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\def\RunningAuthor{Jacques~Britto~N and Kesavi~Durairaj}
\firstPage{3465}
\articleType{Original Article}
\receivedDate{29 Apr 2020}
\acceptedDate{01 Jun 2020}
\revisedDate{30 May 2020}
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\title{\textbf{Phytochemical profile and medicinal potentials of Lannea Coromandelica stem}}
\author{Jacques~Britto~N\textsuperscript{1}, 
            Kesavi~Durairaj\textsuperscript{*}\textsuperscript{2}~\\[5pt]{\textsuperscript{1}Research Scholar, Department of Anatomy, Faculty of Medicine, Sri Ramachandra Institute of Higher Education and Research (DU), Porur, Chennai {\textendash} 600116, Tamil Nadu, India}~\\{\textsuperscript{2}Department of Anatomy, Faculty of Medicine, Sri Ramachandra Institute of Higher Education and Research (DU), Porur, Chennai {\textendash} 600116, Tamil Nadu, India}}

\begin{abstract}
In the history of humanity, great ancestors of Indian sub-continent used plants that possessed unique medicinal properties. They identified the plants from jungles and derived crude drugs out of them for treating infectious diseases. This treatment process was a tradition. In the recent century, this plant-based drug extraction, processing and refining began scientifically by pharmaceutical industries. In this new era, plants with medicinal features are being used in curing broad spectrum of diseases. Lannea Coromandelica (L.C.) is a medicinal plant that belongs to this category. Tribals extensively used it in treating various infectious diseases and common injuries. There have been very few studies on the leaves, barks, flowers, gums and mucilage of this tree. But no preliminary phytochemical composition of L.C. stem has been studied. Therefore, the current work focusses on the screening of the phytochemical profile of the stem of L.C. by the sequential ethanolic extract. Stems of L.C. were procured from Mettur, Tamil Nadu. Thirteen different components were qualitatively analyzed using standard procedures from 100g of L.C. stem extract. In this study, seven components were identified, and their percentage was estimated. Using GC-MS, 50 components were identified of which Pentadecanoic acid, 14-methyl-methyl ester (1.0\%) was the major component. The presence of these components in L.C. stem extract can be used in the treatment of different ailments through their antioxidant, anti-arthritic, anti-diabetic, anti-inflammatory and antimicrobial activities.
\end{abstract}\def\keywordstitle{Keywords}

\begin{keywords}Lannea Coromandelica (L.C.),\newline Phytochemical,\newline Medicinal plants,\newline Ethanolic extract,\newline GCMS
\end{keywords}
\twocolumn[ \maketitle {\printKwdAbsBox}]
\makeatletter{\raggedright\textsuperscript{*}Corresponding Author\par Name:\ Kesavi~Durairaj~\\ Phone:\ ~\\  Email:\ kesavikraja@hotmail.com
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\section{Introduction}
Life on earth predominantly depends upon the natural resources for its survival. Medicinal plants being one of the essential natural resources, its purpose and utility have been well recognized by our great ancestors for many centuries. They used medicinal plants to cure infectious diseases in human beings and animals by traditional methods. Even today in this digital world, medicinal plants contribute extensively for various developments in the form of new drug discovery, drug formulas \unskip~\citep{808536:19339468}  and antibiotics for treating diseases. 

Globally, there are many rare species of plants still unexplored for its medicinal properties. In India, one such rare medicinal plant is Lannea Coromandelica (L.C.) belonging to the Anacardiaceae family \unskip~\citep{808536:19339467,808536:19339442}. 

Traditionally, this plant was used as folk medicine by local tribes in different parts of the India \unskip~\cite{808536:19339444,808536:19339456} and also in other countries like China, Srilanka, Bangladesh \unskip~\citep{808536:19339450,808536:19339463}, Africa \unskip~\citep{808536:19339462,808536:19339456}, Nepal and Pakistan. It is widely known as Indian ash tree \unskip~\citep{808536:19339451,808536:19339458} and by different names based on the growing location of the plant. Tribal people used various parts of this plant for treating pain \unskip~\citep{808536:19339452,808536:19339445}, wounds \unskip~\citep{808536:19339460}, cuts, ulcers \unskip~\citep{808536:19339443,808536:19340919}, gastritis, jaundice \unskip~\citep{808536:19339459}, gout, sprains, diarrhoea \unskip~\citep{808536:19341002}, sore eyes, leprosy, impotence \unskip~\citep{808536:19339467} and dysentery. \unskip~\citep{808536:19339466} in the review article illustrated medicinal importance of Odina, Wodier. She has explained that ethanol extracts of the plant showed to produce a pharmacological response which includes antioxidant activity, antimicrobial, wound healing, anti-diabetic, anti-arthritic activity. 

