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A Generic Method for the

Analysis of Residual Solvents in


Pharmaceuticals Using Static
Headspace-GC-FID/MS
Application Note
Pharmaceuticals

Authors Abstract
Karine Jacq, Frank David, and The determination of residual solvents in pharmaceuticals is one of the most impor-
Pat Sandra tant gas chromatography (GC) applications in quality assurance/quality control
Research Institute for Chromatography, (QA/QC) in the pharmaceutical industry. Sample introduction is normally done using
Pres. Kennedypark 26, B-8500 Kortrijk, static headspace (SHS). In routine QC, GC with flame ionization detection (FID) is pre-
Belgium ferred, while mass spectrometry (MS) can be used for screening and identification. In
this application note, a retention-time locked GC-MS/FID method is presented that
Matthew S. Klee allows the analysis of more than 50 solvents in a single run. The method was opti-
Agilent Technologies, Inc. mized to allow identification and quantification of all three International Conference
2850 Centerville Road on Harmonisation (ICH) classes of solutes at relevant concentration levels.
Wilmington, DE 19808
USA
Introduction glycol, and sulfolane. The analysis of these impurities should
The determination of residual solvents (RS), formerly called be performed using other methods; their analysis is not dis-
Organic Volatile Impurities (OVI), in pharmaceutical products cussed here.
is probably the most important application of gas chromato- The presented retention-time locked method, however, can be
graphy (GC) in pharmaceutical quality control. Recently, meth- considered as generic, since it covers most solvents and can
ods described in U.S. and European Pharmacopoeia have be used both for identification (by MS) and for quantification
been reviewed, updated, and harmonized according to (by FID and/or MS) of residual solvents across a wide con-
International Conference on Harmonisation (ICH) guideline centration range.
Q3C (R3) [1].
In pharmaceutical manufacturing, approximately 60 different Experimental
solvents are in typical use. This set of solvents covers a
rather large range of boiling points and polarities. According Sample Preparation
to the ICH guideline, these solvents are divided into three For the analysis of residual solvents in pharmaceutical prod-
classes. Class 1 includes benzene, carbon tetrachloride, ucts, the drug substance or drug product is typically dissolved
1,1-dichloroethane, 1,2-dichloroethylene, and 1,1,1-trichloro- in a low-volatility (high-boiling) solvent such as dimethyl sul-
ethane; these solvents are toxic and their use should be foxide (DMSO), dimethyl acetamide (DMAC), or 1,3-dimethyl-
avoided. Class 2 solvents are less toxic, but their use should 2-imidazolidinone (DMI). For water-soluble drug substances,
also be limited. Class 1 and Class 2 solvents are preferably dissolution in water can also be used. In this work, 100 mg
being replaced by Class 3 solvents, which have low toxic drug substance was dissolved in 2 mL DMSO or DMSO/water
potential to humans. Taking into account their relative toxici- (1:1). Recently dedicated "GC headspace" -grade solvents
ty, these solvents should be monitored in pharmaceutical were made available from Sigma-Aldrich (NV/SA Bornem,
products, including drug substances (or active pharmaceutical Belgium). DMSO, suitable for GC-HS (cat. no. 51779) was
ingredients [API]) and drug products (formulations) at various used in this work.
levels, ranging from 2 ppm (2 g/g drug substance) for ben- Solvent standard solutions were prepared in DMSO at
zene (Class 1) to 0.05 % (w/w = 5,000 ppm) for Class 3 sol- 15 g/mL concentration for Class 1 solvents and at
vents. Consequently, the analytical method(s) used to monitor 600 g/mL concentration for Class 2 and Class 3 solvents.
these residual solvents in pharmaceutical products needs From these stock solutions, aliquots of 1 to 100 L were
also to cover this range. added in a standard 20 mL HS vial filled with 2 mL DMSO/
For the analysis of residual solvents, gas chromatography in water (1:1, v/v). The concentrations of standards are always
combination with flame ionization detection (GC-FID) is nor- expressed in microgram per gram of drug product or drug sub-
mally used. Sample preparation and introduction is done by stance. So, the concentration of these calibration solutions
static headspace [2]. In this way, the (mostly) volatile sol- ranges from 0.15 to 15 ppm (g/g drug) for Class 1 and from 6
vents are introduced selectively and the analytical system to 600 ppm for Class 2 and Class 3 solvents, if 100 mg product
(inlet, column, and detector) is not contaminated by the is weighed in the HS vial. (The actual concentration of stan-
(mostly) nonvolatile drug substance or drug product. For the dards in the vial, expressed in g per mL solvent, is 20 times
separation, a thick film, medium polar column (for example, lower.)
G43) is selected. Quantification is done versus an external
standard. Instrumental Conditions
Excellent quantitative data, including low limits of detection The samples were analyzed using the SHS-GC-FID/MS con-
(LODs) high repeatability and excellent linearity were figuration presented in Figure 1. Static headspace was per-
obtained using the Agilent G1888 static headspace sampler in formed using a G1888 HS autosampler. The transfer line of
combination with an Agilent 7890 GC [3]. However, since no the headspace sampler is coupled to a standard split/split-
column can guarantee a unique retention time for a given sol- less inlet. Separation was done on a DB-1301 column. The
vent, confirmation analysis by GC-FID on a capillary column column effluent is split using a purged splitter Capillary Flow
coated with a different stationary phase (for example, G16) is Technology device to FID and MS (5975 MSD). The vial pres-
performed. More recently, GC-MS has been successfully used sure is regulated by an AUX EPC channel. The purge at the
for confirmation/identification purposes [4,5]. splitter is also regulated by the AUX EPC (second channel). A
63 cm 0.1 mm id deactivated fused silica capillary was used
In this application note, a system configuration and operation
to connect the splitter to the MSD; a 40 cm 0.1 mm id capil-
conditions are described that allow the analysis of 56 sol-
lary was used to connect the splitter to the FID. Flows in both
vents in a single run. Some solvents listed in the ICH guide-
capillaries are approximately 1.4 mL/min and the retention
line are not volatile (enough) and not amenable to SHS-GC
time is also similar (small offset between FID and MS
analysis. Examples are formic acid, acetic acid, dimethyl sul-
retention times). The analytical and selected ion monitoring
foxide (often used as a method solvent), formamide, ethylene
(SIM) parameters are summarized in Table 1 and Table 2,
respectively.

