Effects of Extraction Technique and Solvent on

httpsbiointerfaceresearchcom 8441

Article

Volume 12 Issue 6 2022 8441 - 8452

httpsdoiorg1033263BRIAC12684418452

Effects of Extraction Technique and Solvent on

Phytochemicals Antioxidant and Antimicrobial Activities

of Cultivated and Wild Rosemary (Rosmarinus officinalis

L) from Taounate Region (Northern Morocco)

Ahmed Zeroual 1 El Hassan Sakar 2 Fatima Mahjoubi 1 Mahdi Chaouch 1

Abdellah Chaqroune 1 Mustapha Taleb 1

1 Engineering Laboratory of Organometallic Molecular Materials and Environment (LIMOME) Department of Chemistry

Faculty of Sciences Dhar Mahraz Fez Sidi Mohamed Ben Abdellah University BP 1796 Fez-Atlas 30003 Fez Morocco

zeroualahmedbiogmailcom (AZ) mahjoubifatimausmbaacma (FM) chaouchmahdiusmbaacma (MC)

chaqrouneabdellahusmbaacma (AC) talebmustaphausmbaacma (MT) 2 Department of Biology Faculty of Sciences of Tetuan Abdelmalek Essaacircdi University BP 2121 Mhannech II 93002

Tetuan Morocco hassanscegmailcom (EHS)

Correspondence zeroualahmedbiogmailcom (AZ) hassanscegmailcom (EHS)

Scopus Author ID 57194743663

Received 25102021 Revised 19112021 Accepted 23112021 Published 12122021

Abstract This work was set up to investigate the effects of domestication solvent and extraction

technique on extracts yield total phenolics content (TPC) flavonoids content (TFC) antimicrobial

(minimal inhibitory concentration MIC) and antioxidant (IC50) activities in rosemary aerial parts at

flowering stage Although solvent was the main variability source in our data all tested factors

significantly impacted yield TPC TFC MIC and IC50 These results were confirmed via principal

component analysis which separated solvents extraction techniques and rosemary types on the first

three components accounting for more than 99 of data variability Better values of yield (417 plusmn 030

ndash2158 plusmn 093 DM ) TPC (1248 plusmn 117ndash3472 plusmn 165 mg GAEg DM) TFC (651 plusmn 179ndash2502 plusmn

153 mg QEg DM) MIC (817 plusmn 104ndash2420 plusmn 098 μgmL) and IC50 (5002 plusmn 008ndash39000 plusmn 100

microgmL) were obtained in the case of wild rosemary with Soxhlet extraction especially when combined

with more polar solvents (ethanol and methanol) It could be concluded that domestication had negative

effects on rosemary phytochemicals and associated antimicrobial and antioxidant activities Rosemary

extracts could serve as important ingredients as food preservatives antimicrobial agents and

nutraceuticals

Keywords wild rosemary cultivated rosemary domestication phytochemicals biological activity

copy 2021 by the authors This article is an open-access article distributed under the terms and conditions of the Creative

Commons Attribution (CC BY) license (httpscreativecommonsorglicensesby40)

1 Introduction

Morocco is among the most important botanic areas in northern Africa thanks to various

factors including the geographical position diverse geology topography climate and

ecoregion [1] The Moroccan flora is estimated to encompass around 978 endemic taxa These

constitute more than half of the endemic species of North African[2] Such an endemic richness

appears to be a result of the presence of well-differentiated and mixed environments [1]

Rosemary known botanically as Rosmarinus officinalis L (R officinalis) grows wild in the

western Mediterranean basin with more than 20 cultivars ecotypes or varieties[3ndash5]

Following these authors R officinalis has been used since ancient times for different



medicinal culinary and ornamental purposes For instance in food science R officinalis is

well known for its essential oil used as a food preservative thanks to its antimicrobial and

antioxidant properties rosemary has many other food applications such as culinary medicinal

and pharmacology uses [46ndash8] Several pharmacological activities of R officinalis have been

outlined and well documented in previous studies as antimicrobial anti-inflammatory

attenuating antitumor antiproliferative inhibitory and antioxidant agent [69ndash12]

Rosemarys nutritional value and its bioactive compounds were reviewed by Ribeiro-

Santos et al (2015) [4] Various minerals fatty acids and vitamins are found in different parts

of R officinalis Also many phytochemicals are reported depending on plant part processing

technique geographical origin among others [41314]

Due to their economic and medicinal values and consumer demand some medicinal

and aromatic plants are domesticated and cultivated to be produced on a large scale In this

context R officinalis became an important industrial crop [1516] It is noteworthy that

cultivation practices along with environmental conditions impact morphological traits

biomass and chemical composition [41516]

The chemical composition and biological activities of rosemary have attracted the

attention of several research works In contrast little is known about the domestication of

rosemary and its influence on nutritional composition as well as secondary metabolites

profiling In the present work we aimed at comparing rosemary growing wild and cultivated

under local conditions of the Taounate region (northern Morocco) regarding phytochemical

screening and mineral composition

As far as we know there is no detailed information regarding phytochemicals and

related biological activities in R officinalis growing in central-northern Morocco Hence the

originality of this paper which had as objectives (i) to assess antioxidant and antimicrobial

activities of R officinalis aerial parts (ii) to investigate the domestication of R officinalis in

terms of phenolics and flavonoids content and related bioactivities and (iii) to compare R

officinalis extracts achieved via various solvents in terms of yield total phenolics total

flavonoids and related antioxidant and antimicrobial activities

2 Materials and Methods

21 Chemicals solvents and reagents

Reagents and solvents used were of high purity 22-diphenyl-1-picrylhydrazyl

(DPPH) aluminum chloride (AlCl3) ascorbic acid methanol n-hexane ethanol ethyl acetate

sodium carbonate sodium sulfate FolinndashCiocalteu dimethyl sulfoxide (DMSO) gallic acid

(GA) quercetin (Q) and potassium acetate were used in this work These chemical reagents

were acquired from Stigma-Aldrich (St Louis MO) All other chemicals and solvents used

were of analytical grade

22 Plant material and preparation of extracts

The plant species was botanically identified and authenticated At the full blooming

stage R officinalis aerial parts (flowering tops) were manually harvested in June from

Taounate province (34deg3148 N 4deg4236 W) Wild R officinalis samples were collected in

Aiumln Mediouna while the sampling of cultivated R officinalis was performed from the

Botanical Garden of the National Agency for Medicinal and Aromatic Plants (ANPAM)



Taounate region belongs to central-northern Morocco characterized by a humid Mediterranean

climate in winter and semi-arid in summer

R officinalis samples were dried under shade to avoid photo-oxidation crushed to a

fine powder via an electric grinder and then subjected to extraction via two extraction

techniques namely Soxhlet and maceration Briefly 25 g of the obtained powder was used for

extraction using different solvents separately (ethanol ethyl acetate n-hexane and methanol)

The extraction period was 72 h for maceration and 6h for Soxhlet The obtained extracts were

then filtered through Whatman The obtained residue was re-extracted twice and then the

combined extracts of every sample were rotary-evaporated at a temperature of 45 degC (Heidolph

Collegiate LV28798826 New Jersey USA) The final extracts were used to investigate their

antioxidant and antimicrobial activities [17]

23 Total phenolics and flavonoids determination

231 Determination of total phenolic content (TPC)

TPC was determined via the Folin-Ciocalteu method [18] with slight modifications In