It has been found by \unskip~\citet{808536:19339465} that L.C. comprised of Flavonoids, tannins, polyphenols, Gallic acid, mucilage, gums and some sterols. The flowers and leaves of L.C. were found to constitute Ellagic acid and Quercetin-3-arabinoside \unskip~\citep{808536:19339464}. No preliminary studies have been reported so far on the stem of Lannea Coromandelica. Therefore, we have delved the phytochemical compositions of L.C. stem for its pharmacological and therapeutic applications.
    
\section{Materials and Methods}
\textbf{Identification and Collection}

Fresh samples of L.C. stems were procured in April 2019 from Siddha Medicinal Plants, Mettur, Tamil Nadu, India with the help of Dr Padma Sorna, Research officer {\textendash} Botanist. The stems of the plant along with leaves, flowers and other plant parts were submitted to Siddha Medicinal Plants Garden, Mettur as herbarium. The sample specimen of L.C. was identified and authenticated by Dr P. Radha, Research officer {\textendash} Botany.

\textbf{Flavonoid estimation}

Aluminium chloride was used to estimate flavonoid found in the L.C. stem extracts in which Quercetin was used as standard described by \unskip~\cite{808536:19339448}. In Ethyl acetate (10 mg/ ml) Quercetin and extracts were prepared. 

In the test tube, 0.9 ml of distilled water was added along with 0.1 ml of L.C. extract and mixed with 75 \ensuremath{\mu}L of 5\% sodium nitrate solution. This mixture was allowed to remain for about 6 minutes, and after that 10 \ensuremath{\mu}L of 10\%, Aluminum chloride was added. After 5 minutes, 0.5 ml of 1M Sodium hydroxide was added to the mixture. To the mix, distilled water was added to make it up to 2.5 ml and thereby the mixture was shaken well. UV-Visible absorbance was measured at 510 nm. The estimation of flavonoid was expressed in terms of Quercetin equivalence (Q.E.) \ensuremath{\mu}g/mg of L.C. extract.

\textbf{Extract preparation}

The stems of Lannea Coromandelica were shade dried for one month and Coarse powdered by using a mixer grinder. The powder was stored at room temperature and covered by plastic wrap to avoid moisture absorption. The finely powdered sample was subjected to sequential extraction using ethanol. The extracts were filtered and concentrated to a dry mass separately, evaporated with the help of rotary evaporator at 35\ensuremath{^\circ} C and stored in the refrigerator for the phytochemical analysis. 

\textbf{Phenol estimation}

Folin Ciocalteu colorimetric method was used to estimate phenol compounds in the L.C. ethanolic extracts. They were with Gallic Acid as standard (10mg/10ml) by \unskip~\cite{808536:19339453}. 

To different test tubes, 20 to 100 \ensuremath{\mu}l of standard solutions were added. 5ml of Folins-Ciocalteu (1:10 dilution) was added to a separate test tube containing extract at a concentration of 10 mg/ml, and the mixture was shaken thoroughly. 

For half an hour, the mixture was incubated after adding 4ml of 0.7 M sodium carbonate. UV-Visible absorbance was measured at 765nm. The estimation of phenol was expressed in terms of Gallic acid equivalence (G.E.) \ensuremath{\mu}g/mg of L.C. extract.


