MACHEREY-NAGEL

Derivatization
CH3 reagents for GC
Chromatography

CO N
Si(CH3)2 CH3 CH3 CF3 CO N
CH3

CF3 CO N Si(CH3)2 CH3

CH3 CH3
CF3 CO N Si(CH3)2 CH3
CF3 CO N
Si(CH3)2 CH3
CH3 Si(CH3)2
CH3 CF CO N
3
CF3 CO N Si(CH3)2 CH3
CH3 Si(CH3)2 CH3
CF3 CO N
Si(CH3)2 CH3 CH3
CH3 CF3 CO N
CF3 CO N Si(CH3)2 CH3

CH3
Si(CH3)2 CH3 CH3
CF3 CO N CF3 CO N
Enhance your GC analysis
Si(CH3)2 CH3
Si(CH

nnUltrapure
nnHighly reactive
nnCost-effective

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Content

A guide to derivatization reagents for GC............................................................4

Silylation.............................................................................................................6

Acylation...........................................................................................................12

Alkylation..........................................................................................................14

Derivatization procedures.................................................................................16

Overview of important functional groups...........................................................20

General reaction mechanisms...........................................................................22

Ordering information.........................................................................................24

Contact............................................................................................................26
A guide to derivatization reagents for GC

Regarding derivatization This is commonly achieved by rinsing

the glass with a silylating agent, e.g.,
Derivatization is one of the most com-
DMCS or HMDS, hence masking all
mon ways to prepare compounds
silanols with non adsorptive methyl
for GC that are otherwise difficult to
groups.
separate. Through derivatization, it is
possible to improve the separation by To achieve a satisfying rate of derivati­
replacing active hydrogens from the zation, it is essential to keep the follow-
analyte with various groups that are ing requirements in mind:
easier to handle. Derivatization gener-  he derivatization reaction needs to
T
nn

ally improves the following GC para­ be complete ≈ 100 %

meters:
 o loss of sample during derivati­
N
nn

nn Chromatographic behavior zation

nn Peak shape  he overall structure of the analyte
T
nn

nn Thermal and chemical stability will not be altered

nn Detectability  roduced derivative will be stable
P
nn

over time
nn Volatility
 o interaction between the reagent
N
nn
It is also important that all the instru-
and the chromatographic system
ments, e.g., laboratory glassware, will
not interfere with the sample.
To make sure that no compounds
containing -OH, -SH or -NH groups
will be adsorbed by present Si-OH on
the surface of the glass, a deactivation
process may be necessary.

H3C CH3 H3C CH3

Si Si
OH OH OH OH O O O O
DMCS
O Si O Si O Si O Si O O Si O Si O Si O Si O
O O O O – HCl O O O O

Glass Silanized glass

4 www.mn-net.com
A guide to derivatization reagents for GC

Naturally, all other components of the Derivatization reagents

sample preparation and handling pro-
There are many reagents in use today
cess need to be contaminant-free and
for derivatization. There are three cate-
in top condition. Since water is, in most
gories they can be allocated to:
cases, a problem, it has to be removed
from the derivatization process, e.g., Silylation
nn

by adding Na2SO4 to the reaction mix- Acylation

nn

ture. Like all reactions, derivatization Alkylation (Methylation)

nn

takes time and a certain amount of

heat to go to completion. As duration Good to know
may vary greatly, dependent on the re-
nn Our derivatization reagents
activity of the analyte, it is often neces- meet the highest demands of
sary to screen several reagents for the purity.
best result. It is also important to real- MADE BY
ize that there is no such thing as the
Highest purity
best derivatization method. There will Certificate / Zertifikat
CERTIFIED
always be several working solutions Silyl 991
Formula / Zusammensetzung
REF
BSTFA : TMCS / 99 : 1
701490

to a chromatographic problem with its

LOT
Expiry date / Haltbarkeit
Date of analysis / Analysendatum
Tester / Prüfer

AC
GE
M

L
own advantages and drawbacks, de-
Specification / Spezifikation Target / Sollwert Result / Messwert
Color colorless* – yellowish 

HER
Farbe farblos* - gelblich

E Y- N A
Content BSTFA 97,0 – 99,5% 98,0%
Gehalt BSTFA
Content TMCS 0,5 – 3,0% 0,8%
Gehalt TMCS
* Slight turbidity may occur and has no impact on reactivity. / Eine leichte Trübung kann auftreten, diese hat keinen Einfluss auf die Reaktivität.

pendent on the equipment or on the This document has been produced electronically and is valid without a signature.
Dieses Dokument wurde elektronisch erstellt und ist ohne Unterschrift gültig.

approach of the chemist.

