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\def\RunningAuthor{Vijey Aanandhi M and Ganesh S}
\firstPage{2549}
\articleType{Original Article}
\receivedDate{04 Nov 2020}
\acceptedDate{13 Dec 2020}
\revisedDate{09 Dec 2020}
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\title{\textbf{Anthelmintic and Antioxidant activity of Aqueous Ethanolic Extract of Erythrina subumbrans (Hassk.) Merr.}}
\author{Ganesh~S\textsuperscript{1}, 
            Vijey~Aanandhi~M\textsuperscript{*}\textsuperscript{2}~\\[5pt]{\textsuperscript{1}Research Scholar, Department of Pharmaceutical Chemistry and Analysis, School of Pharmaceutical Sciences, Vels Institute of Science, Technology and Advanced Studies (VISTAS), Pallavaram, Chennai {\textendash} 600117, Tamil Nadu, India}~\\{\textsuperscript{2}Department of Pharmaceutical Chemistry and Analysis, School of Pharmaceutical Sciences, Vels Institute of Science, Technology and Advanced Studies (VISTAS), Pallavaram, Chennai {\textendash} 600117, Tamil Nadu, India}}

\begin{abstract}
Aqueous Ethanolic extract of vegetative (leaf) part of herbal plant \textit{Erythrina subumbrans (Hassk.)} was assessed for Anthelmintic and Antioxidant activity. The obtained crude extract was prepared in different concentration, i.e., 50 and 100 mg/mL, against the standard Piperazine citrate, i.e., 10mg/mL for anthelmintic activity. Pheretima posthuma test worms were used during the study and the anthelmintic activity was evaluated based on Paralysis and Death time. The average paralysis and death time of standard is 1.4 \& 39.59 min (10mg/mL) compared to sample 5.3 \& 65.09 min (50mg/mL); 2.5 \& 54.08 min (100mg/mL). From the data, aqueous ethanolic extract possesses promising anthelmintic activity compared to piperazine citrate standard. Similarly, antioxidant activity was evaluated for sample at different concentration ranging from 5, 10, 20, 40, 80, 160, and 320 \ensuremath{\mu}g/ml premixed with 2.5mL of 0.0135mM DPPH solution. Ascorbic acid was used as a standard and treated in a similar way as that of a sample. Control samples were also prepared without standard and sample solutions. Prepared solutions were vortexed and kept at benchtop condition for 30 minutes and absorbance was measured at 517 nm in a UV-Vis spectrophotometer. The absorbance pattern of the sample is not comparable with that of the standard solution at the same concentration. Aqueous ethanolic extract with IC\ensuremath{_{50}} (half maximal inhibitory concentration) value is {\textgreater}320 \ensuremath{\mu}g/mL against standard Ascorbic acid with an IC\ensuremath{_{50}} value of 15.5 \ensuremath{\mu}g/ml. Which witness the crude extract possess less antioxidant activity.  
\end{abstract}\def\keywordstitle{Keywords}

\begin{keywords}Erythrina subumbrans (Hassk.),\newline Piperazine citrate,\newline DPPH (2,\newline 2-diphenyl-1-picrylhydrazyl) and Ascorbic acid
\end{keywords}
\twocolumn[ \maketitle {\printKwdAbsBox}]
\makeatletter{\raggedright\textsuperscript{*}Corresponding Author\par Name:\ Vijey~Aanandhi~M~\\ Phone:\ ~\\  Email:\ hodpchemistry@velsuniv.ac.in
        \par}\vspace*{-11pt}\hrulefill\par{\fontsize{12}{14}\selectfont ISSN: 0975-7538}\par% 
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\section{Introduction}
Helminths a well-known parasitic worm infection affecting the wider portion of the world's population and it is one of the most common infections in humans. In well-developed and in developing countries like western countries, India, Africa etc., parasitic worm infection possesses a hefty threat to the health of the public and contributing the incidence of Morbidity, malnutrition, anemia, Pneumonia and eosinophilia. The majority of the infections due to parasitic worms were restricted to tropical zones \unskip~\citep{1012343:21476918}. The world health organization discloses that the affected population due to parasitic infections is closer to 2 billion. Other than humans, crops and livestock are affected by parasitic worms resulting in a slowdown of economic growth, affecting the shortage of food production. These infections influence the majority of the social and economic consequences where the population is high in endemic areas. Although parasitic infection is it is limited to tropical zones, however, this can be developed for travellers who visited those countries, and some can develop in temperate condition. Despite this occurrence, the research of the anthelminthic drug is less significant \unskip~\citep{1012343:21476915}.

