Bhore and Khanvilkar, IJPSR, 2019; Vol. 10(1): 12-22.

E-ISSN: 0975-8232; P-ISSN: 2320-5148

IJPSR (2019), Volume 10, Issue 1 (Review Article)

Received on 07 April 2018; received in revised form, 01 September 2018; accepted, 05 September 2018; published 01 January 2019

SILICA GEL: A KEYSTONE IN CHROMATOGRAPHIC TECHNIQUES

Pranali Bhaskar Bhore * 1 and Vineeta Vivek Khanvilkar 2
Department of Quality Assurance 1, Department of Pharmaceutical Analysis 2, Bharati Vidyapeeth’s
College of Pharmacy, Navi Mumbai - 400614, Maharashtra, India.
Keywords: ABSTRACT: Background: In research, the analyst gets samples for
Silica gel, Sol-gel synthesis, separation ranging from a simple mixture of two or more synthetic
Monolithic silica columns, drugs to complex ones obtained from biological, environmental or
Mesoporous silica, Superficially industrial processes. Chromatography has extensively used in such
porous silica columns, Applications of separations where component separation based upon their affinity
silica gel in chromatography
Correspondence to Author:
towards stationary phase and mobile phase. 90% of the stationary
Ms. Pranali Bhaskar Bhore phases available use silica gel either in its original form or with
advanced surface properties. Objective: To furnish the reader with
Department of Quality Assurance,
Bharati Vidyapeeth’s College of detailed information on silica gel, the functional moiety in
Pharmacy, Navi Mumbai - 400614, chromatography, from its synthesis, properties, types, modification of
Maharashtra, India. surface characteristics and applications. We aim to focus on novel
silica materials evolved recently and getting attention due to their
E-mail:[email protected]
performance.
INTRODUCTION: Silica gel consists of granular, Silica gel patented by chemistry professor Walter
porous form of silicon dioxide synthetically made A. Patrick at Johns Hopkins University, Baltimore,
from sodium silicate and possesses significant Maryland in 1919. In 1959, silica gel was first
characteristics of uniform pore and pore size due to supplied in “closed packages” for used as a
which it becomes highly porous structure 1. buffering agent in museum applications to control
Historically silica gel was known to exist from the relative humidity. Its use as a buffering agent rather
1640s as scientific interest. In World War I silica than as a desiccant was unique during that mid-
gel was used in the adsorption of vapors and gases ranges 2. Silica gel has characteristics of controlled
in gas mask canisters. In World War II, silica gel structure, composition, morphology, and porosity.
used for keeping penicillin dry, protecting military Thus, having hosting and recognition properties, as
equipment from moisture damage. From the years well as their wide-open structures containing many
1914 to 1918 it was popularly used as a fluid easily accessible active sites, make them
cracking catalyst for production of high-octane particularly attractive for chromatographic
gasoline and catalyst support for manufacturing of separations. The purpose of this review article is to
butadiene from ethanol. During this ranges, silica study what is silica gel, how does it get synthesize
gel was also used as a feedstock for synthetic by various methods and how synthesis affects the
rubber program. quality of silica gel, various advancements in silica
QUICK RESPONSE CODE gel to enhance properties and its effectiveness in
DOI:
10.13040/IJPSR.0975-8232.10(1).12-22
chromatographic separations 1-13.
2. Synthesis of Silica Gel:
The article can be accessed online on 2.1. Sol-gel Method for Synthesis of Silica Gel:
www.ijpsr.com
The sol-gel process fabricates glass-like or ceramic
DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.10(1).12-22 materials through subsequent hydrolysis and

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condensation of suitable alkoxides. Tetra-methoxy gelation stage. Alternatively, reagents can be