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Silylation

Silylation is the most versatile method Additionally, silylation improves vola-

of derivatization in GC, i.e. more than tility, so that many compounds that
80 % of all derivatization reactions are are normally considered nonvolatile
actually silylations. Usually the term or thermally instable, can be chroma-
silylation in GC stands for replacement tographed easily. Introducing a silyl
of active hydrogen atoms by a trime­ group may also enhance the GC-MS
thylsilyl group (TMS derivative). Some- properties of the derivative, either
times, however, trialkylsilyl groups or through characteristic ions or more
dimethylalkylsilyl groups with longer favorable diagnostic patterns for struc-
alkyl chains are used for derivatization. ture investigations.
The trialkylsilyl group increases volatili-
ty and enhances thermal stability of the Good to know
sample. nn It is important to mention that
silylated compounds should
As with methylation, the replacement
not be used with WAX or FFAP phases,
of an active hydrogen with a silyl group as the OH groups of the stationary phase
reduces the polarity of the compound, will definitely become derivatized by the
as well as hydrogen bonding. silylating reagent, and this will irreversible
change selectivity of the column.

Silylation can be catalyzed either acid-

ically by the addition of TMCS or basi-
cally by the addition of pyridine or TSIM
(e.g., for sterically hindered molecules,
such as tertiary alcohols).

6 www.mn-net.com
Silylation

Amides N,O-bis-trimethylsilyl-trifluoroacetamide
(BSTFA)
N,O-bis-trimethylsilyl-acetamide (BSA)
M: 257.4 g/mol
M: 203.4 g/mol
Bp: 40 °C (12 mm Hg)
Bp: 71–73 °C (35 mm Hg)
density d20 °/4 ° = 0.96
density d20 °/4 ° = 0.83
O Si(CH3)3
O Si(CH3)3
F3C – C
H3C – C
N Si(CH3)3
N Si(CH3)3

BSA is a strong silylation reagent that BSTFA is a powerful trimethylsilyl do-

forms very stable TMS derivatives with nor with approximately the same donor
a large variety of compounds, e.g., strength as the nonfluorinated analog
non-sterically hindered alcohols, car- BSA.
boxylic acids, phenols, enols, steroids,
(biogenic) amines and alkaloids. Advantage of BSTFA over BSA
Not recommended for use with carbo­ Greater volatility of its derivatives (par-
hydrates or very low molecular weight ticularly useful for GC of some lower
compounds. boiling TMS amino acids). BSTFA will
generally react with all organic material
Good solvent for polar compounds,
present, but may not react with some
but frequently used in combination
amides, secondary amines and hin-
with a solvent (pyridine, DMF etc.), with
dered hydroxyl groups. However, add-
other silylation reagents or catalysts
ing 1 % TMCS (SILYL-991) will solve
such as TFA, HCl or TMBS.
that problem.
Used in combination with DMF, BSA
is the reagent of choice for derivatizing
phenols.

www.mn-net.com 7
Silylation

N-methyl-N-trimethylsilyl-trifluoroacetamide
The already good solution charac-
(MSTFA) teristics can be improved by adding
M: 199.1 g/mol submolar quantities of protic solvents
Bp: 70 °C (75 mm Hg) (e.g., TFA for extremely polar com-
density d20 °/4 ° = 1.11
pounds such as hydrochlorides) or
CH3 pyridine (e.g., for carbohydrates).
F3C CO N
Si(CH3)3 Advantages
 omplete reaction with high reaction
C
nn
MSTFA is the most volatile trimethylsilyl
rates, even without a catalyst
amide available.
(1–2 % TMCS or TSIM)
MSTFA is a very strong TMS donor
 y-product of the reaction
B
nn
that does not cause any noticeable
(N-methyl­trifluoroacetamide) features
FID contamination even after long-time
high volatility and short retention
measuring series. It is one of the most
time.
important silylating reagents. It can be
used, to silylate the hydrochloride salts Reactivity of silylation reagents
(acc. to M. Donike)
of amines or amino acids directly.
TMS amides (e.g., BSA, MSTFA) >
TMS amine = TSIM > Enol-O-TMS ether >
S-TMS ether > O-TMS ether > TMS-O-TMS