Due to the increase in anthelmintic resistance and conventional impact on the environment, it is necessary to develop for alternative plan against the nematodes present in the gastrointestinal tract. Recently, it has been recognized that significant toxicity was observed in human beings that are present in foods that are derived from live stocks, posing a serious threat to living humans. The one common therapy to control these pathogens is phototherapy \unskip~\citep{1012343:21476913}. Regardless of the situation, there is a steep increase in demand of herbal medicine compared to the past period worldwide. The majority of the Phyto plants were not assessed for their quality, efficacy and safety \unskip~\citep{1012343:21476925}. The Discovery and progress of a new chemical entity for controlling parasitic worm infection is greatly needed. Several health agencies are promoting studies of conventional used plants, which are generally considered as a vital source for bioactive substances \unskip~\citep{1012343:21476909}. The key cause for many ailments in humans is free radicals. These irregularities were generated between the neutralization and formation of prooxidants resulting in the condition called oxidative stress condition \unskip~\citep{1012343:21476923}. Nitrogen or oxygen species that are reactive (RNS, ROS) are vital free radicals, which makes our human body function complicated. Radicals like Hydroxy (OH\ensuremath{^{-}}), anion superoxide (O\ensuremath{_{2}}\ensuremath{^{-}}), peroxyl are the reactive oxygen species; Nitric oxide (NO\ensuremath{^{-}}), Peroxynitrite (ONOO\ensuremath{^{-}}), a free radical derived from nitrogen are the metabolites that affect physiologically. These were generated as a resultant of the aerobic organism during respiration; however, the unwarranted level has a direct link to the beginning of many diseases like diabetes, stroke, cancer and other immune deficiency \unskip~\citep{1012343:21476928}.

The recent development on the usage of crude herbal drugs that contain free radical scavengers known for their healing activity \unskip~\citep{1012343:21476919}. The most common natural antioxidants include ascorbic acid, flavonoids, phenolic compounds and carotenoids are highly effective by inhibiting the peroxidation of lipids by lipoxygenase inactivation and to scavenge the oxygen species that are active and free radicals by proliferating the reaction mechanism and by heavy metal ion chelation \unskip~\citep{1012343:21476912}. Traditional use of medicinal plants is considered safer in comparable antibiotics if the synthetic origin and are effective in clinical \unskip~\citep{1012343:21476911}. Carotenoids, tannins and flavonoids that are produced by plants have a very impressive range on avoiding susceptible substrate oxidation \unskip~\citep{1012343:21476914}. Currently, Natural occurring antioxidants have a target for numerous research activities to find the basis drugs are effective, cheap and clinically safe \unskip~\citep{1012343:21476924}. Hence, the present research activity was carried out to search for anthelmintic and antioxidant activity in ethanolic extract of crude \textit{Erythrina subumbrans (Hassk.)}. 
    
\section{Materials and Methods}
\textbf{Material}

A chemical such as Piperazine citrate, DPPH (2, 2-diphenyl-1-picrylhydrazyl), Ascorbic acid and were purchased from SISCO research laboratories Ltd Maharashtra, India. Other chemicals procured from local manufactures (Analytical reagent grade).

\textbf{Plant material}

The leaves of Erythrina subumbrans (Hassk.) Merr were collected from Western Ghats of India and authenticated at Siddha central research institute (Ministry of AYUSH, Government of India), Chennai by Research Officer and Head of the pharmacognosy department Dr K.N. Sunil Kumar and confirmed by Assistant Director in-charge Dr P. Sathiyarajeswaran (Authentication certificate 112.04011901 dated 04 Apr 2019). The harvested vegetative part was dried in a nominal condition and stored in a container made of Kraft paper. 