silane (TMOS) and tetraethoxysilane (TEOS) are added by co-polymerizing tetramethoxysilane
one of the most popular alkoxides for the with organoalkoxysilanes. By this process,
preparation of silicate materials. In a typical matrix stabilizes the entrapped reagent from
procedure, water and TMOS combined with a photodegradation or caustic solution
mutual solvent such as methanol followed by environments 3.
addition of a catalyst such as hydrochloric acid.
During the sol-gel process, the viscosity of the 2.3. Synthesis of Silica Gel by Conventional
solution gradually increases, and sol undergoes Method: Using Sulphuric Acid and Sodium
polycondensation reaction to form an Silicate: Another conventional method for
interconnected rigid, porous network called as a synthesis of silica gel other than sol-gel method
gel. was explained by Ali Hafiz et al. They synthesized
silica gel by acidifying an aqueous solution of
2.2. Advantages of Sol-Gel Method: sodium silicate. The intermediate product formed at
low pH called as a hydrosol which further
1. Silica gel formed by the sol-gel method has undergoes agglomeration to form silica polymers.
found many advantages in chromatographic Governing subsequent washing of hydrosol at
technique; particularly stationary phase relatively low pH, the formation of silica gel occurs
development has been a prime area of interest. with high surface area (>700 m²/g). When the
2. By this method, silica gel in various forms like viscosity of hydrosol increases it is termed as a
films, fibers, monoliths, powders can be readily hydrogel. Silica gel is a colloidal form possessing a
made. continuous three-dimensional network of spherical
3. Sol-gel process readily forms thick, crack-free particles. In the gel structure, both siloxane (-Si-o-
films due to improved flexibility of silica gel. Si-) and silanol (-Si-OH) bonds are present. In-gel
4. In the sol-gel process, functional groups can structure, pores get filled with water from
covalently be attached to the silicon dioxide hydrolysis and condensation reaction.
network which inhibits loss of functional group
in solution. The detailed stepwise process of the formation of
5. Reagents readily incorporated in the matrix by silica gel and process to be performed at each step
simply adding them to the sol before its is tabulated in Table 1.
TABLE 1: STEPS INVOLVED INFORMATION OF SILICA GEL
S. no. Name of the step Process
1 Required materials Slow addition of Liquid Sodium silicate having a specific gravity of 1.43 into the mixture of cobalt
acetate and concentrated sulphuric acid having specific gravity 1.85 forms hydrosol
2 Washing of hydrosol Washing of hydrosol with water is important to step for removing sodium sulphate which formed
during the reaction and for the formation of silica gel with a high surface area
3 Drying After washing of hydrosol, drying is an important step to remove adhered water from the gel
4 Screening Finally, the product screened to get a uniform particle size

Following reaction shows the stepwise structures formed during the synthesis of silica gel.

FIG. 1: REACTION SHOWING SYNTHESIS OF SILICA GEL BY CONVENTIONAL METHOD

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2.4. Parameters Affecting Quality of Silica Gel 2.5. Characterizations: Prepared silica gel is
during Synthesis by Conventional Method: characterized for its physical properties like free
2.4.1. Effect of Temperature: Reaction of moisture contents, water-soluble matter, bulk
sulphuric acid and sodium silicate evolves great density and iodine number using standard
amount of heat, thus indicating that reaction is procedures and the values calculated with the help
exothermic. Performing reaction at low temperature of equations 1 to 6.
produces good quality silica gel.
% Moisture =
2.4.2. Effect of pH: The pH is a major factor for Eq. 1
the formation of granular and fine crystalline silica % of water-soluble matter =
gel. Quality of synthesized product greatly depends
upon the pH at which addition of sodium silicate Eq. 2
will be done into sulphuric acid and hydrosol
allowed to form a gel structure. At low pH, due to
Bulk Density =
the growth of chain length, aggregation of particles Eq. 3
occurs after the creation of three-dimensional gel
networks. But very low pH 2.5 resists to the gel Iodine number expressed as the milliequivalent of
formation. The hydrosol formed at pH 2.5 does not iodine adsorbed per 100 g of silica gel.
get converted into hydrogel even after 24 h. Iodine Number = Millilitre titrating 25 ml original Iodine
solution – Millilitre × Normality of thiosulfate Eq. 4
2.4.3. Effect of Concentration: Concentration of
sodium silicate (Na2SiO3) solution has a profound Water-soluble sulphates/chlorides calculated by
effect on the characteristics of the product. Silica using the equation 5.
gel ready at pH 3 to 3.5 from the hydrosol, using a
relatively concentrated solution of sodium silicate % of water-soluble sulphates / chlorides =
shows better characteristics for moisture absorptive
capacity, bulk density, and iodine number Eq. 5
compared to the less concentrated sodium silicate
solution. The reverse of this observation seen at pH Heat resistance and moisture absorptive capacity of
6 where silica gel ready with a comparatively low silica gel are calculated by equation 6.
concentrated sodium silicate solution shows better Moisture absorptive capacity =
quality product.
2.4.4. Effect of Mixing Rate: By changing stirring Eq. 6
rate, quality of the product and crystal size gets
3. General classification of Silica Gel: By particle
changed. By improper stirring, local coagulation
size, pore size, and shape, silica gel classified in
resulted and after drying the product, silica gel has
three classes. The class and their characteristics
amorphous (powdered) structure 1.
tabulated in Table 2.
2.4.5. Effect of Heat: Heat affects greatly on
4. Properties of Silica Gel:
adsorption property of silica gel. Previous studies
reported that the imbalance in the concentrations of 1. Silica gel is non-toxic, non-flammable and
vicinal and free silanol groups on the silica gel non-reactive, stable product.
surface effects by decreasing adsorption of water 2. It can react with various reagents such as with
by a silica gel. This imbalance occurs due to the hydrogen fluoride, fluorine, oxygen difluoride,
thermal treatment or hydrothermal treatment of the chlorine trifluoride, strong acids, strong bases,
samples. In both cases, the adsorption effectivity of and oxidizers.
water decreases. The high water adsorbing capacity 3. The dried form of silica gel called as silica
can be obtained with a high surface area, a high xerogel which is tough and hard. It is naturally
concentration of silanol groups with a 1:1 ratio of occurring mineral purified and processed into
vicinal and free silanol groups 4. granular or beaded form.