2
Figure 1. SHS-GC-FID/MS system configuration.

Table 1. Analytical Parameters Table 2. SIM Parameters


Group Start time Ions
SHS (G1888 Agilent HS autosampler)
1 0.00 31, 30
Loop size: 1 mL
2 3.50 31, 43, 45, 59
Vial pressure: 14 psig (96.5 kPa)
3 4.60 31, 43, 45, 58, 59, 61, 74, 96
Headspace oven: 80 C
4 5.55 41, 43, 84
Loop temperature: 120 C
5 6.50 57, 61, 73, 96
Transfer line temperature: 120 C
6 7.25 41, 57, 86
Equilibration time: 10 min, high shake
7 7.90 31, 42, 59
Pressurization: 0.15 min
8 8.70 30, 43, 61, 72, 96
Vent (loop fill): 0.5 min
9 9.65 42, 45, 47, 59, 72, 83
Equilibration time: 0.1 min
10 10.30 47, 56, 61, 84, 97, 117
Inject: 0.5 min
11 11.10 43, 45, 49, 61, 62, 74, 76, 78, 90
12 12.02 43, 71
GC (7890A)
13 12.70 31, 56, 95, 130
Inlet: Split/splitless, split 1/10, headspace liner
14 13.40 55, 83
(P/N 5183-4709)
15 13.90 43, 58, 61, 88
Inlet temperature: 250 C
16 14.50 31, 59, 71
Split ratio: 1:10
17 15.60 41, 43, 52, 55, 58, 73, 79, 91
Carrier gas: Helium
18 16.75 42, 55, 70
Inlet pressure: 160 kPa *
19 17.30 43, 58, 100
AUX pressure (at splitter): 60 kPa
20 17.66 43, 56, 73
Column: DB-1301 20 m 0.18 mm 2 m (J & W)
21 18.20 77, 91, 106, 112
Oven: 40 C (5 min) 5 C/min 80 C 10 C/min
22 20.50 44, 78, 87, 105, 108, 120
200 C (2 min)
23 21.30 44, 98, 99
FID: 300 C, 40 mL/min H2, 400 mL/min air
24 26.00 91, 104, 132
MS (5975 C Inert XL MSD)
Transfer line: 300 C
Mode: Simultaneous scan/SIM In this work, a narrow-bore, thick-film column with equivalent
SCAN range: 29 250 m/z
SIM: See Table 2 stationary phase, DB-1301 (equivalent to G43), was selected.
* Retention time locking was applied. The column head pressure was adjusted to elute The lower phase ratio ( = 22) results in better resolution for
toluene at 16.160 min. the first eluting (but frequently detected) solvents, such as
methanol, ethanol, diethylether, acetone, isopropanol, and
Results and Discussion acetonitrile.
Column Selection It is not possible to find a stationary phase that is able to sep-
Many different columns can be used for the analysis of resid- arate all 60 solvents in approximately 30 minutes. In most
ual solvents after static headspace extraction. Typically, a col- practical cases, however, only three to five solvents need to
umn coated with a thick film of a medium polar stationary be determined. The described method allows the analysis of
phase is selected. A classical column for residual solvent 56 solvents and eventually the same column can also be used
analysis is a 30 m 0.53 mm id coated with 1 to 3 m for the analysis of ethylene oxide. For this reason, column
DB-624 ( = 44). On this column, good separation is obtained selection and separation conditions can be considered as
in about 30 minutes of analysis time [3]. generic.