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\caption{\boldmath {The standard calibration curve of Quercetin}}
\label{f-1d312a04465d}
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\egroup

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\caption{\boldmath {The standard calibration curve of Gallic acid}}
\label{f-56c5aec62131}
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\caption{\boldmath {The standard Tannic acid calibration curve}}
\label{f-026ce5d07fa5}
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\egroup

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\caption{\boldmath {Gas chromatogram of L.C. stem}}
\label{f-28b18ec109b6}
\end{figure}
\egroup
\textbf{Tannin estimation}

Folin-Ciocalteu method was used to estimate the tannin found in the ethanolic extracts of L.C. by \unskip~\cite{808536:19339446}. 1 ml of 35 \% Na2CO3, 7.5 ml of distilled water and 0.5 ml of Folin-Ciocalteu reagent was added to the test tube. To this mixture, 0.1 ml of the sample extracts containing 1 mg were added and by adding distilled water to this mixture, it was made up to 10ml. The mixture was kept at room temperature for 30 minutes after being thoroughly shaken. 20, 40, 60, 80 and 100 \ensuremath{\mu }g/ml) were used as standard reference solutions for tannic acid. UV/Visible absorbance was measured at 725 nm against the blank solution. The estimation of tannin was expressed in terms of mg of Tannic acid equivalence (T.E.) \ensuremath{\mu}g/mg of L.C. extract.

\textbf{GC-MS analysis of L.C. stem}

Using Turbo mass software (5.2.0) and Perkin Elmer Clarus 500, the GCMS analysis was performed. A capillary column of 250 \ensuremath{\mu}m column I.D. and 30m length with Elite 5MS (5\% phenyl 95\% dimethyl polysiloxane) was used in the Perkin Elmer Clarus 500. The temperature for the mass analyzer was 8\text{\textordmasculine}C/min to 200\text{\textordmasculine}C. The oven temperature was retained at 50\text{\textordmasculine}C with 1-minute holding time in the initial stage of GC The Helium gas was used as a carrier with flow rate at 1ml/minute and 1:10 as the split ratio, Mass range was 40-600 amu, the energy of electron used was 70ev, Electron Ionization (E.I.) was used in the MS and the source temperature Transfer line was 180\text{\textordmasculine}C, 200\text{\textordmasculine}C, 1.0\ensuremath{\mu}L was injected for the analysis.


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\caption{\boldmath {Chemical structure of compound}}
\label{f-e8a052ddcd2a}
\end{figure}
\egroup

    
\section{Results and Discussion}
The preliminary phytochemical components from the analysis showed that tannins, phenols, flavonoids, alkaloids, saponins, terpenoids and anthraquinones were present in the ethanolic extract of L.C. stems. Table~\ref{tw-7709d43148cc}  shows the results obtained from the qualitative analysis.

The effect of Flavonoids on the cell membrane permeability and inhibiting enzymes found on the membranes like phospholipase A2 and ATPase has been reported by \unskip~\cite{808536:19339447}. The medicinal uses of alkaloids found in the plant stems have been known for centuries, and among them, cytotoxicity is one of their biological properties \unskip~\citep{808536:19339454}. The presence of terpenoid serves as anti-inflammatory and anti-fungal properties. The following inferences have been identified,


  
  \begin{enumerate}
  \item \relax The tannins present in the plant stem L.C. exhibits anti-cancer and anti-inflammation properties.  
  \item \relax The phenols detected from the stems have anti-oxidative properties.
  \item \relax Saponins compound found in L.C. stems known to possess significant anti-cancer properties. 
  \end{enumerate}
  