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4. Silica gel possesses a high specific surface area into the bulk of gel. Due to this mechanism
2 silica gel is useful as a desiccant (drying agent)
(around 800 m /g) allows it to absorb water
1, 2, 3, 5
readily. Silica gel removes moisture by .
adsorbing numerous pores and not absorbing it
TABLE 2: CLASSES OF SILICA GEL
Characteristics Type A Type B Type C
Appearance Clear pellets, drying and moisture Translucent white Translucent, micro-pored
proof properties. Also known as pellets structure
fine pore silica gel
Approximate pore diameter (nm) 2.5 4.5-7.0 -
Applications Catalyst carriers, adsorbents, Liquid adsorbents, Additionally dried and screened form
separators and variable- pressure drier, catalyst (macro-pored silica gel) used as
adsorbent carriers drier, adsorbent and catalyst carrier

5. Advancements in Silica Gel: Advantages of monofunctional silane reaction:

5.1. Modified Silica Gel: Surface polarity can get
dramatically modified by chemically bonding 1. They are reproducible.
functional groups to silica gel, which produces 2. One silanol group reacts with one silane
chromatographic media with distinctive separation molecule, producing predictable structures.
properties. Use of functionalized silica provides 3. Packing made by this route often shows the
better resolution or separation of difficult highest efficiency because of fast diffusion in
compounds 3. and out of the thin stationary phase layer.

5.1.1. Bonded Phases: A previous report suggests Oligomeric Phase: These types of bonded phases
that silica-based bonded phases made by depositing made by doing reaction of silanol surface with
polymeric organic layer on the silica surface. There bifunctional silane.
are three classes of bonded phase and they are “the
brush phase”, “the oligomeric phase” and “the bulk
phase”. There are different classes produced from
the three different types of silane reagents, that is,
the mono-substituted, di-substituted and tri-
substituted silanes.

5.1.1.1. Bonded Phases for Reversed-Phase FIG. 3: FORMATION OF OLIGOMERIC BONDED

Chromatography: PHASES
Brush Phase: It is most widely used packing
Bulk Phase: Some commercial packings made by
produced by silanol surface-reacted with
doing reaction of surface layer resulting from the
monochloro organosilanes as shown in the
reaction of trifunctional silanes with the silica
following the figure.
surface as shown in the following the figure.

FIG. 2: REACTION OF SURFACE SILANOL WITH

CHLORODIMETHYLSILANE

In the above reaction bonded phase packings are FIG. 4: REACTION OF SURFACE SILANOLS WITH
made by reaction of surface silanol with mono- TRIFUNCTIONALSILANE
functional reagents, such as chlorodimethylsilane. Polymeric bonded phases shown in following
Various alkyl and substituted alkyl silicas are made reactions are more stable than monomeric phases at
by this reaction, such as, n-octadecylsilane (ODS or low pH.
C18) bonded phase materials.

