Journal of Chromatography A, 1518 (2017) 70–77

Contents lists available at ScienceDirect

Journal of Chromatography A
journal homepage: www.elsevier.com/locate/chroma

Generic gas chromatography-flame ionization detection method for

quantitation of volatile amines in pharmaceutical drugs and synthetic
intermediates
Gabriel C. Graffius a,∗ , Brandon M. Jocher a , Daniel Zewge a , Holst M. Halsey a , Gary Lee b ,
Frank Bernardoni a , Xiaodong Bu a , Robert Hartman a , Erik L. Regalado a,∗
a
Process Research & Development, MRL, Merck & Co., Inc., Rahway, NJ 07065, USA
b
Agilent Technologies, Incorporated, Wilmington, DE 19808, USA

a r t i c l e i n f o a b s t r a c t

Article history: Volatile amines are among the most frequently used chemicals in organic and pharmaceutical chem-
Received 25 July 2017 istry. Synthetic route optimization often involves the evaluation of several different amines requiring
Received in revised form 16 August 2017 the development and validation of analytical methods for quantitation of residual amine levels. Herein,
Accepted 17 August 2017
a simple and fast generic GC-FID method on an Agilent J&W CP-Volamine capillary column (using either
Available online 23 August 2017
He or H2 as the carrier gas) capable of separating over 25 volatile amines and other basic polar species
commonly used in pharmaceutical chemistry workflows is described. This 16 min method is successfully
Keywords:
applied to the analysis and quantitation of volatile amines in a variety of pharmaceutically-related drugs
Gas chromatography-flame ionization
detector
and synthetic intermediates. Method validation experiments showed excellent analytical performance
Method development in linearity, recovery, repeatability, and limit of quantitation and detection. In addition, diverse examples
Volatile amines for the application of this method to the simultaneous determination of other amine-related chemicals in
Basic compounds reaction mixtures are illustrated, thereby indicating that these GC-FID method conditions can be effec-
Pharmaceutical analysis tively used as starting point during method development for the analysis of other basic polar species
Capillary column beyond the validated list of amines described in this study.
© 2017 Elsevier B.V. All rights reserved.

1. Introduction A wide spectrum of extraction procedures combined with

Volatile amines are among the most frequently used compounds
in pharmaceutical chemistry due to their basic properties and
low boiling point which allow the chemist to control the pH of
reaction mixtures and improve product yield [1–3]. Optimization
of the synthetic route in drug development and manufacturing
involves constant change and the screening of numerous reaction
variables [4–7] including volatile amines. The selection of the “opti-
mal” amine by the synthetic chemist requires a combination of
1) improvement in reaction performance and 2) how effectively
the amine itself can be removed during the subsequent chemical
processes. In this regard, the time spent developing new analytical
methods for quantitation of residual amine content prior to each
analysis session becomes a constraining bottleneck in synthetic
route development.
∗ Corresponding authors.
E-mail addresses: gabriel graffius@merck.com (G.C. Graffius),
erik.regalado@merck.com (E.L. Regalado).
chromatographic techniques have been previously developed for
the quantitation of amines including liquid chromatography-
mass spectrometry (LC–MS) [8], ion chromatography-conductivity
detection (IC-CD) [9], IC-MS [10], gas chromatography-flame ion-
ization detection (GC-FID) [11–15] and GC–MS [16–18] among
others. Derivatization prior to chromatographic separation with
absorbance [19–23], fluorescence [24,25] or MS [26] detection has
also been extensively used. However, most of previous procedures
require detailed sample preparation techniques and specific instru-
mentation. In addition, previous methods are focused on a very
narrow group of volatile amines and do not have the potential
to be used universally when working with the diverse group of
amines commonly used during pharmaceutical research and devel-
opment. Additionally, the highly polar nature of the amines leads
to poor peak shape, limiting the quantitation and linear ranges of
the analyses under most conditions.
In the last decade, the development and validation of generic
or more universal chromatographic methods that cover multiple
related analytes in a single experimental run has gained favor in
both academia and industry [10,18,19,27–33]. This approach can