3
The separation obtained by SHS-GC-FID/MS for a solvent test Column 4 in Table 3 indicates possible coelution of target
mixture in DMSO/water is shown in Figure 2. The concentra- compounds. Most solutes are chromatographically resolved
tion of the solutes was 16 ppm for the five Class 1 solvents and can thus be quantified by either FID or MS when present
and 560 ppm for the Class 2 and Class 3 solvents. With one in the same sample. In some cases, coelution is observed
injection, three data files are obtained. The upper trace shows (labeled with C). In these cases, quantification is still possible
the total ion chromatogram (TIC) obtained by MSD in scan by MS after ion extraction or by using selected ion monitoring
mode, the middle trace shows the corresponding SIM data, (SIM) mode.
and the lower chromatogram is the FID trace. Toluene elutes
In columns 5 through 8, the limit of detection (LOD) for the
at 16.16 minutes in all three chromatograms and the offset
three detection modes, determined from the lowest calibra-
in retention time between FID and MS is less than
tion level at S/N = 3, are compared to the ICH limits. In most
0.03 minute or 2 seconds for all solutes.
cases (43 of the 56 analytes), the LODs were well below the
A detailed view of the separation is shown in Figure 3, show- ICH limits for all three detection modes. Those instances
ing three elution windows of the FID trace. All solutes can be wherein the LOD was above the ICH limit are highlighted in
detected and are labeled on the chromatograms except for Table 3 in boldface. For the Class 1 solvents, it is clear that
tetrachloromethane (trace at 10.93 minutes), 2-methoxy MS in SIM mode is preferred. The benefit of using SIM is
ethanol (co-elutes with benzene, 1,2-dimethoxyethane, and illustrated in Figure 4a for some Class 1 solvents. At 10.6 min-
1,2-dichloroethane), 2-ethoxyethanol (trace at 14.96 minutes), utes, 1,1,1-trichloroethane coelutes with cyclohexane, as
DMAC (co-elutes with cumene), and 1-methyl-2-pyrrolidone seen in the FID trace. However, both solutes can be accurate-
(trace at 25.19 minutes). ly measured with extracted ion chromatograms using m/e =
97 for 1,1,1-trichloroethane and m/e = 56 for cyclohexane.
The same can be observed in Figure 4b for the overlapped
Validation
peaks of benzene, 1,2-dimethoxyethane and 1,2-dichloro-
In a previous application note [3] it was demonstrated that
ethane at 11.5 minutes in the FID trace and the extracted ion
the quantitative data obtained using a G1888 SHS 7890A GC
peaks of m/e = 78 for benzene, m/e 45 for 1,2-dimethoxy-
combination resulted in equal or better quantitative results
ethane, and m/e = 62 for 1,2-dichloroethane (2-methoxy-
than a G1888 6890 GC combination. Electronic pressure
ethanol is not detected at this level).
control of carrier gas and vial pressure results in excellent
repeatability and linearity. An optional pneumatic control Using MS, all compounds, except 2-methoxyethanol can be
module can be used to control the vent pressure by back detected at LOD < ICH limit. Some of the other more polar
pressure regulation (BPR), resulting in increased sensitivity solutes (2-ethoxyethanol, DMF, DMAC, and 1-methyl pyrroli-
(by a factor of 2 to 4) and smaller relative standard deviations done) also give a low response in FID, whereas MS detection
(RSDs) (reduced by a factor of 2). In this study, good quantita- limits are satisfactory. As reported before [3], back-pressure
tive data were obtained even though the BPR approach was regulation of the vent pressure is expected to help decrease
not used. Using BPR would provide higher performance for the LOD by a factor of two, so this an important consideration
those seeking it. if you are using FID for quantitation. In addition, increasing
sample size and/or injection volume (headspace sample loop
Limits of detection, linearity, and repeatability were evaluated
size) would also help if you are doing quantitation with FID.
with the different modes of detection. Repeatability was
checked at 3 ppm level (g/g, equivalent to 0.15 g/mL in The repeatability of the SHS-GC-FID/MS method is excellent.
vial) for Class 1 solvents and at 100 ppm for the others RSDs were mostly below 5 percent, both for GC-FID and GC-
(n = 6). Five-point calibration curves (plus a blank) were mea- MS (SIM and SCAN). The average RSDs were 4.4 percent for
sured between 0.15 and 15 ppm for Class 1 solvents, and SCAN, 3.8 percent for SIM, and 3.0 percent for FID. Again, the
between 6 and 600 ppm for most others. For some solutes values were higher for some more polar solutes. Finally, good
giving low response, linearity was tested in the 500 to linearity was obtained for most compounds. Except for
5,000 ppm range. For coeluting compounds, the experiments 2-methoxyethanol, 2-ethoxyethanol, DMF, DMAC, and
were performed with single compound solutions in order to 1-methyl-2-pyrrolidone (compounds with higher LODs), linear-
allow peak integration with FID. The results are summarized ity was excellent with the three types of detection (R > 0.99).
in Table 3.