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However, packings made in this manner are more Monoliths defined as the United continuous porous
variable concerning retention and selectivity separation media without the interparticular voids
compared with monofunctional phases. and thus it is a promising tool for microscale
separation. Their structure based on the continuous
interconnected skeleton of mesoporous silica.
These materials are assuring a bright future in
separation science which is useful for
pharmaceutical, biological, and food industries.
They were able to give maximum efficiency with
four times less back pressure in liquid
chromatography as compared to columns packed
with 5 μm particulate due to their greatest external
porosity 6.
FIG. 5: LIGHTLY VERTICAL-POLYMERIZED PHASE
6.1. Synthesis of Monolithic Silica Column: For
The following figure shows another type of bonded the preparation of monolithic columns by sol-gel
saline surface, called horizontal polymerization. method monomers are polymerized in a capillary
These materials reported showing superior stability column. The process typically involves polymer
in low and high pH environments. template inducing phase separation and poly-
condensation of sol-gel transition due to the silica
precursor, which leads to a co-continuous silica
network of defined pore structure. Further,
monolithic silica columns with a bimodal pore
structure consisting of macropores and mesopores
in the silica skeleton can be obtained with thermal
treatment. In monolithic silica columns by keeping
mesopores constant, smaller or larger macropores
can be formed this feature makes the monolithic
silica columns unique compared to their packed
counterparts. The group of Tanaka prepared silica-
FIG. 6: HORIZONTAL POLYMERIZED BONDED PHASES
based monolithic rods by hydrolytic polymerization
As the chain length or bulk of the silane increases, of silanes, like tetramethoxysilane, the reaction is
related silanol group percentage decreases. Even catalysed by aqueous acetic acid in the presence of
with the smallest silane (trimethyl or C1), almost polyethylene glycol which forms a sol. This sol is
50% of the silanol groups remain unreacted on the converted to monolithic silica having network
surface. These silanol groups are located under an structures attached to the tube wall in a fused silica
umbrella of organic silane ligands but are still capillary. Then ammonia is introduced for the
available for electrostatic interaction with formation of mesopores (in which analytes diffuse
appropriate solutes. in and out), and thus creates a large specific surface
area and a chromatographic efficiency
6. Monolithic Silica Column: Preparation and corresponding to a 3-5 µm particulate column 7.
development of monolithic silica columns were
first done in the 1990s. The primary feature of Following table shows a comparison between
the monolithic column is they have high and physical and chromatographic characterizations of
variable external porosity. monolithic and conventional packed columns.

TABLE 3: COMPARISON BETWEEN PHYSICAL AND CHROMATOGRAPHIC CHARACTERIZATIONS OF

MONOLITHIC AND CONVENTIONAL PACKED COLUMNS
Characteristics Monolithic column Conventional particle-packed column
Structure The monolithic column consists of a macroporous The particle-packed column contains interparticular
structure voids
Permeability Column back pressure and permeability depends upon Column back pressure depends upon interparticular

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the macroporous structure voids

Performance and In monolithic silica columns, performance and In this type permeability is directly proportional to
permeability permeability are independent of each other particle diameter and inversely proportional to plate
number thus permeability and performance are
dependent on each other
Separation capacity These materials possess higher column efficiency at a Separation efficiency is low as compared to the
high linear velocity monolithic columns
Adsorption capacity Adsorption capacity is 30 to 40% higher than that of Possess 30 to 40% lower adsorption capacity than
particle packed column monolithic columns

Characterization of porous form, flow sol-gel polymerization method which produces

characteristics, and column efficiency revealed the pore diameters adjustable between 20 and 300, with
advantages of a monolithic silica column over a narrow pore-size distributions. Lack of pore size
particle-packed column. Other important distribution makes microporous and macroporous
advantages of monolithic silica columns are listed materials incapable for selecting adsorbing broad
below: spectrum of large organic molecules of technical
interest.
6.2. Advantages:
The pore size of the mesoporous materials allows
 Monolithic silica columns have the advantage not only easy accessibility for molecules but also
of 5 to 10 time’s faster operation leading to possible controllability in functions depending on
shorter analysis times. pore geometries. Recently, the demand for
 They own high linear velocity with high mesoporous materials has triggered due to their
column efficiency per unit column length high specific surface area, large specific pore
which offer a resolution at nearly 1/10 time volume, and pore diameter.
compared with conventional packed columns.
 The high porosity leads to high permeability or There are various methods for functionalization of
low-pressure drop. Due to high permeability, mesoporous silica which is a necessary step for the
monolithic silica columns are suitable for high applications of these materials in chromatographic
throughput analysis. separations
 Monolithic silica columns provide fast
separation especially in combination with 1. Grafting (Post Synthetic Modification): The
Mass spectrometric (MS) detection. grafting method used for modification of
 Monolithic columns effectively separate highly mesoporous silica, and the introduction of various
complex biological mixtures such as tryptic groups like amino, thiol and alkyl groups.
digest 6, 7.
2. Co-condensation: In co-condensation method,
7. Mesoporous Silica: Porous materials with functionalized organosilicates, which have both
regular geometries have been recently paid much organic functional group and condensable group,
attention following their great scientific importance condensed together with silica sources such as
and potentials in separation science. Depending tetra-methyl orthosilicate and tetra-ethyl
upon the pore size, the porous materials are orthosilicate. Therefore, mesoporous silica
classified by IUPAC into three classes: functionalized with organic group obtained directly
in one-pot synthesis, and the functional groups
1. Microporous (Pore diameter in the size range of spread homogeneously in the silica structures.
0.2–2.0 nm).
2. Mesoporous (Pore diameter in the size range of 3. Periodic Mesoporous Organosilicate (PMO):
2.0–50.0 nm). One of the most different approaches in the
3. Macroporous (Pore sizes exceeding 50.0 nm). preparation of mesoporous organic-inorganic
hybrids is a synthesis of periodic mesoporous
Mesoporous silica possesses high surface areas as organosilicates. In this type of silicate materials
1600 m2/g and expected to provide superior created from organic molecules containing multiple
separating ability which is essential for the alkoxysilane groups. These organic components
chromatographic matrix. This silica’s prepared by a play an important role as PMO pore wall.