http://dx.doi.org/10.1016/j.chroma.2017.08.048
0021-9673/© 2017 Elsevier B.V. All rights reserved.
 G.C. Graffius et al. / J. Chromatogr. A 1518 (2017) 70–77 71

greatly enhance the speed at which chemists can generate accu- Table 1
GC-FID operation conditions for the generic amine method.
rate and quality data to support the development of new synthetic
routes. In this study, we report a new systematic approach for the Parameter Setting
analysis of over 25 volatile amines and other basic polar species in a Analytical column
single 16 min chromatographic run using conventional and readily Type Agilent J&W CP-Volamine
available GC-FID instrumentation. The use of this simple and fast Length 30 m
generic GC-FID method using either He or H2 as carrier gas for the Internal diameter 320 ␮m
Film thickness 5 ␮m
analysis of residual amine content in pharmaceuticals is demon-
Carrier gas He (99.999%)
strated with full validation data provided to enable this method to Carrier gas flow rate 2.0 mL/min
be immediately applied in a regulatory setting.
Oven temperature
Initial temperature 40 ◦ C
2. Experimental Initial holding time 2 min
First temperature ramp 11 ◦ C/min
2.1. Chemicals and reagents Second temperature 120 ◦ C
Second holding time 0 min
Second temperature ramp 33 ◦ C/min
Methylamine, dimethylamine, ethylamine, tert-butylamine, Final temperature 250 ◦ C
diethylamine, ethylenediamine, ethanolamine, pyridine, Final holding time 3 min
dimethylformamide (DMF), piperidine, morpholine, 3- Analysis time 16.21 min
(dimethylamino)propylamine, N,N-dimethylacetamide Injector parametersa
(DMAc), N-propylamine, o-anisidine, p-anisidine, m- Pre- Wash 1 3 times with ACN/H2O (50/50, v/v)
anisidine, dimethylcyclohexane-1,2-diamine (DMCHDA), Pre- Wash 2 3 times with Diluent
Post- Wash 1 3 times with Diluent
1,3-dimethyl-3,4,5,6-tetrahydro-2-pyrimidinone (DMPU) and
Post-Wash 2 3 times with ACN/H2O (50/50, v/v)
tetramethylenediamine were obtained from Sigma-Aldrich, Inc. (St Sample wash 3
Louis, MO, USA). Trimethylamine, isopropylamine, n-butylamine, Injector pumps 5
and 2-phenylethylamine were obtained from Tokyo Chemical Type Split
Temperature 200 ◦ C
Industry Co. (Portland, OR). Ethylmethylamine and sec-butylamine
Split ratio 50
were obtained from Acros Organics (Fair Lawn, NJ, USA). Ace- Split flow 100 mL/min
tonitrile and triethylamine were obtained from Fisher Scientific
Detector
(Fair Lawn, NJ, USA). 3-(methylamino)propylamine, N-methyl-2-
Type FID
pyrrolidone (NMP), and 1,3-dimethyl-2-imidazolidinone (DMI) Temperature 260 ◦ C
were obtained from Honeywell Fluka (St. Louis, MO, USA). 1,3- Make-up gas flow 25 mL/min
diaminopropane was obtained from MP Biomedicals (Solon, OH, *
At the end of a sequence, the flow rate decreased to 1.0 mL/min and the split ratio to
USA). 2:1 as a standby method to reduce the total gas consumption. For long-term storage,
lower the temperature and turn off the detector flame.
a
2.2. GC-FID conditions ACN/H2 O (50/50, v/v) used in the washing procedure dramatically reduces carry-
over, yet diluent is used immediately prior to the sampling. The validation contained
here-in also utilized ACN/H2 O (50/50, v/v) as the diluent.
Residual amine analyses were performed primarily on an Agi-
lent 6890N GC-FID with an Agilent 7683 B injector. The system