4
1300000
1200000
1100000
1000000
900000
Abundance

800000
700000
600000
500000
400000
300000
200000
100000

4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00
Time

800000

700000

600000
Abundance

500000

400000

300000

200000

100000

4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00
Time

190000
180000
170000
160000
150000
140000
Abundance

130000
120000
110000
100000
90000
80000
70000
60000

4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00
Time

Figure 2. A 56-component solvent mixture analyzed by SHS-GC-FID/MS.

5
Response
Response Response

60000
80000
100000
120000
140000
160000
180000
200000
220000
240000
260000
280000
60000
80000
100000
120000
140000
160000
180000
200000
220000
240000
260000
280000
300000
320000
60000
80000
100000
120000
140000
160000
180000
200000
220000
240000
260000
280000
300000

19.00
Chlorobenzene

11.00
M-xylene Methanol
P-xylene Isobutyl alcohol

3.00
Benzene +1,2-dimethoxyethane + 1,2-dichloroethane
Isopropyl acetate

20.00
12.00
O-xylene Heptane Pentane

4.00
DMSO Ethanol
1-butanol Ethyl ether

21.00
Anisole Cumene

13.00
Trichloroethene 1,1-dichloroethene

5.00
Acetone
Methylcyclohexane
2-propanol + ethyl formate

22.00
Acetonitrile

14.00
1,4-dioxane Methyl acetate
Propyl acetate
6.00

Dichloromethane

Z-1,2-dichloroethene +

Time
23.00
Time

Time
15.00
tert-butyl methyl ether
7.00

6
Figure 3. Detailed FID chromatogram (16 ppm Class 1 and 560 ppm Class 2 and Class 3 solvents).
Hexane
4-methyl-2-pentanone

