\documentclass[twocolumn,twoside]{article}
\makeatletter\if@twocolumn\PassOptionsToPackage{switch}{lineno}\else\fi\makeatother

\usepackage{amsfonts,amssymb,amsbsy,latexsym,amsmath,tabulary,graphicx,times,xcolor}

%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
% Following additional macros are required to function some 
% functions which are not available in the class used.
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
\usepackage{url,multirow,morefloats,floatflt,cancel,tfrupee}
\makeatletter


\AtBeginDocument{\@ifpackageloaded{textcomp}{}{\usepackage{textcomp}}}
\makeatother
\usepackage{colortbl}
\usepackage{xcolor}
\usepackage{pifont}
\usepackage[nointegrals]{wasysym}
\urlstyle{rm}
\makeatletter

%%%For Table column width calculation.
\def\mcWidth#1{\csname TY@F#1\endcsname+\tabcolsep}

%%Hacking center and right align for table
\def\cAlignHack{\rightskip\@flushglue\leftskip\@flushglue\parindent\z@\parfillskip\z@skip}
\def\rAlignHack{\rightskip\z@skip\leftskip\@flushglue \parindent\z@\parfillskip\z@skip}

%Etal definition in references
\@ifundefined{etal}{\def\etal{\textit{et~al}}}{}


%\if@twocolumn\usepackage{dblfloatfix}\fi
\usepackage{ifxetex}
\ifxetex\else\if@twocolumn\@ifpackageloaded{stfloats}{}{\usepackage{dblfloatfix}}\fi\fi

\AtBeginDocument{
\expandafter\ifx\csname eqalign\endcsname\relax
\def\eqalign#1{\null\vcenter{\def\\{\cr}\openup\jot\m@th
  \ialign{\strut$\displaystyle{##}$\hfil&$\displaystyle{{}##}$\hfil
      \crcr#1\crcr}}\,}
\fi
}

%For fixing hardfail when unicode letters appear inside table with endfloat
\AtBeginDocument{%
  \@ifpackageloaded{endfloat}%
   {\renewcommand\efloat@iwrite[1]{\immediate\expandafter\protected@write\csname efloat@post#1\endcsname{}}}{\newif\ifefloat@tables}%
}%

\def\BreakURLText#1{\@tfor\brk@tempa:=#1\do{\brk@tempa\hskip0pt}}
\let\lt=<
\let\gt=>
\def\processVert{\ifmmode|\else\textbar\fi}
\let\processvert\processVert

\@ifundefined{subparagraph}{
\def\subparagraph{\@startsection{paragraph}{5}{2\parindent}{0ex plus 0.1ex minus 0.1ex}%
{0ex}{\normalfont\small\itshape}}%
}{}

% These are now gobbled, so won't appear in the PDF.
\newcommand\role[1]{\unskip}
\newcommand\aucollab[1]{\unskip}
  
\@ifundefined{tsGraphicsScaleX}{\gdef\tsGraphicsScaleX{1}}{}
\@ifundefined{tsGraphicsScaleY}{\gdef\tsGraphicsScaleY{.9}}{}
% To automatically resize figures to fit inside the text area
\def\checkGraphicsWidth{\ifdim\Gin@nat@width>\linewidth
	\tsGraphicsScaleX\linewidth\else\Gin@nat@width\fi}

\def\checkGraphicsHeight{\ifdim\Gin@nat@height>.9\textheight
	\tsGraphicsScaleY\textheight\else\Gin@nat@height\fi}

\def\fixFloatSize#1{}%\@ifundefined{processdelayedfloats}{\setbox0=\hbox{\includegraphics{#1}}\ifnum\wd0<\columnwidth\relax\renewenvironment{figure*}{\begin{figure}}{\end{figure}}\fi}{}}
\let\ts@includegraphics\includegraphics

\def\inlinegraphic[#1]#2{{\edef\@tempa{#1}\edef\baseline@shift{\ifx\@tempa\@empty0\else#1\fi}\edef\tempZ{\the\numexpr(\numexpr(\baseline@shift*\f@size/100))}\protect\raisebox{\tempZ pt}{\ts@includegraphics{#2}}}}

%\renewcommand{\includegraphics}[1]{\ts@includegraphics[width=\checkGraphicsWidth]{#1}}
\AtBeginDocument{\def\includegraphics{\@ifnextchar[{\ts@includegraphics}{\ts@includegraphics[width=\checkGraphicsWidth,height=\checkGraphicsHeight,keepaspectratio]}}}

\DeclareMathAlphabet{\mathpzc}{OT1}{pzc}{m}{it}

\def\URL#1#2{\@ifundefined{href}{#2}{\href{#1}{#2}}}

%%For url break
\def\UrlOrds{\do\*\do\-\do\~\do\'\do\"\do\-}%
\g@addto@macro{\UrlBreaks}{\UrlOrds}



\edef\fntEncoding{\f@encoding}
\def\EUoneEnc{EU1}
\makeatother
\def\floatpagefraction{0.8} 
\def\dblfloatpagefraction{0.8}
\def\style#1#2{#2}
\def\xxxguillemotleft{\fontencoding{T1}\selectfont\guillemotleft}
\def\xxxguillemotright{\fontencoding{T1}\selectfont\guillemotright}