Brief 100 μL of each extract was added to 500 μL of 110 FolinndashCiocalteau reagent which

was prepared prior to use After 4 min 400 μL of sodium carbonate 75 (mv 75mgmL)

were added The obtained mixture was incubated for 90 minutes at room temperature the

absorbance was then read at 765 nm using a Cary 50 UV-Visible Spectrometer (Varian

Mulgrave Australia) A standard curve was prepared using gallic acid (GA 5 mgmL) with

different concentrations (0-150 μgmL) The results were expressed as mg of GA equivalents

(GAE) per g of sample (dry matter DM)

232 Determination of Total Flavonoids Content (TFC)

TFC was determined by using the aluminum trichloride colorimetric (AlCl3) method

[19] with slight modifications Briefly 250 microL of each extract were mixed with 50 microL of AlCl3

(10 mv) 750 microL of ethanol (absolute) 14 mL of bidistilled water and 50 microL of potassium

acetate (1 M) The obtained mixture was incubated for 30 min at room temperature The

corresponding absorbance was then measured at 415 nm Quercetin was used to perform the

standard curve and the results were expressed as mg of quercetin equivalents (QE) per g of

sample (dry matter DM)

24 Antioxidant activities assessment

The capacity to trap the radical DPPH (22-diphenyl-1- picrylhydrazyl) was evaluated

by the standard method described in Brand-Williams et al (2015) [20] with a slight

modification Briefly 02 mL from different concentrations (30ndash480 μgmL) of R officinalis

extracts were added to 18 mL of DPPH methanolic solution (011 mM) After 30 min of

incubation in darkness at room temperature (23 plusmn 2 degC) the absorbance was measured at 517

nm against a blank DPPH methanolic solution The DPPH radical scavenging activity (also

known as antioxidant activity AA) was calculated according to the following equation

(AA)= [ [(Abscontrol ndash Abssample)

Abscontrol

]times 100

where Abscontrol is the absorbance of the control reaction (containing all reagents excluding the

sample test) and Abssample is the absorbance of extracts at 517 Ascorbic acid was used as a



positive control and the concentration of extracts that inhibits 50 (IC50) of DPPH was

calculated based on the percentage graph inhibition following the extract concentration

25 Determination of antimicrobial activities

Microbial strains used here are of significant concern because they involve health

problems and food spoilage [2122] Extracts obtained from various solvents were tested for

antimicrobial activities against the following strains Escherichia coli ATCC 25922 (E coli)

Staphylococcus aureus ATCC29213 (S aureus) Bacillus subtilis ATCC 3366 (B subtilis) and

Candida albicans ATCC 10231 (C albicans)

To determine minimum inhibitory concentration (MIC) the agar dilution method was

used following the Natural Committee for Clinical Laboratory Standards [23] All tests were

performed in nutrient broth for bacterial strains Different quantities (75 10 125 15 and 20

μg) from dry extracts were added separately to 1 mL nutrient broth tubes containing 105

CFUmL of live microbial cells In order to evenly spread the extracts throughout the broth

the tubes (10 mL broth) were incubated in an incubator shaker The highest dilution (the lowest

concentration) presenting no visible bacterial growth was MIC Thereafter cells from the

tubes displaying no growth were cultured onto nutrient agar plates to determine whether the

inhibition was permanent or not MIC was expressed in μgmL

26 Statistical Analyses

Determinations and measurements were carried out in triplicates Quantitative

differences were assessed by General Linear Procedure followed by LSDs test [24ndash26]

Replicates were nested to solvents Data statistical analyses were performed by the SPSS

package version 23 (IBM Armonk NY USA) Results were expressed as means plusmn standard

deviations (SD) Differences were considered significant at 5 as a probability level Principal

component analysis (PCA) was performed on mean values considering rosemary origins

solvents and extraction techniques as independent variables PCA biplots were by means of

STATGRAPHICS package version XVII (Statpoint Technologies Inc Virginia USA)

3 Results and Discussion

31 Mean comparison

Results regarding TPC TFC MIC and IC50 in various extracts are summarized in

Table 1 As illustrated in these outcomes wide variations were detected among rosemary

origin extraction techniques and solvents in terms of TPC TFC MIC and IC50

Table 1 Mean values of of yield () total phenolics content (TPC mg GAEg DM) and total

flavonoids content (TFC mg QEg DM) minimal inhibitory concentration (μgmL) in various microbial strains

and IC50 (microgmL) for R officinalis wild and cultivated extracts using four solvents and two extraction techniques

(maceration and Soxhlet) M = methanol E = ethanol EA = ethyl acetate and H = hexane Results are

expressed as mean values of three replicates followed by SD Within each column values followed by the same

