PB85-115186
Interim Protocol for the Automated
Analysis of Semivolatile Organic
Compounds by Gas Chroaatography/Fourier
Transform Infrared (GC/FT-IR) Spectrometry
(U.S.)- Environmental Monitoring Systems Lab.
Las Vegas, HV
Oct 84
-------
EPA-600/4-04-C81
October 1984
PB85-115186
INTERIM PROTOCOL FOR THE AUTOMATED ANALYSIS OF SEMIVOLATILE ORGANIC
COMPOUNDS BY GAS CHRCMATOGRAPriY/FOURIER TRANSFORM
INFRARED (GC/FT-IR) SPECTROMETRY
by
Donald F. Gurka
Quality Assurance Division
Environmental Monitoring Systems Laboratory
Las Vegas, Nevada 89114
ENVIRONMENTAL MONITORING SVSTEMS LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
LAS VEGAS, NEVADA 89114
-------
TECHNICAL REPORT DATA
(Pleat rt*d liucntcitom on the retene be ton completing)
I REPORT NO.
EPA-600/4-84-081
i.
3. RECIPIENT'S ACCESSION NO.
5 115186
4.TiTLEAN0suBT.TtE jNTERIM PROTOCOL FOR THE AUTOMATED
ANALYSIS OF SEMIVOLATILE ORGANIC COMPOUNDS BY GAS
CHROMATOGRAPHY/FOURIER TRANSFORM INFRARED (GC/FT-IR)
SPECTRDrtFTRY
REPORT DATE
October 1984
6. PERFORMING ORGANIZATION CODE
?. AUTHORtS)
8. PERFORMING ORGANIZATI" »« REPORT NO.
Donald F. Gurka
9. PERFORMING ORGANISATION NAME AND AOORCSS
Quality Assurance Division
Environmental Monitoring Systems Laboratory
Las Vegas, Nevada 89114
10. PROGRAM ELEMENT NO.
ABSDJA _,
JT/GHANrl
II. CONT»ACT
rHO
1?. SPONSORING AGENCY NAME AND ADDRESS
U.S. Environmental Protection Agency—Las Vegas, NV
Office of Research and Development
Environmental Monitoring Systems Laboratory
Las Vpoa--. Npvada fiQ114
I? TYPE OF REPORT AND PERIOD COVCRED
Proet Reort 1083 - 784
14 SPONSORING AGENCY CODE
EPA/600/07
IS. SUPPLEMENTARY NOTES
16. ABSTRACT
The application of gas chromatography/Fourier transform infrared (GC/FT-IR) data to
regulatory decisions requires the availability of validated analytical protocols.
Such protocols are necessary for tne generation of reliable analytical data. A
GC/FT-IR protocol is described which is applicable to the determination of semivolatile
organic compounds in vnstewater, soils, sediments and solid wastes. The protocol is
designed for the high-throughput automated analysis of multicomponent environmental
and hazardous waste extracts. Wastewater analysis is based upon extracting 1 L of
sample with nwthylene chloride and concentrating the extract to 1 mL. Solid waste
analysis is based upon extracting 50 grams of sample and concentrating the sample
extract to 1.0 me. A gel pesinection option is included to further purify those
extracts which cannot be concentrated to the specified final volume. Using capillary
GC/rr-IR techniques, wastewater identification limits of 150 to 400 ppb can be
achieved with this method while the corresponding identification limits for solid
samples are 3 to 8 pom."xAutomated packed column GC/FT-IR identification limits are
approximately a factor of five highpr than the corresponding capillary GC/FT-IR
values. „.
17.
KEV WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
(•.IDENTIFIERS/OPEN ENO£O TERMS C. COSATI Fldd/GfOUp
18. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19. SECURITY
UNCLASSIFIEU
21. NO. OF PAGES
45
2O. SCCURi rv
UNCLASSIFIED
ZZ. PRICE
-------
NOTICE
This report has been reviewed in accordance with the U.S. Environmental
Protection Agency's peer and adminstrative review policies and approved for
presentation and publication. Mention of trade names or conmercial products
does not constitute endorsement or recommendation for use.