Stability of the TMS derivatives

O-TMS ether > S-TMS ether > Enol-O-TMS
ether > TMS amine > TMS amide

8 www.mn-net.com
Silylation

N-methyl-N-trimethylsilyl-heptafluoro­ N-methyl-N-tert-butyldimethylsilyl-trifluoro­
butyramide (MSHFBA) acetamide (MBDSTFA)
M: 299.1 g/mol M: 241.3 g/mol
Bp: 148 °C (760 mm Hg) Bp: 168–170 °C (760 mm Hg)
density d20 °/4 ° = 1.12
CH3 CH3
F7C3 CO N F3C CO N
Si(CH3)3 Si(CH3)2 C4H9

MSHFBA is similar to MSTFA in reac- MBDSTFA is a silylation reagent that

tivity and chromatography. donates a tert-butyldimethylsilyl group
Used either alone or in combination (TBDMS) for derivatizing active hy-
with a catalyst (TMCS, TSIM) or an- drogens in hydroxyl, carboxyl and
other silylation reagent with or without thiol groups, primary and secondary
solvent. amines, as well as in amino acids.

By-product N-methylheptafluorobutyric Fast reactions (typically 5–20 min) with

amide has a lower retention time than high yields (> 96 %). By-products are
the silylating reagent. neutral and volatile.

Especially useful for FID, because, TBDMS ethers are 104 times more
due to the large 7:1 ratio of fluorine stable than the corresponding TMS
to silicon, the degradation of excess ethers. Chromatographic retention
MSHFBA does not produce SiO2 but times are longer due to the large pro-
volatile, non-corrosive silicon com- tecting group, which may improve
pounds. some separations.
Very useful for GC-MS applications,
because of a high molecular ion con-
centration at M+-57 applications.

www.mn-net.com 9
Silylation

Silanes / silazanes Aprotic solvents, e.g., acetonitrile,

pyridine, dimethylformamide, carbon
Dimethyldichlorosilane (DMCS)
disulfide and dimethylacetamide are
M: 129.06 g/mol
Bp: 70 °C (760 mm Hg)
recommended for use with HMDS.
density d20 °/4 ° = 1.07 Trimethylchlorosilane (TMCS)
CH3 M: 108.7 g/mol
Bp: 57 °C (760 mm Hg)
H3C Si Cl density d20 °/4 ° = 0.86
Cl
CH3

DMCS is used to form dimethylsilyl H3C Si CH3

(DMS) derivatives. Cl

DMS derivatives are much more sus-

TMCS is often used as a catalyst with
ceptible to hydrolysis than TMS de-
other trimethylsilyl reagents. Without
rivatives. Therefore, strictly anhydrous
additives it can be used for preparing
conditions during the reaction are very
TMS derivatives of organic acids.
important.
Together with methanol, TMCS can be
Hexamethyldisilazane (HMDS)
used for Methylation.
M: 161.4 g/mol
Bp: 126 °C (760 mm Hg)
density d20 °/4 ° = 0.77
Silylation reagent mixtures
H3C CH3 SILYL-271 BSA – HMDS – TSIM (2:7:1)
SILYL-271 will derivatize all hydroxyl groups
H3C Si N Si CH3 in any position. Useful in multiderivatization
H
H3C CH3 schemes involving hydroxyl or amine groups.

HMDS is a weak TMS donor. If used SILYL-1139 TSIM – pyridine (11:39)

as sole reagent, it is slow and not very Recommended application: alcohols, phenols,
organic acids, steroids, hormones, glycols,
effective. After addition of catalytic
nucleotides and narcotics.
quantities (e.g., 1 %) of TMCS or as
a mixture with TMCS, it is a fast and
quantitative reagent for trimethylsilyla-
tion of organic compounds.

