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363, Bruce
Chromatography
components when passed through an inert material using chromatographs. This laboratory
technique measures the retention times for analytes, a chemical substance that is being
identified, to pass through a column of absorbents. There are several types of chromatography
techniques that use different phases. The three major methods of chromatography are Column
separation method consists of mainly liquid substances, but gases and solids can be used
depending on the technique, which are referred to as the mobile phases when mixed with a
solvent. A solvent is a liquid which has various components dissolve, forming a solution
(Britannica, 2011). Essentially all of the separation techniques utilize a mobile phase that passes
through a stationary phase that sits in the column of the chromatograph. The stationary phase
History
In the mid 1800’s, a professor and his student by the names of Christian Friedrich
Schonbein and Friedrich Goppelsroeder were the first to publish attempts to study the rate that
different substances travel through a filter paper ("History of chromatography", 2019). Instead
of today’s methods which measure absorption as being responsible for the movement, they
thought capillary action was the reason for the movement of substances. Capillary action is the
movement of liquids through narrow spaces without any external force assisting it (Saig, 2018).
The technique of column chromatography separation was first used in Russia by Mikhail Tsvet
in the early 20th century. He primarily used this technique to separate different plant pigments,
which gave the technique its name. The pigments that Tsvet was separating were chlorophyll
(green pigments), carotenes (orange and yellow pigments), and xanthophylls (yellow pigments)
("Chromatography", 2019).
Later on, in the 20th century new types of this technique were developed. Two scientists
by the name Archer John Port Martin and Richard Laurence Millington Synge further developed
solubilities (International Union, 2006). These two scientists went on to win the Nobel prize in
Chemistry in 1952. From these new principles several other chromatographic techniques were
Column Chromatography
Column Chromatography, one of the most common techniques, separates specific compounds
depending on the type of stationary phase is used to pack the column. The column can separate the
eluent, mobile phase consisting of a solvent that dissolves the sample, based on the molecule’s
hydrophobicity (affinity for water substances), polarity (the “pull” atoms have on their electrons), or
electrostatic charge (either a positive or negative charge of the atom) (Torres, 2016). Different sized
columns can be used when packing the stationary phase depending on the amount of substance you are
attempting to separate. After packing the column, the eluent is placed on top of both the sample and
stationary phase. The bottom end nozzle is then opened to allow the sample to flow through the
column. Depending on molecular properties, the analyte which has a weaker interaction with the
stationary phase will pass through it more rapidly while the analytes with a stronger interaction will take
a longer time to pass (Torres, 2016). The time that it took the desired analyte to pass will then aid the
The above diagram is a depiction of column chromatography. The first column on the left is the
beginning of this technique with only the stationary and sample loaded. The mobile phase is then
placed on top on the sample forming the eluent and then as time passes the different analytes pass
through separately depending on their interactions. The first analyte is then collected as it passes
through, then the second is collected as it passes through.
Gas Chromatography (GC)
compounds. Volatile compounds are organic compounds that, at standard room temperature
of about 23C, have an elevated vapor pressure ("Volatile organic compound", 2019). In this
technique, the sample is mixed with a solvent and then vaporized and passed through the stationary
phase in order to separate. The mobile phase of this technique is compromised of an inert gas, or noble
gas. Unlike the other techniques of separation, the inert gas in the mobile phase does not interact with
the analyte but instead helps carry the molecules of the sample (Libretexts, 2019).
Inlets, also known as injectors, are the entrance attached to the column head for the
sample to be placed into the chromatograph. These inlets introduce the sample into a small
heated chamber from a syringe, the heat breaks down the sample and turns the liquid into a
gas which is then carried to the column by either the entirety, “split-less” injection, or in
portions, “split” injection (Kumar, 2015). Another type of inlet is the “on-column” inlet where the
sample is introduced in its entirety without heat. Programmed Temperature Vaporizing Injector
introduces the sample just below its boiling point through the split line. (Kumar, 2015)
Columns of GC are a long tubing where the stationary phase is held at can come in two
different types: packed columns or capillary columns, which is also known as being open
tubular. Capillary columns come in two different forms usually made of glass (Libretexts, 2019).
One form of capillary columns contains finely divided, inert, solid support material that is
coated on the wall of the column with the stationary phase, also known as wall-coated open
tubular (Libretexts, 2019). The second form is also known as a support-coated open tubular
column. In these the columns are first coated with extremely small layer of adsorbent solid that
is treated with the stationary phase. Of these two capillary columns the, the support-coated
can hold a greater volume of stationary phase while the wall-coated is more efficient in
separation (Libretexts, 2019). Packed columns are usually made of glass or metal with a larger
diameter but do have a shorter length than capillary columns. Due to the greater diameter and
shorter length, the packed columns are way less efficient than wall-coated open tubular
Detectors are used at the end of the chromatogram to give a quantitative (being able to
measure) measurement of the analytes as they pass through with the inert gas (Libretexts,
2019). Regardless of the type of detector, there are two key parts that work together to
transduce (converts a physical amount into an electrical signal) (Libretexts, 2019). The first part is
a sensor at the end of the column and the second is an “electrical equipment used to digitize
the analog signal” for a computer to analyze the collected chromatogram (Libretexts, 2019).
Depending on the type of sample need to be analyzed there are several types of detectors.
Some types of detectors are Mass Spectrometer, Flame Ionization, Thermal Conductivity,
compounds and determine their purity. Just like the other techniques, a stationary phase is used but
composed of a finely ground material such as silica gel. Silica gel is used due its ability to fluoresce (shine
or glow) in Ultra-Violet light. The stationary phase is thinly layered as a “thick slurry” onto a glass place
When the plate is ready, it is then placed into a small amount of the mobile phase in a beaker.
The sample is then elevated up the plate by capillary action. Capillary action is the ability of a liquid to
become elevated depending on its own adhesiveness properties (Dictionary.com, 2019). The After the
experiment, the analyte spots are visualized when a UV light is projected onto the plate ("Thin-layer
chromatography", 2019). The distance traveled by the sample is then measured and compared to the
mobile phase, Retardation factor. If the substance being measured has a similar structure to that of the
mobile phase then the retardation factor will be higher. To keep a consistent measurement for the
substance that is being measured, a known compound is applied to the plate ("Thin-layer
chromatography", 2019).
References
https://www.britannica.com/science/solvent-chemistry
https://www.dictionary.com/browse/capillary--action
https://en.wikipedia.org/wiki/Chromatography
https://en.wikipedia.org/wiki/History_of_chromatography
https://goldbook.iupac.org/html/P/P04436.html
http://www.biologydiscussion.com/biochemistry/chromatography-techniques/top-
12-types-of-chromatographic-techniques-biochemistry/12730
https://chem.libretexts.org/Bookshelves/Analytical_Chemistry/Supplemental_Modules
_(Analytical_Chemistry)/Instrumental_Analysis/Chromatography/Gas_Chromatography
https://davidson.weizmann.ac.il/en/online/askexpert/chemistry/what-capillary-action-
and-how-it-affected-gravity-ariel-michal
9. Thin-layer chromatography. (2019, February 08). Retrieved from
https://en.wikipedia.org/wiki/Thin-layer_chromatography
10. Torres, J. (2016, August 07). The Basics of Running a Chromatography Column. Retrieved
from https://bitesizebio.com/29947/basics-chromatography-column/
https://en.wikipedia.org/wiki/Volatile_organic_compound
Post Write
There are some examples of parenthetical definitions, sentence definitions, graphics, partition,
Acknowledgment
I would like to thank my table partner, Gerardo Sandoval, for being specific on what I