letter are not significantly different at 5 as a probability level

Rosemary Yield TPC TFC E coli S aureus B subtilis C albicans IC50

Wild Soxhlet

M 2158

plusmn

093a

3472

plusmn

165a

2502 plusmn

153a

1283 plusmn

029i

850 plusmn 050k 967 plusmn 126k 817 plusmn 104k 5002 plusmn

008p



Rosemary Yield TPC TFC E coli S aureus B subtilis C albicans IC50 E 1732

plusmn

109d

3088

plusmn

120c

1655 plusmn

206d

1400 plusmn

050h

933 plusmn 029j 1000 plusmn

050j


035k

EA 1427

plusmn

080g

2513

plusmn

111g

1219 plusmn

124g

1500 plusmn

050g

1217 plusmn

029g

1250 plusmn

050h

1183 plusmn

076h

19003 plusmn

045h

H 816 plusmn

066j

1500

plusmn 193j

935 plusmn

234j

1783 plusmn

076e

1417 plusmn

029e

1450 plusmn

050f

1300 plusmn

050f

26500 plusmn

020e

Maceration

M 1906

plusmn 130c

3498

plusmn 033a

2225 plusmn

048b

1683 plusmn

076f

1250 plusmn

050g

1367 plusmn

126g

1217 plusmn

104g

5978 plusmn

056n

E 1331 plusmn

062h

2837 plusmn

153e

1398 plusmn 146f

1783 plusmn 076e

1333 plusmn 029f

1400 plusmn 050f

1367 plusmn 029f

13500 plusmn 030j

EA 762 plusmn

062k

2495

plusmn

332h

1128 plusmn

105h

1883 plusmn

029d

1617 plusmn

029c

1650 plusmn

050d

1583 plusmn

076d

21003 plusmn

025g

H 514 plusmn

014m

1340

plusmn

082l

700 plusmn

182l

2167 plusmn

058b

1833 plusmn

029b

1900 plusmn

050b

1717 plusmn

029c

30052 plusmn

050c

Cultivated Soxhlet

M 2009 plusmn

104b

3322 plusmn

191b

2296 plusmn 188b

1450 plusmn 087h

1050 plusmn 050i

1167 plusmn 126i

1017 plusmn 104i 7000 plusmn 040m

E 1552

plusmn

081f

2944

plusmn

125d

1477 plusmn

158e

1583 plusmn

076g

1150 plusmn

050h

1200 plusmn

050h

1167 plusmn

029h

16000 plusmn

050i

EA 1318

plusmn

102h

2390

plusmn

085i

1069 plusmn

158i

1733 plusmn

029e

1417 plusmn

029e

1450 plusmn

050f

1383 plusmn

076f

24000 plusmn

030f

H 671 plusmn

129l

1420

plusmn 172k

801 plusmn

264k

2017 plusmn

104c

1617 plusmn

029c

1650 plusmn

050d

1433 plusmn

161e

36000 plusmn

020b

Maceration

M 1684

plusmn 172e

3359

plusmn 063b

2080 plusmn

062c

1853 plusmn

084d

1427 plusmn

093e

1567 plusmn

161e

1433 plusmn

076e

8000 plusmn

050l

E 1147 plusmn

053i

2643 plusmn

155f

1263 plusmn 115g

2017 plusmn 101c

1533 plusmn 104d

1550 plusmn 050e

1550 plusmn 050d

16240 plusmn 026i

EA 625 plusmn

077l

2356

plusmn

305i

1014 plusmn

041i

2177 plusmn

015b

1800 plusmn

050b

1850 plusmn

050c

1800 plusmn

132b

27603 plusmn

025d

H 417 plusmn

030n

1248

plusmn

117m

651 plusmn 179m 2420 plusmn

098a

2017 plusmn

076a

2033 plusmn

104a

1950 plusmn

050a

39000 plusmn

100a

Soxhlet performed better in terms of extract yield in the case of wild rosemary for yield

(2158 plusmn 093 ) TPC (3472 plusmn 165 mg GAEg DM) and TFC (2502 plusmn 153 mg QEg DM)

especially when combined with methanol as solvent Likewise combining Soxhlet extraction

with methanol allowed obtaining the smallest values of MIC and IC50 and therefore the most

effective against the studied microbial strains and scavenging DPPH Methanol was followed

by ethanol ethyl acetate and hexane in terms of efficiency for yield TPC TFC as well as

MIC in various microbial strains and IC50 In terms of MIC Gram-negative bacteria namely

B subtilis (967 plusmn 126 μgmL) and S aureus (850 plusmn 050 μgmL) were the most sensitive

strains followed by C albicans (817 plusmn 104 μgmL) and Gram-positive bacterium (Ecoli)

under Soxhlet extraction combined with methanol As for MIC the most effective value of IC50

(5002 plusmn 008 microgmL) was reached under the same conditions Taking all results of extract

characteristics together it appears that rosemary growing wild presented the highest record of

yield TPC TFC MIC and IC50 compared to cultivated rosemary (Table 4)



With respect to antimicrobial activities wide variations in terms of MIC were found

among microbial strains on the one hand and various extracts on the other hand Mean values

found in our work were in agreement with those found in the literature for R officinalis

collected in Morocco and abroad [1627] As highlighted in the results section the most

important values of inhibition zone diameter were recorded in yeast strain (C albicans)

followed by Gram-positive bacteria (B subtilis along with S aureus) and finally the Gram-

negative one (E coli) especially for extracts from Me-OH and Et-OH Such findings agree

with other published works [28ndash30] who found that yeast strains show their superiority over

Gram-positive and Gram-negative bacteria for methanolic extracts

It is widely evidenced that antimicrobial activities of plant extracts demonstrate that

Gram-positive bacteria are more sensitive than Gram-negative bacteria This difference in

sensitivity could be attributed to the presence of an outer membrane This contains hydrophilic

lipopolysaccharides encompassing the bacterial peptidoglycan layer in Gram-negative bacteria

and acts as a barrier for macromolecules as well as hydrophobic compounds and therefore

limits the diffusion of hydrophobic compounds into the bacterium cytoplasm [3132]

As for microbial strains important differences in antimicrobial activities were reported

among solvents with the superiority of Me-OH and Et-OH In accordance with our results

Elshobary et al (2020) [33] found that low polarity solvents like hexane show lower activities

owing to the difference in the solubility of the bioactive compounds [34]

Outcomes of MICs confirmed the effectiveness of different extracts against the studied

microbial strains Regarding solvents low MIC values were reported in Met-OH and Et-OH

extracts This proves that these extracts have strong antimicrobial activity inhibiting microbial

growth even at low concentrations Effects of different solvents and t extraction techniques on

the chemical composition and biological activities have been studied in German chamomile

extracts It was shown that these parameters can significantly modify extract composition as

well as MIC against several pathogen strains [3536] The outcomes of these works

demonstrated that higher polarity solvents are more effective in extracting bioactive

compounds and therefore show the ability to inhibit microbial growth at lower concentrations

(lower MIC) A similar trend was observed for IC50 as compared to n-hexane and ethyl acetate

extracts from Met-OH and Et-OH were able to reduce 50 of DPPH at lesser concentrations

According to Roby et al (2013) [37] IC50 is defined as the moles of phenolic compounds

divided by moles of DPPH necessary to reduce by 50 the absorbance of DPPH and therefore

the lowest the IC50 the higher is the antioxidant activity The difference of antioxidant activities

among solvents observed in our results was consistent with Yazdani et al (2019) [38]

32 Correlation study

Correlation coefficients among the investigated parameters are shown in Table 2 As

can be seen in these outcomes important positive and negative correlations were revealed

Yield of extracts was positively associated with both TPC and TFC and negatively correlated

to minimal inhibitory concentration (MIC) in all studied microbial strains as well as IC50

Similarly TPC and TFC were positively linked to each other and to yield but they were

negatively correlated to MIC in all microbial strains and IC50 This means that extracts of higher

yields contained greater levels of TPC and TFC which in turn were more effective in terms

of antimicrobial activity (MIC) and antioxidant activity (IC50) Similar correlations among

phenolics and both antioxidant and antimicrobial activities were found by other authors

[3940]



Table 2 Correlation coefficients among studied parameters Yield TPC TFC E coli S aureus B subtilis C albicans IC50

Yield

0909 0936 -0871 -0911 -0872 -0868 -0927

TPC

0915 -0720 -0764 -0719 -0671 -0958

TFC

-0730 -0782 -0718 -0743 -0933

E coli

0981 0979 0975 0769

S aureus

0994 0984 0813

B subtilis

0979 0765

C albicans

0740

IC50

33 Principal component analysis (PCA)

PCA is one of the most popular multivariate statistical methods It is used to reduce the

dimensionality in data sets and project them into a reduced space For such a purpose the PCA

approach is widely used in many fields such as food science and agronomy [2441ndash58] The

first three principal components were retained in our work since they explained about 99 of

the total data variability (Figure 1)

Figure 1 Principal component projections on PC1 and PC2 Blue squares plotted are mean values of

mineral elements Blue segments are related to both rosemary (wild and cultivated) Points plotted are dependent

variables mean values M = methanol E = ethanol EA = ethyl acetate and H = hexane

The 16 points plotted in Figure 1A are related to extraction techniques These seem to

be separated along the first (PC2) with a partial variability of about 10 Toward the positive

direction of PC2 were points related to maceration which was associated with higher levels of

MIC in all microbial strains and IC50 On the opposite side of the same component (PC2) the

A B

C



points linked to Soxhlet were distributed with great values of yield TPC and TFC Similarly

Figure 1B presents the distribution of points related to solvents These appear to be separated

along the PC1 with over 87 of data variability both Methanol (M) and Ethanol (E) were

distributed in the positive direction of this component with important levels of yield TPC and

TFC while Hexane (H) along with Ethyl Acetate (EA) were plotted toward the negative side

of PC1 with the greatest records of MIC in the studied microbial strains as well as IC50 Finally

the third component with lesser magnitude appears to be related to rosemary origin (Figure