\textbf{Preparation of plant Extract}

The plant material was washed with distilled water several times and was subjected to air-drying under the shade. After drying, they were ground by an electric mixer until they became a powder. Then the powdered samples were stored in a dark place and subjected to extraction method \unskip~\citep{1012343:21476916}. Extraction of powdered samples was done using aqueous ethanol (80\%). Aliquots of 50 g of the powdered samples were soaked in 250 ml of the solvent for 72 hrs. Later the samples were filtered and concentrated under reduced pressure using a rotary evaporator and keep stored at room temperature (25\ensuremath{^\circ} C). 

\textbf{Test Organism}

Pheretima posthuma (\textit{P. posthuma}), a known Indian adult earthworm were collected from damp soil. The washing was made several times with raw water, followed by a saline solution (0.9\% sodium chloride). The collection of earthworms for the study was made by considering the physical and structural resemblance with roundworms that are present in the human intestine. Length of 3-5 cm and width of 0.15-0.25 cm were selected for anthelmintic study \unskip~\citep{1012343:21476917}. 

\textbf{In Vitro Anthelmintic Activity}

The Anthelmintic activity of the aqueous ethanolic extract of Erythrina subumbrans (Hassk.) was carried out according to the method with minor modification \unskip~\citep{1012343:21476921}. The aqueous ethanolic crude extract was prepared in normal saline solution in two different concentration, i.e., 50 and 100 mg/mL. Piperazine citrate was used as a standard with a 10 mg/mL concentration. Standard drug solution and extract with different concentrations were prepared freshly on the day of use before initiating the experiment. A total of three groups with three earthworms in each group was placed into 10ml of the solution and evaluated for the paralysis and death time of the earthworm. 

\textbf{In Vitro Antioxidant Activity}

The DPPH (2, 2-diphenyl-1-picrylhydrazyl) radical scavenging assay of the aqueous ethanolic extract of Erythrina subumbrans (Hassk.) was carried out according to the method with minor modification \unskip~\citep{1012343:21476926}. In details, the DPPH solution was prepared in methanol at a concentration of 0.0135mM. 2.5mL of the prepared DPPH solution were mixed with different concentration of test sample (i.e., 5, 10, 20, 40, 80, 160 and 320 \ensuremath{\mu}g/ml), standard (Ascorbic acid), and control (without standard/test samples). Then, these sample standard mixtures were vortexed thoroughly and kept at the benchtop (i.e., room temperature) condition for 30 minutes. The absorbance of the mixture was measured at 517 nm by using a UV-visible spectrophotometer. The standard, sample and control sample final solutions were shown in Figure~\ref{f-bbef1cd33028} and Figure~\ref{f-3114b459ae42}. The percentage of Antioxidant activity was measured using the following expression: \% DPPH Inhibition = [(OD of Control - OD of Test)/OD of control] x 100 Where OD of Test is the absorbance of the test and OD of control is the absorbance of the control. All experiments were performed in triplicate. From the data obtained, a curve to be plotted and the IC\ensuremath{_{50}} value will be calculated.


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\caption{\boldmath {Standard solution }}
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\caption{\boldmath {Sample solution}}
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\caption{\boldmath {Statistical curve }}
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\section{Results}
\textbf{Plant Extract}

The percentage yield of the ethanolic extracts was calculated by using the following equation: yield (\%) = (weight of extract/weight of dried plant material) x 100 and the obtained yield was presented in Table~\ref{tw-a92c0a8e0bc2}.