10 www.mn-net.com
Silylation

SILYL-21 HMDS – TMCS (2:1) Imidazoles

SILYL-21 will derivatize amides and many sec- N-Trimethylsilyl-imidazole (TSIM)
ondary amines and hindered hydroxyls that
M: 140.3 g/mol
would not be completely derivatized by HMDS
Bp: 94–96 °C (760 mm Hg)
alone. It can be used without solvent.
density d20 °/4 ° = 0.96
CH3
SILYL-2110 HMDS – TMCS – pyridine (2:1:10)
N
SILYL-319 HMDS – TMCS – pyridine (3:1:9) H3C Si N
SILYL-2110 and SILYL-319 will derivatize alco- CH3
hols, bile acids, phenols, most steroids, ste-
rols, and sugars that would not be completely
derivatized by HMDS alone. SILYL-2110 and
TSIM is the strongest hydroxyl silyla-
SILYL-319 are fast and easy to use, and can tor, the reagent of choice for carbohy-
be used without solvent. drates and most steroids (even highly
hindered steroids).
SILYL-991 BSTFA – TMCS (99:1)
The reagent is unique in that it reacts
BSTFA is a powerful trimethylsilyl donor.
For silylating of fatty acid amides, hindered quickly and smooth with hydroxyl
hydroxyls and other compounds that are dif- (even tert. OH) and carboxyl groups,
ficult to silylate, e.g., secondary alcohols and but not with amines. This character-
amines, we recommend BSTFA + 1 % TMCS,
available under the designation SILYL-991.
istic makes TSIM particularly useful in
multi-derivatization schemes for com-
pounds with different functional groups
Good to know that are to be derivatized differently,
nn Most derivatives are susceptible e.g., -O-TMS / -N-HFB derivatives of
to water and hydrolysis catecholamines.
nn Reactions only in aprotic solvents possible
nn The presence of water does not interfere Summary silylation
with SILYL-1139.
 ilylation can be applied on many
S
nn

compounds
 ilylating reagents are easily
S
nn

prepared
Large variety of reagents available
nn

www.mn-net.com 11
Acylation

Acylation / Benzoylation Acyl halides

Generally, acylation involves the in­ Pentafluorobenzoyl chloride (PFBC)
tro­­­­­duc­tion of an acyl group into a R= C6F5, X=Cl
molecule with a replaceable hydro­­­ M: 230.52 g/mol
gen, or across a double bond. Bp: 158–159 °C (760 mm Hg)
Acylation is used to convert com- density d20 °/4 ° = 1.60

pounds like alcohols, amines and O

thiols into their respective esters, am- R X
ides and thioesters. Additionally, they
enhance the detectability of the com- PFBC will react with hydroxyls, primary
pounds by adding halogenated carbon and secondary amines, amides and
to the compounds. This is achieved thiols.
through the reaction with fluorinated
acyl halides, anhydrides or bisacyl- Anhydrides
amides. While the corresponding acid-
Trifluoroacetic acid anhydride (TFAA)
ic by-products of the reactions with
R=CF3
acyl halides and anhydrides need to be
M: 210.04 g/mol
removed from the system by a suited
Bp: 39.5–40.5 °C (760 mm Hg)
base, e.g., pyridine, to prevent col- density d20 °/4 ° = 1.49
umn damage. By-products of bisacyl­ Heptafluorobutyric acid anhydride (HFBA)
amides are not acidic and normally R=C3F7
do not interfere with the subsequent
M: 410.06 g/mol
analysis. Hence, they are favorable re- Bp: 106–107 °C (760 mm Hg)
agents for acylations. density d20 °/4 ° = 1.665
O
R
O
R
O

Acylation with fluorinated acid anhy-

drides can be used for alcohols, phe-
nols, carboxylic acids, amines, amino
acids and steroids forming volatile,
stable derivatives suited for FID as well
as for ECD detection.