was controlled by EmpowerTM 3 Chromatographic System (Waters, was determined as (a) the concentration that had a minimum sig-
Milford, MA, USA). All amines were separated on an Agilent J&W CP- nal to noise ratio (S/N) of 10:1, and (b) had a maximum of 25%
Volamine (30 m x 0.32 mm, 5.0 ␮m film thickness column, CP7447). RSD for area counts over 6 injections. The LOD was established by
GC-FID experimental conditions are detailed in Table 1. determining the concentration that yielded a minimum signal-to-
noise ratio of 3:1. A summary of the LOD and LOQ is also shown
2.3. Validation experiments as part of Table 2. Specificity: The resolution of all analytes was
established with respect to each other. The method was considered
The GC experiments were designed to satisfy all requirements specific to a particular amine if the resolution factor between it and
for pre-filing and original filing validation. The primary diluent for any other possible interference was greater than 1. Minor adjust-
these experiments was ACN/H2 O (50/50, v/v) as it had the least ments to the recommended temperature gradient program may be
interference and/or contamination with the amines of interest. Sev- made to improve resolution between specific pairs of amines. The
eral other diluents planned for future validation work not included authors note that interactions between amines may occur during
in this paper are THF, DMAC, DMAC/H2 O, DMF, DMSO, DMI, and the sampling and analysis and therefore impact the specificity of
NMP. Precision: Repeatability was evaluated for 6 injections of stan- the method.
dards prepared at 0.1% v/v, an intermediate level (typically 0.01%
v/v), and the LOQ. The intermediate and LOQ concentrations were 2.4. Preparation of standards and samples
selected for each amine based on its linear range. Valid precision
was claimed if the% relative standard deviation (RSD) was below Stock standards and dilutions were prepared following GMP
10% at the intermediate and 0.1% v/v level. Linearity: Linearity for procedures in volumetric flasks using Grade A glass pipettes. A 1%
each amine was established by a minimum of 5 data points, specific v/v stock standard solution was typically prepared by diluting 1 mL
to each amine, ranging from the LOQ to 0.1% v/v, with an additional of the neat compound into a 100 mL volumetric flask and diluted
point taken at 0.2% v/v. For example, linearity points for Pyridine to the mark with diluent. From the stock standard preparation,
included: 0.0002% v/v (LOQ), 0.0004% v/v, 0.001% v/v, 0.002% v/v, serial dilutions were performed to prepare the linearity standards,
0.01% v/v, 0.04% v/v, 0.1% v/v, and 0.2% v/v. A minimum of three specific to the linear range of each amine. Standards used for quan-
injections of each dilution were made and the average was taken. titative analysis are prepared at similar concentration to that of
Acceptable linearity was determined based on correlation coeffi- the target concentration in the sample. For sample analysis, solids
cient (R) ≥ 0.9900. A summary of the linearity for each amine is are weighed and diluted at a concentration necessary to obtain the
shown as part of Table 2. LOQ/LOD: the Limit of Quantitation (LOQ) desired LOQ (see Table 2 for example calculations). To determine
72 G.C. Graffius et al. / J. Chromatogr. A 1518 (2017) 70–77