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Formation of crystalline pore wall in mesoporous separation efficiency and peak symmetry for basic
silicate materials is one of the fantastic applications compounds.
of PMO approach 8.
8.1. Preparation of Second Generation
8. New Generation Silica-based HPLC Columns: Monolithic Columns using Sol-Gel Method:
Second generation silica-based monolithic Tetramethoxysilane (TMOS) acting as silica
columns, columns with sub 2 µm particles, precursor and polyethylene oxide (PEO) acting as a
superficially porous silica columns (core-shell pore-forming agent both dissolved in slightly acidic
packings) these are the new generation columns conditions filled into gelation tube and heated at
providing high throughput, high efficient moderate temperature. After this stage sol forms
separations. silica gel and phase separation occurs separating
silica-rich phase and solvent rich phase. The time
First generation silica-based monolithic column required for phase separation of silica phase from
(Chromolith, Merck Millipore) attracted a lot of solvent phase forms silica skeleton. Removal of
attraction in 2000 because of its novelty. Silica- solvent phase forms the macroporous structure of
based monolithic columns consist of a single piece silica.
of porous silica having higher separation efficiency
(plate count per meter = N/m up-to 200,000) and Further, polycondensation reaction causes
permeability. They provide lower back pressure, shrinkage of the rod which detaches from the inner
higher flow rate, faster analysis, and direct surface of gelation tube allowing its removal. A
application of dirty samples without prior sample better control of this step in combination with
preparation during analysis. New generation silica- smaller domain size and macroporous structure has
based monolithic HPLC columns (Chromolith High led to the second generation of the monolith which
Resolution [HR]) provides more homogeneous shows improved separation on efficiency and peak
porous silica network with well-designed silica symmetry. Monoliths of the second generation
skeleton in combination with bimodal pore were prepared by systematically reducing the
structure than the first generation silica-based macropore size.
monolithic column. These columns have improved
TABLE 4: COMPARISON BETWEEN SILICA-BASED MONOLITHIC HPLC COLUMNS: FIRST GENERATION
vs. SECOND GENERATION
S. no. First generation monolithic column Second generation monolithic column
(Chromolith Merck Millipore) (Chromolith High Resolution [HR])
1 Macropore size of chromolith column is around 2 µm Macropore size of new generation Chromolith HR column is
around 1.1 µm
2 Performance similar to 3.5-5 µm particle packed column Peak symmetry and separation efficiency achieved by
with column back pressure similar to a 1.1 µm particle- symmetrically decreasing macropore size and corresponding
packed column domain size with homogeneous porous silica network
3 Possess separation efficiency of N/m>80,000 Possess much higher separation efficiency than first
generation columns N/m>140,000
4 Separation efficiency increases due to higher column back Chromolith HR shows a column back pressure of around 65
pressure and due to decreased macropore size. 100 mm × bar. Typical length Chromolith HR columns can still be
4.6 mm Chromolith column can be operated at 25 bar employed with low back pressure conventional HPLC
instrumentation

8.2. Particle Packed Columns with Sub 2 µm 8.3. Superficially Porous Silica Columns:
Particles: Particle packed columns filled with sub Nowadays, high-speed and efficiency in HPLC
2 µm silica particles possess high separation analyses are in the demands. Superficially porous
efficiency about 200,000 N/m due to smaller silica columns with particle sizes of 2.5-3 µm
porous particles. There are some disadvantages for possess higher separation efficiencies, column
working with 2 µm particle packed columns permeability, and lower column back pressure. The
Require UHPLC system (instrument working with measure arenas for fast analyses are the
pressure above 1000 bar), frictional heat pharmaceutical and food industries, agrochemical
generation, blockage because of small porosity 9. industries, biological and environmental sciences,

