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\begin{document}


\def\RunningAuthor{Kottai Muthu A and Dinesh Babu J}
\firstPage{1857}
\articleType{Original Article}
\receivedDate{08 Dec 2019}
\acceptedDate{22 Jan 2020}
\revisedDate{20 Jan 2020}
\journalVolume{2020, 11}
\journalIssue{2}
\journalDoi{ijrps.v11i2.2093}
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\title{Studying invitro antioxidant effect of different extract of \textit{Premna tomentosa (Linn)} aerial parts}
\author{Kottai~Muthu~A\textsuperscript{*}\textsuperscript{1}, 
            Dinesh~Babu~J\textsuperscript{2}~\\[5pt]{\textsuperscript{1}Department of Pharmacy, Annamalai University, Annamalai Nagar, Chidambaram-608 002, Tamilnadu, India }~\\{\textsuperscript{2}Scient Institute of Pharmacy, Khanapur Village, Ibrahimpatnam, Rangareddy - 501506, Telangana, India}}

\begin{abstract}
The genus Premna (Verbenaceae) comprises a group of more than 200 different trees, distributed in tropical and subtropical areas of the world. \textit{P. tomentosa} (Verbanaceae) is a well-known medicinal plant used extensively for the treatment of various ailments. In the present study, the entire plant of different concentrates of\textit{\space }aerial parts of \textit{Premna tomentosa }was evaluated for its \textit{in-vitro } anti-oxidant potential by superoxide scavenging activity, hydroxyl radical scavenging taking quercetin and ascorbate as the standard correspondingly. The results of antioxidant activity superoxide radical activity and hydroxyl potential were expressed in terms of \% inhibition of generated free radicals respectively with respect to various concentrations. An IC\ensuremath{_{50}} value was recorded; methanolic concentrate of \textit{Premna }\textit{tomentosa }is more effective in superoxide radical activity, hydroxyl radical than that of EA and petroleum ether concentrate. The methanolic concentrate of \textit{Premna  t}\textit{omentosa }and standard exhibited radical scavenging activity possessing IC\ensuremath{_{50\ }}values 225\ensuremath{\mu}g/mL and 60\ensuremath{\mu}g/mL (Superoxide Scavenging Activity), 288\ensuremath{\mu}g/mL and 65\ensuremath{\mu}g/mL (hydroxyl radical activity) correspondingly. All the above invitro studies clearly indicate that the methanolic concentrate of \textit{Premna }\textit{tomentosa }has better free radical scavenging activity. These \textit{invitro} estimations point out that this methanolic concentrate of \textit{Premna } \textit{tomentosa }is a paramount source of expected antioxidant, which may be supportive in preventing the improvement of a variety of free radical induced diseases. 
\end{abstract}\def\keywordstitle{Keywords}

\begin{keywords}Methanolic concentrate,\newline Premna tomentosa,\newline Superoxide radical,\newline hydroxyl radical
\end{keywords}
\twocolumn[ \maketitle {\printKwdAbsBox}]
\makeatletter{\raggedright\textsuperscript{*}Corresponding Author\par Name:\ Kottai~Muthu~A~\\ Phone:\ 9443171712~\\  Email:\ akottaimuthu@gmail.com
        \par}\vspace*{-11pt}\hrulefill\par{\fontsize{12}{14}\selectfont ISSN: 0975-7538}\par% 
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\section{Introduction}
Free radicals induce lipid peroxidation and provokes damage in cell membrane. Incorporation of antioxidants in fats and oils or foods can prevent the deterioration of lipids in foods. Some of these compounds which retard lipid peroxidation are synthetic antioxidants whereas others occur as natural dietary constituents \unskip~\citep{711514:16871692}. The human body is in constant battle to keep from free radicals. The first line of defence is the preventive antioxidants which quench the free radicals generated in the body. An early stage Augmentation of antioxidant status should either prevent or greatly curtail tissue injury \unskip~\citep{711514:16871693}. Due to the adverse effects of synthetic antioxidants on human health, restrictions have been imposed on their usage. Natural antioxidants have been greatly intensified as synthetic antioxidants exert carcinogenic effect \unskip~\citep{711514:16871694}. Plants are enriched with bioactive ingredients which have most of the free radical scavenging and therapeutic function. 