1C) However there was no clear separation among wild and cultivated rosemary on this

component

4 Conclusions

Our outcomes demonstrate that rosemary aerial parts (leaves) are an important source

of phenolic compounds with large variations between wild and cultivated samples All factors

investigated (rosemary origin solvent and extraction techniques) significantly impacted

extracts yield TPC TFC MIC and IC50 Principal component analysis showed good

separation of three studied factors on the first three components confirming that solvent was

the main variability source in our data with important correlations among investigated

variables Domestication appears to negatively impact yield TPC TFC and related activity

(MIC and IC50) Soxhlet seemed to be more effective in terms of yield TPC and TFC

especially when combined with ethanol or methanol as solvents Based on the obtained

outcomes rosemary domestication resulted in a reduced yield of extracts TPC TFC and

therefore less important antimicrobial and antioxidant activities values Studied extracts could

be of great importance as food preservatives antimicrobial agents as well as nutraceuticals

Funding

This research work received no external funding

Acknowledgments

The authors would like to warmly thank Dr Taha El Kamli from Food and Nutritional

Transition Research Team (ETAN) Laboratory of Biological Tests Ibn Tofail University

Morocco) for his kind technical assistance

Conflicts of Interest

The authors declare no conflict of interest

References

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7 Ahmed HM Babakir-Mina M Investigation of rosemary herbal extracts (Rosmarinus officinalis) and their

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Rep 2007 17 1525ndash1531 httpsdoiorg103892or1761525

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Gerontol 2009 44 383ndash389 httpsdoiorg101016jexger200902015

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Altern Med 2014 14 225 httpsdoiorg1011861472-6882-14-225

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Medicinal Aromatic and Stimulant Plants Handbook of Plant Breeding Springer Cham 2020 12

httpsdoiorg101007978-3-030-38792-1_15

14 Zeroual A Sakar EH Ibourki M Bijla L Ainane A Mahjoubi F Chaouch M Gharby S

Chaqroune A Ainane T phytochemical screening and mineral profiling of wild and cultivated rosemary

(Rosmarinus officinalis l) from Taounate region (northern Morocco) PharmacologyOnLine 2021 2 576ndash

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httpswwwsciencedirectcombook9780857090393handbook-of-herbs-and-spices

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studies under Ankara ecological conditions Ind Crops Prod 2016 88 12ndash16

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officinalis and S Fructicosa) and oregano (Origanum onites and O inderscedens) extracts related to their

phenolic compound content J Sci Food Agr 2002 82 1645ndash1651 httpsdoiorg101002jsfa1240

18 Singleton VL Orthofer R Lamuela-Raventos RM Analysis of total phenols and other oxidation

substrates and antioxidants by means of folin-ciocalteu reagent Methods in Enzymol 1999 299 152ndash178

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extract of Ferula asafoetida and its essential oil composition Grasas Aceites 2009 60 405ndash412

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Activity LWT-Food Sci Technol 1995 28 25ndash30 httpsdoiorg101016S0023-6438(95)80008-5

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extracts_and_essential_oils_from_two_thyme_species_Thymus_vulgaris_and_Thymus_hyemalis_from_no

rthern_Morocco

22 Zeroual A Eloutassi N Chaouch M Chaqroune A Antimicrobial antioxidant activity and chemical

composition of Origanum compactum benth from Taounate province north Morocco Asian J Pharm Clin

Res 2020 13 126ndash131 httpdxdoiorg1022159ajpcr2020v13i336319



23 Prudent D Perineau F Bessiere J M Michel G M Baccou JC Analysis of the essential oil of wild

oregano from Martinique (Coleus aromaticus Benth) evaluation of its bacterioatatic and fungistatic

properties J Essent Oil Res 1995 7 165ndash173 httpsdoiorg1010801041290519959698492

24 Boussakouran A El Yamani M Sakar EH Rharrabti Y Genetic Advance and Grain Yield Stability of

Moroccan Durum Wheats Grown under Rainfed and Irrigated Conditions Int J Agron 2021 2021 5571501

httpsdoiorg10115520215571501

25 Khattabi D Sakar EH Louahlia S Flag leaf tolerance study in Moroccan barley (Hordeum vulgare L)

varieties submitted to a severe salt stress Biointerface Res Appl Chem 2022 12 2787ndash2799


26 Gharby S Hajib A Ibourki M Sakar EH Nounah I El Moudden H Elibrahimi M Harhar H

Induced changes in olive oil subjected to various chemical refining steps A comparative study of quality

indices fatty acids bioactive minor components and oxidation stability kinetic parameters Chem Data

Collect 2021 33 100702 httpsdoiorg101016jcdc2021100702

27 El Yamani M Sakar EH Mansouri F SerghinI-Caid H Elamrani A Rharrabti Y Effect of pigments

and total phenols on oxidative stability of monovarietal virgin olive oil produced in Morocco Riv Ital

Sostanze Gr 2019 96 17ndash24

httpswwwresearchgatenetpublication328265834_Effect_of_pigments_and_total_phenols_on_oxidative

_stability_of_monovarietal_virgin_olive_oil_produced_in_Morocco

28 El Kharraf S El-Guendouz S Farah A Bennani B Mateus M C amp Miguel M G Hydrodistillation

and simultaneous hydrodistillation-steam distillation of Rosmarinus officinalis and Origanum compactum

Antioxidant anti-inflammatory and antibacterial effect of the essential oils Ind Crops Prod 2021 168

113591 httpdxdoiorg101016jindcrop2021113591

29 Zeroual A Sakar EH Eloutassi N Mahjoubi F Chaouch M Chaqroune A Phytochemical Profiling

of Essential Oils Isolated Using Hydrodistillation and Microwave Methods and Characterization of Some

Nutrients in Origanum compactum Benth from Central-Northern Morocco Biointerface Res Appl Chem

2021 11 9358ndash9371 httpsdoiorg1033263BRIAC11293589371

30 Zeroual A Sakar EH Eloutassi N Mahjoubi F Chaouch M Chaqroune A Wild chamomile

[Cladanthus mixtus (L) chevall] collected from central-northern morocco Phytochemical profiling

antioxidant and antimicrobial activities Biointerface Res Appl Chem 2021 11 11440ndash11457


31 Yap PSX Krishnan T Yiap BC Hu CP Chan KG Lim SHE Membrane disruption and

antiquorum sensing effects of synergistic interaction between Lavandula angustifolia (lavender oil) in

combination with antibiotic against plasmidconferred multidrugresistant Escherichia coli J Appl Microb

2014 116 1119ndash1128 httpsdoiorg101111jam12444

32 Yap PSX Krishnan T Chan KG Lim SH Antibacterial mode of action of Cinnamomum verum bark

essential oil alone and in combination with piperacillin against a multi-drug-resistant Escherichia coli strain

J Microbiol Biotechnol 2015 25 1299ndash1306 httpsdxdoiorg104014jmb140707054

33 Elshobary M El-Shenody R Ashour M Zabed HM Qi X Antimicrobial and antioxidant

characterization of bioactive components from Chlorococcum minutum a newly isolated green microalga