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\caption{\boldmath {Extract Yield values} }
\label{tw-a92c0a8e0bc2}
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\caption{\boldmath {Anthelmintic Activity-Paralysistime} }
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\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.2111\linewidth-2\tabcolsep}p{\dimexpr.15729999999999999\linewidth-2\tabcolsep}p{\dimexpr.13929999999999998\linewidth-2\tabcolsep}p{\dimexpr.14200000000000001\linewidth-2\tabcolsep}p{\dimexpr.1697\linewidth-2\tabcolsep}p{\dimexpr.18060000000000002\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Groups & \cAlignHack Dose & \multicolumn{3}{p{\dimexpr(.45099999999999994\linewidth-2\tabcolsep)}}{\cAlignHack Time of paralysis (min)} & \cAlignHack Avg. paralysis \\
\rowcolor{kwdboxcolor}\cAlignHack  & \cAlignHack  & \cAlignHack G-1 & \cAlignHack G-2 & \cAlignHack G-3 & \cAlignHack time in min\\
\tblmidrule 
Piperazine citrate &
  10mg/ml &
  \cAlignHack 1.39 &
  \cAlignHack 1.45 &
  \cAlignHack 1.42 &
  \cAlignHack 1.4\\
\multicolumn{1}{p{\dimexpr(.2111\linewidth-2\tabcolsep)}}{\multirow{2}{\linewidth}{Ethanolic extract}} &
  50mg/ml &
  \cAlignHack 5.45 &
  \cAlignHack 5.3 &
  \cAlignHack 5.25 &
  \cAlignHack 5.3\\
 &
  100mg/ml &
  \cAlignHack 2.50 &
  \cAlignHack 2.45 &
  \cAlignHack 2.54 &
  \cAlignHack 2.5\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Anthelmintic Activity-Deathtime} }
\label{tw-5a866d0490d7}
\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.20569999999999997\linewidth-2\tabcolsep}p{\dimexpr.14060000000000002\linewidth-2\tabcolsep}p{\dimexpr.1531\linewidth-2\tabcolsep}p{\dimexpr.16970000000000002\linewidth-2\tabcolsep}p{\dimexpr.1599\linewidth-2\tabcolsep}p{\dimexpr.171\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Groups & \cAlignHack Dose & \multicolumn{3}{p{\dimexpr(.4827\linewidth-2\tabcolsep)}}{\cAlignHack Death Time (min)} & \cAlignHack Avg. death \\
\rowcolor{kwdboxcolor}\cAlignHack  & \cAlignHack  & \cAlignHack G-1 & \cAlignHack G-2 & \cAlignHack G-3 & \cAlignHack time in min\\
\tblmidrule 
Piperazine citrate &
  10mg/ml &
  \cAlignHack 40.12 &
  \cAlignHack 39.45 &
  \cAlignHack 39.30 &
  \cAlignHack 39.59\\
\multicolumn{1}{p{\dimexpr(.20569999999999997\linewidth-2\tabcolsep)}}{\multirow{2}{\linewidth}{Ethanolic extract}} &
  50mg/ml &
  \cAlignHack 65.12 &
  \cAlignHack 64.45 &
  \cAlignHack 64.50 &
  \cAlignHack 65.09\\
 &
  100mg/ml &
  \cAlignHack 54.25 &
  \cAlignHack 53.45 &
  \cAlignHack 54.55 &
  \cAlignHack 54.08\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Antioxidant Activity-Standard (Absorbance)} }
\label{tw-185ef67f0127}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.16666666666666664\linewidth-2\tabcolsep}p{\dimexpr.1666666666666667\linewidth-2\tabcolsep}p{\dimexpr.1666666666666667\linewidth-2\tabcolsep}p{\dimexpr.16085000000000004\linewidth-2\tabcolsep}p{\dimexpr.16810000000000002\linewidth-2\tabcolsep}p{\dimexpr.17105000000000004\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Type & \cAlignHack Control  & \cAlignHack Conc. & \multicolumn{3}{p{\dimexpr(.50000000000000014\linewidth-2\tabcolsep)}}{\cAlignHack Absorbance}\\
\rowcolor{kwdboxcolor}\cAlignHack  & \cAlignHack Absorbance & \cAlignHack  (\ensuremath{\mu}g/mL) & \cAlignHack Prp-1 & \cAlignHack Prp-2 & \cAlignHack Prp-3\\
\tblmidrule 
\cAlignHack Ascorbic acid &
  \cAlignHack 0.886 &
  \cAlignHack 5 &
  \cAlignHack 0.683 &
  \cAlignHack 0.685 &
  \cAlignHack 0.681\\
\cAlignHack  &
  \cAlignHack  &
  \cAlignHack 10 &
  \cAlignHack 0.597 &
  \cAlignHack 0.591 &
  \cAlignHack 0.595\\
\cAlignHack  &
  \cAlignHack  &
  \cAlignHack 20 &
  \cAlignHack 0.391 &
  \cAlignHack 0.395 &
  \cAlignHack 0.393\\
\cAlignHack  &
  \cAlignHack  &
  \cAlignHack 40 &
  \cAlignHack 0.113 &
  \cAlignHack 0.118 &
  \cAlignHack 0.119\\
\cAlignHack  &
  \cAlignHack  &
  \cAlignHack 80 &
  \cAlignHack 0.081 &
  \cAlignHack 0.088 &
  \cAlignHack 0.085\\
\cAlignHack  &
  \cAlignHack  &
  \cAlignHack 160 &
  \cAlignHack 0.062 &
  \cAlignHack 0.065 &
  \cAlignHack 0.067\\
\cAlignHack  &
  \cAlignHack  &
  \cAlignHack 320 &
  \cAlignHack 0.045 &
  \cAlignHack 0.043 &
  \cAlignHack 0.049\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{Prp.: Preparation, Conc.: Concentration}
\end{tablenotes}
\end{threeparttable}