\textit{Premna tomentosa }(Verbanaceae) is a glowing recognized herb used broadly for the cure of different disorders. \textit{Premna }\textit{tomentosa }was used for the cure the different disorders in India \unskip~\citep{711514:16871681}. \textit{Premna }\textit{tomentosa }bark concentrate was used for treatment of hepatic disorders. \textit{Premna tomentosa }leaves concentrates were used as diuretic \unskip~\citep{711514:16871684}, \textit{Premna tomentosa }was used for the treatment of lipid lowering \unskip~\citep{711514:16871688}, antioxidant \unskip~\citep{711514:16871691}, immune modulatory activities \unskip~\citep{711514:16871683} and mitochondrial dysfunction properties against acetaminophen induced rats \unskip~\citep{711514:16871695}. Flavonoids, triterpenoids and steroids were isolated from \textit{Premna }tomentosa \unskip~\citep{711514:16871696,711514:16871679}. Still, no literature are available on the antioxidant activity of \textit{Premna tomentosa }aerial parts\textit{. }Thus, the present study to assess antioxidant activities of \textit{Premna } \textit{tomentosa aerial} parts\textit{.}
    
\section{Materials and Methods}
\textbf{Gathering \& Identification of Plant }

The aerial parts of \textit{Premna tomentosa } (family Verbenaceae) were gathered form Chenkottai, Tirunelveli District of Tamilnadu India. Plant recognition was made from Botanical investigation of India, Palayamkottai The \textit{Premna tomentosa } were desiccated under shadowy, segregate, crushed through grinder \unskip~\citep{711514:16871689}. 

\textbf{Preparation of Concentrates }

The pulverized materials were packed in muslin cloth and concentrated with pet.ether, ethyl acetate and methanol as solvents respectively according to the increasing order of polarity \unskip~\citep{711514:16871685} through hot constant percolation method in Soxhlet equipment \unskip~\citep{711514:16871690}\ensuremath{^{}}for twenty-four hours. The concentrates were concentrated through rotational evaporator and subjected to solidify drying till dry powder was acquired by using lyophilizer. \unskip~\citep{711514:16871686,711514:16993460}. 

\textbf{Assessment of Antioxidant potential through invitro methods}

The variety of concentrate of \textit{Premna tomentosa }were used as assessment of antioxidant\textit{\space }activity by \unskip~\cite{711514:16871697}  method described for Superoxide radical (O\ensuremath{_{2}}\ensuremath{^{-}}) assay and \unskip~\cite{711514:16871682} method was adopted to determine the OH radical assay.
    
\section{Results and Discussion}
\textbf{Superoxide anion activity}

Super-oxides might be produced in huge amounts by a variety of biological processes. It is recognized to be more harmful to cellular components as creator of the generally ROS, contributing to tissue injure and various disorders \unskip~\citep{711514:16871687}. Superoxide radical potential of PE concentrates of \textit{Premna tomentosa }\textit{\space }\textit{\space }appeared in Table~\ref{tw-6571b22dd1ed}. The more activity of PE concentrates of \textit{Premna tomentosa }and quercetin at 1000\ensuremath{\mu}g/mL were recorded to be 57.68\% \& 99.12\% correspondingly. The IC\ensuremath{_{50}} of PE concentrates of \textit{Premna } \textit{tomentosa } \textit{\&} quercetin were recorded as 762\ensuremath{\mu}g/mL and 60\ensuremath{\mu}g/mL correspondingly. 


\begin{table*}[!htbp]
\caption{\boldmath {Activity of PE concentrate of \textit{Premna tomentosa} on Superoxide radical method} }
\label{tw-6571b22dd1ed}
\centering 
\begin{threeparttable}