\newif\ifmultipleabstract\multipleabstractfalse%
\newenvironment{typesetAbstractGroup}{}{}%

%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
\AtBeginDocument{\let\citet\cite\let\citep\cite}
\usepackage[numbers,sort&compress,numbers,sort&compress]{natbib}



\newif\ifnokeywords\nokeywordsfalse
\def\abbrvJournalTitle{Future J. Pharm. Health. Sci.}
\def\jrnlurlname{fjphs}
\usepackage{balance}

\makeatletter\input{size10-pointfive.clo}\makeatother%
\definecolor{kwdboxcolor}{RGB}{242,242,242}
\usepackage[hidelinks,colorlinks=true,allcolors=blue]{hyperref}
\linespread{1}                 
\def\floatpagefraction{0.8}

\usepackage[paperheight=11.69in,paperwidth=8.26in,top=1in,bottom=1in,left=1in,right=.75in,headsep=24pt]{geometry}

\usepackage{multirow-custom}

\makeatletter

\def\hlinewd#1{%
  \noalign{\ifnum0=`}\fi\hrule \@height #1%
  \futurelet\reserved@a\@xhline}
\def\tbltoprule{\hlinewd{1pt}\\[-14pt]}
\def\tblbottomrule{\noalign{\vspace*{6pt}}\hline\noalign{\vspace*{2pt}}}
\def\tblmidrule{\hline\noalign{\vspace*{2pt}}}



\let\@articleType\@empty
\let\@journalDoi\@empty
\let\@journalVolume\@empty
\let\@journalIssue\@empty
\let\@crossMarkLink\@empty
\let\@receivedDate\@empty
\let\@acceptedDate\@empty
\let\@revisedDate\@empty
\let\@publishedDate\@empty
\let\@copyrightYear\@empty
\let\@firstPage\@empty
\let\@eISSN\@empty
\let\@pISSN\@empty
\def\eISSN#1{\gdef\@eISSN{#1}}
\def\pISSN#1{\gdef\@pISSN{#1}}
\def\articleType#1{\gdef\@articleType{#1}}
\def\journalDoi#1{\gdef\@journalDoi{#1}}
\def\crossMarkLink#1{\gdef\@crossMarkLink{#1}}
\def\receivedDate#1{\gdef\@receivedDate{#1}}
\def\acceptedDate#1{\gdef\@acceptedDate{#1}}
\def\revisedDate#1{\gdef\@revisedDate{#1}}
\def\publishedDate#1{\gdef\@publishedDate{#1}}
\def\copyrightYear#1{\gdef\@copyrightYear{#1}}
\def\journalVolume#1{\gdef\@journalVolume{#1}}
\def\journalIssue#1{\gdef\@journalIssue{#1}}
\def\firstPage#1{\gdef\@firstPage{#1}}


\def\author#1{%
  \gdef\@author{%
    \hskip-\dimexpr(\tabcolsep)\hskip5pt%
    \parbox{\dimexpr\textwidth-1pt}%
    {\fontsize{11}{13}\selectfont\raggedright #1}%
  }%
}


\usepackage{pharmascope-abs}
\usepackage{caption}
\usepackage{lastpage}
\usepackage{fancyhdr}
\usepackage[noindentafter,explicit]{titlesec}
\usepackage{fontspec}

\setmainfont[%
BoldFont=cambriab.ttf,%
ItalicFont=CAMBRIAI.TTF,%
BoldItalicFont=CAMBRIAZ.TTF]{Cambria.ttf}

\usepackage{qrcode}

%\def\abbrvJournalTitle{Future J. Pharm. Health. Sci.}
\def\jshort{fjphs}

\def\title#1{%
  \gdef\@title{%
%      \vspace*{-20pt}%
       \vspace*{-48pt}
       \ifx\@articleType\@empty\else{\fontsize{10}{12}\scshape\selectfont\hspace{8pt}\@articleType\hfill\mbox{}\par\vspace{2pt}}\fi%
       \minipage{\linewidth}
         \hrulefill\\[-0.7pt]%
         \parbox{.1\linewidth}{\includegraphics[width=4.2cm]{pharmasprings_logo.png}}\hfill
%         \fcolorbox{kwdboxcolor}{kwdboxcolor}{\parbox{.792\linewidth}{%
         \fcolorbox{kwdboxcolor}{kwdboxcolor}{\parbox{.729\linewidth}{%
          \begin{center}
          \vskip-4pt 
          \scshape\fontsize{18}{20}\selectfont\journalTitle\hfill\end{center}%
          \vspace*{-16pt}{\vspace*{-3.1pt}\fontsize{8}{9}\selectfont Published by Pharma Springs Publication\hfill  Journal Home Page: \href{https://pharmasprings.com/index.php/\jrnlurlname}{\color{blue}\underline{\smash{https://pharmasprings.com/\jrnlurlname}}}}\hspace*{4pt}\mbox{}}}%
       \par\rule{\linewidth}{1.3pt}%
       \endminipage%
    \par\vspace*{9.2pt}\parbox{.98\linewidth}{\linespread{.9}\raggedright\fontsize{14}{17}\selectfont #1}%
    \vspace*{-8pt}%
  }
}