24.00
Isoamyl alcohol + pyridine

16.00
Toluene
8.00

1-propanol
(Octane)
Isobutyl acetate

25.00
1-pentanol

17.00
9.00

Nitromethane
E-1,2-dichloroethene
2-butanone
2-hexanone Ethyl acetate
Butyl acetate 2-butanol

26.00
THF

18.00
-
10.00

Chloroform
1,1,1-trichloroethane
Tetralin Cyclohexane
Solvent Influence tial with this method, some SHS-GC-FID/MS analyses were
The influence of the solvent used to dissolve the drug sub- performed on solutions of 100 mg drug substance (prometh-
stance or drug product was also evaluated. In general, equally azine) in the solvent (2 mL DMSO/H2O). A blank (not spiked)
good quantitative data in terms of linearity and repeatability and two spiking levels (for example, 3 and 8 ppm for Class 1
are obtained when using water, DMSO, DMAC, DMI, or mix- solvents, and 110 and 280 ppm for Class 2 and 3 solvents)
tures thereof. The LOD (and slope of the calibration curve), were analyzed in triplicate.
however, depends on the solvent used. In general, for apolar Both repeatability (RSDs) and linearity (R) were comparable
solvents in the most polar matrix (water), the lowest LODs with the data obtained without active pharmaceutical ingredi-
will be obtained. For the (apolar) Class 1 solvents, the LOD ent. In general, the responses for the spiked samples were
can be up to 10 lower (more sensitive) if static headspace is between 80 and 105 percent of the response obtained with-
performed in DMSO/water versus pure DMSO. out active pharmaceutical compound (API). This recovery is
well within the limits generally accepted for trace impurity
Matrix Influence analysis.
The presence of a relatively high concentration of drug sub-
stance in the sample solution (100 mg/2 mL) might influence
the absolute response of the solutes. To evaluate that poten-

Table 3. Validation Results for 56 Solutes

RSD r2
Class 1: _ 3 ppm Class 1: 0.1515 ppm
ICH RT Co- LOD (ppm, rel. 100 mg API) Class 2 and 3: _ 100 ppm Class 2 and 3: 6600 ppm
Compound class (min) elution1 ICH SCAN SIM FID SCAN SIM FID SCAN SIM FID
Methanol 2 2.77 No 3000 14.2 0.29 132 8.4 3.5 6.0 0.997 0.996 0.999
n-pentane 3 3.88 No 5000 3.3 0.09 6.2 3.7 2.8 1.1 0.994 0.994 0.998
Ethanol 3 4.11 No 5000 18.2 0.80 132 3.6 3.0 8.3 0.995 0.995 0.996
Ethyl ether 3 4.35 No 5000 1.4 0.06 6.0 3.5 3.5 1.3 0.996 0.995 1.000
1,1-dichloroethene 1 4.86 No 8 1.9 0.02 7.4 15.9 4.0 7.3 0.992 0.996 0.990
Acetone 3 5.01 No 5000 4.7 0.13 20.1 4.9 4.0 2.4 0.994 0.995 1.000
2-propanol 3 5.37 C 5000 17.0 0.12 230 4.0 3.3 2.2 0.997 0.997 1.000
Ethyl formate 3 5.38 C 5000 14.1 0.33 27 3.1 3.3 0.8 0.998 0.997 1.000
Acetonitrile 2 5.71 No 410 11.1 0.46 73 4.1 3.5 4.2 0.995 0.996 0.999
Methyl acetate 3 5.84 No 5000 2.8 0.16 40 2.5 3.8 2.8 0.997 0.997 1.000
Dichloromethane 2 6.06 No 600 1.7 0.06 24 4.6 4.0 2.4 0.997 0.996 0.999
Z-1,2-dichloroethene 2 6.74 C 1870 0.9 0.01 9.0 4.1 4.0 1.3 0.996 0.996 1.000
t-butyl methyl ether 3 6.73 C 5000 2.4 0.02 7.5 3.2 3.3 1.4 0.996 0.996 1.000
n-hexane 2 7.42 No 290 1.2 0.03 4.4 2.5 3.2 0.9 0.994 0.994 0.999
1-propanol 3 8.11 No 5000 23 0.45 126 6.6 3.6 9.1 0.995 0.995 0.998
Nitromethane 2 9.13 No 50 104 1.0 214 7.8 1.5 11.3 0.995 0.997 0.992
E -1,2-dichloroethene 2 9.24 Partial 1870 3.9 0.09 16 4.0 4.0 2.6 0.997 0.996 1.000
2-butanone 3 9.33 Partial 5000 11.1 0.32 40 3.3 3.4 3.1 0.997 0.997 0.999
Ethyl acetate 3 9.50 No 5000 12.4 0.5 19.1 3.4 4.3 3.8 0.997 0.997 0.999
2-butanol 3 9.82 No 5000 18.4 1.6 64 7.0 3.9 2.0 0.993 0.995 0.999

