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\def\RunningAuthor{Zineb Aliat et al.}
\firstPage{3075}
\articleType{Original Article}
\receivedDate{26 Mar 2020}
\acceptedDate{26 Apr 2020}
\revisedDate{15 Apr 2020}
\journalVolume{2020, 11}
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\title{Phytochemical screening, antioxidant and antilithiasis activities of \textit{Juncus acutus}}
\author{Zineb~Aliat\textsuperscript{*}\textsuperscript{1},  
            Lamyae~Yachi\textsuperscript{1},  
            Houda~Attjioui\textsuperscript{1},  
            Amine~Cheikh\textsuperscript{2},  
            Miloud~EL~Karbane\textsuperscript{1},  
            Rachid~Nejjari\textsuperscript{3}, 
            Mustapha~Bouatia\textsuperscript{1}~\\[5pt]{\textsuperscript{1}Department of Medicine and Pharmacy, Laboratory of Analytical Chemistry Mohammed V University- Rabat, Morocco}~\\{\textsuperscript{2}Department of Pharmacy Abulcasis University-Rabat, Morocco}~\\{\textsuperscript{3}Department of Medicine and Pharmacy, Team of Formulation and Quality Control of Health Products Mohammed V University- Rabat, Morocco}}

\begin{abstract}
The objective of this work was to perform a phytochemical screening of Juncus acutus and to evaluate its antioxidant activity and antilithiasis activity. Phytochemical screening has demonstrated the presence of catechin, tannins and alkaloids in the various extracts studied. The antioxidant activity of the methanolic and ethyl acetate extracts by ultrasound and maceration was evaluated by the method of DPPH, FRAP and phosphomolybdate. The ethyl acetate extract by ultrasound, the methanolic extract by maceration and ultrasound respectively showed IC 50 with DPPH of 1,449mg / ml, 1,535mg / ml and 4,771mg / ml and a reducing power was observed at concentrations of 419,561mgBHA /100g, 207,143mgBHA /100g and 142,20mgBHA /100g by FRAP and at concentrations of 839,470mgAAE/100g, 283,260mgAAE/100g and 141,018mgAAE/100g by the phosphomolybdate test. The determination of the total polyphenols of the ethyl acetate extract by ultrasound, the methanolic extract by maceration and by ultrasound presented respectively contents of 2,428 mg GAE / g, 1,960mg GAE / g and 1,172mg GAE / g, likewise, the flavonoid assay showed respectively concentrations of 14,469mg EQ/g, 6,466 mg EQ/g and 3,143 mg EQ/g. Antilithiasis activity was evaluated on carbapatite, cystine and uric acid stones. The mass loss of uric acid, carbapatite and cystine stones was respectively 42\%, 7\% and 16\%. 
\end{abstract}\def\keywordstitle{Keywords}

\begin{keywords}Juncus acutus,\newline Antioxydant activity,\newline Antilithiasis activity,\newline flavoinoids,\newline polyphenols
\end{keywords}
\twocolumn[ \maketitle {\printKwdAbsBox}]
\makeatletter{\raggedright\textsuperscript{*}Corresponding Author\par Name:\ Zineb~Aliat~\\ Phone:\ +212673919725~\\  Email:\ zineb.aliat@gmail.com
        \par}\vspace*{-11pt}\hrulefill\par{\fontsize{12}{14}\selectfont ISSN: 0975-7538}\par% 
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\section{Introduction}
Medicinal plants are the main human remedy for the manufacture of pharmaceutical remedies. Indeed, natural substances from plants have multiple interests put to use in the traditional care system and in different industries. Among these compounds, we find to a large extent the secondary metabolites which are especially illustrated in therapeutics\unskip~\citep{793109:18894255}. In our study, we will focus on antioxidant and antilithiasis activities of \textit{Juncus acutus}. 