\setlength{\parindent}{0pt}
\setlength{\parskip}{0.4pc plus 1pt minus 1pt}


\fancypagestyle{headings}{%
  \renewcommand{\headrulewidth}{0pt}%
  \renewcommand{\footrulewidth}{0.3pt}
  \fancyhf{}%
  \fancyhead[R]{%
    \fontsize{9.12}{11}\selectfont\RunningAuthor,\ \abbrvJournalTitle\ \@copyrightYear;\space\ifx\@journalVolume\@empty X\else\@journalVolume\fi%
    \ifx\@journalIssue\@empty\else(\@journalIssue)\fi%
    :\ \ifx\@firstPage\@empty 1\else\@firstPage\fi-\pageref*{LastPage}%
  }%
  \fancyfoot[RE]{\fontsize{9.12}{11}\selectfont\textcopyright\ Pharma Springs Publication\ $|$\space \journalTitle}%
  \fancyfoot[LO]{\fontsize{9.12}{11}\selectfont\textcopyright\ Pharma Springs Publication\ $|$\space \journalTitle}%
  \fancyfoot[RO,LE]{\fontsize{9.12}{11}\selectfont\thepage}
}\pagestyle{headings}


                                                         
\fancypagestyle{plain}{%
  \renewcommand{\headrulewidth}{0pt}%
  \renewcommand{\footrulewidth}{0.3pt}%
  \fancyhf{}%
  \fancyhead[R]{%
    \fontsize{9.12}{11}\selectfont\RunningAuthor,\ \abbrvJournalTitle\ \@copyrightYear;\space\ifx\@journalVolume\@empty X\else\@journalVolume\fi%
    \ifx\@journalIssue\@empty\else(\@journalIssue)\fi%
    :\ \ifx\@firstPage\@empty 1\else\@firstPage\fi-\pageref*{LastPage}%
  }%
  \fancyfoot[RE]{\fontsize{9.12}{11}\selectfont\textcopyright\ Pharma Springs Publication\ $|$\space \journalTitle}%
  \fancyfoot[LO]{\fontsize{9.12}{11}\selectfont\textcopyright\ Pharma Springs Publication\ $|$\space \journalTitle}%
  \fancyfoot[RO,LE]{\fontsize{9.12}{11}\selectfont\thepage}
  \ifx\@firstPage\@empty\else\setcounter{page}{\@firstPage}\fi
}


\def\NormalBaseline{\def\baselinestretch{1.1}}
\usepackage{textcase}
\setcounter{secnumdepth}{0}

\titleformat{\section}[block]{\bfseries\boldmath\NormalBaseline\filright\fontsize{10.5}{13}\selectfont}
{\thesection}
{6pt}
{\MakeTextUppercase{#1}}
[]
\titleformat{\subsection}[block]{\bfseries\boldmath\NormalBaseline\filright\fontsize{10.5}{12}\selectfont}
{\thesubsection}
{6pt}
{#1}
[]
\titleformat{\subsubsection}[block]{\NormalBaseline\filright\fontsize{10.5}{12}\selectfont}
{\thesubsubsection}
{6pt}
{#1}
[]
\titleformat{\paragraph}[block]{\NormalBaseline\filright\fontsize{10.5}{10}\selectfont}
{\theparagraph}
{6pt}
{#1}
[]
\titleformat{\subparagraph}[block]{\NormalBaseline\filright\fontsize{10.5}{12}\selectfont}
{\thesubparagraph}
{6pt}
{#1}
[]

\titlespacing{\section}{0pt}{.5\baselineskip}{.5\baselineskip}  
\titlespacing{\subsection}{0pt}{.5\baselineskip}{.5\baselineskip}  
\titlespacing{\subsubsection}{0pt}{.5\baselineskip}{.5\baselineskip}  
\titlespacing{\paragraph}{0pt}{.5\baselineskip}{.5\baselineskip}  
\titlespacing{\subparagraph}{0pt}{.5\baselineskip}{.5\baselineskip}

\captionsetup[figure]{skip=1.4pt,font=bf,labelsep=colon,justification=raggedright,singlelinecheck=false}
\captionsetup[table]{skip=1.4pt,font=bf,labelsep=colon,justification=raggedright,singlelinecheck=false}