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\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack  S.No & \cAlignHack Concentrate & \multicolumn{2}{p{\dimexpr(.5814\linewidth-2\tabcolsep)}}{\cAlignHack \% of activity(\ensuremath{\pm}SEM)*}\\
\rowcolor{kwdboxcolor} & \cAlignHack  (\ensuremath{\mu}g/mL) & \cAlignHack PE concentrate  & \cAlignHack Quercetin \\
\tblmidrule 
\cAlignHack 1 &
  \cAlignHack 125 &
  \cAlignHack 29.56\ensuremath{\pm}0.024 &
  \cAlignHack 72.76\ensuremath{\pm}0.012\\
\cAlignHack 2 &
  \cAlignHack 250 &
  \cAlignHack 42.62\ensuremath{\pm}0.038 &
  \cAlignHack 90.29\ensuremath{\pm}0.014\\
\cAlignHack 3 &
  \cAlignHack 500 &
  \cAlignHack 48.46\ensuremath{\pm}0.052 &
  \cAlignHack 96.89\ensuremath{\pm}0.010\\
\cAlignHack 4 &
  \cAlignHack 1000 &
  \cAlignHack 57.68\ensuremath{\pm}0.069 &
  \cAlignHack 99.12\ensuremath{\pm}0.018\\
\multicolumn{2}{p{\dimexpr(.4186\linewidth-2\tabcolsep)}}{} &
  \cAlignHack IC\ensuremath{_{50}}  =762 \ensuremath{\mu}g/mL &
  \cAlignHack IC\ensuremath{_{50}}  = 60 \ensuremath{\mu}g/mL\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{* Every value was articulated as mean \ensuremath{\pm} SEM for 3 experimentation}
\end{tablenotes}
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\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Activity of EA concentrate of \textit{Premna tomentosa} on superoxide radical method} }
\label{tw-7293b3ff369c}
\centering 
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\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack S.No & \cAlignHack Concentrate & \multicolumn{2}{p{\dimexpr(.5832\linewidth-2\tabcolsep)}}{\cAlignHack \% of activity(\ensuremath{\pm}SEM)*}\\
\rowcolor{kwdboxcolor} & \cAlignHack  (\ensuremath{\mu}g/mL) & \cAlignHack EA concentrate  & \cAlignHack Quercetin \\
\tblmidrule 
\cAlignHack 1 &
  \cAlignHack 125 &
  \cAlignHack 23.54\ensuremath{\pm}0.024 &
  \cAlignHack 72.76\ensuremath{\pm}0.012\\
\cAlignHack 2 &
  \cAlignHack 250 &
  \cAlignHack 37.25\ensuremath{\pm}0.045 &
  \cAlignHack 90.29\ensuremath{\pm}0.014\\
\cAlignHack 3 &
  \cAlignHack 500 &
  \cAlignHack 56.80\ensuremath{\pm}0.052 &
  \cAlignHack 96.89\ensuremath{\pm}0.010\\
\cAlignHack 4 &
  \cAlignHack 1000 &
  \cAlignHack 75.66\ensuremath{\pm}0.034 &
  \cAlignHack 99.12\ensuremath{\pm}0.018\\
\multicolumn{2}{p{\dimexpr(.4167999999999999\linewidth-2\tabcolsep)}}{} &
  \cAlignHack IC\ensuremath{_{50}}  = 375\ensuremath{\mu}g/mL &
  \cAlignHack IC\ensuremath{_{50}}  = 60 \ensuremath{\mu}g/mL\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{* Every value was articulated as mean \ensuremath{\pm} SEM for 3 experimentation}
\end{tablenotes}
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\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Activity of methanol concentrate of \textit{Premna tomentosa} on Superoxide radical method } }
\label{tw-b0a500658d92}
\centering 
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\tblmidrule 
\cAlignHack 1 &
  \cAlignHack 125 &
  \cAlignHack 40.34\ensuremath{\pm}0.045 &
  \cAlignHack 72.76\ensuremath{\pm}0.012\\
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  \cAlignHack 250 &
  \cAlignHack 54.42\ensuremath{\pm}0.056 &
  \cAlignHack 90.29\ensuremath{\pm}0.014\\
\cAlignHack 3 &
  \cAlignHack 500 &
  \cAlignHack 69.44\ensuremath{\pm}0.031 &
  \cAlignHack 96.89\ensuremath{\pm}0.010\\
\cAlignHack 4 &
  \cAlignHack 1000 &
  \cAlignHack 78.68\ensuremath{\pm}0.022 &
  \cAlignHack 99.12\ensuremath{\pm}0.018\\
\multicolumn{2}{p{\dimexpr(.36309999999999995\linewidth-2\tabcolsep)}}{} &
  \cAlignHack IC\ensuremath{_{50}}  =225 \ensuremath{\mu}g/mL &
  \cAlignHack IC\ensuremath{_{50}}  = 60 \ensuremath{\mu}g/mL\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{* Every value was articulated as mean \ensuremath{\pm} SEM for 3 experimentation}
\end{tablenotes}
\end{threeparttable}