Food Bioscience 2020 35 100567 httpsdoiorg101016jfbio2020100567

34 Patil AG Koli SP Patil DA Phatak AV Evaluation of extraction techniques with various solvents to

determine extraction efficiency of selected medicinal plants Int J Pharm Sci Res 2012 3 2607ndash2612

httpsijpsrcombft-articleevaluation-of-extraction-techniques-with-various-solvents-to-determine-

extraction-efficiency-of-selected-medicinal-plants

35 Cvetanović A Švarc-Gajić J Mašković P Savić S Nikolić L Antioxidant and biological activity of

chamomile extracts obtained by different techniques perspective of using superheated water for isolation of

biologically active compounds Ind Crops Prod 2015 65 582ndash591


36 Metrouh-Amir H Duarte CM Maiza F Solvent effect on total phenolic contents antioxidant and

antibacterial activities of Matricaria pubescens Ind Crops Prod 2015 67 249ndash256


37 Roby MHH Sarhan MA Selim KAH Khalel KI Antioxidant and antimicrobial activities of essential

oil and extracts of fennel (Foeniculum vulgare L) and chamomile (Matricaria chamomilla L) Ind Crops

Prod 2013 44 437ndash445 httpsdoiorg101002ffj1929



38 Yazdani E Talebi M Zarshenas MM Moein M Evaluation of possible antioxidant activities of barberry

solid formulation a selected formulation from traditional persian medicine (tpm) via various procedures

Biointerface Res Appl Chem 2019 9 4517ndash1521 httpsdoiorg1033263BRIAC96517521

39 Chepel V Lisun V Skrypnik L Changes in the Content of Some Groups of Phenolic Compounds and

Biological Activity of Extracts of Various Parts of Heather (Calluna vulgaris (L) Hull) at Different Growth

Stages Plants 2020 9 926 httpsdoiorg103390plants9080926

40 Karalija E Ćavar Zeljković S Dahija S Bešta-Gajević R Parić A Phenolics of Aerial Parts

of Gentiana lutea L and Their Biological Activity Agronomy 2021 11 1442

httpsdoiorg103390agronomy11071442

41 Sakar EH El Yamani M Rharrabti Y Variability of oil content and its physico-chemical traits from five

almond (Prunus dulcis) cultivars grown in northern Morocco J Mater Environ Sci 2017 8 2679ndash2686

httpswwwresearchgatenetpublication316634449_Variability_of_oil_content_and_its_physico-

chemical_traits_from_five_almond_Prunisdulcis_cultivars_grown_in_Northern_Morocco

42 Sakar EH El Yamani M Rharrabti Y Geometrical Traits in Almond Fruit as Affected by Genotypic and

Environmental Variations in Northern Morocco Erwerbs-Obstbau 2019 61 103ndash112

httpsdoiorg101007s10341-018-0401-y

43 Boussakouran A Sakar EH El Yamani M Rharrabti Y Morphological Traits Associated with Drought

Stress Tolerance in Six Moroccan Durum Wheat Varieties Released Between 1984 and 2007 J Crop Sci

Biotechnol 2019 22 345ndash353 httpsdoiorg101007s12892-019-0138-0

44 Bijla L Aissa R Ait Bouzid H Sakar EH Ibourki M Laknifli A Gharby S Spent Coffee Ground

Oil as a Potential Alternative for Vegetable Oil Production Evidence from Oil Content Lipid Profiling and

Physicochemical Characterization Biointerface Res Appl Chem 2021 12 6308ndash6320


45 El Yamani M Sakar E H Boussakouran A Benali T Rharrabti Y Antioxidant activity of phenolic

extracts from olive mill wastewater and their influence on virgin olive oil stability Mor J Chem 2019 7 211ndash

223

httpswwwresearchgatenetpublication331988832_Antioxidant_activity_of_phenolic_extracts_from_oli

ve_mill_wastewater_and_their_influence_on_virgin_olive_oil_stability_published_in_Moroccan_Journal_

of_Chemistry

46 El Yamani M Sakar EH Boussakouran A Rharrabti Y Physiological and biochemical responses of

young olive trees (Olea europaea L) to water stress during flowering Arch Biol Sci 2019 71 123ndash132

httpsdoiorg102298ABS181001054E

47 Sakar EH El Yamani M Rharrabti Y Fruit Gravimetrical Traits in Almond [Prunus dulcis (Mill) DA

Webb] Combined Effects of Genetic Control and Environmental Drivers Erwerbs-Obstbau 2020 62 37ndash

46 httpsdoiorg101007s10341-019-00457-x

48 El Yamani M Sakar EH Boussakouran A Rharrabti Y Leaf water status physiological behavior and

biochemical mechanism involved in young olive plants under water deficit Sci Hortic 2020 261 108906

httpsdoiorg101016jscienta2019108906

49 El Yamani M Sakar EH Boussakouran A Rharrabti Y Activity of two natural additives in improving

the stability of virgin olive oil quality during storage OCL 2020 27 44 httpsdoiorg101051ocl2020039

50 El Yamani M Sakar EH Boussakouran A Rharrabti Y Influence of ripening index and water regime

on the yield and quality of Moroccan Picholine virgin olive oil OCL 2020 27 19

httpsdoiorg101051ocl2020015

51 El Yamani M Sakar E H Boussakouran A Ghabbour N Rharrabti Y Physicochemical and

Microbiological Characterization of Olive Mill Wastewater (OMW) from Different Regions of Northern

Morocco Environ Technol 2020 41 3081ndash3093 httpsdoi1010800959333020191597926

52 Boukyoud Z Gharby S Ibourki M Sakar E H Bijla L Atifi H Matthaumlus B Laknifli A Charrouf

Z Can the water quality influence the chemical composition sensory properties and oxidative stability of

traditionally extracted argan oil Med J Nutrition Metab 2021 httpsdoiorg103233MNM-210005

53 Sakar EH El Yamani M Boussakouran A Rharrabti Y Genotypic and Environmental Variations in

Kernel Color Indices in the Main Almond [Prunus dulcis (Mill) DA Webb] Cultivars Grown in North-

eastern Morocco Scientifica 2021 2021 9970223 httpsdoiorg10115520219970223

54 Sakar EH El Yamani M Boussakouran A Ainane A Ainane T Gharby S Rharrabti Y Variability

of oil content and its physicochemical traits from the main almond [Prunus dulcis Mill DA Webb] cultivars



grown under contrasting environments in north-eastern Morocco Biocatal Agric Biotechnol 2021 32

101952 httpsdoiorg101016jbcab2021101952

55 Boussakouran A El Yamani M Sakar EH Rharrabti Y Genetic Progress in Physiological and

Biochemical Traits Related to Grain Yield in Moroccan Durum Wheat Varieties from 1984 to 2007 Crop Sci

2021 httpsdoiorg101002csc220643

56 Ibourki M Azouguigh F Jadouali S Sakar EH Bijla L Majourhat K Gharby S Laknifli A

Physical fruit traits nutritional composition and seed oil fatty acids profiling in the main date palm (Phoenix

dactylifera L) varieties grown in Morocco J Food Qual 2021 2021 5138043

httpsdoiorg10115520215138043

57 El Yamani M Sakar EH Boussakouran A Rharrabti Y Physicochemical and Microbiological

Characterization of Olive Mill Wastewater (OMW) from Different Regions of Northern Morocco Biocatal

Agric Biotechnol 2022

58 Ibourki M Ait Bouzid H Bijla L Sakar EH Asdadi A Laknifli A El Hammadi A Gharby S

Mineral profiling of twenty wild and cultivated aromatic and medicinal plants growing in Morocco