12 www.mn-net.com
Acylation

Bisacylamides Summary acylation

N-methyl-bis(trifluoroacetamide) (MBTFA)  ddition of halogenated carbons
A
nn

R=CF3 enhances detectability by ECD

M: 223.08 g/mol Derivatives are hydrolytically stable
nn

Bp: 123–124 °C (760 mm Hg)

density d20 °/4 ° = 1.55 Increased sensitivity by adding
nn

N-methyl-bis(heptafluorobutyramide) molecular weight

(MBHFBA)
R=C3F7
M: 423.1 g/mol
Bp: 165–166 °C (760 mm Hg)
density d20 °/4 ° = 1.67
O
R
N CH3
R
O

Acylation with fluorinated acid amides

is recommended for alcohols, primary
and secondary amines as well as for
thiols under mild, neutral conditions.
MBTFA also forms very volatile deri­
vatives with carbohydrates.

Good to know
nn Acylation reagents are moisture
sensitive
nn Reaction products (acidic by-products)
often have to be removed before analysis

www.mn-net.com 13
Alkylation

Alkylation (methylation) /  Trimethylsulfonium compounds

esterification Trimethylsulfonium hydroxide (TMSH, 0.2 M
in methanol)
Alkylation is a derivatization method
used to replace an acidic hydrogen with M: 94.06 g/mol

an alkyl or methyl group. It is generally H 3C +

–
restricted to amines or hydroxy groups S CH3 OH
H 3C
like in amino or carboxylic acids. The
resulting derivatives are ethers, esters,
methylamines or –amides and less Methylation with TMSH is recom-
polar than the original compounds. mended for free acids, chlorophenoxy-
Therefore, less hydrogen bonding oc- carboxylic acids, their salts and deriva-
curs. The acidity of the hydrogen to tives as well as for phenols and
be replaced significantly determines chlorophenols. Lipids or triglycerides
the conditions needed to perform the can be converted to the corresponding
alkylation. The less acidic, the more fatty acid methyl esters (FAMEs) by a
vigorous the conditions. simple transesterification.
This reaction is very elegant and con-
Methylation reagents venient, because it is just necessary to
add the reagent (0.2 M in methanol) to
Dialkylacetals the sample solution. Removal of ex-
N,N-dimethylformamide dimethylacetal cess reagent is not required, since in
(DMF-DMA) the injector of the gas chromatograph,
M: 119.17 g/mol at 250 °C, pyrolysis to volatile meth-
Bp: 106–107 °C (760 mm Hg)
density d20 °/4 ° = 0.89
anol and dimethylsulfide will occur.
Due to the high reactivity, complete
H3C CH3
derivatization is often obtained at am-
N
H3C CH3 bient temperature. However, heating
O O (e.g., 10 min at 100 °C) in
a closed sample vial may
DMF-DMA is recommended for steri-
be necessary to complete
cally hindered carboxylic acids, alde-
the reaction.
hydes, phenols and amines.

14 www.mn-net.com
Alkylation

Esterification reagents
Methylation
with methanol / TMCS
An 1M solution of TMCS in methanol is
suited for the esterification of free car-
boxylic acids and transesterification of
glycerides. Formation of HCl catalyzes
the reaction. TMCS and silyl ether re-
move water and thus drive the reaction
to completion. The mixture should be
freshly prepared.

Summary alkylation
(methylation)
nn  ethylation derivatives are generally
M
stable
nn  ide range of reaction conditions
W
(from strongly acidic to strongly
basic)
nn  ome reactions can be achieved
S
with water present

Good to know
nn Reactions are limited to acidic
hydrogens or amines
nn Reaction conditions may be extreme


www.mn-net.com 15
Derivatization procedures

Silylation

with BSA, BSTFA or SILYL-991 (BSTFA + 1 % TMCS)

BSA MN Appl. No. 213091 · BSTFA MN Appl. No. 213092 · SILYL-991
MN Appl. No. 213093

Add 0.5 mL of the silylation reagent to 1–10 mg sample; if necessary, add some
solvent (normally pyridine or DMF [dimethylformamide]). Heat to 60–80 °C for
20 min to increase the reaction rate. 1–2 drops of TMCS (trimethylchlorosilane)
or TSIM will also speed up the reaction.

with BSA in combination with other silylation reagents · MN Appl. No. 213100

BSA alone silylates all sterically unhindered hydroxyl groups of the steroid skel-
eton; addition of TMCS will enable reaction of moderately hindered OH groups
(reaction time 3–6 h at 60 °C). After addition of TSIM even strongly hindered
hydroxyl groups will react (reaction time 6–24 h at 60 °C).