7
Table 3. Validation Results for 56 Solutes (continued)
RSD r2
Class 1: _ 3 ppm Class 1: 0.1515 ppm
ICH RT Co- LOD (ppm, rel. 100 mg API) Class 2 and 3: _ 100 ppm Class 2 and 3: 6600 ppm
Compound class (min) elution1 ICH SCAN SIM FID SCAN SIM FID SCAN SIM FID
THF 3 9.96 No 5000 3.5 0.22 20 3.3 3.6 3.0 0.998 0.997 1.000
chloroform 2 10.07 Partial 60 1.6 0.04 34 3.0 3.7 3.8 0.998 0.997 0.995
1,1,1-trichloroethane 1 10.51 C 1500 2.1 0.03 18.8 4.7 3.3 6.0 0.997 0.997 0.993
Cyclohexane 2 10.62 C 3880 1.9 0.03 2.90 3.3 3.5 1.0 0.995 0.996 1.000
Tetrachloromethane 1 10.93 No 4 0.71 0.04 45 5.7 3.3 6.0 0.998 0.997 0.994
Isobutyl alcohol 3 11.32 No 5000 18 1.7 64 4.9 3.2 7.3 0.998 0.996 0.998
Benzene 1 11.48 C 2 0.70 0.02 4.8 6.2 3.5 2.4 0.994 0.997 1.000
2-methoxyethanol 2 11.48 C 50 835 248 2650 20 9.5 16 1 point 0.988 0.868
1,2-dimethoxyethane 2 11.46 C 100 13.2 < 3.5 65 4.5 3.2 5.5 0.995 0.995 0.999
1,2-dichloroethane 1 11.54 C 5 8.2 0.70 30 2.3 4.0 1.6 0.999 0.997 0.999
Isopropyl acetate 3 11.74 No 5000 1.4 0.07 13.5 3.2 4.3 1.3 0.996 0.997 1.000
n-heptane 3 12.18 No 5000 1.3 0.06 3.5 3.3 3.6 1.8 0.995 0.995 0.999
1-butanol 3 13.05 C 5000 21.1 1.1 178 6.9 5.1 1.1 0.994 0.996 1.000
Trichloroethene 2 13.15 C 80 1.2 0.03 16 3.3 4.0 1.6 0.998 0.996 0.999
Methylcyclohexane 2 13.61 No 1180 0.6 0.02 3.8 3.7 3.8 1.1 0.996 0.996 1.000
1,4-dioxane 2 14.08 No 380 30 0.41 141 3.5 3.5 4.4 0.992 0.997 0.994
Propyl acetate 3 14.22 No 5000 1.3 0.20 16.2 3.6 3.9 1.8 0.997 0.997 1.000
2-ethoxyethanol 2 14.96 No 160 576 115 1000 5.0 14.1 3.0 0.977 0.956 0.989
4-methyl-2-pentanone 3 15.80 No 5000 5.8 0.16 11.9 3.0 3.3 0.9 0.996 0.997 1.000
Isoamyl alcohol 3 15.94 No 5000 73 1.6 76 5.6 4.3 3.6 0.995 0.996 0.999
Pyridine 2 15.95 No 200 46 3.1 63 3.7 3.3 4.3 0.993 0.995 0.999
Toluene 2 16.16 No 890 0.4 0.02 3.6 4.2 3.9 1.1 0.997 0.997 1.000
Isobutyl acetate 3 16.50 No 5000 5.4 0.44 10.1 3.4 4.1 0.9 0.997 0.997 0.999
1-pentanol 3 17.01 No 5000 19.3 0.94 84 4.1 3.9 4.5 0.995 0.995 0.998
2-hexanone 2 17.55 No 50 1.8 0.10 15.0 2.1 3.4 1.0 0.997 0.997 1.000
n-butyl acetate 3 17.80 No 5000 0.9 0.08 12.0 4.2 3.6 0.8 0.997 0.997 0.999
DMF 2 18.59 No 880 962 192 2885 12.2 10.9 17.8 0.970 0.977 0.937
Chlorobenzene 2 19.03 No 360 0.7 0.03 8.5 3.5 3.9 1.3 0.996 0.996 0.998
Xylene 2 19.23 No 2170 2.2 0.15 29 4.6 3.8 2.2 0.996 0.997 0.999
Xylene 2 19.45 No 2170 0.5 0.03 5.9 3.8 3.8 1.2 0.997 0.997 0.999
Xylene 2 20.22 No 2170 2.4 0.16 25 3.7 3.8 2.7 0.997 0.996 0.999
DMAC 2 20.92 C 1090 154 18.8 3200 20 17.3 19.5 0.914 0.921 0.935
Cumene 3 20.92 C 5000 0.2 0.03 <1 4.0 3.7 1.8 0.997 0.997 0.999
Anisole 3 21.07 No 5000 2.7 0.13 <1 2.7 3.3 1.5 0.995 0.997 0.998
1-methyl-2-pyrrolidone 2 25.19 No 4840 6150 117 6150 ND 12.1 ND 1 point 0.972 1 point
Tetralin 2 26.72 No 100 1.7 0.12 12.5 4.2 3.5 1.3 0.995 0.992 0.997
ND = Not detected
1 C = Coelution, but resolved by MS; Partial = partial overlap
2 Calculated at 2,500 ppm
3 Calculated in the 500 to 5,000 ppm range