Biol Trace Elem Res 2022



medicinal culinary and ornamental purposes For instance in food science R officinalis is

well known for its essential oil used as a food preservative thanks to its antimicrobial and

antioxidant properties rosemary has many other food applications such as culinary medicinal

and pharmacology uses [46ndash8] Several pharmacological activities of R officinalis have been

outlined and well documented in previous studies as antimicrobial anti-inflammatory

attenuating antitumor antiproliferative inhibitory and antioxidant agent [69ndash12]

Rosemarys nutritional value and its bioactive compounds were reviewed by Ribeiro-

Santos et al (2015) [4] Various minerals fatty acids and vitamins are found in different parts

of R officinalis Also many phytochemicals are reported depending on plant part processing

technique geographical origin among others [41314]

Due to their economic and medicinal values and consumer demand some medicinal

and aromatic plants are domesticated and cultivated to be produced on a large scale In this

context R officinalis became an important industrial crop [1516] It is noteworthy that

cultivation practices along with environmental conditions impact morphological traits

biomass and chemical composition [41516]

The chemical composition and biological activities of rosemary have attracted the

attention of several research works In contrast little is known about the domestication of

rosemary and its influence on nutritional composition as well as secondary metabolites

profiling In the present work we aimed at comparing rosemary growing wild and cultivated

under local conditions of the Taounate region (northern Morocco) regarding phytochemical

screening and mineral composition

As far as we know there is no detailed information regarding phytochemicals and

related biological activities in R officinalis growing in central-northern Morocco Hence the

originality of this paper which had as objectives (i) to assess antioxidant and antimicrobial

activities of R officinalis aerial parts (ii) to investigate the domestication of R officinalis in

terms of phenolics and flavonoids content and related bioactivities and (iii) to compare R

officinalis extracts achieved via various solvents in terms of yield total phenolics total

flavonoids and related antioxidant and antimicrobial activities

2 Materials and Methods

21 Chemicals solvents and reagents

Reagents and solvents used were of high purity 22-diphenyl-1-picrylhydrazyl

(DPPH) aluminum chloride (AlCl3) ascorbic acid methanol n-hexane ethanol ethyl acetate

sodium carbonate sodium sulfate FolinndashCiocalteu dimethyl sulfoxide (DMSO) gallic acid

(GA) quercetin (Q) and potassium acetate were used in this work These chemical reagents

were acquired from Stigma-Aldrich (St Louis MO) All other chemicals and solvents used

were of analytical grade

22 Plant material and preparation of extracts

The plant species was botanically identified and authenticated At the full blooming

stage R officinalis aerial parts (flowering tops) were manually harvested in June from

Taounate province (34deg3148 N 4deg4236 W) Wild R officinalis samples were collected in

Aiumln Mediouna while the sampling of cultivated R officinalis was performed from the

Botanical Garden of the National Agency for Medicinal and Aromatic Plants (ANPAM)









































Abscontrol

]times 100
































31 Mean comparison











Wild Soxhlet

M 2158

plusmn

093a

3472

plusmn

165a

2502 plusmn

153a

1283 plusmn

029i


008p




plusmn

109d

3088

plusmn

120c

1655 plusmn

206d

1400 plusmn

050h


050j


035k

EA 1427

plusmn

080g

2513

plusmn

111g

1219 plusmn

124g

1500 plusmn

050g

1217 plusmn

029g

1250 plusmn

050h

1183 plusmn

076h

19003 plusmn

045h

H 816 plusmn

066j

1500

plusmn 193j

935 plusmn

234j

1783 plusmn

076e

1417 plusmn

029e

1450 plusmn

050f

1300 plusmn

050f

26500 plusmn

020e

Maceration

M 1906

plusmn 130c

3498

plusmn 033a

2225 plusmn

048b

1683 plusmn

076f

1250 plusmn

050g

1367 plusmn

126g

1217 plusmn

104g

5978 plusmn

056n

E 1331 plusmn

062h

2837 plusmn

153e

1398 plusmn 146f

1783 plusmn 076e

1333 plusmn 029f

1400 plusmn 050f

1367 plusmn 029f

13500 plusmn 030j

EA 762 plusmn

062k

2495

plusmn

332h

1128 plusmn

105h

1883 plusmn

029d

1617 plusmn

029c

1650 plusmn

050d

1583 plusmn

076d

21003 plusmn

025g

H 514 plusmn

014m

1340

plusmn

082l

700 plusmn

182l

2167 plusmn

058b

1833 plusmn

029b

1900 plusmn

050b

1717 plusmn

029c

30052 plusmn

050c

Cultivated Soxhlet

M 2009 plusmn

104b

3322 plusmn

191b

2296 plusmn 188b

1450 plusmn 087h

1050 plusmn 050i

1167 plusmn 126i


E 1552

plusmn

081f

2944

plusmn

125d

1477 plusmn

158e

1583 plusmn

076g

1150 plusmn

050h

1200 plusmn

050h

1167 plusmn

029h

16000 plusmn

050i

EA 1318

plusmn

102h

2390

plusmn

085i

1069 plusmn

158i

1733 plusmn

029e

1417 plusmn

029e

1450 plusmn

050f

1383 plusmn

076f

24000 plusmn

030f

H 671 plusmn

129l

1420

plusmn 172k

801 plusmn

264k

2017 plusmn

104c

1617 plusmn

029c

1650 plusmn

050d

1433 plusmn

161e

36000 plusmn

020b

Maceration

M 1684

plusmn 172e

3359

plusmn 063b

2080 plusmn

062c

1853 plusmn

084d

1427 plusmn

093e

1567 plusmn

161e

1433 plusmn

076e

8000 plusmn

050l

E 1147 plusmn

053i

2643 plusmn

155f

1263 plusmn 115g

2017 plusmn 101c

1533 plusmn 104d

1550 plusmn 050e

1550 plusmn 050d

16240 plusmn 026i

EA 625 plusmn

077l

2356

plusmn

305i

1014 plusmn

041i

2177 plusmn

015b

1800 plusmn

050b

1850 plusmn

050c

1800 plusmn

132b

27603 plusmn

025d

H 417 plusmn

030n

1248

plusmn

117m


098a

2017 plusmn

076a

2033 plusmn

104a

1950 plusmn

050a

39000 plusmn

100a




























































[3940]




