (MGI: Conclusion: RSM optimization was successfully applied for UAE from S. parviflora. The considerable antioxidant and biological properties were attributed to the phenolic compounds.


1.Rechinger KH. Flora Iranica. No.150; 1982.

2.Said A, Abuotabl EA, Abdel Raoof GF, Huefner A, Nada SA. Phenolic contents and bioactivities of pericarp and seeds of Pleiogynium solandri (Benth.) Engl. (Anacardiaceae). Iran J Basic Med Sci 2015; 18:164-171.

3.Geng-Ruei Chang GR, Chen PL, Hou PH, Mao FC. Resveratrol protects against diet-induced atherosclerosis by reducing low-density lipoprotein cholesterol and inhibiting inflammation in apolipoprotein E-deficient mice. Iran J Basic Med Sci 2015; 18:1063-1071.

4.Laggoune S, Zeghib A, Kabouche A, Kabouche Z, Maklad YA, Leon F, et al. Components and antioxidant, anti-inflammatory, anti-ulcer and antinociceptive activities of the endemic species Stachys mialheside Noé. Arab J Chem 2011; 1-7.

5.Khanavi M, Manayi A, Lotfi M, Abbasi R, Majdzadeh M, Ostad SN. Investigation of cytotoxic activity in four Stachys species from Iran. Iran J Pharm Res 2012; 11:589-593.

6.Wang X, Wu Y, Chen G, Yue W, Liang Q, Wu Q. Optimisation of ultrasound assisted extraction of phenolic compounds from sparganii rhizoma with response surface methodology. Ultrason Sonochem 2012; 20:846-854.

7.Ma Y-Q, Chen J-C, Liu D-H, Ye X-Q. Simultaneous extraction of phenolic compounds of citrus peel extracts: Effect of ultrasound. Ultrason Sonochem 2009; 16:57–62.

8.Salar Bashi D, Mortazavi SA, Rezaei K, Rajaei A, Karimkhani MM. Optimization of ultrasound-assisted extraction of Phenolic Compounds from Yarrow (Achillea beibrestinii) by Response Surface Methodology. Food Sci Biotech 2012; 21:1005-1011.

9.Nwabueze TU. Basic steps in adapting response surface methodology as mathematical modeling for bioprocess optimisation in the food systems. Int J Food Sci Tech 2010; 45:1768–1776.

10.Rodrigues S, Pinto GAS, Fernandes FAN. Optimization of ultrasound extraction of phenolic compounds from coconut (Cocos nucifera) shell powder by response surface methodology. Ultrason Sonochem 2008; 15:95–100.

11.Morelli LLL, Prado MA. Extraction optimization for antioxidant phenolic compounds in red grape jam using ultrasound with a response surface methodology. Ultrason Sonochem 2012; 19:1144–1149.

12.Muniz-Marquez DB, Martinez-Avila GC, Wong-Paz JE, Belmares-Cerda R, Rodriguez-Herrera R, Aguilar CN. Ultrasound-assisted extraction of phenolic compounds from Laurus nobilis L. and their antioxidant activity. Ultrason Sonochem 2013; 20:1149-1154.

13.Teng H, Choi YH. Analytical Methods, Optimization of ultrasonic-assisted extraction of bioactive alkaloid compounds from rhizoma coptidis (Coptis chinensis Franch.) using response surface methodology. Food Chem 2014; 142:299–305.

14.Ahmad VU, Arshad S, Bader S, Iqbal S, Khan A, Suleman Khan S, et al. New terpenoids from Stachys parviflora Benth. Magn Reson Chem 2008; 46:986-989.

15.Hajdari A, Novak J, Mustafa B, Franz C. Essential oil composition and antioxidant activity of Stachys sylvatica L. (Lamiaceae) from different wild populations in Kosovo. Nat Prod Res 2012; 26:1676–1681.

16.Lakhal H, Boudiar T, Kabouche A, Laggoune S, Kabouche Z, Topcu G. Antioxidant Activity and Flavonoids of Stachys ocymastrum. Chem Nat Compd 2011; 46:964-965.

17.Hashemi Sohi SM, Eghdami A, Sadeghi F. Antioxidant activity and high performance liquid chromatography analyzation of methanolic and aqueous extract of Stachys inflata. Org Chem J 2011; 1:36-43.

18.Eghdami A, Sadeghi F, Eradatmand Asli D, Houshmandfar A. Antioxidant activity of methanolic and aqueous extract of Stachys Inflate. Adv. Environ Biol 2011; 5:1256-1259.

19.Salar Bashi D, Emami SA, Iranshahy M, Asili J. Essential Oil Composition of Aerial Parts of Stachys parviflora L. from Iran. J Ess Oil Bear Plants 2013; 16:261-264.

20.Salar Bashi D, Fazly Bazzaz BS, Sahebkar AH, Karimkhani MM, Ahmadi A. Investigation of optimal extraction, antioxidant, and antimicrobial activities of Achillea biebersteinii and A. wilhelmsii. Pharm Biol 2012b; 50:1168-1176.

21.Salarbashi D, Fazly Bazzaz BS, Karimkhani MM, Sabeti Noghabi Z, Khanzadeh F, Sahebkar AH. Oil stability index and biological activities of Achillea biebersteinii and A. wilhelmsii extracts as influenced by various ultrasound intensities. Ind Crop Prod 2014; 55:163-172.

22.Siger A, Nogala-Kalucka M, Lampart-Szczapa E. The content and antioxidant activity of phenolic compounds in cold-pressed plant oils. J Food Lipids 2007; 15:137–149.

23.Patra M, Shahi SK, Midgely G, Dikshit A. Utilization of essential oil as natural antifungal against nail-infective fungi. Flavour Frag J 2002; 17:91-94.

24.Chemat S, Lagha A, AitAmar HV, Bartels P, Chemat F. Comparison of conventional and ultrasound-assisted extraction of carvone and limonene from caraway seeds. Flavour Frag J 2004; 19:188-195.

25.Myers RH, Montgomery DC. Response Surface Methodology: Process and product optimization using design experiments, 2nd ed. John Wiley & Sons, New York, 2002. pp. 235–377.

26.Heydari Majd M, Rajaei A, Salar Bashi D, Mortazavi SA, Bolourian S. Optimization of ultrasonic-assisted extraction of phenolic compounds from bovine pennyroyal (Phlomidoschema parviflorum) leaves using response surface methodology. Ind Crop Prod 2014; 57:195-202.

27.Acikara OB, Çitoğlu GS, Çoban T. Phytochemical Screening and Antioxidant Activities of Selected Scorzonera Species. Turk J Pharm Sci 2013; 10:453-462.

28.Saeedi M, Morteza-Semnani K, Mahdavi MR, Rahimi F, Antimicrobial studies on extracts of four species of Stachys. Indian J Pharm Sci 2008; 70:403-406.

29.Dulger G, Aki C. Antimicrobial activity of the leaves of endemic Stachys pseudopinardii in Turkey. Trop J Pharm Res 2009; 8:371-375.

30.Ebrahimabadi AH, Ebrahimabadi EH, Djafari-Bidgoli Z, Jookar Kashi F, Mazoochi A, Batooli H. Composition and antioxidant and antimicrobial activity of the essential oil and extracts of Stachys inflata