The most popular antioxidant are ascorbic acid (Vitamin C),  -carotene (Provitamin A), tocopherol (Vitamine E) and phenolic compounds. Indeed, most synthetic or naturally occurring antioxidants have hydroxyphenolic groups in their structures and antioxidant properties are attributed in part to the ability of these natural compounds to trap free radicals such as hydroxyl radicals and superoxides \unskip~\citep{793109:18894262,793109:18894248,793109:18894254}. 

Several methods are used to evaluate, in vitro and in vivo, the antioxidant activity by trapping different radicals, such as peroxides by the ORAC (Oxygen Radical Absorbance capacity) and TRAP (Total Radical Trapping Antioxidant Parameter) \unskip~\citep{793109:18894247} methods; Ferric ions by the FRAP method (Ferric ion Reducing Antioxidant Parameter)\unskip~\citep{793109:18894349}; or the ABTS (ammonium salt of 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid) radicals \unskip~\citep{793109:18894242}, as well as the method using the free radical DPPH (Diphenyl-picrylhydrazyl) \unskip~\citep{793109:18894254,793109:18894260}. 

In our study, three methods will be used to evaluate this activity: FRAP, DPPH and phosphomolybdate test.

In Morocco as reported by \unskip~\citep{793109:18894259}, the preponderant type of stone was calcium oxalate monohydrate (56.4\%), followed by uric acid (18,1\%) , carbapatite (7,9\%) and cystine (0,6\%).

In this study, we chose to study the effect of \textit{Juncus acutus} extracts on renal carbapatite, uric acid and cystine stones.

This study has set as main objectives,
  
  \begin{enumerate}
  \item \relax Performing a phytochemical screening to search for certain phytochemical groups. 
  \item \relax Determination of the total polyphenols and total flavonoids contained in the plant. 
  \item \relax Study of the antioxidant activity of methanolic extracts and ethyl acetate of Juncus acutus. 
  \item \relax Study of the antilithiasis activity of methanolic extracts and ethyl acetate of Juncus acutus.
  \end{enumerate}
  
    
\section{Experimental section}
\textbf{Plant material}

The sample collected comes from the southern region of Morocco.In the laboratory we grind the seeds of \textit{Juncus acutus} with a grinder until a fine powder is obtained.

\textbf{Preparation of the extracts}

We used two types of extraction,  maceration with stirring for 24 hours and extraction with ultrasound for 60 minutes.

\textbf{Maceration }

40 g of \textit{Juncus acutus} powder are extracted into 100 ml of ethyl acetate by maceration with stirring for 24 hours at room temperature, after filtration and drying of the same powder, a second extraction in 100 ml of methanol has been carried out in same conditions as the previous one.

\textbf{Ultrasound }

40 g of \textit{Juncus acutus} powder are extracted into 100 ml of ethyl acetate under ultrasound for 60 minutes at 39 \ensuremath{^\circ}C. After filtration and drying of the same powder, a second extraction in 100 ml of methanol was carried out in the same conditions than the previous one. The 4 filtrates are concentrated in an oven at 40 \ensuremath{^\circ} C in order to get rid of the solvent and to recover the extracts. The 4 extracts thus recovered are stored in sterile refrigerator bottles.


\begin{table*}[!htbp]
\caption{\boldmath {Screening of some phytochemical groups of 4 extracts of Juncus acutus} }
\label{tw-dfb946efd362}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
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\centering 
\begin{tabulary}{\linewidth}{LLLLL}
\tbltoprule \rowcolor{kwdboxcolor}Constituents & \multicolumn{2}{p{\dimexpr(\mcWidth{2}+\mcWidth{3})}}{\cAlignHack Maceration} & \multicolumn{2}{p{\dimexpr(\mcWidth{4}+\mcWidth{5})}}{\cAlignHack Under ultrasound}\\
\rowcolor{kwdboxcolor} & Ethylacetate Extract & Methanol Extract & Ethylacetate Extract & Methanol Extract\\
\tblmidrule 
Sterols and polyterpenes &
  + &
  + &
  - &
  -\\
Polyphenols &
  - &
  - &
  - &
  -\\
Flavonoids &
  - &
  - &
  - &
  -\\
Catechictanins &
  + &
  + &
  + &
  +\\
Gallic tannins &
  - &
  - &
  - &
  -\\
Alkaloids &
  + &
  + &
  + &
  +\\
Saponosides  &
  - &
  - &
  - &
  -\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{(+) indicates pr{\'e}sence ; (-) indicates absence}
\end{tablenotes}
\end{threeparttable}