\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Antioxidant Activity-Standard (\% Assay)} }
\label{tw-90f680711d20}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.11595555555555556\linewidth-2\tabcolsep}p{\dimexpr.12502222222222223\linewidth-2\tabcolsep}p{\dimexpr.12333333333333332\linewidth-2\tabcolsep}p{\dimexpr.1178222222222222\linewidth-2\tabcolsep}p{\dimexpr.12146666666666665\linewidth-2\tabcolsep}p{\dimexpr.12706666666666667\linewidth-2\tabcolsep}p{\dimexpr.13995555555555555\linewidth-2\tabcolsep}p{\dimexpr.12937777777777779\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Type & \cAlignHack Conc. & \multicolumn{3}{p{\dimexpr(.3626222222222222\linewidth-2\tabcolsep)}}{\cAlignHack \% Inhibition} & \cAlignHack Mean & \cAlignHack Std. Dev & \cAlignHack IC\ensuremath{_{50}} value\\
\rowcolor{kwdboxcolor}\cAlignHack  & \cAlignHack (\ensuremath{\mu}g/mL) & \cAlignHack Prp-1 & \cAlignHack Prp-2 & \cAlignHack Prp-3 & \cAlignHack  & \cAlignHack  & \cAlignHack \\
\tblmidrule 
\multicolumn{1}{p{\dimexpr(.11595555555555556\linewidth-2\tabcolsep)}}{\multirow{7}{\linewidth}{\cAlignHack Ascorbic acid }} &
  \cAlignHack 5 &
  \cAlignHack 22.9699 &
  \cAlignHack 22.7443 &
  \cAlignHack 23.1954 &
  \cAlignHack 22.9699 &
  \cAlignHack 0.2255 &
  \multicolumn{1}{p{\dimexpr(.12937777777777779\linewidth-2\tabcolsep)}}{\multirow{7}{\linewidth}{\cAlignHack 15.5 \ensuremath{\mu}g/mL}}\\
 &
  \cAlignHack 10 &
  \cAlignHack 32.6691 &
  \cAlignHack 33.3458 &
  \cAlignHack 32.8947 &
  \cAlignHack 32.9699 &
  \cAlignHack 0.3445 &
  \\
 &
  \cAlignHack 20 &
  \cAlignHack 55.9022 &
  \cAlignHack 55.4511 &
  \cAlignHack 55.6766 &
  \cAlignHack 55.6766 &
  \cAlignHack 0.2255 &
  \\
 &
  \cAlignHack 40 &
  \cAlignHack 87.2556 &
  \cAlignHack 86.6917 &
  \cAlignHack 86.5789 &
  \cAlignHack 86.8421 &
  \cAlignHack 0.3625 &
  \\
 &
  \cAlignHack 80 &
  \cAlignHack 90.8646 &
  \cAlignHack 90.0751 &
  \cAlignHack 90.4135 &
  \cAlignHack 90.4511 &
  \cAlignHack 0.3960 &
  \\
 &
  \cAlignHack 160 &
  \cAlignHack 93.0075 &
  \cAlignHack 92.6691 &
  \cAlignHack 92.4436 &
  \cAlignHack 92.7067 &
  \cAlignHack 0.2838 &
  \\
 &
  \cAlignHack 320 &
  \cAlignHack 94.9248 &
  \cAlignHack 95.1503 &
  \cAlignHack 94.4736 &
  \cAlignHack 94.8496 &
  \cAlignHack 0.3445 &
  \\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{Prp: Preparation, Conc.: Concentration}
\end{tablenotes}
\end{threeparttable}