with MSTFA, MSHFBA or MBDSTFA

MSTFA MN Appl. No. 213111 · MSHFBA MN Appl. No. 213112 · MBDSTFA
MN Appl. No. 213113

Dissolve 10–15 mg sample in 0.8 mL solvent, then add 0.2 mL of the silylation
reagent. The reaction mixture can be heated to 60–70 °C for up to 1 h and
can be analyzed directly. If TFA is used as a solvent, proceed as follows [20]:
dissolve 1–2 mg sample in 100 µL TFA. Dropwise add 0.9 mL of the silylating
reagent. After cooling the sample can be chromatographed directly.

with TSIM or SILYL-1139 (TSIM – pyridine 11:39)

TSIM MN Appl. No. 213121 SILYL-1139 MN Appl. No. 213122

Dissolve 10–15 mg sample in 0.8 mL solvent, then add 0.2 mL of the silylation
reagent. The reaction mixture can be heated to 60–70 °C for up to 1 hour
and can be analyzed directly. Recommended solvent is pyridine. When using
SILYL-1139, the presence of water does not interfere.

16 www.mn-net.com
Derivatization procedures

Silylation

with SILYL-21 or SILYL-2110 or SILYL-319 · SILYL-21 MN Appl. No. 213131 ·

SILYL-2110 · MN Appl. No. 213132

Carefully add SILYL-21, SILYL-2110 or SILYL-319 to 1–10 mg of the sample.

Precipitated ammonium chloride does not interfere. If the sample should not
dissolve within 5 min, heat to 75–85 °C. If no mutarotation is to be expected,
you may dissolve the sugar in warm pyridine first and then add the silylation re-
agent. In some cases it may be advantageous to use a different solvent instead
of pyridine. For derivatization of 3-ketosteroids we recommend to use DMF
(dimethyl­forma­mide).

O-trimethylsilylation with MSTFA followed by N-trifluoroacetylation with MBTFA

MN Appl. No. 213140

Completely silylate 2 mg of the sample with 0.3 mL MSTFA. After addition of

0.3 mL MBTFA the N-trimethylsilyl group is replaced by the N-trifluoroacetyl
group. The mixture can be analyzed directly.

www.mn-net.com 17
Derivatization procedures

Acylation

with fluorinated acid anhydrides

TFAA MN Appl. No. 213041 · HFBA MN Appl. No. 213042

Dissolve 0.1 to 1 mg sample in 0.1 mL solvent, add 0.1 mL of the anhydride and
heat to 60–70 °C for 1–2 h. If the sample needs not be concentrated prior to the
analysis and if there is no danger of catalytically induced side reactions, pyridine
is used as solvent. The reaction solution can be injected directly into the gas
chromatograph. Otherwise, use a volatile solvent and evaporate solvent, excess
reagent and free acid in a stream of nitrogen. Dissolve residue in 50 μL hexane,
chloroform etc. and inject aliquot portions.

with fluorinated acid amides

MBTFA MN Appl. No. 213051 · MBHFBA MN Appl. No. 213052

Add 0.5 mL MBTFA or MBHFBA to about 2 mg sample. If there is no reaction at

ambient temperature, heat the reaction mixture to 120 °C. Compounds difficult
to dissolve, can be trifluoroacetylated in suitable solvent mixtures. It is recom-
mended to use a ratio of solvent to MBTFA or MBHFBA of 4:1. The reaction
mixture is chromatographed directly.

18 www.mn-net.com
Derivatization procedures

Alkylation (Methylation)

with TMSH · MN Appl. No. 213060

Dissolve 100 mg sample (e.g., butter) in 5 mL of a solvent (e.g., tert.-butyl meth-

yl ether). Add 50 μL reagent to 100 μL of this solution. The mixture is injected
directly. The temperature of the injector must be at least 250 °C.

with DMF-DMA · MN Appl. No. 213070

Add 1 mL of a mixture of DMF-DMA and pyridine (1:1) to 1–50 mg fatty acids.