\def\bibyear#1{#1}
\def\bibjtitle#1{#1}
\def\bibauand{}
\setlength\bibsep{3pt}
\setlength\bibhang{8pt}
\lefthyphenmin = 3
\makeatother
\date{}
\makeatletter
\usepackage{mdframed}
%\usepackage{balance}
\definecolor{unitednationsblue}{RGB}{222,234,246}
\definecolor{cpboxcolor}{RGB}{211,211,211}
\definecolor{citeboxcolor}{RGB}{242,242,242}
\def\printBio{%
  %\noindent\begin{mdframed}[innerbottommargin=7pt,backgroundcolor=unitednationsblue,hidealllines=true,frametitle={ABOUT AUTHORS},innerleftmargin=7pt,leftmargin=2pt,skipabove=1pt,innertopmargin=2pt,innerbottommargin=1pt,skipbelow=1pt]%
  \noindent\begin{mdframed}[innerbottommargin=7pt,backgroundcolor=unitednationsblue,hidealllines=true,frametitle={ABOUT AUTHORS},innerrightmargin=20pt,rightmargin=4,innerleftmargin=7pt,leftmargin=2pt,skipabove=1pt,innertopmargin=2pt,innerbottommargin=1pt,skipbelow=1pt]%
   \@tempcnta=0
    \loop
      \advance \@tempcnta by 1
      \def\aubioCnt{\the\@tempcnta}
      \vskip1pt
%      \noindent\hspace{-2pt}%\fbox{
%     \vspace*{1pt}
      \raisebox{62pt}%{\begin{minipage}[t]{.35\linewidth}%
      \expandafter\ifx\csname authorImage\aubioCnt\endcsname\relax%
       \else%
         \includegraphics[height=60pt,width=60pt]{\csname authorImage\aubioCnt\endcsname}%height=145pt
       \fi%
      %\end{minipage}
      %}%}
      \hskip 4pt%\fbox{
      \raisebox{52pt}{\begin{minipage}[t]{.74\linewidth}%
        {\fontsize{8}{10}\selectfont{\csname authorName\aubioCnt\endcsname\\[5pt]}}
        \fontsize{8}{10}\selectfont\csname authorDetails\aubioCnt\endcsname\\
      \end{minipage}}
%      }
%     \vskip10pt%
    \ifnum\@tempcnta < \theaubio
    \repeat
%     \vspace*{-16.5pt}%
  \end{mdframed}
}


\makeatother


\def\citeauthor{~Manapati~Jyothi,
              ~Varaprasad~A,
              ~Vandana~B,
              ~Nikitha~G,
              ~Sandhya~S,
              ~Sukanya~U}
\def\citetitle{RP-HPLC Method Designed for Determining Charantin in its Capsule Dosage Form}
\def\copyrightstmt{This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial- ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.}
\publishedDate{}
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
% Feature enabled:
%citation-style: unsrt
%citation-type: numbered
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%

\usepackage{float}

\begin{document}


\def\RunningAuthor{Manapati Jyothi et al.}
\firstPage{118}
\articleType{Original Article}
\receivedDate{20 May 2021}
\acceptedDate{01 Jun 2021}
\revisedDate{28 May 2021}
\journalVolume{1}
\journalIssue{3}
\journalDoi{fjphs.v1i3.219}
\copyrightYear{2021}
\eISSN{2583-116X}
\journalDoi{fjphs.v1i3.219}
\def\authorCount{6}
\def\affCount{2}

\def\journalTitle{Future Journal of Pharmaceuticals and Health Sciences}

\title{RP-HPLC Method Designed for Determining Charantin in its Capsule Dosage Form}
\author{Manapati~Jyothi\textsuperscript{*}\textsuperscript{1},  
            Varaprasad~A\textsuperscript{2},  
            Vandana~B\textsuperscript{2},  
            Nikitha~G\textsuperscript{2},  
            Sandhya~S\textsuperscript{2}, 
            Sukanya~U\textsuperscript{2}~\\[5pt]{\textsuperscript{1}Department of Pharmaceutical Analysis, Sun College of Pharmaceutical Education and Research, Kakupalli, Nellore-524346, Andhra Pradesh, India}~\\{\textsuperscript{2}Sun College of Pharmaceutical Education and Research, Kakupalli, Nellore-524346, Andhra Pradesh, India}}

\begin{abstract}
Charantin, a steroidal saponin found in the plant \textit{Momordica} \textit{charantia} and functions like insulin. It is combined with natural insulin, increasing insulin release and inhibiting gluconeogenesis. Charantin improves blood glucose levels by enhancing glucose absorption and glycogen synthesis in the liver, muscles, and fat cells, according to a large body of research. The great majority of the time, it is taken in the form of a capsule. Because of the increased availability of these newly released formulations, a new way of calculating the amount of medications included in the formulations is now required. The current work's RP-HPLC technique is aimed to estimate Charantin as well as confirm that Charantin is being calculated. The proposed RP-HPLC technology for assessing Charantin in capsule dosage form was found to be precise, specific, accurate, quick, and cost-effective. Label Claims (also known as labelling claims) are representations of the actual product attributes as stated on the product label. The sample recoveries from all formulations were consistent with their respective Label Claims, and this method is suitable for routine analysis. It is suitable for normal laboratory analysis and may be used to control the quality of raw materials, formulations, dissolution tests, and bioequivalence studies for the same formulation. 
\end{abstract}\def\keywordstitle{Keywords}

