7/24/2019 Thin Layer Chromatography and Column Chromatography Results and Discussion

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RESULTS AND DISCUSSIONIn this experiment, two types of chromatography

based on solid-liquid chromatographic methods

were used for the purpose of separation and

purity determination. Column chromatography,

which runs a solvent that moves down slowly

through a glass column with packed adsorbent,

was used to separate the colored components of

malunggayleaves. On the other hand, thin layer

chromatography !"C# which utili$es a stationary

phase composed of a thin layer of adsorbent

material on a glass plate or plastic sheet, was

used to separate the components of each

collected eluate to overall determine their purity

based on the retention values %f values#

obtained.

&tarting with column chromatography for

separating the colored components of the

malunggay leaves, hexane-acetone was

determined to be the suitable solvent system to

be used as it run through the glass column with

packed adsorbent without dissolving the solid.

!his reagent was continuously added up on the

column to prevent it from running dry, as thepure extract of malunggay leaves was prepared

with the same reagent, to be added next to

perform the chromatography process.

's the pure extract slowly moved down the

column, (.) m" each of the solvents, specifically

hexane-acetone, acetone, and acetone-methanol

respectively, were added continously at the same

time while collecting different colors of eluates in

separate test tubes. *or this, four different-

colored eluates were collected, of which two were

mixed to provide three distinct-colored eluates,

particularly, pale yellow, green, and yellow green.

!he following table presents the data collected

under column chromatography.

Table 1. Characteristic Color and Volume o

Collected Eluate !Column Chromato"ra#h$%

Color o Com#onentVolume o Eluate

!Dro#s%

+ ale yellow

reen

( /ellow reen

!hese eluates were then used as individua

samples for spotting the !"C plates which were

then put inside the developing chamber forcompleting the thin layer chromatography

%etention factors were then determined by

measuring the distance of each component from

the origin and by dividing this to the solvent

front, represented by the formula0

Distance of sample

Distance of solvent

Rf=origin

origin

Table &.Characteristic Color and Distance o

Com#onent rom Ori"in !'% (ith Determined

R Value

Color o

Com#onent

Distance o

com#onent

rom ori"in !'%

in cm

R Value

+ ale /ellow 1.23 cm ).45

reen 1.43 cm ).41

( /ellow reen ).1 cm ).1

attach explanation why retention factor is

important for purity determination or how thin

layer chromatograhy is related to the purity of

the sample6now this explanation correlates with

that of the reuslts obtained from the

chromatogram as seen from the distance

between each eluate, and as shown from the

actual chromatogram presented below etc etc#

7/24/2019 Thin Layer Chromatography and Column Chromatography Results and Discussion

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)i"ure 1.

Chromato"ram under UV li"ht

)i"ure &. Dried Chromato"ram (ith

*ar+in"s

In conclusion6