\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Activity of PE concentrate of \textit{Premna tomentosa} hydroxyl radical method} }
\label{tw-6631c61268a6}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.1391\linewidth-2\tabcolsep}p{\dimexpr.264\linewidth-2\tabcolsep}p{\dimexpr.3054\linewidth-2\tabcolsep}p{\dimexpr.2915\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack S.No & \cAlignHack Concentrate & \multicolumn{2}{p{\dimexpr(.5969\linewidth-2\tabcolsep)}}{\cAlignHack \% of activity(\ensuremath{\pm}SEM)*}\\
\rowcolor{kwdboxcolor}\cAlignHack  & \cAlignHack  (\ensuremath{\mu}g/mL) & \cAlignHack  PE concentrate  & \cAlignHack  Ascorbate\\
\tblmidrule 
\cAlignHack 1 &
  \cAlignHack 125 &
  \cAlignHack 20.22\ensuremath{\pm}0.064 &
  \cAlignHack 70.34\ensuremath{\pm}0.38\\
\cAlignHack 2 &
  \cAlignHack 250 &
  \cAlignHack 29.22\ensuremath{\pm}0.034 &
  \cAlignHack 85.54\ensuremath{\pm}0.15\\
\cAlignHack 3 &
  \cAlignHack 500 &
  \cAlignHack 38.68\ensuremath{\pm}0.022 &
  \cAlignHack 90.23\ensuremath{\pm}0.24\\
\cAlignHack 4 &
  \cAlignHack 1000 &
  \cAlignHack 49.55\ensuremath{\pm}0.064 &
  \cAlignHack 96.50\ensuremath{\pm}0.16\\
\multicolumn{2}{p{\dimexpr(.4031\linewidth-2\tabcolsep)}}{} &
  \cAlignHack IC\ensuremath{_{50}}  = 1038 \ensuremath{\mu}g/mL  &
  \cAlignHack IC\ensuremath{_{50}}  = 65 \ensuremath{\mu}g/mL \\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{* Every value was articulated as mean \ensuremath{\pm} SEM for 3 experimentation}
\end{tablenotes}
\end{threeparttable}

