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02 ABRANTES, THEA CLARRIESSE B.

2A BIOCHEMISTRY
CHEM 203

THIN LAYER CHROMATOGRAPHY

It is observed that detection limits obtained after uses of


PDBIQ are very low in both cases before heating (0.1 - 2.0
g) and after heating (0.05 - 1.0 g) and various
distinguishable colors were produced. Sometimes the
detection limit is same before and after heating and in other
cases it is somewhat different. It should be noted that
identification of amino acids by ninhydrin is in practice
difficult, in spite of the high sensitivity of ninhydrin So the
new spray reagent unable to differentiate the amino
acids by color reaction. The mechanism of the color
formation is still uncertain, but we may assume that
carboxylic group of aminoacids first condensed with PDBIQ
(heating at 90C for 10 min) to form a carbamide type
intermediates which form charge transfer complexes with
ninhydrin.

REFERENCE: Sen, S., Sarkar, S., Kundu, P., Laskar, S., (2012). Separation of Amino Acids Based on Thin-Layer
Chromatography by a Novel Quinazoline Based Anti-Microbial Agent. American Journal of Analytical Chemistry. 3,
669-674

COLUMN CHROMATOGRAPHY

The multiple aspects of this experiment make it a useful


introduction to the interdisciplinary nature of neuroscience
research. Dissecting out specific regions of the brain allows
the students to review their neuroanatomy and provides
them with a hands-on opportunity to visualize familiar
structures. The use of unilateral lesions for behavioral and
pharmacological studies is presented in the context of the
current experiment. This type of
experimental protocol provides a within-subject control
value (the contralateral or unlesioned hemisphere) with
which to carry out statistical comparisons

REFERENCE: Vhurch, W., (2005). Column Chromatography Analysis of Brain Tissue: An


advanced Laboratory Exercise for Neuroscience Majors. The Journal of Undergraduate Neuroscience Education.
3(2), A36-A41
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Very good and rapid separations of tocopherols were also


obtained using nano-HPLC with a capillary column (100 m
ID) filled with 3 m RP18 particles6. Monolithic column
were shown to improve the determination of vitamin E and
A in human serum in shortening the time of analysis four
times in comparison with using traditional particulate
columns. An example of simultaneous determination of -
tocopherol, -carotene and retinol by HPLC with diode-
array detection may be found in the work7,8. The precise
analysis of the carboxyethyl-hydroxychroman metabolites
of -and -tocopherol found in plasma or urine is possible
when using a combination of gas chromatography with
mass spectrometry.

REFERENCE: Edison, B., (2009). Analysis of Tocopherols by High Performance Liquid Chromatography. E-Journal of
Chemistry. 6(2), 395-398

GAS CHROMATOGRAPHY

Identification of 19 fatty acids in human ES cells was


possible using this method, demonstrating that GC/MS is
effective for determination of fatty acids. It is believed that
the method introduced here can be applied effectively to
fatty acid composition analysis for various types of cultured
cells.

REFERENCE: Suzuki, T., Miyoshi, M., Nakagawa, K., Suemori, H., Chumas, S., Nakatsuji, N. (2013). Determination of
Fatty-acid Composition of Lipids in Human Embryonic Stem Cells Using GC-MS. Shimadzu Corporation. 3655-
08332.