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\def\citeauthor{~Banothu~Srikanth,
              ~Aaysha~Firdose,
              ~Muna,
              ~Mubarak~Ibrahim~Ahmed~Mohamed}
\def\citetitle{Analytical Validation and Simultaneous Estimation of Cefixime and Ornidazole by RP-HPLC Method}
\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.}
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\def\RunningAuthor{Banothu Srikanth et al.}
\firstPage{42}
\articleType{Original Article}
\receivedDate{12 May 2021}
\acceptedDate{27 May 2021}
\revisedDate{22 May 2021}
\journalVolume{1}
\journalIssue{2}
\copyrightYear{2021}
\eISSN{2583-0953}
\journalDoi{10.26452/ijcpms.v1i2.196}
\def\authorCount{4}
\def\affCount{2}

\def\journalTitle{International Journal of Clinical Pharmacokinetics and Medical Sciences}

\title{Analytical Validation and Simultaneous Estimation of Cefixime and Ornidazole by RP-HPLC Method}
\author{Banothu~Srikanth\textsuperscript{*}\textsuperscript{1},  
            Aaysha~Firdose\textsuperscript{2},  
            Muna\textsuperscript{2}, 
            Mubarak~Ibrahim~Ahmed~Mohamed\textsuperscript{2}~\\[5pt]{\textsuperscript{1}Department of Pharmaceutical Analysis \& Quality Assurance, St. Mary's Pharmacy College, Deshmukhi (Village), Pochampally (Mandal), Yadadri Bhuvanagiri (Dist), Telangana-508284, India}~\\{\textsuperscript{2}St. Mary's Pharmacy College, Deshmukhi (Village), Pochampally (Mandal), Yadadri Bhuvanagiri (Dist), Telangana-508284, India}}

\begin{abstract}
Suitability, linearity, Precision, Accuracy method perform as the Analytical validation and simultaneous estimate consisting of the Cefixime and Ornidazole. The retention time \& \% RSD of Cefixime and Ornidazole find up to be 5.4 \ensuremath{\pm} 0.2 min, 3.2 \ensuremath{\pm}0.4 min, and 0.60\% and 0.65\%.  The \% RSD Obtained for Cefixime and Ornidazole were 0.60\% and 0.65\%. LOD, LOQ \& Assay encounter ultimate integrity will be acquired from the Cefixime and Ornidazole were 0.80 ppm, 0.81 ppm, 2.25 ppm \& 2.22 ppm, and 101.04\% and 100.41\% respectively.
\end{abstract}\def\keywordstitle{Keywords}

\begin{keywords}Analytical Validation,\newline Simultaneous Estimation,\newline RP-HPLC Method,\newline Limit of Detection \& Quantification
\end{keywords}
\twocolumn[ \maketitle {\printKwdAbsBox}]
\makeatletter{\raggedright{\Large$^\ast$}Corresponding Author\par Name:\ Banothu~Srikanth~\\ Phone:\ +91 7893557404~\\  Email:\ srikanth007banoth@gmail.com
        \par}\vspace*{-13pt}\hrulefill~\\{\fontsize{12}{14}\selectfont\ifx\@eISSN\@empty\else eISSN: \@eISSN\fi\ifx\@pISSN\@empty\else\hfill pISSN: \@pISSN\fi}\par% 
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\section{Introduction}
The current written report used to be interested in an attempt to develop as well as validating a simple, accurate, precise, and economical RP-HPLC approach to the simultaneous estimate of Cefixime and Ornidazole \unskip~\cite{1115274:22308727}.  Cefixime is utilized in the overall treatment of nonimmune diseases including gonorrhea, otitis, throat infection, chronic bronchitis, etc. Ornidazole is an antimicrobial agent utilized in the treatment of unresisting protozoa diseases. The brochure study exhibits there will be methods since the estimate of Cefixime and Ornidazole  together or surrendered or with other medicine victimization UV, HPLC \unskip~\cite{1115274:22308728}. HPLC approach is a lot of photosensitive in comparison to UV, however, the RP-HPLC approach revealed have been with an admixture of 3 solvents as mobile phase. There's an object to originate the RP-HPLC approach for the simultaneous estimate of Cefixime and Ornidazole  using 2 solvent that is more cost-effective \unskip~\cite{1115274:22308729}. 
    
\section{MATERIALS AND METHODS }
The Cefixime and Ornidazole  had been acquired freely given samples from Chandra labs (Hyderabad, India). Acetonitrile, ammonium acetate, glacial acetic acid, tetrahydrofuran, potassium dehydrogenate phosphate buffer, Orthophosphoric acid, Triethylamine have been procured from S.D. Fine Chemicals Ltd, Mumbai. Methanol \& water (HPLC grade) used to be acquired from Merck specialities private limited, Mumbai.