\begin{keywords}Charantin,\newline RP-HPLC,\newline Precision,\newline Accuracy,\newline Stability
\end{keywords}
\twocolumn[ \maketitle {\printKwdAbsBox}]
\makeatletter{\raggedright{\Large$^\ast$}Corresponding Author\par Name:\ Manapati~Jyothi~\\ Phone:\ +91 9908286276~\\  Email:\ mjyothipharmacy@gmail.com
        \par}\vspace*{-13pt}\hrulefill\par\ifx\@eISSN\@empty\else{\fontsize{12}{14}\selectfont eISSN: \@eISSN\hfill pISSN: \@pISSN}\par\fi% 
        \textsc{DOI:}\ \href{https://doi.org/10.26452/\@journalDoi}{\textcolor{blue}{\underline{\smash{https://doi.org/10.26452/\@journalDoi}}}}\par%
        \vspace*{-12pt}\hrulefill\\[5pt]\hypersetup{allcolors=black}\qrcode[version=4,height=1.5cm]{https://doi.org/10.26452/\@journalDoi}\hypersetup{allcolors=blue} \hspace*{10.5pt}\parbox[t]{5cm}{\raisebox{19pt}{\fontsize{9.12}{10.12}\selectfont Production and Hosted by}\\[-13pt]{{\fontsize{12}{14}\selectfont  Pharmasprings.com \\[-1pt]{\fontsize{9.12}{10.12}\selectfont\textcopyright\ \@copyrightYear\ $|$ All rights reserved.}\\[-8pt]\hspace*{-1pt}\rule{.762\columnwidth}{1.2pt}}}}\hypersetup{allcolors=blue}%{\fontsize{12}{14}\selectfont Pharmasprings.com}\par%
        \makeatother
        
    
\section{Introduction}
Charantin is a steroidal saponin derived from \textit{Momordica} \textit{charantia}  that shows insulin-like action by boosting insulin release and inhibiting gluconeogenesis \unskip~\cite{1116378:22316697}. A strong basic chemical like Charantin is almost insoluble (in water) and, as a result, it is nearly impossible to dissolve in dilute acids or bases (based on its pKa). It is a whitish crystalline material, which is neutral and tasteless, melting at around 266{\textendash}268 \ensuremath{^\circ}C. As far as the water-soluble compounds are concerned, it is soluble in highly polar solvents like hexane, but is insoluble in apolar solvents like ether, ethanol, and methanol \unskip~\cite{1116378:22316706}. Naftidrofuryl is the IUPAC Name IUPAC name 2-(diethylamino)ethyl 3-(naphthalen-1-yl)-2-oxopropanoate. Charantin has been shown to enhance blood sugar levels by enhancing glucose uptake and glycogen synthesis in the liver, muscles, and fat cells [Figure~\ref{f-6e7d8682a7b3}]. Additionally, it improves insulin secretion from pancreatic beta cells, and aids or helps in the creation of new beta cells that secrete insulin. K-3's alcoholic extract proved to be more efficient in reducing the frequency of attacks of diabetes than did tolbutamide, which was also utilized on occasion in treating diabetes. In most cases, it is used in capsule form. HPLC has been employed to determine various drugs. These newly released formulations necessitate a fresh analysis to estimate the medicine in the formulations. The current analysis methodologies for such medications are accessible in the scientific literature, however not all techniques are stable and cost-effective. Other procedures are frequently time-consuming. HPLC has been employed to determine various drugs \unskip~\cite{1116378:22316701}. There was no documentation of stability-indicating and time-consuming computational procedures \unskip~\cite{1116378:22316708}. In the current research, RPHPLC method was developed to estimate the Charantin and validating the procedure.


\bgroup
\fixFloatSize{images/303965d3-357f-4dca-bf4b-c069c81e466e-upicture1.png}
\begin{figure}[!htbp]
\centering \makeatletter\IfFileExists{images/303965d3-357f-4dca-bf4b-c069c81e466e-upicture1.png}{\includegraphics{images/303965d3-357f-4dca-bf4b-c069c81e466e-upicture1.png}}{}
\makeatother 
\caption{\boldmath {Structure of Charantin}}
\label{f-6e7d8682a7b3}
\end{figure}
\egroup

    
\section{MATERIALS AND METHODS}
\textbf{Instruments used}

For the research, the Shimadzu LC-2010, CHT autosampler coupled with A PDA-SPD-M20A-Prominence-Diode Array (PDA) detector was used. A PhenomenexKromasil column with 5\ensuremath{\mu } 100 RC18 (250 mm x 4.6 mm, i.e. 5 microns) dimensions was paired to the HPLC system. UV-Visible Spectrophotometer T60 developed by SHIMADZU was used. LC-solution is tools used to analyze peaks.

\textbf{Mobile Phase used}

A standard Charantin medication and pills were purchased from a retail vendor in Bengaluru. Using a software-controlled pump, acetonitrile, methanol, and buffer solutions are blended uniformly and balanced. The degassing process is complete, and HPLC grade solvents are employed. Various solvents were used to store solvent schemes \unskip~\cite{1116378:22316704}.

\textbf{Methodology}

\textbf{Optimization}

After performing the trials in triplicates the optimized parameters were developed and proceeded for further experiments maintaining the same conditions.

\textbf{Assay}

25mg of Charantin working standard was weighed and transferred to a 25ml clean, dry volumetric flask, 5ml of diluent-1 was added, sonicated to dissolve at room temperature, and diluted to volume with diluent-2 was added and mixed. 5ml of this solution was diluted to 25ml with diluent -3 and mixed \unskip~\cite{1116378:22316703}. The solution was diluted using a membrane filter (milliposepvf 0.45m/ nylon 0.45m or Whatmann GF/C 1.2 m or appropriate). Along with the capsules shells, two capsules were weighed and placed to a 500ml clean and dry volumetric flask, together with roughly 50 ml of diluents-3, and sonicated for about 15 minutes at room temperature with occasional shaking to thoroughly scatter the capsule. Add about 300 ml of diluent-1 and sonicate for approximately 20 minutes at room temperature with occasional shaking, then dilute with diluent -3 up to 1cm below the mark, leave to cool at room temperature, and mix with diluent -3. Centrifuge the solutions for 5 minutes at 10,000 RPM. 5ml of this solution should be diluted to 50ml with dilutent -3 and mixed. Using a 0.45 m membrane filter, filter the solution.20 L of the aforesaid solutions were injected into the HPLC system, and the area of the peaks was measured, as well as the percentage of drug, according to the standard formula.