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proteomic and genomic research. To meet these graphy. The stationary phase is a substance
requirements, reduction in the column length, the fixed in place for the chromatographic
linear velocity of the mobile phase and column procedure.
particle size done respectively, but these are not  Silica-based packings are compatible with
economically feasible as they need costly UPLC water and all organic solvents, and no
machines. Due to a reduction in the particle, dimensional variation (e.g., swelling) in silica
thermal deterioration of the column bed occurs packings occurs with the change in solvents.
which further resulted in efficiency losses. This feature permits the formation of
stabilization packed beds that are stable during
Additionally, small particle columns need severe use with various solvent types and during
filtration of mobile phase to avoid blockage of gradient elution.
0.5µm frits of the column. These problems resulted  The hydroxy (OH) groups on the surface of
in an advanced HPLC column technology, i.e., the silica can be functionalized to afford specialty
development of superficially porous silica particle silica gels that exhibit unique stationary phase
columns (SPSPCs). This column contains particles parameters.
of sub 3.0 µm and has a solid, nonporous core
 It possesses high mechanical strength which
along with a shell of constant thickness made up of
permits the formation of efficiently packed beds
a porous silica gel. These SPSPCs are more
that are stable under high operating pressures
efficient and reduce run time by 70%.
for long periods.
Generally, SPSPCs have twice the number of  The silica-based column provides the highest
theoretical plates (>200 000 per meter) and column efficiency of any of the materials used
resolving ability with moderate back pressure. The to produce columns that exhibit lower
unretained solutes eluted quickly from these backpressures and longer lifetime.
columns in comparison to monolithic columns.  The stronger spherical particles are more easily
SPSPCs are suitable for separating small as well as and reproducibly packed into efficient columns.
large molecules with good sample loading  The surface of silica support gets chemically
capacities. The most commonly used mobile phases modified with a large variety of bonded phases
are water/acetonitrile, water/methanol, buffer/ having different functionalities.
acetonitrile, buffer/methanol, etc.  The hydrated silica gel surface has three kinds
of silanol groups: free silanol, geminal, and
There are seven manufacturers in the market for
associated silanols. Nonhydrogen-bonded free
supplying these columns, i.e. Advanced Material
silanol is more acidic and can cause strong and
Technology, USA (Halo), Sigma-Aldrich, USA
deterious binding of basic solutes because of
(Ascentis Express), Agilent Technologies, USA
their highly acidic nature. Therefore, silica gel
(Pro shell 120), Phenomenex, USA (Kinetex),
with a higher concentration of free and highly
Chromanik Tech., Japan (Sunshell), Macherey-
acidic silanols often shows increased retention
Nagel, Germany (Nucleoshell), and Thermo
and broad peak tailing for basic compounds.
Scientific, USA (Accucore). These new generation
columns give super-fast separation by using Fortunately, free silanols generally occur in a
conventional HPLC machine. Moreover, these low concentration on the silica gel surface. The
columns need a low amount of solvents and surface of silica gel with the highest
handling time making them economical and less concentration of geminal and associated
hazardous to the environment. These features are silanols favored most for the chromatography
attracting industrialists, academicians, scientists, of basic compounds because these silanols are
and regulatory authorities 10. less acidic 1-12.
9. Applications of Silica Gel in Chromatographic types based on separation
Chromatography: mechanism and uses of silica gel in all these types
of chromatography tabulated in Table 4.
 Silica gel is most popular packing material
which used as a stationary phase in chromato-

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TABLE 5: APPLICATIONS OF THE SILICA GEL IN DIFFERENT TYPES OF CHROMATOGRAPHY

S. no. Types of chromatography based Uses of silica gel in chromatography
on the separation mechanism
1 Partition Chromatography In Partition Chromatography, silica gel used as a stationary phase
2 Adsorption Chromatography Silica gel is used as an adsorbent and remains dominant stationary phase in
Adsorption Chromatography
3 Ion exchange Chromatography In ion Exchange Chromatography, silica gel used as a stationary phase
which contains ionizable functional groups

