In Paper Chromatography wide variety of papers are available commercially in different sizes, shapes, porosities, thickness and chemical treatments.

1. Ubaid Tariq Bhat
M. Pharm (Pharmaceutical Biotechnology)
Delhi Pharmaceutical Science and Research University
Email: bhatubaid55@gmail.com
PAPERS FOR
CHROMATOGRAPHY

2. Wide variety of papers are available commercially in different sizes,
shapes, porosities, thickness and chemical treatments.
Types of papers
A. Pure cellulose papers (Whatman papers)
B. Modified cellulose papers
1. Chemically modified papers
a. Carboxyl papers
b. Acetylated papers
2. Impregnated papers
3. Loaded papers
a. Paper loaded with adsorbent powder
b. Papers loaded with Ion-Exchange celluloses
c. Papers loaded with synthetic organic Ion-Exchange resins
d. Papers loaded with inorganic Ion-Exchangers
C. Glass fibre type papers

3. A. PURE CELLULOSE PAPER
These types of papers are prepared from cotton linters selected
to be especially low in organic and inorganic impurities and
uniform in physical characteristics. Whatman papers have
approximately the following percentage composition.
components percentage
α Cellulose 98-99
β Cellulose 0.3-1.0
Pentosans 0.4-0.8
Ether soluble matter 0.015-0.03
Ammonia 0.001-0.06
Organic nitrogen <0.01
Inorganic material 0.008-0.06

4. Various types of Whatman chromatography papers

5. Mostly used whatmann chromatographic filter papers are:
Whatmann 31ET (coarser and faster)used for separation of
substances having sufficiently wide apart Rf.
Whatmann 3MM (fine paper) is generally used for preparative
purposes.
Whatmann 20 (slow paper) (MN 261, EDROL 208, S and S
2045, S and S 2071)
 Troublesome inorganic impurities include copper and iron,
which form complexes with amino acids and peptides,
leading to tailing zones or multiple spots.
 These interfering metal impurities are removed by washing
with dilute HCl or metal complexing agent EDTA followed
washing with distilled water.

6. B. MODIFIED CELLULOSE PAPERS
Cellulose paper can be modified in several ways to alter its
behaviour.
1. Chemically modified papers
a. Carboxyl papers
• The exchange capacity of the paper is increased by increasing
the carboxyl content (1.4%) by partial oxidation.
• Used for efficient separation of polar substances, Cationic
separation of potential amines and amino acids.
b. Acetylated papers
• Whatmann no 1, S&S 602, ED papers are partially acetylated by
treating with glacial acetic acid containing 5% of acetic
anhydride.

7. • After standing at room temperature overnight the solution is
then replaced by 22.5% acetic anhydride in amyl acetate and
then perchloric acid is added slowly and kept on standby for 2-
3 hours.
• Paper is removed, washed with cold water, rinsed with
methanol and dried at room temperature.
• The acetyl content is then about 26% by weight which is
sufficient for separation of hydrophobic substances.
• Uses: Reverse phase chromatography of lipophilic substances
like steroids, insecticides, pigments and also metal cations.
2. Impregnated papers
• Paper sheets are impregnated with oils to separate hydrophobic
substances such as amines, lipids, steroids, vitamins, and
pesticides (reverse phased chromatography)

8. 3. Loaded papers
• Loaded papers consist of dispersion of powder, fibre, or resin in
a normal cellulosic paper
a. Paper loaded with adsorbent powder
• E.g. Silica papers, alumina papers, zirconia papers, kieselguhr
papers.
• These papers are used to separate less polar and nonpolar
substances (such as lipids, sterols, steryl esters, steroids,
terpenes, dyes, insectcides, chloroplast pigments and inorganic
ions.
b. Papers loaded with Ion-Exchange celluloses
• Chromatographic papers are coated by anionic or cationic
exchangers

9.  Cellulose phosphate paper (Whatmann P 81) (bifunctional
cationic exchanger)
 Carboxymethylcellulose paper (Whatmann CM 82)
(momofunctional cationic exchanger)
Aminoethylcellulose paper (Whatmann AE 81) (monofuntional
anionic exchanger)
Diethylaminoethylcellulose paper (whatmann DE 81)
 Ecteola cellulose paper (Whatmann ET 81)

11. c. Papers loaded with synthetic organic Ion-Exchange
resins:
• coated with micro pulverised ion-exchange resin incorporated
into α-cellulose pulp (reeve angle papers)
d. Papers loaded with inorganic Ion exchangers:
e.g. zirconium phosphate, hydrous oxide, and ammonium
molybdophosphate
C. GLASS FIBER TYPE PAPERS
•Manufactured from fine borosilicate glass fibres and can be
employed with detection reagents that are too corrosive
for cellulose based papers (e.g. chromic-sulphuric acid
charring solution)
•Also used in extreme conditions of temperature and acidity
where cellulose papers are unstable.