Yield

0909 0936 -0871 -0911 -0872 -0868 -0927

TPC

0915 -0720 -0764 -0719 -0671 -0958

TFC

-0730 -0782 -0718 -0743 -0933

E coli

0981 0979 0975 0769

S aureus

0994 0984 0813

B subtilis

0979 0765

C albicans

0740

IC50














A B

C











component

4 Conclusions













Funding


Acknowledgments






References

















416641-700077-8



















httpsdoiorg101007978-3-030-38792-1_15















httpsdoiorg101016S0076-6879(99)99017-1











rthern_Morocco











httpsdoiorg10115520215571501


































































httpsdoiorg101007s10341-018-0401-y










223



of_Chemistry






46 httpsdoiorg101007s10341-019-00457-x






























httpsdoiorg10115520215138043















































Abscontrol

]times 100
































31 Mean comparison











Wild Soxhlet

M 2158

plusmn

093a

3472

plusmn

165a

2502 plusmn

153a

1283 plusmn

029i


008p




plusmn

109d

3088

plusmn

120c

1655 plusmn

206d

1400 plusmn

050h


050j


035k

EA 1427

plusmn

080g

2513

plusmn

111g

1219 plusmn

124g

1500 plusmn

050g

1217 plusmn

029g

1250 plusmn

050h

1183 plusmn

076h

19003 plusmn

045h

H 816 plusmn

066j

1500

plusmn 193j

935 plusmn

234j

1783 plusmn

076e

1417 plusmn

029e

1450 plusmn

050f

1300 plusmn

050f

26500 plusmn

020e

Maceration

M 1906

plusmn 130c

3498

plusmn 033a

2225 plusmn

048b

1683 plusmn

076f

1250 plusmn

050g

1367 plusmn

126g

1217 plusmn

104g

5978 plusmn

056n

E 1331 plusmn

062h

2837 plusmn

153e

1398 plusmn 146f

1783 plusmn 076e

1333 plusmn 029f

1400 plusmn 050f

1367 plusmn 029f

13500 plusmn 030j

EA 762 plusmn

062k

2495

plusmn

332h

1128 plusmn

105h

1883 plusmn

029d

1617 plusmn

029c

1650 plusmn

050d

1583 plusmn

076d

21003 plusmn

025g

H 514 plusmn

014m

1340

plusmn

082l

700 plusmn

182l

2167 plusmn

058b

1833 plusmn

029b

1900 plusmn

050b

1717 plusmn

029c

30052 plusmn

050c

Cultivated Soxhlet

M 2009 plusmn

104b

3322 plusmn

191b

2296 plusmn 188b

1450 plusmn 087h

1050 plusmn 050i

1167 plusmn 126i


E 1552

plusmn

081f

2944

plusmn

125d

1477 plusmn

158e

1583 plusmn

076g

1150 plusmn

050h

1200 plusmn

050h

1167 plusmn

029h

16000 plusmn

050i

EA 1318

plusmn

102h

2390

plusmn

085i

1069 plusmn

158i

1733 plusmn

029e

1417 plusmn

029e

1450 plusmn

050f

1383 plusmn

076f

24000 plusmn

030f

H 671 plusmn

129l

1420

plusmn 172k

801 plusmn

264k

2017 plusmn

104c

1617 plusmn

029c

1650 plusmn

050d

1433 plusmn

161e

36000 plusmn

020b

Maceration

M 1684

plusmn 172e

3359

plusmn 063b

2080 plusmn

062c

1853 plusmn

084d

1427 plusmn

093e

1567 plusmn

161e

1433 plusmn

076e

8000 plusmn

050l

E 1147 plusmn

053i

2643 plusmn

155f

1263 plusmn 115g

2017 plusmn 101c

1533 plusmn 104d

1550 plusmn 050e

1550 plusmn 050d

16240 plusmn 026i

EA 625 plusmn

077l

2356

plusmn

305i

1014 plusmn

041i

2177 plusmn

015b

1800 plusmn

050b

1850 plusmn

050c

1800 plusmn

132b

27603 plusmn

025d

H 417 plusmn

030n

1248

plusmn

117m


098a

2017 plusmn

076a

2033 plusmn

104a

1950 plusmn

050a

39000 plusmn

100a




























































[3940]




Yield

0909 0936 -0871 -0911 -0872 -0868 -0927

TPC

0915 -0720 -0764 -0719 -0671 -0958

TFC

-0730 -0782 -0718 -0743 -0933

E coli

0981 0979 0975 0769

S aureus

0994 0984 0813

B subtilis

0979 0765

C albicans

0740

IC50














A B

C











component

4 Conclusions













Funding


Acknowledgments






References

















416641-700077-8



















httpsdoiorg101007978-3-030-38792-1_15















httpsdoiorg101016S0076-6879(99)99017-1











rthern_Morocco











httpsdoiorg10115520215571501


































































httpsdoiorg101007s10341-018-0401-y










223



of_Chemistry






46 httpsdoiorg101007s10341-019-00457-x






























httpsdoiorg10115520215138043




































31 Mean comparison











Wild Soxhlet

M 2158

plusmn

093a

3472

plusmn

165a

2502 plusmn

153a

1283 plusmn

029i


008p




plusmn

109d

3088

plusmn

120c

1655 plusmn

206d

1400 plusmn

050h


050j


035k

EA 1427

plusmn

080g

2513

plusmn

111g

1219 plusmn

124g

1500 plusmn

050g

1217 plusmn

029g

1250 plusmn

050h

1183 plusmn

076h

19003 plusmn

045h

H 816 plusmn

066j

1500

plusmn 193j

935 plusmn

234j

1783 plusmn

076e

1417 plusmn

029e

1450 plusmn

050f

1300 plusmn

050f

26500 plusmn

020e

Maceration

M 1906

plusmn 130c

3498

plusmn 033a

2225 plusmn

048b

1683 plusmn

076f

1250 plusmn

050g

1367 plusmn

126g

1217 plusmn

104g

5978 plusmn

056n

E 1331 plusmn

062h

2837 plusmn

153e

1398 plusmn 146f

1783 plusmn 076e

1333 plusmn 029f

1400 plusmn 050f

1367 plusmn 029f

13500 plusmn 030j

EA 762 plusmn

062k

2495

plusmn

332h

1128 plusmn

105h

1883 plusmn

029d

1617 plusmn

029c

1650 plusmn

050d

1583 plusmn

076d

21003 plusmn

025g

H 514 plusmn

014m

1340

plusmn

082l

700 plusmn

182l

2167 plusmn

058b

1833 plusmn

029b

1900 plusmn

050b

1717 plusmn

029c

30052 plusmn

050c

Cultivated Soxhlet

M 2009 plusmn

104b

3322 plusmn

191b

2296 plusmn 188b

1450 plusmn 087h

1050 plusmn 050i

1167 plusmn 126i


E 1552

plusmn

081f

2944

plusmn

125d

1477 plusmn

158e

1583 plusmn

076g

1150 plusmn

050h

1200 plusmn

050h

1167 plusmn

029h

16000 plusmn

050i

EA 1318

plusmn

102h

2390

plusmn

085i

1069 plusmn

158i

1733 plusmn

029e

1417 plusmn

029e

1450 plusmn

050f

1383 plusmn

076f

24000 plusmn

030f

H 671 plusmn

129l

1420

plusmn 172k

801 plusmn

264k

2017 plusmn

104c

1617 plusmn

029c

1650 plusmn

050d

1433 plusmn

161e

36000 plusmn

020b

Maceration

M 1684

plusmn 172e

3359

plusmn 063b

2080 plusmn

062c

1853 plusmn

084d

1427 plusmn

093e

1567 plusmn

161e

1433 plusmn

076e

8000 plusmn

050l

E 1147 plusmn

053i

2643 plusmn

155f

1263 plusmn 115g

2017 plusmn 101c

1533 plusmn 104d

1550 plusmn 050e

1550 plusmn 050d

16240 plusmn 026i

EA 625 plusmn

077l

2356

plusmn

305i

1014 plusmn

041i

2177 plusmn

015b

1800 plusmn

050b

1850 plusmn

050c

1800 plusmn

132b

27603 plusmn

025d

H 417 plusmn

030n

1248

plusmn

117m


098a

2017 plusmn

076a

2033 plusmn

104a

1950 plusmn

050a

39000 plusmn

100a




























































[3940]