\textbf{Methodology}

\textbf{Linearity }

Linearity solutions will be prepared as Stock solutions of Cefixime and Ornidazole go down excited 6 abundant volumetric flasks and diluted to 10ml in addition to diluents along with the drugs \unskip~\cite{1115274:22308730}. 


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\caption{\boldmath {Calibration curve of Cefixime}}
\label{f-db8cf11d99c8}
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\caption{\boldmath {Calibration curve of Ornidazole}}
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\caption{\boldmath {Chromatogram of Cefixime and Ornidazole Linearity method}}
\label{f-73f77604bed3}
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\caption{\boldmath {LOD Chromatogram of Cefixime and Ornidazole method}}
\label{f-bfa8ca3e2781}
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\caption{\boldmath {LOQ Chromatogram of Cefixime and Ornidazole method}}
\label{f-61f23417abeb}
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\caption{\boldmath {Assay of Tablet}}
\label{f-9072b6e19c39}
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\begin{table*}[!htbp]
\caption{\boldmath {System suitability studies of Drugs} }
\label{tw-7f8617784a77}
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\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Property & \cAlignHack Cefixime & \cAlignHack Ornidazole\\
\tblmidrule 
\cAlignHack Retention time (tR) &
  \cAlignHack 5.4 \ensuremath{\pm} 0.2 min &
  \cAlignHack 3.2 \ensuremath{\pm}0.4 min\\
\cAlignHack Theoretical plates (N) &
  \cAlignHack 2921 \ensuremath{\pm} 174.51 &
  \cAlignHack 4832 \ensuremath{\pm} 172.38\\
\cAlignHack Tailing factor (T) &
  \cAlignHack 2.39 \ensuremath{\pm} 0.226 &
  \cAlignHack 2.42\ensuremath{\pm} 0.238\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Calibration data of Cefixime and Ornidazole method} }
\label{tw-afe806454b8f}
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\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.11380000000000003\linewidth-2\tabcolsep}p{\dimexpr.26659999999999997\linewidth-2\tabcolsep}p{\dimexpr.1922\linewidth-2\tabcolsep}p{\dimexpr.22740000000000002\linewidth-2\tabcolsep}p{\dimexpr.20\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack S.No & \cAlignHack Concentration \mbox{}\protect\newline Cefixime (\ensuremath{\mu}g/ml) & \cAlignHack Response & \cAlignHack Concentration \mbox{}\protect\newline Ornidazole (\ensuremath{\mu}g/ml) & \cAlignHack Response\\
\tblmidrule 
\cAlignHack 1 &
  \cAlignHack 0 &
  \cAlignHack 0 &
  \cAlignHack 0 &
  \cAlignHack 0\\
\cAlignHack 2 &
  \cAlignHack 25 &
  \cAlignHack 74516 &
  \cAlignHack 60 &
  \cAlignHack 130553\\
\cAlignHack 3 &
  \cAlignHack 50 &
  \cAlignHack 130300 &
  \cAlignHack 120 &
  \cAlignHack 255624\\
\cAlignHack 4 &
  \cAlignHack 75 &
  \cAlignHack 224982 &
  \cAlignHack 180 &
  \cAlignHack 379671\\
\cAlignHack 5 &
  \cAlignHack 100 &
  \cAlignHack 277837 &
  \cAlignHack 260 &
  \cAlignHack 507321\\
\cAlignHack 6 &
  \cAlignHack 125 &
  \cAlignHack 372858 &
  \cAlignHack 320 &
  \cAlignHack 646451\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}
\textbf{Standard Preparation}

Appropriately visually inspected along with transmitted 15mg of Cefixime and 30mg of Ornidazole operating requirements right into a 10ml transfer into a volumetric flask, add 3/4\ensuremath{^{th}} of diluents, sonicated for five mins final volume with diluents \unskip~\cite{1115274:22308724}.

\textbf{Sample Preparation}

15 pills were weighed (equivalent to 600 milligrams of Cefixime and 1350mg of Ornidazole) used to be conveyed right into a 100mL volumetric flask, 75mL of diluent additional furthermore sonicated for 25 minutes \unskip~\cite{1115274:22308731}. Further, the volume retract diluents and screened. 