Table 2
Summary of Validation Results for the Generic Amine Method.

No Analyte Rt (min)±RSDa Density (g/mL) 50 mg/mL dilution

LOD [2] LOQ [3] Linear range

Weight% (s/n) Weight% (s/n) Weight% (R2 )

1 Methylamine 2.62 ± 0.05% 0.662 0.013 0.026 0.026–1.3

(40% wt in water) (46) (164) (0.999)
2 Dimethylamine 3.49 ± 0.09% 0.706 0.014 0.028 0.028–1.4
(40% wt in water) (50) (203) (0.999)
3 Ethylamine 3.63 ± 0.06% 0.689 0.014 0.028 0.028–1.4
(2.0 M in THF) (44) (121) (0.999)
4 Trimethylamine 3.78 ± 0.05% 0.630 0.006 0.013 0.013–1.3
(28% in water) (85) (365) (0.999)
5 Isopropylamineb 4.58 ± 0.02% 0.689 0.003 0.007 0.007–1.4
(99%) (22) (56) (0.999)
6 Ethylmethylamine 4.95 ± 0.05% 0.688 0.014 0.028 0.028–1.4
(97%) (14) (49) (0.999)
7 tert-Butylamine 5.32 ± 0.04% 0.696 0.003 0.006 0.006–1.4
(99.50%) (7) (27) (0.999)
8 n-Propylamine 5.67 ± 0.02% 0.719 0.007 0.014 0.014–1.5
(99%) (10) (55) (0.999)
9 Diethylamine 6.60 ± 0.02% 0.707 0.007 0.014 0.014–1.4
(99.5%) (15) (60) (0.999)
10 sec-Butylamine 6.90 ± 0.04% 0.724 0.003 0.007 0.007–1.4
(99%) (17) (84) (0.999)
11 Tetrohydrofuran 7.85 ± 0.01% 0.883 0.002 0.004 0.004–1.8
(99%) (17) (34) (1.000)
12 n-Butylamine 7.99 ± 0.01% 0.733 0.007 0.015 0.015–1.5
(99.0%) (63) (278) (0.999)
13 Ethylenediamine 8.39 ± 0.01% 0.898 0.036 0.072 0.072–1.8
(99.5%) (74) (155) (0.999)
14 Ethanolamine 8.40 ± 0.08% 1.012 0.020 0.040 0.040–2.0
(99%) (11) (34) (0.999)
15 Triethylamine 9.28 ± 0.02 0.726 0.001 0.003 0.003–1.5
(99%) (7) (17) (0.999)
16 Pyridine 10.11 ± 0.02% 0.978 0.002 0.004 0.004–2.0
(99.80%) (32) (72) (0.999)
17 1,3-Diaminopropane 10.47 ± 0.02% 0.888 0.018 0.036 0.036–1.8
(99%) (10) (180) (0.999)
18 DMF 10.38 ± 0.01% 0.944 0.002 0.004 0.004–1.9
(99.90%) (9) (26) (1.000)
19 Piperidine 10.54 ± 0.02% 0.862 0.003 0.009 0.009–1.7
(99%) (14) (98) (0.999)
20 Morpholine 10.84 ± 0.02% 0.996 0.002 0.004 0.004–2.0
(99.0%) (9) (30) (0.999)
21 3-(methylamino)propylamine 11.21 ± 0.02% 0.844 0.017 0.034 0.034–1.7
(97%) (20) (140) (0.999)
22 3-(dimethylamino)propylamine 11.40 ± 0.02% 0.812 0.016 0.032 0.032–1.6
(99%) (42) (184) (0.999)
23 DMAc 11.53 ± 0.02% 0.937 0.002 0.004 0.004–1.9
(99%) (21) (50) (0.999)
24 Tetraethylenediamine 11.77 ± 0.01% 0.877 0.009 0.018 0.018–1.8
(99%) (5) (37) (0.999)
25 NMP 13.13 ± 0.01% 1.027 0.002 0.004 0.004–2.0
(99.9%) (37) (81) (1.000)
26 DMI 13.67 ± 0.01% 1.052 0.002 0.004 0.004–2.1
(99%) (28) (60) (0.999)
27 2-Phenylethylamine 13.72 ± 0.01% 0.964 0.002 0.004 0.004–1.9
(98.0%) (19) (68) (0.999)
28 DMPU 14.71 ± 0.01% 1.064 0.004 0.011 0.011–2.1
(98%) (27) (57) (0.999)
*
Stability of the column stationary phase is limited when using diluent with high water content.
a
Repeatability in RT was performed at 0.01% v/v.
b
Isopropylamine was validated using DMI as diluent due to the ACN diluent interference.