\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Antioxidant Activity -Sample (Absorbance)} }
\label{tw-c374a0d057c3}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.1531\linewidth-2\tabcolsep}p{\dimexpr.17659999999999997\linewidth-2\tabcolsep}p{\dimexpr.1503\linewidth-2\tabcolsep}p{\dimexpr.1669\linewidth-2\tabcolsep}p{\dimexpr.1725\linewidth-2\tabcolsep}p{\dimexpr.1806\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Type & \cAlignHack Control  & \cAlignHack Conc. & \multicolumn{3}{p{\dimexpr(.52\linewidth-2\tabcolsep)}}{\cAlignHack Absorbance}\\
\rowcolor{kwdboxcolor}\cAlignHack  & \cAlignHack Absorbance & \cAlignHack (\ensuremath{\mu}g/mL) & \cAlignHack Prp-1 & \cAlignHack Prp-2 & \cAlignHack Prp-3\\
\tblmidrule 
\multicolumn{1}{p{\dimexpr(.1531\linewidth-2\tabcolsep)}}{\multirow{7}{\linewidth}{Erythrina subumbrans (ES)}} &
  \multicolumn{1}{p{\dimexpr(.17659999999999997\linewidth-2\tabcolsep)}}{\multirow{7}{\linewidth}{\cAlignHack 0.886}} &
  \cAlignHack 5 &
  \cAlignHack 0.856 &
  \cAlignHack 0.854 &
  \cAlignHack 0.855\\
 &
   &
  \cAlignHack 10 &
  \cAlignHack 0.824 &
  \cAlignHack 0.826 &
  \cAlignHack 0.823\\
 &
   &
  \cAlignHack 20 &
  \cAlignHack 0.801 &
  \cAlignHack 0.808 &
  \cAlignHack 0.805\\
 &
   &
  \cAlignHack 40 &
  \cAlignHack 0.772 &
  \cAlignHack 0.775 &
  \cAlignHack 0.778\\
 &
   &
  \cAlignHack 80 &
  \cAlignHack 0.732 &
  \cAlignHack 0.738 &
  \cAlignHack 0.735\\
 &
   &
  \cAlignHack 160 &
  \cAlignHack 0.621 &
  \cAlignHack 0.626 &
  \cAlignHack 0.628\\
 &
   &
  \cAlignHack 320 &
  \cAlignHack 0.413 &
  \cAlignHack 0.416 &
  \cAlignHack 0.418\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{Prp: Preparation, Conc.: Concentration}
\end{tablenotes}
\end{threeparttable}