\textbf{Accuracy}

Accuracy solutions will be prepared as Stock solutions of Cefixime and Ornidazole go down excited 6 abundant volumetric flasks and diluted to 10ml in addition to diluents along with the drugs \unskip~\cite{1115274:22308733}. 

\textbf{Standard Preparation }

Appropriately visually inspected along with transmitted 15mg of Cefixime and 30mg of Ornidazole operating requirements right into a 10ml transfer into a volumetric flask, add 3/4\ensuremath{^{th}} of diluents, sonicated for five mins final volume with diluents \unskip~\cite{1115274:22308725}.

\textbf{Method Validation}

The developed method was validated for Suitability, Precision \unskip~\cite{1115274:22308726}, Assay \unskip~\cite{1115274:22308732}, LOD, and LOQ \unskip~\cite{1115274:22308723} as per the ICH guidelines.


\begin{table*}[!htbp]
\caption{\boldmath {Repeatability results for drugs} }
\label{tw-7894e940c1ca}
\centering 
\begin{threeparttable}

\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.286\linewidth-2\tabcolsep}p{\dimexpr.3618\linewidth-2\tabcolsep}p{\dimexpr.3522\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack S. No & \cAlignHack Cefixime & \cAlignHack Ornidazole\\
\tblmidrule 
\cAlignHack 1 &
  \cAlignHack 305211 &
  \cAlignHack 507191\\
\cAlignHack 2 &
  \cAlignHack 304821 &
  \cAlignHack 503640\\
\cAlignHack 3 &
  \cAlignHack 300582 &
  \cAlignHack 498908\\
\cAlignHack 4 &
  \cAlignHack 302957 &
  \cAlignHack 495368\\
\cAlignHack 5 &
  \cAlignHack 303650 &
  \cAlignHack 496621\\
\cAlignHack 6 &
  \cAlignHack 301165 &
  \cAlignHack 494450\\
\cAlignHack Mean &
  \cAlignHack 302878.6 &
  \cAlignHack 494510.9\\
\cAlignHack Std. Dev. &
  \cAlignHack 2058.5 &
  \cAlignHack 3068.21\\
\cAlignHack \%RSD &
  \cAlignHack 0.70 &
  \cAlignHack 0.66\\
\tblbottomrule 
\end{tabulary}\par 
\begin{tablenotes}\footnotesize 
    
\item{*Average of six determinations}
\end{tablenotes}
\end{threeparttable}

\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Inter day precision results for Cefixime and Ornidazole} }
\label{tw-679cc9bd126e}
\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.2909\linewidth-2\tabcolsep}p{\dimexpr.3691\linewidth-2\tabcolsep}p{\dimexpr.34\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack S. No & \cAlignHack Cefixime & \cAlignHack Ornidazole\\
\tblmidrule 
\cAlignHack 1 &
  \cAlignHack 307885 &
  \cAlignHack 499644\\
\cAlignHack 2 &
  \cAlignHack 307232 &
  \cAlignHack 498109\\
\cAlignHack 3 &
  \cAlignHack 306596 &
  \cAlignHack 499877\\
\cAlignHack 4 &
  \cAlignHack 308616 &
  \cAlignHack 499919\\
\cAlignHack 5 &
  \cAlignHack 307070 &
  \cAlignHack 499117\\
\cAlignHack 6 &
  \cAlignHack 304394 &
  \cAlignHack 499009\\
\cAlignHack Mean &
  \cAlignHack 306856.6 &
  \cAlignHack 498847.5\\
\cAlignHack Std. Dev. &
  \cAlignHack 1639.7 &
  \cAlignHack 602.802\\
\cAlignHack \%RSD &
  \cAlignHack 0.60 &
  \cAlignHack 0.65\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Accuracy results of Cefixime and Ornidazole } }
\label{tw-70705413460c}
\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.196\linewidth-2\tabcolsep}p{\dimexpr.204\linewidth-2\tabcolsep}p{\dimexpr.20\linewidth-2\tabcolsep}p{\dimexpr.20\linewidth-2\tabcolsep}p{\dimexpr.20\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack Sample & \cAlignHack Amount added (\ensuremath{\mu}g/ml) & \cAlignHack Amount Recovered (\ensuremath{\mu}g/ml) & \cAlignHack Recovery \mbox{}\protect\newline (\%) & \cAlignHack \% RSD\\
\tblmidrule 
\cAlignHack Cefixime &
  \cAlignHack 100 &
  \cAlignHack 100.15 &
  \cAlignHack 100.15 &
  \cAlignHack 0.09\\
\cAlignHack  &
  \cAlignHack 150 &
  \cAlignHack 162.50 &
  \cAlignHack 113.77 &
  \cAlignHack 0.30\\
\cAlignHack Ornidazole &
  \cAlignHack 250 &
  \cAlignHack 250.70 &
  \cAlignHack 100.45 &
  \cAlignHack 0.05\\
\cAlignHack  &
  \cAlignHack 375 &
  \cAlignHack 385.70 &
  \cAlignHack 110.17 &
  \cAlignHack 0.80\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