the weight% of residual amine in an actual sample, the following Dslvnt = density of solvent to be determined (g/mL)
equation can be used: Astd = area counts of the standard
  Msmpl = mass of the sample (g)
(Asmpl ) (VPstd ) Vsmpl (Dslvnt )
WP =  
(Astd ) Msmpl 2.5. Accuracy study (Spike and recovery)

WP = weight percent of the amine in the drug The amines utilized in the synthesis of four separate MSD pro-
Asmpl = area counts for the sample grams and four generic Active Pharmaceutical Ingredients (API)
VPstd = volume percent of the standard were spiked into the sample matrix of the API at levels ranging
Vsmpl = volume of sample diluent used (mL) from a minimum of the filed specification. Grazoprevir, Sitagliptin,
 G.C. Graffius et al. / J. Chromatogr. A 1518 (2017) 70–77
Verubecestat, Ceftolozane (Merck & Co., Inc., Rahway, NJ, USA),
epinephrine, ibuprofen, 4-flavanone and quercetin (Sigma-Aldrich,
Inc.) were all prepared individually at approximately 20 mg/mL.
ACN/H2O (50/50, v/v) was used as diluent for all drugs, except
for Sitagliptin, Verubecestat and Ceftolozane (DMI). Each analyte
was spiked into the sample matrix at three concentrations levels
(0.002, 0.01 and 0.1%). Duplicate injections of two sample prepara-
tions were analyzed, and the experimental result was compared to
the theoretical spiked value to assess recovery using the following
equation:
% Recovery
Wt% of Solvent Analyte Determined Experimentally
= × 100
Theoretical Wt% Based on Spike
3. Results and discussion
The development and validation of a generic GC-FID method
for chromatographic separation and analysis of multiple volatile
amines in a single experimental run is vital for a fast and reli-
able turnaround of analytical results. All the analytes selected in
this investigation (Table 2) form part of a comprehensive list of
volatile amines and other basic polar species that have been used by
medicinal and process chemists in pharmaceutical laboratories for
drug research and development during the last decade. Fig. 1 illus-
trates a new optimized method on a CP-Volamine capillary column
using a gradient temperature program capable of separating over
25 amines and other basic polar species in less than 16 min using
conventional GC-FID instrumentation. This is a fused silica column,
coated with a base deactivated non-polar siloxane type station-
ary phase (SP) which creates a highly inert coating surface with a
minimum degree of adsorption for difficult basic compounds that
generally tail on traditional gas chromatography stationary phases.
High-boiling-point solvents (e.g. DMAc, DMSO, DMF, NMP and
DMI) are typically the preferred choice of diluents for GC analysis
because they often afford excellent solubility and minimal inter-
ference with earlier eluting analytes in the GC assay. However,
GC-FID and GC–MS analyses of some of these diluents clearly show
a number of high-boiling point impurities, interfering with some
of the amines targeted in this method. Interestingly, this method
is capable to separate a lot of impurities from the diluent peak
indicating that it can be used to test the quality of diluents used
in synthetic reactions, which is very useful to help establish the
source of unexpected impurities that may be observed in the final
API and synthetic intermediates. Overall, 50/50 ACN/H2 O (v/v) was
found to be a good diluent choice to demonstrate this method as it
reduced carryover (see pre/post-wash cycles in Table 1) and mini-
mized potential coelution with strongly retained amines that elute
close to the high-boiling point diluents. Isopropylamine (5) is the
only component that elutes too closely to the ACN diluent peak. For
this particular case, a high-boing point diluent (DMI) was selected
for method validation.
With the selection of diluent and GC-FID conditions, we next
focused on the validation experiments. Table 2 summarizes the lin-
earity range, precision, and LOQ/LOD experiments performed for
all amines and some additional low and high boiling point solvents
typically used as diluents during GC-FID analysis. Response factors
of standards were determined to be linear within the range stud-
ied, having a correlation coefficient (R) ≥0.990. The% RSD for all