\begin{table*}[!htbp]
\caption{\boldmath {Phytochemical Qualitative analysis} }
\label{tw-7709d43148cc}
\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.0945\linewidth-2\tabcolsep}p{\dimexpr.16939999999999998\linewidth-2\tabcolsep}p{\dimexpr.2914\linewidth-2\tabcolsep}p{\dimexpr.2629\linewidth-2\tabcolsep}p{\dimexpr.1818\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}S.No &  Test & Procedure & Observation & Ethanolic extraction of L.C Stems\\
\tblmidrule 
1. &
  Tannins &
  Extract + 5\% ferric chloride  &
  Dark blue or greenish black &
  \cAlignHack + \\
 &
   &
   &
   &
  \\
2. &
  Saponins &
  Extract + distilled H2O &
  Formation of 1cm layer of foam &
  \cAlignHack + \\
 &
   &
   &
   &
  \\
3. &
  Flavonoids &
  Extract + NaOH &
  Yellow color &
  \cAlignHack + \\
 &
   &
   &
   &
  \\
4. &
  Alkaloids &
  Extract + HCL, \mbox{}\protect\newline Mayer's reagent  &
  Green color or white precipitate &
  \cAlignHack + \\
 &
   &
   &
   &
  \\
5. &
  Quinones &
  Extract+ Conc. H2SO4 &
  Red color &
  \cAlignHack - \\
 &
   &
   &
   &
  \\
6. &
  Glycosides &
  Extract + chloroform + 10\% NH3 &
  Pink color &
  \cAlignHack - \\
 &
   &
   &
   &
  \\
7. &
  Terpenoids &
  Extract + chloroform + Conc. H2SO4 &
  Red brown color &
  \cAlignHack + \\
 &
   &
   &
   &
  \\
8. &
  Phenols &
  Phenol Ciocalteau's reagent \mbox{}\protect\newline 15\% Na2CO3 &
  Blue or green color &
  \cAlignHack + \\
 &
   &
   &
   &
  \\
9. &
  Coumarin &
  Extract + 10\% NaOH &
  Yellow color &
  \cAlignHack - \\
 &
   &
   &
   &
  \\
10. &
  Steroids &
  Chloroform + Conc. H2SO4 &
  Appearance of brown ring &
  \cAlignHack - \\
 &
   &
   &
   &
  \\
11. &
  Phlobatannins &
  Extract + 2\% HCL &
  Red color precipitate &
  \cAlignHack - \\
 &
   &
   &
   &
  \\
12. &
  Anthraquinones &
  Extract + 10\% NH3 &
  Pink color precipitate &
  \cAlignHack + \\
 &
   &
   &
   &
  \\
13. &
  Cardiac Glycosides &
  Extract + glacial acid + 5\% ferric chloride  \mbox{}\protect\newline Conc.H2SO4 &
  Formation of brown ring interface &
  \cAlignHack - \\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Phytochemical Quantitative analysis} }
\label{tw-7939b41cadc0}
\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.2798\linewidth-2\tabcolsep}p{\dimexpr.38019999999999996\linewidth-2\tabcolsep}p{\dimexpr.34\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}S.No & Estimation & Stem \%\\
\tblmidrule 
1. &
  Flavonoids (Quercetin) &
  0.30\%\\
2. &
  Phenols (Gallic acid) &
  3.02\%\\
3. &
  Tannins(Tannic acid) &
  23.7\%\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