\begin{table*}[!htbp]
\caption{\boldmath {Assay of Tablet} }
\label{tw-98b5a7e03df6}
\def\arraystretch{1.1}
\ignorespaces 
\centering 
\begin{tabulary}{\linewidth}{p{\dimexpr.3031\linewidth-2\tabcolsep}p{\dimexpr.35449999999999996\linewidth-2\tabcolsep}p{\dimexpr.3424\linewidth-2\tabcolsep}}
\tbltoprule \rowcolor{kwdboxcolor}\cAlignHack S. No. & \cAlignHack Cefixime (\%) & \cAlignHack Ornidazole (\%)\\
\tblmidrule 
\cAlignHack 1 &
  \cAlignHack 110.65 &
  \cAlignHack 109.07\\
\cAlignHack 2 &
  \cAlignHack 108.79 &
  \cAlignHack 110.32\\
\cAlignHack 3 &
  \cAlignHack 106.30 &
  \cAlignHack 108.32\\
\cAlignHack 4 &
  \cAlignHack 105.10 &
  \cAlignHack 107.65\\
\cAlignHack 5 &
  \cAlignHack 106.40 &
  \cAlignHack 108.89\\
\cAlignHack AVG &
  \cAlignHack 107.44 &
  \cAlignHack 108.85\\
\cAlignHack STDEV &
  \cAlignHack 0.70 &
  \cAlignHack 0.65\\
\cAlignHack     \%RSD   &
  \cAlignHack 0.69 &
  \cAlignHack 0.65\\
\tblbottomrule 
\end{tabulary}\par 
\end{table*}

    
\section{RESULTS AND DISCUSSION}
\textbf{System Suitability}

The entire suitability probabilities will be within just range and satisfactory given that according to ICH guidelines [Table~\ref{tw-7f8617784a77}].

\textbf{Linearity}

Six concentrations of Cefixime and Ornidazole will be prepared plus injected. The calibration curve \& regression coefficient of the drugs as displayed Table~\ref{tw-afe806454b8f}  \& Figures~\ref{f-db8cf11d99c8}, \ref{f-6333568ae0b1} and~\ref{f-73f77604bed3}.

\textbf{Precision: Intraday precision (Repeatability)}

The \% RSD for combined drugs encounter impending 0.70\% and 0.66\% respectively Table~\ref{tw-7894e940c1ca}.

\textbf{Inter Day Precision}

It used to be carried out with 24 hours time lag and also the percentage RSD procured as Cefixime and Ornidazole was 0.60\% and 0.65\% Table~\ref{tw-679cc9bd126e}.

\textbf{Accuracy}

2 levels 100\%, 150\%, were injected during a triplicate way, and also the amount Recovered and \% Recovery was displayed in Table~\ref{tw-70705413460c}.

\textbf{Limit of Detection }

The Cefixime and Ornidazole approach encounter to be 0.80 ppm, 0.81 ppm respectively [Figure~\ref{f-bfa8ca3e2781}].

\textbf{Limit of Quantification }

The Cefixime and Ornidazole find to be 2.25 ppm \& 2.22 ppm respectively [Figure~\ref{f-61f23417abeb}].

\textbf{Assay}

The typical percentage assay used to be calculated and found impending 101.04\% plus 100.41\% for Cefixime and Ornidazole respectively [Table~\ref{tw-98b5a7e03df6}  \& Figure~\ref{f-9072b6e19c39}]
    
\section{CONCLUSION}
The analytical method validation consisting of Cefixime and Ornidazole in tablet dosage form by way of encounter impending RP-HPLC was found to be satisfactory and could use as routine pharmaceutical analysis. The process used to be validated as specified in ICH guidelines like system suitability, accuracy, precision, linearity, LOD \& LOQ, and solution stability, therefore the RP-HPLC method can be employed as routine analysis of those drugs in pharmaceutical formulations.
    
\section{Acknowledgement}
I would like to thanks Principal sir St. Mary's Pharmacy College, Deshmukhi (Village), Pochampally (Mandal), Yadadri Bhuvanagiri (Dist), Telangana-508284, India.

\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.
    
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