\end{table*}
\textbf{Phytochemical characterization}

We carried out qualitative reactions separately on our different extracts of juncus acutus for the screening of certain phytochemical groups using the methods of \unskip~\citep{793109:18894243,793109:18894245} thus reported by \unskip~\citep{793109:18894261,793109:18894249}. 

\textbf{Determination of total polyphenols}

The total phenolic compound content was determined using the FolinCiocalteu method \unskip~\citep{793109:18894241}. 1.25 ml of Folin reagent (ten times diluted) was added to 250 \ensuremath{\mu }l of extract or standard (prepared in methanol) with concentrations ranging from 0.3 to 3 mg / ml. After 4 min, 1 ml of a sodium carbonate solution (7.5g/100 ml) is added to the preceding mixture. The whole is incubated in the dark at room temperature for 30 min. The absorbance was measured at 765 nm. The concentration of the total polyphenols is expressed in milligrams of gallic acid equivalent per gram of extract (mg GAE/g extract).

\textbf{Dosage of total flavonoids}

The acetic trichloride method \unskip~\citep{793109:18894253} is used to estimate the total flavono{\"{\i}}ds in the extracts. 1.5 ml of the AlCl\ensuremath{_{3}} solution (2\%) is added to 1.5 ml of the sample or standard. After 1 hour of reaction in the absence of light, the optical densitywas read at 415 nm. The flavono{\"{\i}}d concentration was expressed as milligram equivalent of quercetin per gram of extract (mg EQ/g extract).


\begin{table*}[!htbp]
\caption{\boldmath {: Total polyphenol contents expressed in milligrams equivalent of gallic acid per gram of Juncus acutus powder} }
\label{tw-d9aa5763e9b2}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
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\begin{tabulary}{\linewidth}{p{\dimexpr.60230000000000004\linewidth-2\tabcolsep}p{\dimexpr.39769999999999996\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}Extract of Juncus acutus  & Total Polyphenol contents (mg GAE/g) \\
\tblmidrule 
Extraction by maceration \mbox{}\protect\newline - MethanolExtract &
  1,960\\
Extraction by ultrasound \mbox{}\protect\newline - Ethylacetate Extract \mbox{}\protect\newline - Methanol Extract &
  2,428 \mbox{}\protect\newline 1,172\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{The ethyl acetate extract by ultrasound shows the highest content of phenolic compounds 2,428 mg GAE /g.}
\end{tablenotes}
\end{threeparttable}

\end{table*}
\textbf{Antioxidant activity}

\textbf{Free radical scavenging capacity}

\textbf{DPPH}

DPPH, a free radical of violet color, is reduced to a yellow compound in the presence of anti-free radical compounds\unskip~\citep{793109:18894252} . In the test tubes, 500 \ensuremath{\mu }l of each extract or standard of different concentrations (2.5, 5, 10 and 15 \ensuremath{\mu }g / ml) and 2 ml of DPPH methanol solution (50 \ensuremath{\mu }M) were introduced, after vortexing, the tubes are incubated in the dark at room temperature for 30 minutes. The measurement was carried out using a UV-V spectrophotometer at a wavelength of 517 nm.

IC50 values calculated to determine the 50\% inhibition of DPPH radicals. The BHT was used as standard.

