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Thiols Analysis Service

Thiols, also named as mercaptan, are a class of organic compounds containing a sulfhydryl group (SH). They are similar to the alcohols and phenols but with the oxygen atom replaced by a sulfur atom.

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17 views

Thiols Analysis Service

Thiols, also named as mercaptan, are a class of organic compounds containing a sulfhydryl group (SH). They are similar to the alcohols and phenols but with the oxygen atom replaced by a sulfur atom.

Uploaded by

Steven4654
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Thiols Analysis Service

Thiols, also named as mercaptan, are a class of organic compounds


containing a sulfhydryl group (SH). They are similar to the alcohols and
phenols but with the oxygen atom replaced by a sulfur atom. Actually, thiols
are responsible for high volatility and unpleasant odor resembling that of garlic
or rotten eggs. Thiols also contribute to the telltale odor skunks sprays in self-
defense and the scent of freshly chopped onions. A concentration of 10 ppb
can be recognized by human nose. Because of their disagreeable odours, it is
easy to say whether thiols exist in petroleum and natural gas. They are also
the same compounds responsible for the smell associated with bad breath
and flatulence in humans. However, not the odors properties of all thiols are
offensive. The thiols in many botanicals lead to their aromatic properties. For
example, thioterpineol, a thiol in grapefruit, contributes to the distinctive yet
not unpleasant fragrance of grapefruit. Thiols also are involved in heady
aroma familiar to those brew beer or wine at home.
Different types of proteins like antibodies, receptors, hormones, and enzymes
often include one or more Cys residues, existing as free thiols or as intra- and
inter-molecular disulfide bond(s). The free Cys residue of some enzymes is
part of the catalytic activity site, while a disulfide bond in peptides and proteins
enhance the conformational rigidity on a protein. The formation of non-native
intramolecular disulfide bond(s) during the folding process may cause protein
misfolding, which in turn may lead to precipitation and aggregation. Native
disulfide bonds in proteins are formed during the oxidative folding process in
the ER lumen in eukaryotes and in the periplasm in prokaryotes. This
oxidative folding process in eukaryotes is assisted by a series of enzymes like
Erv2, Ero1 (endoplasmic reticulum oxireductin-1) and PDI (protein disulfide
isomerases). Through a series of thiol-disulfide exchange reactions, PDI
works together with Ero1 or Erv2 to oxidize thiol groups in a protein directly.
The Ero1 enzyme inside the ER, which is associated with the ER membrane,
can oxidize PDI and form a disulfide between Ero1 and PDI. PDI will catalyze
the reduction of incorrectly formed disulfide bonds and, then will re-oxidize the
protein substrate to form the correct disulfide bond. The protein disulfide
oxidoreductase (PDO), which contributing to oxidization of proteins from the
cysteines to disulfides, mainly exists in cytoplasm. Therefore, generally
speaking, proteins in cytoplasm contain free thiols, while proteins found in
other compartments like the ER usually including disulfide bonds.
HPLC-UV is widely used for the quantitative determination of thiols in
proteins, because thiols can react with different reagents and show maximum
absorption at different wavelength. For example, in the reaction with DTNB,
the earliest reagent widely used for thiol group quantification, thiol group(s)
produces an equivalent amount of 5-thio-2-nitrobenzoic acids, with maximum
absorbance at 412 nm.
Platform
 HPLC-UV
Summary
 Identification & Quantification of Thiols
Sample Requirement
 Normal Volume: 400ul plasma
 Minimal Volume: 200ul plasma
Report
 A detailed technical report will be provided at the end of the whole project,
including the experiment procedure, MS instrument parameters
 Analytes are reported as uM/ml, while CV's are generally 10%.
 The name of the analytes, abbreviation, formula, molecular weight and CAS#
would also be included in the report.

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