\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Antioxidant Activity-Sample (\% Assay)} }
\label{tw-980e8ab95ccc}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.11373333333333335\linewidth-2\tabcolsep}p{\dimexpr.12653333333333332\linewidth-2\tabcolsep}p{\dimexpr.1211111111111111\linewidth-2\tabcolsep}p{\dimexpr.13222222222222221\linewidth-2\tabcolsep}p{\dimexpr.13035555555555554\linewidth-2\tabcolsep}p{\dimexpr.13408888888888887\linewidth-2\tabcolsep}p{\dimexpr.12493333333333332\linewidth-2\tabcolsep}p{\dimexpr.1170222222222222\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Type & \cAlignHack Conc. & \multicolumn{3}{p{\dimexpr(.3836888888888889\linewidth-2\tabcolsep)}}{\cAlignHack \% Inhibition} & \cAlignHack Mean & \cAlignHack Std. Dev & \cAlignHack IC\ensuremath{_{50}}\\
\rowcolor{kwdboxcolor}\cAlignHack  & \cAlignHack (\ensuremath{\mu}g/mL) & \cAlignHack Prp-1 & \cAlignHack Prp-2 & \cAlignHack Prp-3 & \cAlignHack  & \cAlignHack  & \cAlignHack value\\
\tblmidrule 
\multicolumn{1}{p{\dimexpr(.11373333333333335\linewidth-2\tabcolsep)}}{\multirow{7}{\linewidth}{\cAlignHack Erythrina subumbrans (ES)}} &
  \cAlignHack 5 &
  \cAlignHack 3.4586 &
  \cAlignHack 3.6842 &
  \cAlignHack 3.5714 &
  \cAlignHack 3.5714 &
  \cAlignHack 0.1127 &
  \multicolumn{1}{p{\dimexpr(.1170222222222222\linewidth-2\tabcolsep)}}{\multirow{7}{\linewidth}{\cAlignHack {\textgreater} 320 \ensuremath{\mu}g/mL}}\\
 &
  \cAlignHack 10 &
  \cAlignHack 7.0676 &
  \cAlignHack 6.8421 &
  \cAlignHack 7.1804 &
  \cAlignHack 7.0300 &
  \cAlignHack 0.1722 &
  \\
 &
  \cAlignHack 20 &
  \cAlignHack 9.6616 &
  \cAlignHack 8.8721 &
  \cAlignHack 9.2105 &
  \cAlignHack 9.2481 &
  \cAlignHack 0.3960 &
  \\
 &
  \cAlignHack 40 &
  \cAlignHack 12.9323 &
  \cAlignHack 12.5939 &
  \cAlignHack 12.2556 &
  \cAlignHack 12.5939 &
  \cAlignHack 0.3383 &
  \\
 &
  \cAlignHack 80 &
  \cAlignHack 17.4436 &
  \cAlignHack 16.7669 &
  \cAlignHack 17.1052 &
  \cAlignHack 17.1052 &
  \cAlignHack 0.3383 &
  \\
 &
  \cAlignHack 160 &
  \cAlignHack 29.9624 &
  \cAlignHack 29.3984 &
  \cAlignHack 29.1729 &
  \cAlignHack 29.5112 &
  \cAlignHack 0.4066 &
  \\
 &
  \cAlignHack 320 &
  \cAlignHack 53.4210 &
  \cAlignHack 53.0827 &
  \cAlignHack 52.8571 &
  \cAlignHack 53.1203 &
  \cAlignHack 0.2838 &
  \\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{Prp: Preparation, Conc.: Concentration}
\end{tablenotes}
\end{threeparttable}

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

Anthelmintic activity of Erythrina subumbrans (Hassk.) was evaluated by time, recorded during Paralysis and death of the earthworm. The paralysis time of the worms was concluded when there is no movement of worms and death time is concluded when there is a change in body texture and completely lost its motility \unskip~\citep{1012343:21476922}. Erythrina subumbrans (Hassk.) at the concentration of 50 mg/ml exhibits death and paralysis time of 65.09 and 5.30 minutes; 100 mg/mL concentration exhibits 54.08 and 2.50 minutes, respectively. Similarly, piperazine citrate exhibits death and paralysis time of 39.59 and 1.42 minutes at 10mg/ mL concentration. Details of the data were presented in Table~\ref{tw-9684f05613e9} and Table~\ref{tw-5a866d0490d7}.