\textbf{Ferric reducing antioxidant power assay}

\textbf{FRAP}

The reducing power of our extracts has been determined by using the method of Florentine Marie-Chantal \unskip~\citep{793109:18894258}. The volume of 250\ensuremath{\mu }l of each extract or the standard of concentrations (0.01 to 0.1mg/ml) was added to the mixture of 1.25ml of phosphate buffer(0.2M, pH6.6) and 1.25ml of potassium ferricyanide (1\%). After incubation for 20 minutes in a water bath at 50\ensuremath{^\circ}C, 1.25ml of the acetic trichloride acid (10\%) was added to the mixture and then centrifuged at 3000 rpm for 10 minutes. The supernatant (1.25ml) was added to a mixture of 1.25ml of distilled water and 250\ensuremath{\mu }l of ferric chloride (0.1\%). The absorbance was measured at the spectrophotometer at 700nm. Increased absorbance of the reaction mixture suggests a high reducing power. The results were expressed as milligram equivalent of Butylhydroxyanisole(BHA) per 100 grams of extract (mg BHA/100g extract).

\textbf{Phosphomolybdate}

The test is based on the reduction of Mo(VI) to Mo(V) by the analyte of the sample and subsequent formation of a green Mo(V) phosphate complex at acid pH. The volume of 0.3ml of the extract solution is added to 3ml of the reagent (0.6M sulfuric acid, 28mM sodium phosphate and 4mM ammonium molybdate). The tube is plugged and incubated in a water bath at 95 \ensuremath{^\circ} C for 90 minutes. After cooling to room temperature, the absorbance of the solution is measured at 695nm against a blank with a UV-Visible spectrophotometer. Ascorbic acid was used as standard. The results were expressed as milligram equivalent of Ascorbic acid per 100 grams of extract (mg AAE/100g extract).

\textbf{Antilithiasis activity}

Twelve kidney stones of carbapatite; cystine and pure uric acid from southern Morocco were selected after Fourier transform infrared spectrophotometry (FTIR) analysis. Their mass was 330\ensuremath{\pm}12mg. An extract of the plant was prepared by infusion of 3g of the dry plant in 100ml of boiling physiological solution for 30 minutes. The NaCl solution of 9g/l being used by elsewhere as a control medium to appreciate the mass modifications of the calculations. After filtration, the extract was partitioned (60mL) into glass Erlenmeyer flasks. The calculation were putin a porous bag of braided fibers and suspended in the extract \unskip~\citep{793109:18894251}.

The loss of mass of the kidney stones was evaluated by weighing the calculation after drying at 40\ensuremath{^\circ}C for 18hours. Each test has been performed 3 times.

The evaluation of the activity of the extract was determined by calculating the rate of dissolution of the stones after stay in the experimental medium by comparing the residual weight of the stones with their initial weight before incubation with our extract. The percentage of dissolution was determined by the formula,
\begin{eqnarray*}a\%=\frac{\left(W_{initial}-W_{final}\;\right)}{W_{initial}}\times100 \end{eqnarray*}
Where: a\% is the dissolution rate of the calculation and W\ensuremath{_{initial}} and W\ensuremath{_{final}} are the calculation weights before and after incubation with plant extracts. 

\textbf{Statistical analysis}

The results are expressed as mean \ensuremath{\pm} standard deviation (SD) and all analyses were performed in triplicate. The results were compared by one-way ANOVA with bonferoni'spost test. A difference is significant if p\ensuremath{\leq }0,05.


\begin{table*}[!htbp]
\caption{\boldmath {Total flavonoid content expressed in equivalent milligrams of Quercetin per gram of Juncus acutus powder} }
\label{tw-14f99142d710}
\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{LL}
\tbltoprule \rowcolor{kwdboxcolor}Extractof Juncusacutus  & Total flavonoid content (mgQE/g) \\
\tblmidrule 
Extraction by maceration \mbox{}\protect\newline - Methanol Extract &
  6,466\\
Extraction by ultrasound \mbox{}\protect\newline -Ethylacetate Extract \mbox{}\protect\newline - Methanol Extract &
  14,694 \mbox{}\protect\newline 3,143\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

    
\section{Results and Discussion}
\textbf{Extraction yields}

The yield was 0.517\% for the maceration carried out at the first stage with ethyl acetate on the \textit{Juncus acutus }powder, the second extraction carried out with methanol gave a higher yield which is 1.62\%.