\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Activity of EA concentrate of \textit{Premna tomentosa} hydroxyl radical method} }
\label{tw-44dbf9d2912e}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.1445\linewidth-2\tabcolsep}p{\dimexpr.2482999999999999\linewidth-2\tabcolsep}p{\dimexpr.30530000000000005\linewidth-2\tabcolsep}p{\dimexpr.30189999999999998\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack S.No & \cAlignHack Concentrate & \multicolumn{2}{p{\dimexpr(.6072\linewidth-2\tabcolsep)}}{\cAlignHack \% of activity(\ensuremath{\pm}SEM)*}\\
\rowcolor{kwdboxcolor}\cAlignHack  & \cAlignHack  (\ensuremath{\mu}g/mL) & \cAlignHack  EA concentrate  & \cAlignHack  (Ascorbate)\\
\tblmidrule 
\cAlignHack 1 &
  \cAlignHack 125 &
  \cAlignHack 20.32\ensuremath{\pm}0.045 &
  \cAlignHack 70.34\ensuremath{\pm}0.38\\
\cAlignHack 2 &
  \cAlignHack 250 &
  \cAlignHack 44.89\ensuremath{\pm}0.042 &
  \cAlignHack 85.54\ensuremath{\pm}0.15\\
\cAlignHack 3 &
  \cAlignHack 500 &
  \cAlignHack 53.78\ensuremath{\pm}0.024 &
  \cAlignHack 90.23\ensuremath{\pm}0.24\\
\cAlignHack 4 &
  \cAlignHack 1000 &
  \cAlignHack 62.88\ensuremath{\pm}0.016 &
  \cAlignHack 96.50\ensuremath{\pm}0.16\\
\multicolumn{2}{p{\dimexpr(.3927999999999999\linewidth-2\tabcolsep)}}{} &
  \cAlignHack IC\ensuremath{_{50}}  =464 \ensuremath{\mu}g/mL  &
  \cAlignHack IC\ensuremath{_{50}}  = 65 \ensuremath{\mu}g/mL  \\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{* Every value was articulated as mean \ensuremath{\pm} SEM for 3 experimentation}
\end{tablenotes}
\end{threeparttable}

\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Activity of Methanolic concentrate of \textit{Premna tomentosa} hydroxyl radical method } }
\label{tw-64f2252393a1}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.1167\linewidth-2\tabcolsep}p{\dimexpr.2498\linewidth-2\tabcolsep}p{\dimexpr.32800000000000004\linewidth-2\tabcolsep}p{\dimexpr.3055\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack S.No & \cAlignHack Concentrate & \multicolumn{2}{p{\dimexpr(.6335000000000001\linewidth-2\tabcolsep)}}{\cAlignHack \% of activity(\ensuremath{\pm}SEM)*}\\
\rowcolor{kwdboxcolor}\cAlignHack  & \cAlignHack  (\ensuremath{\mu}g/mL) & \cAlignHack Methanolic concentrate  & \cAlignHack Ascorbate\\
\tblmidrule 
\cAlignHack 1 &
  \cAlignHack 125 &
  \cAlignHack 43.66\ensuremath{\pm}0.032 &
  \cAlignHack 70.34\ensuremath{\pm}0.38\\
\cAlignHack 2 &
  \cAlignHack 250 &
  \cAlignHack 49.28\ensuremath{\pm}0.046 &
  \cAlignHack 85.54\ensuremath{\pm}0.15\\
\cAlignHack 3 &
  \cAlignHack 500 &
  \cAlignHack 67.46\ensuremath{\pm}0.048 &
  \cAlignHack 90.23\ensuremath{\pm}0.24\\
\cAlignHack 4 &
  \cAlignHack 1000 &
  \cAlignHack 70.34\ensuremath{\pm}0.056 &
  \cAlignHack 96.50\ensuremath{\pm}0.16\\
\multicolumn{2}{p{\dimexpr(.3665\linewidth-2\tabcolsep)}}{} &
  \cAlignHack IC\ensuremath{_{50}}  = 288 \ensuremath{\mu}g/mL  &
  \cAlignHack IC\ensuremath{_{50}}  = 65 \ensuremath{\mu}g/mL \\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{* Every value was articulated as mean \ensuremath{\pm} SEM for 3 experimentation}
\end{tablenotes}
\end{threeparttable}

\end{table*}
Superoxide radical potential of EA concentrates of \textit{Premna tomentosa }appeared in Table~\ref{tw-7293b3ff369c}. The more activity of EA concentrates of \textit{Premna t}\textit{omentosa }and standard at 1000 \ensuremath{\mu}g/mL was found to be 75.66\% and 99.12\% correspondingly. IC\ensuremath{_{50}} EA concentrates of \textit{Premna } \textit{tomentosa } \textit{and} standard was recorded as 375\ensuremath{\mu}g/mL and 60\ensuremath{\mu}g/mL correspondingly. 