1. Partition Chromatography: Partition hydrated silica gel, silica gel G, silica gel H, silica
chromatography classified as: gel S, silica gel N, silica gel GF254, silica gel HF254,
silica gel PF254.
1.1. Liquid-Liquid Chromatography: Liquid-
liquid chromatography is a part of partition Adsorption chromatography can further be
chromatography and is further classified as normal classified as:
phase chromatography and reversed phase
chromatography. 2.1. Column Chromatography: Column
Chromatography consists of separation of each
1.1.1. Normal Phase Chromatography: In this component from a mixture by loading the mixture
type of chromatography, stationary phase (silica into a column which packed with finely powdered
support) is a polar and mobile phase is non-polar. adsorbent and then developing the column with a
Due to silica gel's polarity, non-polar components solvent, i.e. eluting the individual component in the
tend to elute before more polar components, hence mixture by passing a solvent through the column.
the name normal phase chromatography. The individual components collected as
consecutive eluent fractions. Silica gel used as an
1.1.2. Reversed Phase Chromatography: In this adsorbent in column chromatography. In this type
type of chromatography, stationary phase (silica of chromatography, the stationary phase is most
support) made non-polar by attaching C18 groups often composed of silica gel particles with 40-63
and polar solvent used as a mobile phase. When μm. For achieving the desired separation of certain
hydrophobic C18 groups attached to the silica gel, molecular sizes, different particle sizes are used 10,
then polar components elute first, and the method 11
.
referred to as reverse phase chromatography.
2.1.1. Silica gel in High-Performance Liquid
1.2. Gas-Liquid Chromatography: In this type of Chromatography (HPLC):
chromatography the substance to be separated is 2.1.1.1. Column Packing Particles in HPLC:
moved by an inert gas along with tube filled with Particle type and size of the silica affects separation
the finely divided inert solid support of silica with a in the HPLC system. Several particle types
diatomaceous structure which further coated with available for HPLC application are as follows:
non-volatile oil or waxy polymer.

2. Adsorption Chromatography or Liquid-Solid

Chromatography: As an adsorbent silica gel has
the following characteristics:

 Insoluble in the mobile phase.

 Inert to solute.
 Colorless especially when working with the
color mixture. FIG. 7: POROUS MICROSPHERES
 Suitable particle size is enough to give good
separation and a reasonable flow rate. 2.1.1.1.1. Porous Microspheres: Porous micro-
spheres are most commonly used because of the
In adsorption chromatography, various forms of favorable compromise of desired properties:
silica gel can be used as an adsorbent such as: efficiency, sample loading, a variety of diameter,

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pore sizes, and surface areas so that all types of 2.2. Thin Layer Chromatography (TLC): The
HPLC methods can be developed with these silica gel used as an adsorbent for TLC plates has a
materials. mean particle size of approximately 12μm. The
pore diameter is mainly 60 Å, and the surface area
2.1.1.1.2. Micropellicular Particles: Micro- is approximately 500 m2/g. The most frequently
pellicular particles have a solid core with the thin used thicknesses of silica-based thin layer for
outer skin of interactive stationary phase (silica analytical procedures are approximately 200-250
gel). This silica particle, usually available in 1.5 to μm. The thin layer for preparative purposes is
2.5 µm sizes, displays outstanding efficiency for available with thicknesses of up to 2 mm. The thin
macromolecules because of fast mass transfer layer of the adsorbent can also contain a
kinetics. Pellicular particles of sizes 1.5 µm are fluorescence indicator with a particle size less than
useful for extremely rapid separations of the sorbent. The advantage of these indicators lies
macromolecules such as proteins. in the fact that all substances with a conjugated p-
electron system (e.g., aromatic compounds) appear
as dark spots on a bright emitting background when
the chromatogram viewed under UV light.
An extension of TLC is High-Performance Thin
Layer Chromatography (HPTLC). It is a robust,
simplest, rapid, and efficient tool in the quantitative
analysis of compounds. HPTLC is an analytical
technique based on TLC, but with enhanced
FIG. 8: MICROPELLICULAR PARTICLES resolution of the compounds to be separated and to
allow quantitative analysis of the compounds. The
2.1.1.1.3. Perfusion Particles: Perfusion particles difference between TLC and HPTLC plate is
contain very large pores (e.g., 4000 to 8000 Å) particle size of coated material, which is 5 to 20
throughout the support and also include a network mm of TLC and 4 to 8 mm for HPTLC. Plates are
of smaller interconnecting pores solutes can enter produced from 4 to 5 mm silica gel with an inert
(and leave) this pore structure through a binder to form a 200 mm layer. Plates of 20 × 20
combination of convective (flow) and diffusion. cm and 5 × 7.5 cm used. Silica gel F254 having a
This effect minimizes band broadening so that pore size of 6 mm with a fluorescent indicator is a
large porous particles resemble smaller particles coating material 11, 12.
regarding column efficiency but with a fraction of
the pressure drop. 3. Ion-Exchange Chromatography: In this type
of chromatography separation of molecules based
on their respective charged groups and retention of
analyte molecules based on ionic interactions.
Mainly, molecules undergo electrostatic
interactions with opposite charges bonded with
stationary phase matrix. The stationary phase
consists of an immobile matrix that bonded with
charged ionizable functional groups. The silica-
based stationary phase surface exhibit ionic
functional groups (R-X) that interact with
FIG. 9: PERFUSION PARTICLES
oppositely charged analyte ions. To achieve
Particle size is very important in HPLC. A particle electroneutrality, these inert charges couple with
diameter of about 5 µm represents a good counterions in the solution. Ionizable molecules
compromise for analytical columns regarding that are to be purified compete with these
column efficiency, backpressure, and lifetime. counterions for binding to the immobilized charges
Smaller porous particles (e.g., 3 µm) are available on the stationary phase. These ionizable molecules
for faster separations 5. are retained based on their charges 11, 13.