\begingroup
\makeatletter\if@twocolumn\@ifundefined{theposttbl}{\gdef\TwoColDocument{true}\onecolumn\onecolumn}{}\fi\makeatother \setlength\LTcapwidth{\textwidth}
\begin{longtable}{p{\dimexpr.1077\linewidth-2\tabcolsep}p{\dimexpr.10780000000000001\linewidth-2\tabcolsep}p{\dimexpr.16840000000000003\linewidth-2\tabcolsep}p{\dimexpr.6161\linewidth-2\tabcolsep}}
\caption{Chemical compounds identified by GC-MS}
\label{tw-b6a8dc40f91c}
\def\arraystretch{1.1}\\\endfirsthead \hline \noalign{\vskip3pt} \noalign{\textit{Table \thetable\ continued}} \noalign{\vskip3pt} 
\tbltoprule \rowcolor{kwdboxcolor}S.No  & RT & Area \%  &  Compound name\\
\tblmidrule \endhead \hline \noalign{\vskip3pt} \noalign{\textit{\hfill Continued on next page}} \noalign{\vskip3pt} \endfoot \endlastfoot 
\tbltoprule \rowcolor{kwdboxcolor}S.No  & RT & Area \%  &  Compound name\\
\tblmidrule 
1 &
  3.013 &
  0.369 &
  1,4-Benzenediol, 2,6-bis(1,1-dimethylethyl)-\\
2 &
  3.153 &
  0.475 &
  6-Amino-5-cyano-4-(5-cyano-2,4-dimethyl-1Hpyrrol- \mbox{}\protect\newline 3-yl)-2-methyl-4H-pyran-3-carboxylic acid \mbox{}\protect\newline ethyl ester \\
3 &
  3.194 &
  0.257 &
  Coumarin-6-ol, 3,4-dihydro-4,4-dimethyl-5,7-dinitro- \\
4 &
  10.066 &
  0.229 &
  Carda-4,20(22)-dienolide, 3-[(6-deoxy-3-O-methyl\`{a}- \mbox{}\protect\newline L-mannopyranosyl)oxy]-14-hydroxy-, (3\'{a})- \\
5 &
  11.98 &
  0.343 &
  Phenol, 3,5-bis(1,1-dimethylethyl)- \\
6 &
  13.99 &
  0.307 &
  d-Mannitol, 1-decylsulfonyl- \\
7 &
  18.20 &
  0.340 &
  Octadecane, 3-ethyl-5-(2-ethylbutyl)- \\
8 &
  18.7 &
  0.229 &
  1-Benzazirene-1-carboxylic acid, 2,2,5a-trimethyl- \mbox{}\protect\newline 1a-[3-oxo-1-butenyl] perhydro-, methyl ester \\
9 &
  18.79 &
  0.243 &
  18-Norcholest-17(20),24-dien-21-oic acid, 16- \mbox{}\protect\newline acetoxy-4,8,14-trimethyl-3,11-dioxo-, methyl ester \\
10 &
  20.72 &
  1.022 &
  Pentadecanoic acid, 14-methyl-, methyl ester \\
11 &
  21.42 &
  0.333 &
  Urea, N-[5-(ethylsulfonyl)-1,3,4-thiadiazol-2-yl]-N, N'-dimethyl-\\
12 &
  21.47 &
  0.327 &
  Taxa-4,11-diene\\
13 &
  21.54 &
  0.229 &
  \`{a}-d-Xylopyranoside, methyl-2,3,4-tris-O-[9- \mbox{}\protect\newline borabicyclo[3.3.1]non-9-yl]- \\
14 &
  21.626 &
  0.302 &
  16-Nitrobicyclo[10.4.0]hexadecan-1-ol-13-one \\
15 &
  22.191 &
  0.329 &
  Milbemycin B, 6,28-anhydro-15-chloro-25- \mbox{}\protect\newline isopropyl-13-dehydro-5-O-demethyl-4-methyl- \\
16 &
  23.72 &
  0.243 &
  Fumaric acid, tetradec-3-enyl tridecyl ester \\
17 &
  23.74 &
  0.354 &
  Pregn-4-ene-3,20-dione, 17,21-dihydroxy-, bis(Omethyloxime) \\
18 &
  24.02 &
  0.354 &
  Milbemycin B, 6,28-anhydro-15-chloro-25- \mbox{}\protect\newline isopropyl-13-dehydro-5-O-demethyl-4-methyl- \\
19 &
  24.13 &
  0.399 &
  Aspidospermidine, 1,2-didehydro-, (5\`{a},12\'{a},19\`{a})- \\
20 &
  24.18 &
  0.229 &
  (5\'{a})Pregnane-3,20\'{a}-diol, 14\`{a},18\`{a}-[4-methyl-3-oxo- \mbox{}\protect\newline (1-oxa-4-azabutane-1,4-diyl)]-, diacetate \\
21 &
  24.24 &
  0.25 &
  Echitamine \\
22 &
  24.53 &
  0.42 &
  Strychane, 1-acetyl-20\`{a}-hydroxy-16-methylene- \\
23 &
  28.68 &
  0.283 &
  2-(Ethyl)oxybenzylidene acetophenone \\
24 &
  29.72 &
  0.289 &
  Carbonic acid, (1,2,3,4-tetrahydro-6- \mbox{}\protect\newline methoxycarboniloxy-3-methyl-1- \mbox{}\protect\newline naphthylidenamino)-, methyl diester \\
25 &
  29.83 &
  0.246 &
  1-Benzazirene-1-carboxylic acid, 2,2,5a-trimethyl- \mbox{}\protect\newline 1a-[3-oxo-1-butenyl] perhydro-, methyl ester \\
26 &
  29.86 &
  0.228 &
  2-Azatricyclo[3.2.0.0(2,4)]hept-6-ene-5-carboxylic \mbox{}\protect\newline acid, 1,6,7-tri-t-butyl-4-phenyl-, t-butyl ester \\
27 &
  29.92 &
  0.283 &
   Milbemycin B, 6,28-anhydro-15-chloro-25- \mbox{}\protect\newline isopropyl-13-dehydro-5-O-demethyl-4-methyl- \\
28 &
  29.99 &
  0.338 &
  Heptasiloxane, 1,1,3,3,5,5,7,7,9,9,11,11,13,13- \mbox{}\protect\newline tetradecamethyl- \\