$\style{font-family:'Times New Roman'}{\style{font-size:8px}{\%\;Assay=\frac{(AT\times WS\times DT\times P\times Wt)}{AS\times DS\times WT\times LC}\times100}} $

\textbf{System Suitability}

Three injections of standard preparation (50 mg/mL for Charantin) were used to test system appropriateness criteria such as resolution, tailing factor, peak purity, and theoretical plates \unskip~\cite{1116378:22316698}  estimated the percent RSD of three injections. The initial mobile stage was used to calibrate the HPLC. Six tests were performed using conventional procedures to determine the device's appropriateness for the research \unskip~\cite{1116378:22316699}. 

\textbf{Linearity of the estimation}

Weigh and transfer around 25mg of Charantin working standards into a 25ml clean dry volumetric flask, then add about 5ml of diluents-1 and sonicate to dissolve at room temperature before topping up with diluent -2 and mixing. 1ml of each stock solution was pipetted into a 10ml volumetric flask and the full volume was made up with diluent. The peak regions were measured after injecting 50g/ml, 100g/ml, 150g/ml, 200g/ml, 250g/ml, and 300g/ml into the chromatographic apparatus \unskip~\cite{1116378:22316700}. The slope regression constant was calculated.

\textbf{Accuracy of the estimation}

Recovery tests were used to determine the method's accuracy. The drug's reference standards were added to the formulation at 50 percent, 100 percent, and 150 percent concentrations. The recovery studies were completed and the percentage recovery and percentage relative standard deviation were computed \unskip~\cite{1116378:22316702}.

\textbf{Precision of the procedure}

Weigh and transfer around 25mg of Charantin working standards into a 25ml clean dry volumetric flask, then add about 5ml of diluent-1 and sonicate to dissolve at room temperature before topping up with diluent -2 and mixing. 1ml of each stock solution was pipetted into a 10ml volumetric flask and the full volume was made up with diluent. The solution was injected three times, and the area of each injection was measured in HPLC \unskip~\cite{1116378:22316705}. The region of six duplicate injections' percent RSD was determined to be within the prescribed limits.

\textbf{Robustness of the procedure}

Robustness was measured by making modest changes to the flow rates, temperature, and wavelength. To determine robustness, the column temperature and flow rates were both changed to a limited extent. It was tested at three different concentrations, and the results were represented as a percentage of the total \unskip~\cite{1116378:22316707}. This system's identification limits and quantification limits were visually estimated by trial and error.
    
\section{RESULTS \& DISCUSSION}
The purpose of optimizing chromatographic conditions was to take into account the many technique development goals, to appropriately weigh each goal (theoretical plates, runtime, sensitivity, peak symmetry, etc.) and to pick an acceptable technique for the estimation of Charantin in Capsules dosage form. The RP-HPLC method's sensitivity is determined by the wavelength used. 249 nm was used as the detecting wavelength. Charantin had a retention time of 2.37 minutes as measured by the chromatogram. Trial 1 has a rounded summit. The chromatogram revealed a retention period of 9.12min for Charantin. In trial 2, the wide peak failed to meet the system appropriateness requirements. The chromatogram showed that Charantin has a retention period of 3.54min. The peak is sharp, and the tailing was determined to be 1.25 within acceptable limits. The theoretical plates had a limit of 7864, which was greater than the limit of the final optimised experiment [Table~\ref{tw-760d66adca6b}]. In trial 3, the produced peak meets all system suitability requirements [Table~\ref{tw-04ee9848eac9}].


\bgroup
\fixFloatSize{images/5b33a9d0-cc04-4adc-b33a-b1fe88f193a4-upicture2.png}
\begin{figure}[!htbp]
\centering \makeatletter\IfFileExists{images/5b33a9d0-cc04-4adc-b33a-b1fe88f193a4-upicture2.png}{\includegraphics{images/5b33a9d0-cc04-4adc-b33a-b1fe88f193a4-upicture2.png}}{}
\makeatother 
\caption{\boldmath {Assay of Chromatogram; A. Blank; B. Standard; C.Sample}}
\label{f-75db2c730dc7}
\end{figure}
\egroup