tested standard solutions was ≤5%, with the majority ≤3%. These
validation experiments serve as the basis for the method used in
the initial stages of drug development, prior to expanded valida-
tion of the method for the specific product. The quantitation ranges
(0.007–0.013% w/w) and the weight% (wt%) calculations for a the-
oretical 50 mg/mL sample preparation are also listed in Table 2.
73
However, the detection range for a particular amine is usually
dependent upon the solubility of the drug substance in the dilu-
ent. For example, the quantitation limit of residual pyridine in a
drug substance that is soluble at 50 mg/mL is 5 times lower than
the quantitation limit of pyridine in a drug substance that is soluble
at only 10 mg/mL.
The reliability of the method was confirmed by recovery exper-
iments performed in multiple pharmaceuticals containing diverse
functional groups (Table 3): MSD APIs (Grazoprevir, Sitagliptin,
Verubecestat and Ceftolozane), as well as generic drugs and nat-
ural products (epinephrine, ibuprofen, 4-flavanone and quercetin).
Three different concentrations across the linear range (0.1, 0.01 and
0.002% v/v) were spiked into each drug. Analytes spiked into MSD
APIs are the same used during drug synthesis and manufactur-
ing, e.g. triethylamine (TEA) and N,N-dimethylacetamide (DMAc)
in Ceftolozane, 1,3-dimethyl-3,4,5,6-tetrahydro-2-pyrimidinone
(DMPU) in Verubecestat, isopropylamine in Sitagliptin, and pyri-
dine in Grazoprevir. However, a diverse group of amines were
randomly selected for recovery experiments in the case of generic
drugs and natural products, e.g. sec-butylamine and TEA in
epinephrine, DMAc and pyridine in ibuprofen, diethylamine, n-
butylamine and n-propylamine in 4-flavanone, and DMAc and
pyridine in quercetin. The average recovery (%) was calculated from
duplicate injections of two sample preparations as described in the
experimental section. The results depicted in Table 3 show recover-
ies that meet the GMP requirements (75–125%) across all different
spiking levels in all drugs, with overall relative standard deviation
below 5%. It is important to point out that spike and recovery is a
very important criteria in method validation and it is recommended
to perform these experiments in cases where this method is used
for quantitation of volatile amines in any other pharmaceuticals. In
case of low recovery values and/or analyte interference, the use of
head-space sampling prior to GC-FID analysis can help minimize
potential matrix effects. Finally, determination of pyridine, THF,
isopropylamine, ethylendiamine, DMAc and TEA content (% w/w)
in MSD drugs and synthetic intermediate show results that meet
the specifications set for batch release, as can be seen in Table 3.
This method covers a diverse set of volatile amines and high-
boiling point diluents commonly used by medicinal and process
chemists in the synthesis of new synthetic intermediates and phar-
maceutical ingredients. These GC-FID conditions can also serve as
a starting point for method development in order to analyze other
challenging polar amines not represented in Table 2. Fig. 2 illus-
trates numerous cases where minor modifications of this method
(using other diluent instead of ACN/H2 O) allowed us to solve
difficult analytical separations for quantitative analyses of other
amine-related chemicals used during process development.
In the example shown in Fig. 2a, the same method conditions
using DMAc as the diluent are effectively employed to the anal-
ysis of 1,3-dimethyl-3,4,5,6-tetrahydro-2-pyrimidinone (DMPU),
a polar aprotic solvent used as an additive to minimize aggre-
gation of lithiated species [34] for the synthesis of intermediate
3 via flow chemistry [35]. Fig. 2b provides an example from a
recent project of how this method can also be used for simul-
taneous determination of ethylenediamine (EDA) and the ligand
(dimethylcyclohexane-1,2-diamine) levels in the copper catalyzed
coupling of an amide (intermediate 5) with an aryl bromide inter-
mediate 4 [36]. In another example (Fig. 2c), a method was required
for starting material testing [37]. Once again, the exact method
conditions delivered baseline resolution and excellent peak shape
for the separation of anisidine isomers which also happen to be