For extraction under ultrasound, the yields were 2.9\% for the ethyl acetate extract and 1.16\% for the methanol extract.

Extraction by methanol from the powder of Juncus acutus seeds by ultrasound showed a higher yield than that achieved by maceration, and almost identical yields for extraction with ethyl acetate for both methods.

\textbf{Phytochemical characterization }

The results of the qualitative characterization of our extracts is summarized in Table~\ref{tw-dfb946efd362}.


\bgroup
\fixFloatSize{images/40604cf4-6f8c-4089-a1f9-137517e5a364-upicture1.png}
\begin{figure*}[!htbp]
\centering \makeatletter\IfFileExists{images/40604cf4-6f8c-4089-a1f9-137517e5a364-upicture1.png}{\includegraphics{images/40604cf4-6f8c-4089-a1f9-137517e5a364-upicture1.png}}{}
\makeatother 
\caption{\boldmath {Radical scavenging and antioxidant activities of different extracts of Juncus acutus using DPPH assay}}
\label{f-f9da536d2f2e}
\end{figure*}
\egroup
The phytochemical screening of the seeds of our sample showed the presence of catechin tannins and alkaloids in all the extracts. Sterols and polyterpenes were present only in the maceration extracts. A study of the aqueous extracts of \textit{Juncus subulatusForsak} revealed the presence of alkaloids, glycosides, saponisides, flavonoids and terpenoids \unskip~\citep{793109:18894250}.

\textbf{Total Polyphenol content}

UV / Visible spectrophotometry made it possible to quantify the level of the polyphenols present in the 3 extracts of \textit{Juncus acutus}.

The calibration curve established using different concentrations of gallic acid made it possible to estimate the phenolic content of each of the 3 extracts. The concentration of the phenolic compounds was then calculated from the calibration curve and expressed in mg GAE/g.

The total polyphenol content in the various extracts of \textit{Juncus acutus} is given in Table~\ref{tw-d9aa5763e9b2}.


\bgroup
\fixFloatSize{images/f55a2e61-09e4-4502-8e9c-aefb1871404c-upicture2.png}
\begin{figure*}[!htbp]
\centering \makeatletter\IfFileExists{images/f55a2e61-09e4-4502-8e9c-aefb1871404c-upicture2.png}{\includegraphics{images/f55a2e61-09e4-4502-8e9c-aefb1871404c-upicture2.png}}{}
\makeatother 
\caption{\boldmath {The DPPH radical scavenging capacity (IC\ensuremath{_{50}}) values of different extracts of Juncus acutus}}
\label{f-8ba98bdfb0c1}
\end{figure*}
\egroup
The ethyl acetate extract by ultrasound shows the highest content of phenolic compounds 2,428 mg GAE /g. 

\textbf{Total flavonoid content}

The calibration curve is plotted using different concentrations of Quercetin.The concentration of flavonoids in \textit{Juncus acutus} extracts is determined by reference to the calibration curve.

The content of flavonoids is expressed in milligrams equivalent of quercetin per gram of extract(mg QE/g extract).

The content of total flavonoids in the various extracts of \textit{Juncus acutus} is given in Table~\ref{tw-14f99142d710}.


\bgroup
\fixFloatSize{images/b3029be5-4142-4dae-9e69-d592a7ca8ce4-upicture3.png}
\begin{figure*}[!htbp]
\centering \makeatletter\IfFileExists{images/b3029be5-4142-4dae-9e69-d592a7ca8ce4-upicture3.png}{\includegraphics{images/b3029be5-4142-4dae-9e69-d592a7ca8ce4-upicture3.png}}{}
\makeatother 
\caption{\boldmath {Reducing power of extracts of Juncus acutus}}
\label{f-379076226c11}
\end{figure*}
\egroup
The ultrasound ethyl acetate extract shows the highest flavonoide content with a rate of 14,694 mg QE / g.