29 &
  30.02 &
  0.724 &
  Propanoic acid, 2-(3-acetoxy-4,4,14- \mbox{}\protect\newline trimethylandrost-8-en-17-yl)- \\
30 &
  30.09 &
  0.37 &
  Acetic acid, 17-acetoxy-4,4,10,13-tetramethyl-7- \mbox{}\protect\newline oxo-2,3,4,7,8,9,10,11,12,13,14,15,16,17- \mbox{}\protect\newline tetradecahydro-1H-cyclopenta[a]phenanthren-3-yl \mbox{}\protect\newline (ester) \\
31 &
  30.164 &
  0.233 &
  Corynan-17-ol, 18,19-didehydro-10-methoxy- \\
32 &
  30.504 &
  0.234 &
  2-Nonadecanone 2,4-dinitrophenylhydrazine \\
33 &
  30.69 &
  0.404 &
  2-[2-(4-Chlorophenyl)-3-morpholin-4-yl-3-thioxopropenylamino] \mbox{}\protect\newline Acetamide \\
34 &
  30.74 &
  0.265 &
  Propanoic acid, 2-(3-acetoxy-4,4,14- \mbox{}\protect\newline trimethylandrost-8-en-17-yl)- \\
35 &
  30.84 &
  0.594 &
  2-(Ethyl)oxybenzylidene acetophenone \\
36 &
  30.95 &
  0.275 &
  Curan-17-oic acid, 2,16-didehydro-20-hydroxy-19- \mbox{}\protect\newline oxo-, methyl ester \\
37 &
  31.01 &
  0.338 &
  1H-Cyclopropa[3,4]benz[1,2-e]azulene-5,7b,9,9atetrol, \mbox{}\protect\newline 1a,1b,4,4a,5,7a,8,9-octahydro-3- \mbox{}\protect\newline (hydroxymethyl)-1,1,6,8-tetramethyl-, 5,9,9atriacetate, \mbox{}\protect\newline [1aR-(1a\`{a},1b\'{a},4a\'{a},5\'{a},7a\`{a},7b\`{a},8\`{a},9\'{a}, \mbox{}\protect\newline 9a\`{a})]- \\
38 &
  31.09 &
  0.246 &
  Methyl glycocholate, 3TMS derivative \\
39 &
  31.18 &
  0.253 &
  Butanoic acid, 4-chloro-, 1,1a,1b,4,4a,5,7a,7b,8,9- \mbox{}\protect\newline decahydro-4a,7b-dihydroxy-3-(hydroxymethyl)-1, \mbox{}\protect\newline 1,6,8-tetramethyl-5-oxo-9aH-cyclopropa[3, \mbox{}\protect\newline 4]benz[1,2-e]azulene-9,9a-diyl ester, [1ar-(1a\`{a}, \mbox{}\protect\newline 1b\'{a},4a\'{a},7a\`{a},7b\`{a},8\`{a},9\'{a},9a\`{a})]- \\
40 &
  31.42 &
  0.457 &
  Aspidofractinine-1-carboxaldehyde, 3-oxo-, (2\`{a},5\`{a})- \\
41 &
  31.62 &
  0.299 &
  5-Methyl-2-N-methylaminobenzophenone \mbox{}\protect\newline Semicarbazone \\
42 &
  31.73 &
  0.553 &
  Strychane, 1-acetyl-20\`{a}-hydroxy-16-methylene- \\
43 &
  32.03 &
  0.348 &
  Methyl glycocholate, 3TMS derivative \\
44 &
  32.21 &
  0.353 &
  6-Amino-5-cyano-4-(5-cyano-2,4-dimethyl-1Hpyrrol- \mbox{}\protect\newline 3-yl)-2-methyl-4H-pyran-3-carboxylic acid \mbox{}\protect\newline ethyl ester \\
45 &
  32.66 &
  0.254 &
  1,4-Benzenediol, 2,6-bis(1,1-dimethylethyl)- \\
46 &
  33.49 &
  0.364 &
  2,4,6,8,10-Tetradecapentaenoic acid, 9a- \mbox{}\protect\newline (acetyloxy)-1a,1b,4,4a,5,7a,7b,8,9,9a-decahydro- \mbox{}\protect\newline 4a,7b-dihydroxy-3-(hydroxymethyl)-1,1,6,8- \mbox{}\protect\newline tetramethyl-5-oxo-1H-cyclopropa[3,4]benz[1,2- \mbox{}\protect\newline e]azulen-9-yl ester, [1aR-(1a\`{a},1b\'{a},4a\'{a},7a\`{a},7b\`{a},8\`{a}, \mbox{}\protect\newline 9\'{a},9a\`{a})]- \\
47 &
  33.65 &
  0.233 &
  5,7,9(11)-Androstatriene, 3-hydroxy-17-oxo- \\
48 &
  34.61 &
  0.228 &
  1H-Cyclopropa[3,4]benz[1,2-e]azulene-5,7b,9,9atetrol, \mbox{}\protect\newline 1a,1b,4,4a,5,7a,8,9-octahydro-3- \mbox{}\protect\newline (hydroxymethyl)-1,1,6,8-tetramethyl-, 5,9,9atriacetate, \mbox{}\protect\newline [1aR-(1a\`{a},1b\'{a},4a\'{a},5\'{a},7a\`{a},7b\`{a},8\`{a},9\'{a},9a\`{a})]-\\
49 &
  34.69 &
  0.894 &
  (22S)-6\'{a},11\'{a},21-Trihydroxy-16\`{a},17\`{a}propylmethylenedioxypregna-1,4-diene-3,20-dione\\
50 &
  34.77 &
  0.269 &
  Butanoic acid, 1a,2,5,5a,6,9,10,10a-octahydro-5ahydroxy-4-(hydroxymethyl)-1,1,7,9-tetramethyl-6, \mbox{}\protect\newline 11-dioxo-1H-2,8amethanocyclopenta[ \mbox{}\protect\newline a]cyclopropa[e]cyclodecen-5-yl ester, [1aR-(1a\`{a},2\`{a},5\'{a},5a\'{a},8a\`{a},9\`{a},10a\`{a})]-\\
\tblbottomrule 
\end{longtable}
\endgroup
\makeatletter\@ifundefined{TwoColDocument}{}{\twocolumn}\makeatother 
In this present work, different compounds showed various percentages which are mainly due to the geographical location of the plant.

