Optimized PAH Analysis Using the

Agilent Self-Cleaning Ion Source and

the Enhanced PAH Analyzer

Application Note
PAH, environmental, gas chromatography, mass spectrometry,
semivolatiles
Author Abstract
Mike Szelewski and The Agilent Enhanced PAH Analyzer was used for the analysis of polycyclic
Bruce D. Quimby aromatic hydrocarbons (PAHs) with the Self-Cleaning Ion Source in Continuous
Agilent Technologies, Inc. Cleaning Mode. Both the Agilent 5977A Series GC/MSD System and Agilent 7000B
2850 Centerville Road Triple Quadrupole GC/MS versions of the analyzer were used. All instrument para-
Wilmington, DE 19808 meters including inlet, column, and MS were investigated and optimized. Linearity
and ISTD reproducibility, across a calibration range of 1–1,000 pg, were improved
while maintaining sensitivity.
Introduction Experimental
PAHs are monitored as trace contaminants in food such as The recommended instrument operating parameters are listed
shellfish and seafood, and in the environment including air, in Table 1. These conditions have been optimized for PAH
water, and soil. PAHs originate from three sources: analysis.

• Petrogenic – Derived from petroleum inputs associated Splitless injections are used due to the sensitivity require-
with fossil fuels ments. An inlet temperature of 310–330 °C is necessary to
vaporize the late eluters. A pressure pulse of 50 psi is used to
• Pyrogenic – Derived from combustion sources
rapidly transfer PAHs onto the column. The pressure pulse is
• Biogenic – Formed from natural biological processes held for the full splitless time, 0.5–1.0 minutes. Cold splitless
They are toxic to aquatic life and are suspected human or solvent vent modes may show worse performance for
carcinogens. heavy PAHs. Once they deposit in the inlet they are difficult to
vaporize.
PAHs are usually analyzed by HPLC/UV, GC/FID, GC/MS or
GC/MS/MS. The straight bore 4.0-mm liner with glass wool is a must. The
wool transfers heat to the PAHs and blocks the line of sight to
This application note focuses on GC/MS in SIM mode and the inlet base. If the PAHs condense on the inlet base, they
GC/MS/MS in MRM mode. A common calibration range is are difficult to vaporize and sweep back onto the column.
from 1–1,000 pg with an acceptable linearity of R2 > 0.99.
Internal Standard (ISTD) area reproducibility is typically speci- The oven program allows separation of the isomers in a
fied at ± 20% with calibration standards and ± 30%–50% with reasonable run time.
samples. The 30 m × 0.25 mm, 0.25 µm DB EUPAH column is a new
A number of issues arise with the analysis due to the proper- dimension, and was developed during this study. This column
ties of PAHs. They span wide molecular weight and BP allows 1–2 µL injections while maintaining peak shapes com-
ranges. Although not considered active or subject to degrada- pared to the previous analyzer column, 20 m × 0.18 mm,
tion, they are sticky and readily adhere to surfaces. PAHs are 0.14 µm. The first column is connected from the inlet to the
subject to desublimation (deposition) and are difficult to purged ultimate union (PUU). The column is run in constant
vaporize. High temperatures are important along with mini- flow mode. A second column is configured from the PUU to
mizing surface contact. Peak tailing is often seen on the later the MS. This column is configured as a segmented column
eluters, which means manual integration. There have been and also runs in constant flow mode, so the entire system is
cases where the ISTD response is inconsistent across the in constant flow. The PUU is used for backflushing the first
calibration range. This leads to linearity that does not meet column at the end of every run.
expectations. The system was RTLocked to 1-methylphenanthrene at
The Self-Cleaning Ion Source (SCIS) includes a specially 12.500 minutes. After clipping the column, a rerun and analy-
engineered aux module that introduces small amounts of sis of the locking standard is all that is needed to restore
hydrogen directly into the source. A low hydrogen flow, shifted peak times. Times for MRM or SIM groups, quantita-
0.075–0.2 mL/min, is introduced continuously during data tion database entries, and integration events do not have to
acquisition. The source is kept clean from run to run. Linearity be changed.
is improved and ISTD response is consistent, while The standard 3-mm drawout (or extractor) lens is replaced
maintaining sensitivity. with a 9-mm lens when the analyzer is built. This wider lens
The SCIS is included with the Enhanced PAH Analyzer used provides better linearity and less surface area for deposition,
for this application note. while maintaining sufficient sensitivity across the calibration
range of 1–1,000 pg.