\bgroup
\fixFloatSize{images/0d280c7d-62d9-412b-923f-8e411f627e4c-upicture3.png}
\begin{figure}[!htbp]
\centering \makeatletter\IfFileExists{images/0d280c7d-62d9-412b-923f-8e411f627e4c-upicture3.png}{\includegraphics[width=.82\linewidth]{images/0d280c7d-62d9-412b-923f-8e411f627e4c-upicture3.png}}{}
\makeatother 
\caption{\boldmath {Calibration curve graph for Charantin}}
\label{f-a8f35db971b1}
\end{figure}
\egroup

\begin{table*}[!htbp]
\caption{\boldmath {Optimized chromatographic conditions} }
\label{tw-760d66adca6b}
\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.4878\linewidth-2\tabcolsep}p{\dimexpr.5122\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Item & \cAlignHack Condition\\
\tblmidrule 
\cAlignHack Mobile Phase &
  \cAlignHack MeoH:ACN: H\ensuremath{_{2}}O (40:40:20)\\
\cAlignHack Flow rate (ml/min) &
  \cAlignHack 1.0ml/min\\
\cAlignHack Volume of injection loop (ml) &
  \cAlignHack 20 (ml)\\
\cAlignHack Detection wavelength (nm) &
  \cAlignHack 249nm\\
\cAlignHack Column temperature (\ensuremath{^\circ}C) &
  \cAlignHack 40\ensuremath{^\circ}C\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Results for system suitability and Precision} }
\label{tw-04ee9848eac9}
\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.18240000000000002\linewidth-2\tabcolsep}p{\dimexpr.17259999999999998\linewidth-2\tabcolsep}p{\dimexpr.245\linewidth-2\tabcolsep}p{\dimexpr.20\linewidth-2\tabcolsep}p{\dimexpr.20\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Drug & \cAlignHack RT & \cAlignHack Area & \cAlignHack Plate Count & \cAlignHack USP-Tailing\\
\tblmidrule 
\cAlignHack System suitability &
  \cAlignHack 3.7\ensuremath{\pm}0.021 &
  \cAlignHack 4013351\ensuremath{\pm}39899.57 &
  \cAlignHack 6328\ensuremath{\pm}17.45 &
  \cAlignHack 0.99\ensuremath{\pm}0.021\\
\cAlignHack \% RSD &
  \cAlignHack 0.583 &
  \cAlignHack 0.99 &
  \cAlignHack 0.275 &
  \cAlignHack 2.182\\
\cAlignHack Precision  &
  \cAlignHack - &
  \cAlignHack 4028770\ensuremath{\pm}30833.88 &
  \cAlignHack - &
  \cAlignHack -\\
\cAlignHack \% RSD &
  \cAlignHack - &
  \cAlignHack 0.765 &
  \cAlignHack - &
  \cAlignHack -\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Results of Accuracy (recovery of Charantin)} }
\label{tw-6de72455c4d3}
\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.18939999999999998\linewidth-2\tabcolsep}p{\dimexpr.14860000000000001\linewidth-2\tabcolsep}p{\dimexpr.14200000000000001\linewidth-2\tabcolsep}p{\dimexpr.16\linewidth-2\tabcolsep}p{\dimexpr.16\linewidth-2\tabcolsep}p{\dimexpr.20\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Concentration & \cAlignHack Peak \mbox{}\protect\newline Response & \cAlignHack Average \mbox{}\protect\newline Peak area & \cAlignHack Amount Added & \cAlignHack Amount Found & \cAlignHack Mean Recovery\\
\tblmidrule 
\cAlignHack 50\% &
  \cAlignHack 5613784 &
  \cAlignHack 5616405 &
  \cAlignHack 300 &
  \cAlignHack 305.11 &
  \cAlignHack \\
\cAlignHack 100\% &
  \cAlignHack 9928623 &
  \cAlignHack 9924074 &
  \cAlignHack 400 &
  \cAlignHack 397.13 &
  \cAlignHack 100.36\%\\
\cAlignHack 150\% &
  \cAlignHack 15384336 &
  \cAlignHack 15340588 &
  \cAlignHack 500 &
  \cAlignHack 504.07 &
  \cAlignHack \\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Observation of show Temperature variations} }
\label{tw-f8c269cf1f93}
\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.1518\linewidth-2\tabcolsep}p{\dimexpr.15839999999999998\linewidth-2\tabcolsep}p{\dimexpr.16979999999999997\linewidth-2\tabcolsep}p{\dimexpr.16\linewidth-2\tabcolsep}p{\dimexpr.16\linewidth-2\tabcolsep}p{\dimexpr.20\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\multicolumn{2}{p{\dimexpr(.31019999999999996\linewidth-2\tabcolsep)}}{\cAlignHack Robustness} & \cAlignHack Retention Time & \cAlignHack Area & \cAlignHack USP Tailing & \cAlignHack Plate count\\
\tblmidrule 
\multicolumn{6}{p{\dimexpr(1\linewidth-2\tabcolsep)}}{\cAlignHack Flow rate}\\
\cAlignHack 1 &
  \cAlignHack 0.8 ml/min &
  \cAlignHack 3.62 &
  \cAlignHack 3796285 &
  \cAlignHack 1.02 &
  \cAlignHack 6321\\
\cAlignHack 2 &
  \cAlignHack 1.0 ml/min &
  \cAlignHack 3.65 &
  \cAlignHack 4306191 &
  \cAlignHack 0.98 &
  \cAlignHack 6388\\
\cAlignHack 3 &
  \cAlignHack 1.2 ml/min &
  \cAlignHack 3.51 &
  \cAlignHack 3041944 &
  \cAlignHack 0.98 &
  \cAlignHack 6311\\
\multicolumn{6}{p{\dimexpr(1\linewidth-2\tabcolsep)}}{\cAlignHack Wave Length}\\
\cAlignHack 1 &
  \cAlignHack 244nm &
  \cAlignHack 3.33 &
  \cAlignHack 5555265 &
  \cAlignHack 1.02 &
  \cAlignHack 6322\\
\cAlignHack 2 &
  \cAlignHack 249nm &
  \cAlignHack 3.67 &
  \cAlignHack 4306196 &
  \cAlignHack 0.98 &
  \cAlignHack 6389\\
\cAlignHack 3 &
  \cAlignHack 254nm &
  \cAlignHack 3.32 &
  \cAlignHack 5561285 &
  \cAlignHack 0.97 &
  \cAlignHack 6313\\
\multicolumn{6}{p{\dimexpr(1\linewidth-2\tabcolsep)}}{\cAlignHack Temperature}\\
\cAlignHack 1 &
  \cAlignHack 380c &
  \cAlignHack 3.35 &
  \cAlignHack 5555262 &
  \cAlignHack 1.00 &
  \cAlignHack 6320\\
\cAlignHack 2 &
  \cAlignHack 400c &
  \cAlignHack 3.69 &
  \cAlignHack 4306194 &
  \cAlignHack 0.99 &
  \cAlignHack 6387\\
\cAlignHack 3 &
  \cAlignHack 420c &
  \cAlignHack 3.31 &
  \cAlignHack 5561286 &
  \cAlignHack 0.97 &
  \cAlignHack 6313\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}
The chromatogram for Charantin's blank, standard, and sample [Figure~\ref{f-75db2c730dc7}]. The assay limits for Charantin were 90-110 percent, whereas the findings for Charantin were 100.52 percent. As a consequence, the findings were within the parameters. The accuracy studies were presented as percent recovery for Charantin at 50 percent, 100 percent, and 150 percent at limitations of percent recovered should be in the range of 98=102 percent the findings obtained for Charantin were determined to be within limitations. As a result, the approach was discovered to be accurate [Table~\ref{tw-6de72455c4d3}]. 