The total Flavonoid content from stems of L.C. by the ethanolic extract was 3.05\ensuremath{\mu}g/mg (0.30\%) expressed in Quercetin equivalence. Figure~\ref{f-1d312a04465d} shows the standard calibration curve of Quercetin. In the stems of L.C., the phenolic content was estimated to be 30.2 \ensuremath{\mu}g/mg (3.02\%) expressed in Gallic acid equivalence, and the calibration curve is shown in Figure~\ref{f-56c5aec62131}. The tannin content in this study was found to be higher 237.5 \ensuremath{\mu}g/mg (23.7\%) expressed in tannic acid equivalence. The results of tannin standard calibration curve are shown in Figure~\ref{f-026ce5d07fa5} and given in the Table~\ref{tw-7939b41cadc0}. \textbf{\space }

\textbf{Gas Chromatography-Mass Spectroscopy (GCMS)}

The GCMS analysis (Figure~\ref{f-28b18ec109b6}) from L.C. stem illustrates peaks showing a wide range of compounds. An inceptive inquiry of the stem revealed the identification of fifty compounds. Results of the compounds are given in Table~\ref{tw-b6a8dc40f91c}. The primary component was Pentadecanoic acid, 14-methyl-methyl ester (1.0\%) (Figure~\ref{f-e8a052ddcd2a}) and other compounds were present in traces. The different compounds present in the L.C. stem is mainly due to climatic and geographical location of the plant \unskip~\citep{808536:19339449}.
    
\section{Conclusion}
The presence of these chemical (Flavonoid, alkaloid, saponin, terpenoid, phenol and tannin) components from phytochemical screening in L.C. stem extract can be used in the different ailments through their anti-inflammatory, anti-cancer, anti-arthritic, antioxidant and antimicrobial properties. Further exploration of the L.C. stem could be achieved in the future by isolation and detecting its active compounds.
\section*{Acknowledgement}J.B. would like to heartily thank his supervisor Prof Kesavi Durairaj for guidance and support in all stages of work. J.B. extends his sincere gratitude to Prof. S. P. Thyagarajan, Professor of Eminence \& Dean (Research), Sri Ramachandra Institute of Higher Education and Research for providing facilities and financial support. J.B. also acknowledges assistance rendered by Dr Padma Sorna and Dr P. Radha in the identification and collection of plant materials.

\textbf{Funding Support}

Nil

\textbf{Conflict of Interest}

Nil
    

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