potential mutagenic impurities (PMIs 30-32). Interestingly, all tar-
get analytes presented in Fig. 2a-c are baseline resolved from one
another without the need of method optimization. It is impor-
tant to point out that having many of these species in the same
reaction mixture is highly unlikely. However, having a validated
74 G.C. Graffius et al. / J. Chromatogr. A 1518 (2017) 70–77
Fig. 1. GC-FID method for analysis and separation of over 25 amines and other basic polar species. Method conditions are detailed in Table 1 (Experimental section).
Table 3
Quantitation of amine in pharmaceutical drugs and synthetic intermediates.
generic method in place capable of resolving complex multicom-
ponent mixtures of volatile basic species increases the simplicity
and speed of troubleshooting process development.
The cumulative solvent use and waste from chromatographic
systems have been a very important topic targeted in recent green
chemistry investigations [38–46] including the replacement of
helium with hydrogen as the carrier gas for routine GC analysis
[47–50]. Fig. 3 illustrates the GC-FID profile of separation of all 27
volatile amines using H2 instead of He as the carrier gas. H2 carrier
gas was able to provide excellent separation of the amines with no
modifications to the chromatographic parameters used with He.
Overall, the H2 method elution is one minute faster and provides
better chromatographic performance including sharper peaks, less
tailing, and better separation of critical pairs. It is important to note
that the H2 method demonstrated improved separation of amines
13 (ethylenediamine) and 14 (ethanolamine), two components that
coelute using He as carrier gas. The overall better chromatographic
performance of H2 over He in terms of speed and separation helps
to illustrate the power of H2 as carrier gas for high linear velocity
methods. Full method validation using H2 as the carrier gas while
following standard operating procedures (SOPs) and good manu-
facturing practice (GMP) requirements will be carried out either by
this or another laboratory in the future.
4. Conclusions
Process chemistry investigations in the pharmaceutical industry
often involve optimization of numerous reaction variables, includ-
ing volatile amines. Consequently, the analytical chemists have to
spend time and resources developing and validating chromato-
graphic methods for determination of residual amine content prior
to each analysis session. To better meet these needs, a simple
and fast generic method using conventional GC-FID technology
(using either He or H2 as carrier gas) capable of separating over 25
volatile amines and other basic polar compounds commonly used in
pharmaceutical chemistry workflows was developed and validated.
Validation experiments showed excellent sensitivity, precision, lin-
ear correlation, and accuracy for all of the amines. This method has
 G.C. Graffius et al. / J. Chromatogr. A 1518 (2017) 70–77 75

Fig. 2. Direct application of GC-FID method to the separation and analysis of other volatile basic compounds. a) Analysis of 1,3-dimethyl-3,4,5,6-tetrahydro-2-pyrimidinone
(DMPU: 28) in a synthetic intermediate using DMAc as diluent. b) Simultaneous monitoring of ethylenediamine (EDA: 13) and the ligand (dimethylcyclohexane-1,2-diamine:
29) levels in a copper catalyzed reaction mixture using DMAc as diluent. c) Separation of potential mutagenic impurities (PMIs 30-32).

Fig. 3. GC-FID method using hydrogen as carrier gas for the separation of volatile amines. Method conditions are equivalent to those detailed in Table 1 (Experimental
section).
76 G.C. Graffius et al. / J. Chromatogr. A 1518 (2017) 70–77
been successfully implemented for quantitation of volatile amines
in pharmaceutical drugs and synthetic intermediates. In addition,
diverse examples of the use of this method for simultaneous deter-
mination of other amine-related chemicals in reaction mixtures
were illustrated indicating that these GC-FID conditions can be
effectively used as a starting point for solving challenging sepa-
rations and analysis of volatile polar species beyond the validated
list described in this study.
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