The sample can be injected as soon as a clear solution has formed. It is recom-
mended, however, to heat the solution to 60–100 °C for 10–15 min.

with methanol – TMCS · MN Appl. No. 213080

Add 1 mL methanol – TMCS to about 50 mg carboxylic acid or glyceride and

heat. Then evaporate in a stream of nitrogen and dissolve again for injection in,
e.g., n-heptane.

www.mn-net.com 19
Overview of important functional groups

Functional Silylation* Acylation /  Alkylation

Group Benzoylation
Primary BSA, MSTFA, MSHFBA, TSIM, SILYL-2110, TFAA, HFBA, TMSH
alcohols SILYL-319, SILYL-21, SILYL-1139 MBTFA,
MBHFBA, PFBC
Secondary BSA, MSTFA, MSHFBA, TSIM, SILYL-2110, TFAA, HFBA, TMSH
alcohols SILYL-319, SILYL-21, SILYL-1139 MBTFA,
MBHFBA, PFBC
Tertiary (and TSIM, BSTFA, SILYL-991 TFAA, HFBA, PFBC
sterically hin-
dered) alcohols
Thiols BSA, MSTFA, MSHFBA, TSIM, SILYL-2110, MBTFA, MBHFBA, TMSH
SILYL-319, SILYL-21, SILYL-1139 HFBA, TFAA
Phenols BSA, MSTFA, MSHFBA, TSIM, SILYL-2110, TFAA, HFBA, DMF-DMA,
SILYL-319, SILYL-21, SILYL-1139 MBTFA, TMSH
MBHFBA, PFBC
Glycols BSA, MSTFA, MSHFBA, TSIM, SILYL-2110, TFAA, HFBA, TMSH
SILYL-319, SILYL-21, SILYL-1139 MBTFA,
MBHFBA, PFBC
Aldehydes BSA, MSTFA, MSHFBA, TSIM, SILYL-2110, TFAA, HFBA, DMF-DMA,
SILYL-319, SILYL-21, SILYL-1139 MBTFA, TMSH,
MBHFBA, PFBC MeOH/TMCS
Ketones BSA, MSTFA, MSHFBA, TSIM, SILYL-2110, TFAA, HFBA, DMF-DMA,
SILYL-319, SILYL-21, SILYL-1139 MBTFA, TMSH,
MBHFBA, PFBC MeOH/TMCS
Carboxylic BSA, MSTFA, MSHFBA, TSIM, SILYL-2110, TFAA, HFBA, DMF-DMA,
acids SILYL-319, SILYL-21, SILYL-1139 MBTFA, TMSH,
MBHFBA, PFBC MeOH/TMCS
Carbohy- MSTFA, TSIM, SILYL-2110, SILYL-319, TFAA, MBTFA,
drates  /Sugars SILYL-991, HMDS PFBC
Acid anhydrides MeOH/TMCS
α-hydroxy acids MSTFA MBTFA

20 www.mn-net.com
Overview of important functional groups

Functional Silylation* Acylation /  Alkylation

Group Benzoylation
Primary amines BSA, MSTFA, MSHFBA, SILYL-991 TFAA, HFBA, DMF-DMA
MBTFA, MBHFBA,
PFBC
Secondary BSA, MSTFA, MSHFBA, SILYL-991 TFAA, HFBA, DMF-DMA
amines MBTFA, MBHFBA,
PFBC
Amides Silylamides are not stable TFAA, HFBA,
MBTFA, MBHFBA,
PFBC
Amino acids BSA, BSTFA, MSTFA, MSHFBA TFAA, HFBA, MeOH/TMCS,
MBTFA, MBHFBA, TMSH
PFBC
Amino sugars BSA, MSTFA, MSHFBA, SILYL-991 TFAA, HFBA,
MBTFA, MBHFBA,
PFBC
Imino acids BSA, MSTFA, MSHFBA, SILYL-991 TFAA, HFBA,
MBTFA, MBHFBA,
PFBC
Carbamides Silylamides are not stable TFAA, HFBA, MBT-
FA, MBHFBA
Alkylamides Silylamides are not stable PFBC
Amino alcohols MSTFA MBTFA
* (Avoid polar stationary phases containing active protons, e.g., WAX or FFAP)

www.mn-net.com 21
General reaction mechanisms

Silylation
CH3 CH3
Analyte X H + H3C Si Y Analyte X Si CH3 + HY
CH3 CH3

X = e.g., O, S, COO, etc.
Y = rest of silylation reagents; stuctures see page 7-11

