“Chromatography is used to separate mixtures of substances into their components”

CHROMATOGRAPHYCHROMATOGRAPHY

A physical method of separationComponents partition between two phases:

Stationary phase ~ does not move Mobile phase ~ does move

- stationary phase: a solid, or a liquid supported on a solid- mobile phase: a liquid or a gas- The mobile phase flows through the stationary phase and

carries the components of the mixture with it. - Different components travel at different rates. We'll look at

the reasons for this further down the page.

“Solutes are separated due to differences in how they interact with the two phases”

Sifat Fisik yang Terlibat dalam Kromatografi

Kelarutan : Kecenderungan molekul larut dalam cairan

Adsorpsi : Kecenderungan molekul untuk melekat pada permukaan halus

Keatsirian : Kecenderungan molekul untuk menguap

PEMAKAIAN PEMAKAIAN KROMATOGRAFIKROMATOGRAFI

Kualitatif: Senyawa apa yang ada?Kuantitatif: Berapa banyak yang ada?Preparatif: Bagaimana memperoleh

komponen murni?

Kromatografi Cair

Paper Chromatography/ Kromatografi Kertas (PC/ KKT)

Thin Layer Chromatography/ Kromatografi Lapis Tipis (TLC/ KLT)

Column Chromatography/ Kromatografi Kolom

High Performance Liquid Chromatography/ Kromatografi Cair Kinerja Tinggi (HPLC/ KCKT)

Paper Chromatography

Batas pelarut

pelarut

For example, if one component of a mixture travelled 9.6 cm from the base line while the solvent had travelled 12.0 cm, then the Rf value for that component is:

THIN LAYER CHROMATOGRAPHY – TLC THIN LAYER CHROMATOGRAPHY – TLC

Thin layer chromatography: using a thin, uniform layer of silica gel or alumina coated onto a piece of glass, metal or rigid plastic.

The silica gel (or the alumina) is the stationary phase. The stationary phase for thin layer chromatography also often contains a substance which fluoresces in UV light - for reasons you will see later.

The mobile phase is a suitable liquid solvent or mixture of solvents.

The diagram shows the plate after the solvent has moved about half way up it

As the solvent slowly travels up the plate, the different components of the dye mixture travel at different rates and the mixture is separated into different colored spots

COLUMN CHROMATOGRAPHY COLUMN CHROMATOGRAPHY

used to purify individual chemical compounds from mixtures of compounds.

The stationary phase or adsorbent in column chromatography is a solid. The most common stationary phase for column chromatography is silica gel or alumina.

The mobile phase or eluent is either a pure solvent or a mixture of different solvents

The classical preparative chromatography column is a glass tube with a diameter from 5 to 50 mm and a height of 50 cm to 1 m with a tap at the bottom.

Slurry is prepared of the eluent with the stationary phase powder and then carefully poured into the column. Care must be taken to avoid air bubbles.

A solution of the organic material is pipetted on top of the stationary phase. This layer is usually topped with a small layer of sand or with cotton or glass wool to protect the shape of the organic layer from the velocity of newly added eluent.

Eluent is slowly passed through the column to advance the organic material.

“High performance liquid chromatography is basically a highly improved form of column chromatography”

HIGH PERFORMANCE LIQUID CHROMATOGRAPHY - HIGH PERFORMANCE LIQUID CHROMATOGRAPHY - HPLC HPLC

Waktu retensi (tR): Waktu yang diperlukan oleh gas pembawa untuk membawa komponen sepanjang kolom.

GAS CHROMATOGRAPHY GAS CHROMATOGRAPHY

type of chromatography in which the mobile phase is a carrier gas, usually an inert gas such as helium or an unreactive gas such as nitrogen,

the stationary phase is a microscopic layer of liquid or polymer on an inert solid support, inside glass or metal tubing, called a column.