2
Table 1. Gas Chromatograph and Mass Spectrometer Conditions Front injector Agilent 7693A Automatic Liquid Sampler
Injection volume 2.0 µL
GC conditions
Syringe size 10 µL
GC Agilent 7890A CG System or Agilent 7890B GC
System Plunger speed Fast
Inlet Multimode Wash volume 3 µL

Mode Pulsed splitless Sample washes 1

Sample pumps 2
Heater 320 °C
PreInj Solv A washes 0
Pressure 16.857 psi
PreInj Solv B washes 1
Total flow 54.106 mL/min
PostInj Solv A washes 3
Septum purge flow 3 mL/min
PostInj Solv B washes 2
Septum purge flow mode Switched
Viscosity delay 1 second
Injection pulse pressure 50 psi until 0.9 minutes
PreInjection dwell 0 minutes
Purge flow to split vent 50 mL/min at 0.9 minutes
PostInjection dwell 0 minutes
Gas saver Off Thermal Aux 1 (MSD transfer line)
Inlet liner Agilent 4-mm, single taper, with glass wool, Heater On 320 °C
p/n 5190-2293
Aux EPC 1 He Hydrogen is connected here for SCIS
Oven 120 V Pressure program On
Oven ramp Ramp °C/min Next °C Hold (min) Pressure program 50 psi
Initial 80 1.00 Run time 29 minutes
Ramp 1 25 200 0.00
Aux EPC 2 He Helium is connected here – used when Aux 1 is
Ramp 2 8 335 6.325
off
Total run time 29 minutes
Pressure program Off
Equilibration time 0.5 minutes
Pressure program 30 psi
Oven max temperature 340 °C
Run time 29 minutes
Columns
Aux EPC 3 He This is the bleeder for either Aux 1 or 2
Column 1 Agilent J&W DB-EUPAH UI 122-9632
Pressure program On
Dimensions 30 m × 250 µm, 0.25 µm
Pressure program 3 psi
In Front MM Inlet He
Run time 29 minutes
Out Aux EPC 4
APC
Pressure 16.857 psi
Aux EPC 4 He Supplies Column 2
Flow 1.1063 mL/min
Pressure program 4.124 psi initial
Mode Constant flow
Run time 29 minutes
Column 2 Agilent J&W 160-7625-5 Aux EPC 5 He Off
Silcotek deactivated tubing
Aux EPC 6 He Off
Overall dimensions 1.36 m × 150 µm, 0 µm
Acquisition mode SIM (MSD) or MRM (MS/MS)
Main segment 1.19 m, Heated by oven
Solvent delay 5.00 minutes
Segment 2 0.17 m, Heated by thermal Aux 1
Tune file atune.u or etune.u for MSD
In Aux EPC 4 He Atunes.eiex.tune.xml for MS/MS
Out MSD
EM Setting mode gain 1.0
(Initial) 80 °C
Number of SIM groups 11
Pressure 4.1242 psi
Flow 1.5 mL/min MSZones
Mode Constant flow MS source 320 °C maximum 350 °C
RTL System retention time locked to MS quad 180 °C maximum 200 °C
1-methylphenanthrene at 12.500 minutes

3
The ion source temperature is run in the range of 320–350 °C. Results and Discussion
Running a lower temperature results in peak tailing. The
quads should be run between 180–200 °C. The performance differences of stock systems compared to
their respective optimized analyzers are similar. Examples
Standards were diluted from Accustandard and UltraScientific using either GC/MS or GC/MS/MS based
stock solutions. Dilutions were made in toluene (preferred) or configurations are discussed.
isooctane. ISTD concentrations were constant at 500 ppb.
The standards used in this study (28–34 PAHs) were more A sensitivity comparison for the MSD is shown in Figure 1.
extensive than those provided in the analyzer checkout mix The SIM target ion for the latest eluting PAH studied is shown
(27 PAHs). with and without the SCIS in continuous clean mode (CCM).
The height differences are due to tuning. There is a small
increase in noise when running hydrogen, therefore the
signal-to-noise (S/N) is slightly worse. Each set of runs was
separated by six runs not shown. The displayed level is 500 fg,
two times lower than the lowest calibration level. This
demonstrates that sufficient signal is still present for the
typical 1–1,000 pg calibration

110

100

90

80
Abundance

75 µL/min H2 added
70

60

50 No H2 added

40

30

24.5 25.0
Time

Figure 1. S/N comparison using 500 fg Dibenzo(a,l)pyrene, 3 runs each with and without hydrogen added.