Yield

0909 0936 -0871 -0911 -0872 -0868 -0927

TPC

0915 -0720 -0764 -0719 -0671 -0958

TFC

-0730 -0782 -0718 -0743 -0933

E coli

0981 0979 0975 0769

S aureus

0994 0984 0813

B subtilis

0979 0765

C albicans

0740

IC50














A B

C











component

4 Conclusions













Funding


Acknowledgments






References

















416641-700077-8



















httpsdoiorg101007978-3-030-38792-1_15















httpsdoiorg101016S0076-6879(99)99017-1











rthern_Morocco











httpsdoiorg10115520215571501


































































httpsdoiorg101007s10341-018-0401-y










223



of_Chemistry






46 httpsdoiorg101007s10341-019-00457-x






























httpsdoiorg10115520215138043










plusmn

109d

3088

plusmn

120c

1655 plusmn

206d

1400 plusmn

050h


050j


035k

EA 1427

plusmn

080g

2513

plusmn

111g

1219 plusmn

124g

1500 plusmn

050g

1217 plusmn

029g

1250 plusmn

050h

1183 plusmn

076h

19003 plusmn

045h

H 816 plusmn

066j

1500

plusmn 193j

935 plusmn

234j

1783 plusmn

076e

1417 plusmn

029e

1450 plusmn

050f

1300 plusmn

050f

26500 plusmn

020e

Maceration

M 1906

plusmn 130c

3498

plusmn 033a

2225 plusmn

048b

1683 plusmn

076f

1250 plusmn

050g

1367 plusmn

126g

1217 plusmn

104g

5978 plusmn

056n

E 1331 plusmn

062h

2837 plusmn

153e

1398 plusmn 146f

1783 plusmn 076e

1333 plusmn 029f

1400 plusmn 050f

1367 plusmn 029f

13500 plusmn 030j

EA 762 plusmn

062k

2495

plusmn

332h

1128 plusmn

105h

1883 plusmn

029d

1617 plusmn

029c

1650 plusmn

050d

1583 plusmn

076d

21003 plusmn

025g

H 514 plusmn

014m

1340

plusmn

082l

700 plusmn

182l

2167 plusmn

058b

1833 plusmn

029b

1900 plusmn

050b

1717 plusmn

029c

30052 plusmn

050c

Cultivated Soxhlet

M 2009 plusmn

104b

3322 plusmn

191b

2296 plusmn 188b

1450 plusmn 087h

1050 plusmn 050i

1167 plusmn 126i


E 1552

plusmn

081f

2944

plusmn

125d

1477 plusmn

158e

1583 plusmn

076g

1150 plusmn

050h

1200 plusmn

050h

1167 plusmn

029h

16000 plusmn

050i

EA 1318

plusmn

102h

2390

plusmn

085i

1069 plusmn

158i

1733 plusmn

029e

1417 plusmn

029e

1450 plusmn

050f

1383 plusmn

076f

24000 plusmn

030f

H 671 plusmn

129l

1420

plusmn 172k

801 plusmn

264k

2017 plusmn

104c

1617 plusmn

029c

1650 plusmn

050d

1433 plusmn

161e

36000 plusmn

020b

Maceration

M 1684

plusmn 172e

3359

plusmn 063b

2080 plusmn

062c

1853 plusmn

084d

1427 plusmn

093e

1567 plusmn

161e

1433 plusmn

076e

8000 plusmn

050l

E 1147 plusmn

053i

2643 plusmn

155f

1263 plusmn 115g

2017 plusmn 101c

1533 plusmn 104d

1550 plusmn 050e

1550 plusmn 050d

16240 plusmn 026i

EA 625 plusmn

077l

2356

plusmn

305i

1014 plusmn

041i

2177 plusmn

015b

1800 plusmn

050b

1850 plusmn

050c

1800 plusmn

132b

27603 plusmn

025d

H 417 plusmn

030n

1248

plusmn

117m


098a

2017 plusmn

076a

2033 plusmn

104a

1950 plusmn

050a

39000 plusmn

100a




























































[3940]




Yield

0909 0936 -0871 -0911 -0872 -0868 -0927

TPC

0915 -0720 -0764 -0719 -0671 -0958

TFC

-0730 -0782 -0718 -0743 -0933

E coli

0981 0979 0975 0769

S aureus

0994 0984 0813

B subtilis

0979 0765

C albicans

0740

IC50














A B

C











component

4 Conclusions













Funding


Acknowledgments






References

















416641-700077-8



















httpsdoiorg101007978-3-030-38792-1_15















httpsdoiorg101016S0076-6879(99)99017-1











rthern_Morocco











httpsdoiorg10115520215571501


































































httpsdoiorg101007s10341-018-0401-y










223



of_Chemistry






46 httpsdoiorg101007s10341-019-00457-x






























httpsdoiorg10115520215138043





















































[3940]




Yield

0909 0936 -0871 -0911 -0872 -0868 -0927

TPC

0915 -0720 -0764 -0719 -0671 -0958

TFC

-0730 -0782 -0718 -0743 -0933

E coli

0981 0979 0975 0769

S aureus

0994 0984 0813

B subtilis

0979 0765

C albicans

0740

IC50














A B

C











component

4 Conclusions













Funding


Acknowledgments






References

















416641-700077-8



















httpsdoiorg101007978-3-030-38792-1_15















httpsdoiorg101016S0076-6879(99)99017-1











rthern_Morocco











httpsdoiorg10115520215571501


































































httpsdoiorg101007s10341-018-0401-y










223



of_Chemistry






46 httpsdoiorg101007s10341-019-00457-x






























httpsdoiorg10115520215138043










Yield

0909 0936 -0871 -0911 -0872 -0868 -0927

TPC

0915 -0720 -0764 -0719 -0671 -0958

TFC

-0730 -0782 -0718 -0743 -0933

E coli

0981 0979 0975 0769

S aureus

0994 0984 0813

B subtilis

0979 0765

C albicans

0740

IC50














A B

C











component

4 Conclusions













Funding


Acknowledgments






References

















416641-700077-8



















httpsdoiorg101007978-3-030-38792-1_15















httpsdoiorg101016S0076-6879(99)99017-1











rthern_Morocco











httpsdoiorg10115520215571501


































































httpsdoiorg101007s10341-018-0401-y










223



of_Chemistry






46 httpsdoiorg101007s10341-019-00457-x






























httpsdoiorg10115520215138043

















component

4 Conclusions













Funding


Acknowledgments






References

















416641-700077-8



















httpsdoiorg101007978-3-030-38792-1_15















httpsdoiorg101016S0076-6879(99)99017-1











rthern_Morocco











httpsdoiorg10115520215571501


































































httpsdoiorg101007s10341-018-0401-y










223



of_Chemistry






46 httpsdoiorg101007s10341-019-00457-x






























httpsdoiorg10115520215138043













416641-700077-8



















httpsdoiorg101007978-3-030-38792-1_15















httpsdoiorg101016S0076-6879(99)99017-1











rthern_Morocco











httpsdoiorg10115520215571501


































































httpsdoiorg101007s10341-018-0401-y










223



of_Chemistry






46 httpsdoiorg101007s10341-019-00457-x






























httpsdoiorg10115520215138043














httpsdoiorg10115520215571501


































































httpsdoiorg101007s10341-018-0401-y










223



of_Chemistry






46 httpsdoiorg101007s10341-019-00457-x






























httpsdoiorg10115520215138043
























httpsdoiorg101007s10341-018-0401-y










223



of_Chemistry






46 httpsdoiorg101007s10341-019-00457-x






























httpsdoiorg10115520215138043

















httpsdoiorg10115520215138043







Documents

Effects of Extraction Technique and Solvent on