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% Document Info
\author{UmeshJagtap}
\pdfinfo{
  /Title (tlc-separation-of-amino-acids.pdf)
  /Creator (Cheatography)
  /Author (UmeshJagtap)
  /Subject (TLC:Separation of Amino Acids Cheat Sheet)
}

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\noindent
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    {\parbox{\dimexpr\textwidth-2\fboxsep\relax}{\noindent
        \hspace*{-6pt}\includegraphics[width=5.8cm]{/web/www.cheatography.com/public/images/cheatography_logo.pdf}}
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\begin{tabulary}{11cm}{L}
    \vspace{-2pt}\large{\bf{\textcolor{DarkBackground}{\textrm{TLC:Separation of Amino Acids Cheat Sheet}}}} \\
    \normalsize{by \textcolor{DarkBackground}{UmeshJagtap} via \textcolor{DarkBackground}{\uline{cheatography.com/186232/cs/42100/}}}
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  \mymulticolumn{2}{p{5.377cm}}{\bf\textcolor{white}{Cheatographer}}  \\
  \vspace{-2pt}UmeshJagtap \\
  \uline{cheatography.com/umeshjagtap} \\
  \end{tabulary}
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  \mymulticolumn{1}{p{5.377cm}}{\bf\textcolor{white}{Cheat Sheet}}  \\
   \vspace{-2pt}Published 21st January, 2024.\\
   Updated 17th January, 2024.\\
   Page {\thepage} of \pageref{LastPage}.
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  Measure your website readability!\\
  www.readability-score.com
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\begin{document}
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\begin{multicols*}{2}

\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Aim}}  \tn
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\mymulticolumn{1}{x{8.4cm}}{To separate and identify amino acids in a mixture using thin layer chromatography (TLC).% Row Count 2 (+ 2)
} \tn 
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\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Introduction}}  \tn
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\mymulticolumn{1}{x{8.4cm}}{{\bf{Chromatography}} \newline % Row Count 1 (+ 1)
Chromatography is a technique used for separating compounds in a mixture based on differences in their distribution between a stationary phase (like silica) and a mobile phase (solvent mixture). \newline % Row Count 5 (+ 4)
{\bf{Thin Layer Chromatography (TLC)}} \newline % Row Count 6 (+ 1)
TLC separates compounds based on  \newline % Row Count 7 (+ 1)
\{\{fa-check\}\}solubility, \newline % Row Count 8 (+ 1)
\{\{fa-check\}\} interaction with the stationary phase, and \newline % Row Count 10 (+ 2)
\{\{fa-check\}\} molecular size.  \newline % Row Count 11 (+ 1)
It's useful for qualitative and quantitative analysis. \newline % Row Count 13 (+ 2)
{\bf{Chromatographic Separation of Amino Acids}} \newline % Row Count 14 (+ 1)
Amino acids, having different R groups, interact variably with silica, affecting their movement on a TLC plate. Ninhydrin is used to visualize amino acids, forming purple spots on reaction.% Row Count 18 (+ 4)
} \tn 
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\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Materials Required}}  \tn
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Plant Material}} \newline % Row Count 1 (+ 1)
\{\{fa-square-o\}\} Green Mung Beans (Vigna radiata) extract. \newline % Row Count 3 (+ 2)
{\bf{Reagents}} \newline % Row Count 4 (+ 1)
\{fa-square-o\}\} Standard solutions of individual amino acids (Known Concentrations). \newline % Row Count 6 (+ 2)
\{\{fa-square-o\}\} Solvent mixture (butanol:acetic acid:water in 12:3:5 ratio). \newline % Row Count 8 (+ 2)
\{\{fa-square-o\}\} Ninhydrin reagent (2\% in acetone). \newline % Row Count 10 (+ 2)
\{\{fa-square-o\}\} Silica gel for TLC \newline % Row Count 11 (+ 1)
{\bf{Equipments}} \newline % Row Count 12 (+ 1)
\{\{fa-square-o\}\} Glass Plates for TLC / Readymade TLC plate. \newline % Row Count 14 (+ 2)
\{\{fa-square-o\}\} TLC chamber. \newline % Row Count 15 (+ 1)