4
ISTD response consistency is shown in Figure 2. Response consistency for ISTDs run by GC/MS/MS is shown
A seven-level calibration was run of a 30 PAH mix with five in Figure 3. The calibration levels and mix are the same as
ISTDs. The ISTD responses are plotted normalized to the those used for MSD. Here, the No Hydrogen runs were made
response from the highest calibration level. With hydrogen with the standard 3-mm extractor lens. The CCM runs were
added to the source, the ISTD responses are within 5%. In a made with a 9-mm extractor lens and 200 µL/min of
stock system, the ISTD responses drop with decreasing stan- hydrogen. The CCM runs were repeated 3 and 12 days later.
dard concentration. The performance is independent of the The ISTD consistency has improved from a ~ 35% loss to
order in which the standards are run. This drop would make < 8% difference.
accurate quantitation difficult and dependent on the amount
of other analytes in the sample. A 9-mm drawout lens was
used for all data shown in Figure 2.

1.150
Naphthalene-d8
Acenaphthene-d10
1.100
Phenanthrene d10
H 2 added continuous H 2 not added
Chrysene-d12
1.050
Normalized response

Perylene-d12
1.000

0.950

0.900

0.850

0.800

0.750
0
0
0
50
10
1

0
0
0
50
10
1

0
0
0
50
10
1

0
0
0
50
10
1

0
0
0
50
10
1

0
0
0
50
10
1
00
50
10

00
50
10

00
50
10

00
50
10

00
50
10

00
50
10
1,

1,

1,

1,

1,

1,

pg injected
Figure 2. PAH ISTD response from different calibration levels, normalized. 9-mm drawout lens, cal range 0.5–500 pg to MSD, SIM.

1.4 Naphthalene-d8
Acenaphthene-d10
1.3 Phenanthrene d10
Chrysene-d12
Perylene-d12
1.2
No H 2 CCM Day 1 CCM Day 3 CCM Day 12
1.1
Normalized response

1.0

0.9

0.8

0.7

0.6

0.5

0.4
00

0
0
50
10
5
1

00

0
0
50
10
5
1

00

0
0
50
10
5
1

00

0
0
50
10
5
1
50
10

50
10

50
10

50
10
0

0
1,

1,

1,

1,

pg injected
Figure 3. PAH ISTD response from a standard system compared to an Agilent 7000B Triple Quadrupole GC/MS System in Continuous Cleaning Mode.
Cal range 1–1,000 pg, MRM.

5
Peak shape is important for integration, accurate quantitation,
and reproducibility. Figure 4 shows MRM TICs for a portion of
the PAH run, using slightly different mixes and oven ramps.
The peak shapes using the SCIS in CCM are significantly
improved over those in a stock system. Figure 5 shows an
expanded view of the latest eluter, dibenzo(a,l)pyrene.

System 1, stock configuration

×10 6
9 10 10 11
7
6
5
Counts

4
3
2
1
0
13.9 14 14.1 14.2 14.3 14.4 14.5 14.6 14.7 14.8 14.9 15 15.1 15.2 15.3 15.4 15.5 15.6 15.7 15.8 15.9 16 16.1 16.2
Acquisition time (min)

System 2, Optimized PAH Analyzer, SCIS CCM

×10 6
9 10 10 11
3

2.5

2
Counts

1.5

0.5

15 15.2 15.4 15.6 15.8 16 16.2 16.4 16.6 16.8 17 17.2 17.4 17.6 17.8 18 18.2 18.4 18.6 18.8 19 19.2 19.4
Acquisition time (min)
Figure 4. PAH peak shape improvement.