The accuracy examinations revealed that the percent recovery of the Charantin is 100.36 percent. The limits of percent recovery of medicines were 98-102 percent, indicating that the procedure was accurate and that the widely used excipients found in pharmaceutical formulations did not interfere in the suggested process. Charantin's linearity range was determined to be 150-600 (g/ml). The calibration curve was drawn, and the drug's correlation co-efficient was determined to be 0.999 [Figure~\ref{f-a8f35db971b1}]. 

As a consequence, the findings obtained were within the parameters. The chromatograms of linearity were recorded. The detection and quantification limits are 0.507 g/ml and 1.536 g/ml, respectively. The robustness of the experiment was determined by adjusting specific parameters such as mobile phase, flow rate, column, and sample volume injection. The procedure was carried out at doses of 0.8, 1.0, and 1.2 g/ml [Table~\ref{tw-f8c269cf1f93}].
    
\section{CONCLUSION}
The suggested RP-HPLC technique for estimating Charantin in capsule dose form was found to be exact, specific, accurate, fast, and cost-effective. The sample recoveries in all formulations were consistent with their individual Label Claims, and this approach is suitable for routine analysis. It is ideal for normal laboratory analysis and may be used for quality control of raw materials, formulations, dissolution studies, and bioequivalence investigations for the same formulation.
    
\section{ACKNOWLEDGEMENT}
Authors are thankful to all who have extended their constant support for the completion of the work.

\textbf{Funding Support}

The authors declare no funding support for this study.

\textbf{Conflict of Interest}

The authors declare no Conflict of interest, financial or otherwise.
    

\bibliographystyle{unsrt}

\bibliography{\jobname}\vspace*{20pt} 
\begin{mdframed}[backgroundcolor=cpboxcolor,hidealllines=true,innerbottommargin=7pt,innerrightmargin=1pt,rightmargin=1pt,innerleftmargin=1pt,leftmargin=2pt,skipabove=1pt,innertopmargin=2pt,innerbottommargin=1pt,skipbelow=1pt]%
{\fontsize{8.5}{10}\selectfont\textbf{Copyright:}~\copyrightstmt}	
\end{mdframed}
        
\begin{mdframed}[backgroundcolor=citeboxcolor,hidealllines=true,innerbottommargin=7pt,innerrightmargin=4pt,rightmargin=4pt,innerleftmargin=2pt,leftmargin=2pt,skipabove=1pt,innertopmargin=2pt,innerbottommargin=4pt,skipbelow=2pt]%
{\fontsize{8.5}{10}\selectfont\textbf{Cite~this~article:}~\citeauthor.~\citetitle.\space\makeatletter\abbrvJournalTitle\ \@copyrightYear;\ \ifx\@journalVolume\@empty X\else\@journalVolume\fi\ifx\@journalIssue\@empty\else(\@journalIssue)\fi:\ \ifx\@firstPage\@empty 1\else\@firstPage\fi-\pageref*{LastPage}.\makeatother}	
\end{mdframed}
\vskip-1.1pc

\vskip.5pc
{\includegraphics[width=5cm]{pharmasprings_logo.png}}
~\\
{\fontsize{9.12}{10.12}\selectfont\textcopyright\ \makeatletter\@copyrightYear\makeatother\ Pharma Springs Publication.}

\end{document}