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CONCLUSION: The review is a systematic 2. Weintraub and Steven: Demystifying silica gel. Object
Specialty Group 2002.
compilation of data related to silica gel. In this 3. Collinson MM: Recent trends in analytical applications of
review we have elaborated the sol-gel process organically modified silicate materials. Trends in
which allows the association of mineral phases Analytical Chemistry 2002; 21(1): 31-39.
4. Christy and Alfred A: Effect of heat on the adsorption
with organic or biological systems. Sol-gel process properties of silica gel. International Journal of
has been the cornerstone in the development of Engineering and Technology 2012; 4(4): 484.
monolith columns and functionalized mesoporous 5. Snyder LR, Kirkland JJ and Glajch JL: Practical HPLC
method Development. Wiley India Pvt. Ltd., Edition 2nd,
silica particles providing wide open structures and 2011: 189-192, 175-181.
gaining the popularity in chromatographic 6. Cabrera and Karin: Applications of silica‐based monolithic
separations today. The article also describes the HPLC columns. Journal of Separation Science. J Sep Sci
2004; 27: 10‐11, 843-852.
new generation columns packed with superficially 7. Ikegami T and Tanaka N: Monolithic columns for high-
porous silica particles providing stationary phases efficiency HPLC separations. Current Opinion in
with moderate sample loading capacity and low Chemical Biology 2004; 8(5): 527-533.
8. Vinu A, Hossain KZ and Ariga K: Recent advances in
back pressure useful in ultra-fast HPLC functionalization of mesoporous silica. Journal of
separations. Nanoscience and Nanotechnology. J Nanosci Nanotchnol
2005; 5(3): 347-371.
ACKNOWLEDGEMENT: I am highly indebted 9. Cabrera and Karin: A new generation of silica-based
monolithic HPLC columns with improved performance.
to my guide Mrs. Vineeta V. Khanvilkar for her LC GC Magazine-North America-Solutions for Separation
support and constant supervision as well as for Scientists 2012; 69: 56.
providing necessary information regarding the 10. Ali I, AL‐Othman ZA, Nagae N, Gaitonde VD and Dutta
KK: Recent trends in ultra‐fast HPLC: New generation
review article and also for her support in superficially porous silica columns. Journal of Separation
completing the manuscript. I would like to express Science 2012; 35(23): 3235-3249.
my gratitude towards my parents for their kind co- 11. Chatwal GR and Anand SK: Instrumental methods of
chemical analysis. Himalaya Publishing House Pvt. Ltd.,
operation and encouragement which help me in the Edition 5th, 2011; 2.599-2.700.
completion of this review article. 12. Gocan and Simion: Stationary phases for thin-layer
chromatography. Journal of Chromatographic Science
CONFLICT OF INTEREST: Nil 2002; 40(10): 538-549.
13. Ohta K: Application of pure silica gel as the cation-
exchange stationary phase in ion chromatography with
REFERENCES: indirect photometric detection for common mono-and
1. Asghar AH, Arshad C and Sattar A: Synthesis of quality divalent cations using aromatic monoamines as eluents.
silica gel; Optimization of parameters. Journal of Faculty Chromatographia 2002; 55(1-2): 95-100.
of Engineering & Technology 2010, 16(1): 19-32.

How to cite this article:

Bhore PB and Khanvilkar VV: Silica gel: a keystone in chromatographic techniques. Int J Pharm Sci & Res 2019; 10(1): 12-22. doi:
10.13040/IJPSR.0975-8232.10(1).12-22.
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