×10 3 *19.944 minutes ×10 5 *24.945 minutes

6.5
6 Dibenzo(a,l)pyrene, a late eluter 1
Optimized PAH Analyzer
5.5 0.9
5 Stock configuration 0.8
4.5
4 0.7
Counts

3.5
Counts

0.6
3 0.5
2.5
2 0.4
1.5 0.3
1 0.2
0.5
0 0.1
-0.5 0
-1 -0.1
19.6 19.7 19.8 19.9 20 20.1 20.2 20.3 20.4 20.5 20.6 20.7 24.6 24.7 24.8 24.9 25 25.1 25.2 25.3 25.4 25.5 25.6 25.7
Acquisition time (min) Acquisition time (min)

Figure 5. PAH peak shape improvement for the latest eluter, Agilent 7000B GC/MS/MS Systems, MRM.

6
Figure 6 shows the improved linearity using a completely opti- Table 3. PAHs on an Agilent 7000B Triple Quad GC/MS System, R2 Values
mized GC/MS/MS Enhanced PAH Analyzer (top plot). The of Raw Area Calibration 1–1,000 pg, MRM Continuous Cleaning
Mode
bottom plot shows the typical response from a stock system.
Initial data R 2 CCM R 2
Table 3 compares the R2 values from a standard triple quad
GC/MS to those from an Enhanced Analyzer. The linearity Naphthalene 0.9982 0.9999
demonstrated here 0.9997–1.0000 exceeds the lab 1-Methyl naphthalene 0.9981 1.0000
requirements of 0.99 by a wide margin.
2-Methyl naphthalene 0.9977 1.0000

1,2-Dimethyl naphthalene 0.9974 1.0000

Dibenzo(a,l)pyrene - 7 Levels, 7 Levels Used, 7 Points, 7 Points Used, 0 QCs 1,6-Dimethyl naphthalene 0.9976 1.0000
1.5 y = 0.282750*x – 0.001979
1.4 Acenaphthylene 0.9975 0.9999
R 2 = 0.99998466
1.3
1.2 Acenaphthene 0.9983 1.0000
1.1
Fluorene 0.9976 1.0000
Relative responses

1
0.9
0.8 Phenanthrene 0.9972 0.9999
0.7
0.6 Anthracene 0.9959 0.9999
0.5
0.4 2-Methyl phenanthrene 0.9846 0.9999
0.3
0.2 2-Methyl anthracene 0.9846 0.9999
0.1
0 1-Methyl phenanthrene 0.9969 1.0000
-0.1
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 3,6-Dimethyl phenanthrene 0.9851 1.0000
Relative concentration
2,3-Dimethyl anthracene 0.9648 0.9999
Standard Configuration
Fluoranthene 0.9978 0.9999
2
9,10 Dimethyl anthracene 0.9726 1.0000
Relative responses

1.6

1.2
Pyrene 0.9846 1.0000

0.8 1-Methyl pyrene 0.9927 0.9997

0.4 Benz (a) anthracene 0.9976 0.9998
0 Chrysene 0.9976 0.9999
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5
Relative concentration 6-Methyl chrysene 0.9690 0.9998

Figure 6. Dibenzo(a,l)pyrene, calibration from 1–1,000 pg, R2 = 0.99998 PAH Benzo (k) fluoranthene 0.9954 1.0000
MS/MS Analyzer in continuous clean mode, MRM.
Benzo (a) pyrene 0.9576 1.0000

Dibenz (a,h) anthracene 0.9581 0.9999

Indeno (1,2,3-c,d) pyrene 0.9642 0.9999

Benzo (ghi) perylene 0.9965 1.0000

Dibenzo (a,l) pyrene 0.9788 1.0000

R2 values for 5977A Series GC/MSD SIM with continuous cleaning are nearly
identical to an Agilent 7000B Triple Quadrupole GC/MS, across the same
1–1,000 pg calibration range.

7
Conclusions
The Enhanced PAH Analyzer exceeds the requirements that
most labs have for PAH analysis. The optimized system, in
either GC/MS or GC/MS/MS configuration, demonstrates
ISTD response consistency, less peak tailing, similar sensitiv-
ity, and improved linearity over a standard system. The
Agilent Self-Cleaning Ion Source contributes to these
improvements and maintains system performance without
venting and downtime.

For More Information

These data represent typical results. For more information on
our products and services, visit our Web site at
www.agilent.com/chem.

www.agilent.com/chem
Agilent shall not be liable for errors contained herein or for incidental or consequential
damages in connection with the furnishing, performance, or use of this material.

Information, descriptions, and specifications in this publication are subject to change

without notice.

© Agilent Technologies, Inc., 2013

Printed in the USA
September 12, 2013
5991-3003EN