The total polyphenol and total flavonoids contents of our extracts were higher than those found in other studies of Juncus acutus\unskip~\citep{793109:18894246} and Juncus effuses \unskip~\citep{793109:18894257}. 


\bgroup
\fixFloatSize{images/0e60fe96-e6f7-42d9-a9dd-d25a98778aa3-upicture4.png}
\begin{figure*}[!htbp]
\centering \makeatletter\IfFileExists{images/0e60fe96-e6f7-42d9-a9dd-d25a98778aa3-upicture4.png}{\includegraphics{images/0e60fe96-e6f7-42d9-a9dd-d25a98778aa3-upicture4.png}}{}
\makeatother 
\caption{\boldmath {Phosphomolybdate test of Juncus acutus}}
\label{f-17d97146e614}
\end{figure*}
\egroup
\textbf{DPPH }

The different concentrations of the ethyl acetate extract by ultrasound (2.5; 5; 10 and 15\ensuremath{\mu }g/ml) showed dose-dependent antioxidant activities with 52,30\%; 59,.14\%; 71.83\%; and 83,82\% inhibition.As for the methanolic extract, it showed rates of 50.46\%; 57,35\%, 66,02\% and 75,35\% inhibition for maceration extraction, and rates of 48,30\%; 50,45\%; 53,36\% and 56,65\% for extraction by ultrasonics (Figure~\ref{f-f9da536d2f2e}).


\bgroup
\fixFloatSize{images/f31752ea-01cc-4fc8-9edd-1e99fd774764-upicture5.png}
\begin{figure*}[!htbp]
\centering \makeatletter\IfFileExists{images/f31752ea-01cc-4fc8-9edd-1e99fd774764-upicture5.png}{\includegraphics{images/f31752ea-01cc-4fc8-9edd-1e99fd774764-upicture5.png}}{}
\makeatother 
\caption{\boldmath {Dissolution rate of cystine stones in Juncus acutus extract and control solution}}
\label{f-ec3f0a8d4969}
\end{figure*}
\egroup
The antioxidant capacity of the various extracts was determined from the IC50, the concentration necessary to reduce 50\% of the DPPH radical. The smaller the value if IC50, the greater the activity of the extract tested \unskip~\citep{793109:18894255}. The IC50 values found for extracts of  \textit{Juncus acutus }are shown in Figure~\ref{f-8ba98bdfb0c1}. The most important antioxidant activity was noted for the ultrasound ethyl acetate extract of \textit{Juncus acutus} with an IC 50 of 1,459 mg / ml.


\bgroup
\fixFloatSize{images/3569e0b7-cdf0-4394-8f36-797a38232972-upicture6.png}
\begin{figure*}[!htbp]
\centering \makeatletter\IfFileExists{images/3569e0b7-cdf0-4394-8f36-797a38232972-upicture6.png}{\includegraphics{images/3569e0b7-cdf0-4394-8f36-797a38232972-upicture6.png}}{}
\makeatother 
\caption{\boldmath {Dissolution rate of uric acid stones in Juncus acutus extract and control solution}}
\label{f-786e539d949a}
\end{figure*}
\egroup
\textbf{FRAP (Reducing power assay)}

Ferric reducing power can serve as significant indicator of the potential antioxidant. Reduction capacities of the accumulated \textit{Juncus acutus }extracts were determined by reference to the calibration curve of the reference anti-oxidant BHA. The most important reducing power was noted for the ultrasound ethyl acetate extract with a concentration of 419,561mg BHAE / 100g. (Figure~\ref{f-379076226c11})