8
Application
The method was applied to a number of available drug sub-
stances. In samples of penicillin V and levamisol
(tetramisole), traces of residual solvent were detected. As an
illustration, the FID chromatograms for the two samples (for
each analysis, 100 mg sample was dissolved in 2 mL
DMSO/water, 1:1) are shown in Figures 5 and 6. In the peni-
cillin sample (Figure 5), a trace amount of n-butyl acetate was
detected. The concentration, measured by external calibra-
tion, was 52 ppm well below the ICH limit (5,000 ppm). In
the levamisol sample (Figure 6), a trace amount of toluene
was detected. The concentration was 66 ppm, well below the
890 ppm ICH limit. It is interesting to note that a trace amount
of dimethyl sulfide was detected in both chromatograms.
Figure 4a. Comparison of overlapped peaks in FID and individual SIM ion chromatograms
DMS is an impurity in the DMSO solvent used to dissolve the for 1,1,1-trichloroethane (m/e = 97) and cyclohexane (m/e = 56).
samples. In addition, an "unknown" was detected in lev-
amisol. Through MS spectral library searching, the peak was
identified as 2-chloropropane. This solvent is not included in
the ICH solvent list. However, the ability to unequivocally
identify this unknown in the sample clearly demonstrates the
advantage of using parallel MS detection.