Superoxide radical potential of Methanol concentrates of \textit{Premna tomentosa }appeared in Table~\ref{tw-b0a500658d92}. The more activity of Methanol concentrates of \textit{Premna }\textit{tomentosa }\textit{and} standard at 1000 \ensuremath{\mu}g/mL was found to be 78.68\% \& 99.12\% correspondingly. The IC\ensuremath{_{50}} EA concentrates of \textit{Premna t} \textit{omentosa } \textit{and} standard was recorded as 225\ensuremath{\mu}g/mL and 60\ensuremath{\mu}g/mL correspondingly.

IC\ensuremath{_{50}} values and percentage scavenging capacity, it was recorded that methanolic concentrates of \textit{Premna tomentosa }had better activity in superoxide radical when compared EA and PE concentrates.

\textbf{Hydroxyl radical scavenging activity}

Hydroxyl radical activity was determined by generating the OH radicals using ascorbic acid. OH radicals were produced by the oxidation reaction with the Dimethylsulfoxide to give in HCHO, which provides an appropriate method to identify OH radicals by added with Nash reagent \unskip~\citep{711514:16871680}. 

Hydroxyl radical activity of PE concentrate of \textit{Premna tomentosa appeared }in Table~\ref{tw-6631c61268a6}. The hydroxyl radical activity of PE concentrate of \textit{Premna tomentosa and }ascorbic acid \textit{at} 1000 \ensuremath{\mu}g/mL was found to be 49.55\% \& 96.50 \% correspondingly. The IC\ensuremath{_{50}} of PE concentrate of \textit{Premna t} \textit{omentosa and} ascorbic acid were found to be 1038\ensuremath{\mu}g/mL and 65\ensuremath{\mu}g/mL correspondingly.

Hydroxyl radical is the most ROS and causes severe injure to adjacent biomolecule. Hydroxyl radical activity of EA concentrate of \textit{Premna tomentosa }and ascorbic acid were showed\textit{\space }in Table~\ref{tw-44dbf9d2912e}. The hydroxyl radical activity of EA concentrate of \textit{Premna tomentosa and }ascorbic acid \textit{at} 1000 \ensuremath{\mu}g/mL was found to be 62.88\% \& 96.50 \% correspondingly. The IC\ensuremath{_{50}} of EA concentrate of \textit{Premna } \textit{tomentosa and} ascorbic acid were found to be 464\ensuremath{\mu}g/mL and 65\ensuremath{\mu}g/mL correspondingly.

Hydroxyl radical scavenging potency of methanolic concentrate of \textit{Premna tomentosa }and ascorbic acid were\textit{\space }showed in Table~\ref{tw-64f2252393a1}. The hydroxyl radical activity of methanol concentrate of \textit{Premna tomentosa and }ascorbic acid \textit{at} 1000 \ensuremath{\mu}g/mL was found to be 70.34\% \& 96.50 \% correspondingly. The IC\ensuremath{_{50}} of methanol concentrate of \textit{Premna tomentosa }and ascorbic acid were found to be 288\ensuremath{\mu}g/mL and 65\ensuremath{\mu}g/mL correspondingly.

Based on the above report the methanolic concentrate of \textit{Premna tomentosa }(IC\ensuremath{_{50}} = 288 \ensuremath{\mu }g/ml) was found more effective than that of PE(IC\ensuremath{_{50}} = 1038 \ensuremath{\mu }g/ml) and EA concentrates (IC\ensuremath{_{50}} = 464 \ensuremath{\mu}g/mL). The methanolic concentrate of \textit{Premna }\textit{tomentosa} \textit{ exhibited} better hydroxyl radical scavenging activity compared to other concentrates. The scavenging of the hydroxyl radicals possibly may be presence of hydrogen donating ability flavonoids from methanolic concentrates. 
    
\section{Conclusion }
From this investigation, it is concluded that aerial parts of methanolic concentrate of \textit{Premna tomentosa} is established to have valuable potential than PE and EA concentrates. Thus, present data suggest that methanolic concentrate of \textit{Premna tomentosa} is a remarkable resource of natural antioxidant, which might be helpful in preventing the improvement of a variety of free radical induced diseases. 
    

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