\bgroup
\fixFloatSize{images/942b4b57-7b24-4930-b3b7-38ccea6cd88c-upicture7.png}
\begin{figure*}[!htbp]
\centering \makeatletter\IfFileExists{images/942b4b57-7b24-4930-b3b7-38ccea6cd88c-upicture7.png}{\includegraphics{images/942b4b57-7b24-4930-b3b7-38ccea6cd88c-upicture7.png}}{}
\makeatother 
\caption{\boldmath {Dissolution rate of carbapatite stones in Juncus acutus extract and control solution}}
\label{f-c6026b4f9eab}
\end{figure*}
\egroup
\textbf{Phosphomolybdate test}

The test is based on the reduction of Mo(VI) to Mo(V) by the analyte of the sample and subsequent formation of a green Mo(V) phosphate complex at acid pH. The reduction capacities of \textit{Juncus acutus} extracts were determined by reference to the calibration curve of vitamin C.The most important reducing power was noted for the ultrasound ethyl acetate extract with a rate of 839,470 mg AAE/100g (Figure~\ref{f-17d97146e614}).

Our results showed greater antioxidant activity than those found in other studies performed on Juncus Bufonius\unskip~\citep{793109:18894244}, Juncus acutus\unskip~\citep{793109:18894246}\textit{. }It is also noted that the highest total polyphenol content as well as total flavonoids content and the most important antioxidant activity was observed in the ethyl acetate extract by ultrasound.\textit{}

\textbf{Antilithiasis activity}

The mass loss of the cystine stones was 60 mg \ensuremath{\pm} 5 mg for \textit{Juncus acutus} and 20 mg \ensuremath{\pm} 2 mg for the control solution (16\% vs 4\% for the control solution (Figure~\ref{f-ec3f0a8d4969}).

This mass loss was 158 mg \ensuremath{\pm} 35 mg for \textit{Juncus acutus} and 61 mg \ensuremath{\pm} 3 mg for the control solution (42\% vs 20\% for the control solution (Figure~\ref{f-786e539d949a}) for uric acid calculations. 

And it was 25 mg \ensuremath{\pm} 4 mg for \textit{Juncus acutus} and 19 mg \ensuremath{\pm} 2 mg for the control solution (7\% vs 5\% for the control solution (Figure~\ref{f-c6026b4f9eab}) for carbapatite calculations.

The mass of the three stones during the incubation with the extract of \textit{Juncus acutus} and the control solution during the first three weeks was little changed, in fact the dissolution rate remained low (p{\textgreater} 0.05) and started to increase between fourth and sixth week (p = 0.01).

Our results did not show a significant difference between the extract and the control solution with respect to the loss of mass of the cystine, carbapatite and uric acid stones over the chosen experimental period. However, the slope of the dissolution curve obtained for the Cystine and uric acid calculations increased sharply in the last weeks compared to the curves observed with the carbapatite calculations. Suggesting that this plant is probably more effective at dissolving Cystine and uric acid stones, but probably requires more time for the dissolution of stones.
    
\section{Conclusions}
This work enabled us first of all to carry out a phytochemical screening, in order to look for some phytochemical groups such as sterols and polyterpenes, polyphenols, flavonoids, catechin and gallic tannins, alkaloids and saponosides; in extracts of \textit{Juncus acutus}. It also allowed us to quantify total flavonoids and total polyphenols. The application of the Folain-Ciocalteu method and the aluminum trichloride method have shown that the total polyphenol content is between 1,172 and 2,428 mg EAG/g; and that the total flavonoids content is between 3,143 and 14.469mgEQ/g.

Regarding the antioxidant activity, we carried out 3 tests: the DPPH test, the FRAP and the phosphomolybdate test. The highest total polyphenol content as well as total flavonoids content and the most imporants antioxidant activity was observed in the ethyl acetate extract by ultrasound. The study of the antioxidant activity of the methanolic extracts and of ethyl acetate of \textit{Juncus  acutus}, showed that they possess a non-negligible antioxidant activity. The study of antilithiasis activity demonstrate that Juncus acutus is probably more effective at dissolving Cystine and uric acid stones, but probably requires more time for the dissolution of stones.

\textbf{ Declaration of interests}

The authors declare no conflict of interest.

\textbf{Funding Support}

None.
    

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