Solvent Backflush
Capillary column backflushing is starting to be routinely
implemented as a means of improving analysis cycle times
and data quality. The simplicity and benefits of implementing
backflush for residual solvents analysis were evaluated. To
reduce the analysis time, late-eluting solvents (DMSO and
DMAC) can be backflushed by increasing the pressure at the
column outlet (AUX pressure controlling the purged splitter) Figure 4b. Comparison of overlapped peaks in FID and individual SIM ion chromatograms
for benzene (m/e = 78), dimethoxyethane (m/e = 45), and 1,2-dichloroethane
and lowering the inlet pressure. This is demonstrated in (m/e = 62).
Figures 7A through 7C. In a normal run of a solvent mixture,
DMSO elutes at 20.8 minutes, and the standard run continues Conclusions
to 200 C to ensure that there is no carry-over of sample com- More than 50 residual solvents can be determined in pharma-
ponents into the next run. If no analytes of interest elute after ceutical products in a single run using a static headspace
DMSO, backflushing can be used to reduce cycle times. GC-FID/MS configuration. Quantification can be performed
Figure 7B shows the same analysis as in Figure 7A, only with routinely by FID for most target compounds, while MS is
a backflush initiated at 20 minutes (just before elution of especially suited for trace level determination of Class 1 sol-
DMSO). The outlet pressure was increased from 60 to vents and for identification of unknowns. Mass spectrometry
200 kPa and the GC was held at 150 C for 10 minutes. No also excels for determination of coeluting peaks through use
peaks are observed after the backflush was initiated. Next, a of extracted ion or SIM ion chromatograms, thereby eliminat-
blank run (without backflush, original method) was performed ing the need for additional analyses on dissimilar columns.
(Figure 7C). It clearly shows that DMSO solvent was totally The quantitative data, including repeatability, linearity, and
removed. The ability to implement backflush was very simple LOD are excellent, meeting or exceeding ICH guidelines.
because of the purged-split configuration of FID and MSD.
Thermal stress on the column was certainly reduced and the
cooldown time from 150 C instead of the 200 C original end-
ing temperature was faster. Further improvement in cycle
time can be achieved by reducing backflush time to the mini-
mum time required to fully backflush DMSO.

9
105000

100000

95000

90000

85000
Response

80000

75000
n-butyl
70000 acetate
65000

60000
dimethyl
55000
sulfide
50000
2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00
Time

Figure 5. Analysis of commercial penicillin sample. n-butyl acetate was determined to be present at 52 ppm well below the ICH limit of 5,000 ppm.

105000

100000 toluene
95000

90000

85000
Response

80000

75000 2-chloropropane

70000

65000
dimethyl
60000 sulfide
55000

50000
2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00
Time

Figure 6. Analysis of levamisol sample. Toluene was determined to be present at 66 ppm well below the ICH limit of 890 ppm.

10
360000

340000

320000

300000

280000

260000

240000
Response

220000

200000

180000

160000

140000

120000

100000

80000

60000

0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00
Time

Figure 7A. Standards analysis without backflush.

260000

240000

220000

200000

180000
Response

160000

140000

120000

100000

80000

60000

0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00
Time

Figure 7B. Standards analysis with backflush at 20 minutes (just before DMSO elution).

11
280000

260000

240000

220000

200000

180000
Response

160000

140000

120000

100000

80000

60000

0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00
Time

Figure 7C. Blank run after the backflush run demonstrates that the backflush completely removed DMSO.

References
1. ICH Harmonised Tripartite Guideline, Q3C(R3),
http://www.ich.org/LOB/media/MEDIA423.pdf
2. R. L. Firor, The Determination of Residual Solvents in Pharmaceuticals Using
the Agilent G1888 Network Headspace Sampler, Agilent Technologies publica-
tion 5989-1263EN, 2004.
3. A. E. Gudat, R. L. Firor, and U. Bober, Better Precision, Sensitivity, and Higher
Sample Throughput for the Analysis of Residual Solvents in Pharmaceuticals
Using the Agilent 7890A GC System with G1888 Headspace Sampler in Drug
Quality Control, Agilent Technologies publication 5989-6023EN, 2007.
www.agilent.com/chem
4. R. L. Firor and A. E. Gudat, The Determination of Residual Solvents in
Pharmaceuticals Using the Agilent G1888 HS/6890GC/5975 inert MSD Agilent shall not be liable for errors contained herein or
System, Agilent Technologies publication 5989-3196EN, 2005. for incidental or consequential damages in connection
with the furnishing, performance, or use of this material.
5. A. E. Gudat and R. L. Firor, The Determination of Extractables and Leachables
Information, descriptions, and specifications in this
in Pharmaceutical Packaging Materials Using Headspace/GC/MS, Agilent publication are subject to change without notice.
Technologies publication 5989-5494EN, 2006.
Agilent Technologies, Inc., 2008
Printed in the USA
For More Information September 18, 2008
5989-9726EN
For more information on our products and services, visit our Web site at
www.agilent.com/chem.