\textbf{DPPH radical scavenging Activity}

The average control (i.e., without standard/test samples) absorbance value is 0.886 AU. The absorbance value and \% assay of standard ascorbic acid was calculated using the formula and details of the calculation was summarized in Table~\ref{tw-185ef67f0127} \& Table~\ref{tw-90f680711d20}. Similarly, absorbance value and \% inhibition of aqueous ethanolic extract of Erythrina subumbrans (Hassk.) Merr sample preparation was calculated using the formula and the details of the calculation was summarized in Table~\ref{tw-c374a0d057c3} \& Table~\ref{tw-980e8ab95ccc}. From the data, a curve was plotted, and the Assay concentration (IC\ensuremath{_{50}}) value was calculated. IC\ensuremath{_{50}} is defined as the concentration of the samples required for a 50\% Assay of the enzyme. The IC\ensuremath{_{50}} value was calculated from the graph by plotting concentration on X-Axis and \% Assay on Y-axis and the statistical curve was displayed in Figure~\ref{f-10ccac1b76ef}. The IC\ensuremath{_{50}} value of the ethanolic extract of \textit{Erythrina subumbrans (Hassk.) Merr} was found to be {\textgreater}320 \ensuremath{\mu}g/ml and the standard drug (Ascorbic acid) was 15.5 \ensuremath{\mu}g/ml, respectively. 
    
\section{Discussion}
Traditional medicinal plants possess active metabolites that are bioactive in nature and exhibits promising activity in anthelmintic and antioxidant activity in the Human system \unskip~\citep{1012343:21476927}. The most known compounds, like flavonoids and phenol, are widely dispersed in plants that are used as medicine. These compounds own significant Anthelmintic, antioxidant, Anti-ulcer, anti-diabetic, antioxidant, hepatoprotective and anti-carcinogen properties. Piperazine citrate, a known anthelmintic agent, works by blocking the response of muscle worms to acetylcholine by producing hyperpolarization of nerve endings, resulting in paralysis and removed from the human intestinal lumen \unskip~\citep{1012343:21476920}.

Radical scavenging activity (DPPH) a traditional mechanism used for the evaluation of antioxidant activity in the crude extract. The absorbance of the solution decreases when the color of the solution changes to yellow from purple. Traditional medicinal plants can able to lower the free radical level to yellow color 1,1 diphenyl -2-picrlhydrazyl \unskip~\citep{1012343:21476910}. Hydrogen donation property is a key element for producing DPPH scavenging activity by antioxidants. When free radicals react with reducing agents, the solution becomes color less and losses it's color depending on the electrons used during the reaction mechanism \unskip~\citep{1012343:21476929}. 

It was found that crude extract of \textit{Erythrina subumbrans (Hassk.) Merr} possess promising anthelmintic activity with observed with a sample concentration of 50 \& 100 mg/mL compared to standard piperazine citrate at the concentration of 10 mg/mL. The crude extract does not possess antioxidant activity with IC\ensuremath{_{50}} values of 320 \ensuremath{\mu}g/ml as compared to the positive control, Ascorbic acid (IC\ensuremath{_{50}}: 15.5 \ensuremath{\mu}g/ml). When there is a rise in absorbance value of the mixture, it an indicative of reducing inability, which serves as a substantial indication of anti-scavenging/ oxidant activity. However, this was not observed in the sample during the analysis and the absorbance of the standard and sample does not possess a similar pattern which is evident that it has less antioxidant activity.
    
\section{Conclusion}
In this study, leaves of \textit{Erythrina subumbrans (Hassk.) Merr} was collected from the Western Ghats of India and authenticated at Siddha central research institute, Chennai. Extraction was carried out using aqueous Ethanol (80\%) and the obtained extract was subjected for anthelmintic and antioxidant activity. A promising anthelmintic activity was observed with a sample concentration of 50 and 100mg/ml against standard piperazine citrate 10mg/mL, whereas less antioxidant activity observed from crude extract\textit{\space } (IC\ensuremath{_{50}}: {\textgreater}320g/ml) compared against standard Ascorbic acid (IC\ensuremath{_{50}}: 15.5mg/ml). Further activities to be carried out on isolation of bioactive compounds for the crude extract to evaluate its efficiency on the various disorder. 
\section*{Acknowledgement}The authors are thankful to the School of Pharmaceutical Sciences, Vels Institute of Science, Technology, and Advanced Studies, and its management for providing research facilities and encouragement. 

\textbf{Funding Support}

The authors declare that they have no funding support for this study.

\textbf{Conflict of Interest}

The authors declare that they have no conflict of interest for this study.
    

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