\{\{fa-square-o\}\} Capillary tubes. \newline % Row Count 16 (+ 1)
\{\{fa-square-o\}\} Reagent spray bottle. \newline % Row Count 17 (+ 1)
\{\{fa-square-o\}\} Spreader for silica gel. \newline % Row Count 18 (+ 1)
\{\{fa-square-o\}\} Conical flasks, \newline % Row Count 19 (+ 1)
\{\{fa-square-o\}\} beakers. \newline % Row Count 20 (+ 1)
\{\{fa-square-o\}\} Conical flasks \newline % Row Count 21 (+ 1)
\{\{fa-square-o\}\} Measuring Cylinder \newline % Row Count 22 (+ 1)
\{\{fa-square-o\}\} Weighing Balance \newline % Row Count 23 (+ 1)
\{\{fa-square-o\}\} Oven \newline % Row Count 24 (+ 1)
\{\{fa-square-o\}\} Ruler% Row Count 25 (+ 1)
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\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Procedure}}  \tn
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\mymulticolumn{1}{x{8.4cm}}{{\bf{1. TLC Plate Preparation:}} \newline % Row Count 1 (+ 1)
\{\{fa-check\}\}Clean the glass plates thoroughly. \newline % Row Count 2 (+ 1)
\{\{fa-check\}\}Prepare a slurry of silica gel with water (1:2) in a beaker. \newline % Row Count 4 (+ 2)
\{\{fa-check\}\}Pour the silica gel slurry onto the glass plate and spread evenly using a spreader. \newline % Row Count 6 (+ 2)
\{\{fa-check\}\}Allow the silica layer to dry and then activate it by heating in an oven at 110°C for 1 hour. \newline % Row Count 9 (+ 3)
{\bf{2. Solvent Preparation:}} Pour solvent mixture into TLC chamber and let it saturate for 30 minutes. \newline % Row Count 12 (+ 3)
{\bf{3. Sample Application:}} Use capillary tubes to spot amino acid solutions onto the baseline (approximately 2 cm from the bottom of the plate). Allow to dry. \newline % Row Count 16 (+ 4)
{\bf{4. Development:}} Place the plate in the TLC chamber ensuring the baseline is above the solvent. Let the solvent ascend to about 1 cm from the top. \newline % Row Count 19 (+ 3)
{\bf{5. Drying:}} Remove the plate and mark the solvent front with a pencil. Dry the plate under a hood. \newline % Row Count 22 (+ 3)
{\bf{6. Detection:}} Spray the dry plate with ninhydrin and dry in an oven at 105°C for 5 minutes. \newline % Row Count 24 (+ 2)
{\bf{7. Analysis:}} Measure the distances moved by the solute and solvent, and calculate Rf values. \newline % Row Count 26 (+ 2)
Rf Value Calculation: \newline % Row Count 27 (+ 1)
Rf = Distance moved by solute / Distance moved by solvent X 100% Row Count 29 (+ 2)
} \tn 
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Observation Table}}  \tn
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\mymulticolumn{1}{x{8.4cm}}{**S.N.} \tn 
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\mymulticolumn{1}{x{8.4cm}}{Std AA -Standard Amino acid \newline Sample Band 1- Separated Amino acid in Plant sample numbered accordingly as Band 1, Band 2 etc}  \tn 
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\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Results}}  \tn
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\mymulticolumn{1}{x{8.4cm}}{Calculate and record the Rf values for each amino acid to identify them in the mixture.% Row Count 2 (+ 2)
} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
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\mymulticolumn{1}{x{8.4cm}}{Perform the experiment and fill in the Rf values and the identified amino acids.}  \tn 
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\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Figure TLC Chromatogram of Amino Acids}}  \tn
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\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/umeshjagtap_1705490247_TLC Chromatogram.jpeg}}} \tn 
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\mymulticolumn{1}{x{8.4cm}}{MET- Methionine; TRY-Tyrosine; PA- Phenyl alanine, VAL-Valine \newline MIX- Mixure (Sample) \newline M1-Mixture Band 1 \newline M2- Mixure Band 2}  \tn 
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% That's all folks
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