Acylation
O O
Analyte X H + Y R Analyte X R + HY

X = e.g., O, S, NH, etc.
Y = rest of acylation reagents; stuctures see page 12, 13

22 www.mn-net.com
General reaction mechanisms

Alkylation (Methylation)  example TMSH

+
CH3
-
Analyte X H + TMSH OH Analyte X CH3 + S + H2O
CH3

X = e.g., O, S, COO, etc.
Alkylation reagents; stuctures see page 14

References
1. MN Chromatography catalog
2. Blau, K., Halket, J., Handbook of Derivatives for Chromatography, Second Edition; John Wiley &
Sons; Chichester, 1993
3. Knapp, D.R. Handbook of Analytical Derivatizations Reactions; John Wiley & Sons; New York, 1979
4. Butte, W., J. Chromatogr. A, 261, 1983

www.mn-net.com 23
Ordering information

Packing unit
Substance 10 x 1 mL 20 x 1 mL 1 x 10 mL 5 x 10 mL
Silylation reagents*
BSA 701210.110 701210.510
BSTFA 701220.201 701220.110 701220.510
DMCS**
HMDS 701240.510
TMCS** 701280.201
TSIM 701310.201 701310.110 701310.510
MSHFBA 701260.201 701260.110 701260.510
MSTFA 701270.201 701270.110 701270.510
MBDSTFA 701440.101 701440.201
SILYL-271 701450.201 701450.110 701450.510
(BSA – HMDS – TSIM 2:7:1)
SILYL-1139 701460.201
(TSIM – pyridine 11:39)
SILYL-21 (HMDS – TMCS 2:1) 701470.201
SILYL-2110 701480.201
(HMDS – TMCS – pyridine 2:1:10)
SILYL-319 701241.201
(HMDS – TMCS – pyridine 3:1:9)
SILYL-991 701490.201
(BSTFA – TMCS 99:1)
Acylation reagents*
HFBA 701110.201 701110.110 701110.510
MBTFA 701410.201 701410.110 701410.510
MBHFBA 701420.101 701420.201
PFBC 701120.101
TFAA 701130.110 701130.510
Alkylation reagents*
DMF-DMA 701430.201 701430.110
TMSH 701520.101 701520.201 701520.110 701520.510

Due to their purpose, derivatization reagents are very reactive chemicals. For this reason, they should
be stored cool and protected from moisture. For easy access with a syringe, our derivatization reagents
are supplied in vials with crimp caps. Vials with pierced sealing disks have limited stability and should be
used up soon.

24 www.mn-net.com
Ordering information

1 x 50 mL 6 x 50 mL 1 x 100 mL 6 x 100 mL 12 x 100 mL

701210.150

701230.650
701240.650
701280.650

701260.1100 701260.6100
701270.650 701270.1100 701270.6100 701270.12100

701490.150 701490.1100

*  These products contain harmful chemicals which must be specially labeled as hazardous.
For detailed information please see SDS.
** Vials with screw caps. Further screw caps on request.

www.mn-net.com 25
Contact

MACHEREY-NAGEL GmbH & Co. KG

Neumann-Neander-Str. 6–8 · 52355 Düren · Germany
DE / International:
Tel.: +49 24 21 969-0
Fax: +49 24 21 969-199
E-mail: info@mn-net.com
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Tel.: +41 62 388 55 00
Fax: +41 62 388 55 05
E-mail: sales-ch@mn-net.com
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Tel.: +33 388 68 22 68
Fax: +33 388 51 76 88
E-mail: sales-fr@mn-net.com
US:
Tel.: +1 484 821 0984
Fax: +1 484 821 1272
E-mail: sales-us@mn-net.com
www.mn-net.com
m
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© ymgerman · Worytko Pawel · Angel Simon · Mara Zemgaliete – fotolia.com
KATEN200144 Brochure Derivatization reagents en2 / 2 / 0 / 10.2016 DD · Printed in Germany