ii
-------
ABSTRACT
The application of gas chromatography/Fourier transform infrared
(GC/FT-IR) data tc regulatory decisions requires the availability of validated
analytical protocols. Such protocols are necessary for the generation of
reliable analytical data. A GC/FT-IR protocol is described which is applicable
to the determination of semivolatile organic compounds in wastewater, soils,
sediments and solid wastes. The protocol is designed for the high-throughput
automated analysis of multicomponent environmental and hazardous waste
extracts. Wastewater analysis for semi volatile organic compounds is based upon
extracting 1 I of sample with methylene chloride and concentrating the extract
to I mL. The analysis of the semivolatile fraction derived from solid waste
analysis is based upon extracting 50 grams of sample and concentrating the
sample extract to 1.0 ml. A gel permeation option is included to further
purify those extracts which cannot be concentrated to the specified .final
volume. Using capillary GC/FT-IR techniques, wastewater identification limits
of 150 to 400 ppb can be achieved with this method while the corresponding
Identification limits for solid samples are 3 to 8 ppm. Automated packed
column GC/FT-IR identification limits are approximately a factor of five higher
than the corresponding capillary GC/FT-IR values. The roost frequent obstacle
to achieving these identification limits is expected to be the presence of
large quantities of interfering high boiling co-extractants. These co-
extractants would raise the identification limits by preventing the concentra-
tion of extracts to the desired final volume, thereby necessitating gel
permeation cleanup, and/or by decreasing the spectral signal-to-noise of GC-
volatile analytes by raising the spectral background intensity.
111
-------
CONTENTS
Page
Abstract ill
Tables vi
Abbreviations and Symbols vii
Acknowledgment viii
Introduction 1
Conclusions and Recommendations 2
Results and Discussion 3
References 10
Appendix
A-l Interim Protocol for the Automated Analysis of Semivolatile
Organic Compounds by Gas Chromatography/Fourier Transform
Infrared (GC/FT-IR) Spectrometry A-l
Preceding page blank
-------
TABLES
Page
Fused Silica Capillary Column Gas Chromatographic/Fourler
Transform Infrared On-Hne Automated Identification Limits
for Base-Neutral Extractables 4
2 Fused Silica Capillary Column Gas Chromatographic/Fourier
Transform Infrared On-line Automated Identification Limits
for Acidic Extractables 6
3 GC/FT-IR Sensitivity Parameters 6
4 Reproducibility of Software-Determined GC/FT-IR Spectral
Frequencies 7
v1
-------
ABBREVIATIONS AND SYMBOLS
A/D
AOAC
ASTM
D*
EMSL-LV
EPA
FSCC
FT-1 P.
GC
GC/FT-IR
GC/FT-IR/MS
GC/MS
GIFTS
GM
GPC
i.d.
IR
IRC
K-D
ppb
ppm
L
MIQ
mL
mm
MS
N
NEP
uL
QA/QC
RRT
S/N
Area of Detector Element
Analog/Digital
The Association of Official Analytical Chemists
American Society for Testing Materials
Detector Specific Detectivity
The Environmental Systems Laboratory at Las Vegas
Environmental Protection Agency
Fused Silica Capillary Column
Fourier Transform Infrared
Gas Chromatography
Gas Chromatography/Fourier Transform Infrared Spectrometry
Directly Linked Gas Chromatography/Fourier Transform
Infrared/Mass Spectrometry
Gas Chromatography/'Mass Spectrometry
Gas Infrared Fourier Transform Software
Gram
Gel Permeation Chromatography
inside diameter
Infrared
Infrared Reconstructed Chromatogram
Kuderna-Danish
parts per billion
parts per million
Liter
Minimum Identifiable Quantity
Milliliter
Millimeter
Mass Spectral
Normal
Noise-Equivalent-Power
Microliter
Quality Assurance/Quality Control
Relative Retention Time
S i ngle-to-Noi se-Rati o
vii
-------
ACKNOWLEDGMENT
The author would like to thank Professor Peter Griffiths of the University
of California at Riverside, Dr. James Brasch of the Battelle Columbus Labora-
tories, Columbus, Ohio, Dr. Leo Azarraga of the U.S. Environmental Protec-
tion Agency, Athens, Georgia, Professor James de Haseth of the University of
Georgia, and Dr. Jeanette Grasselli and her group from Standard Oil of Ohio for
reviewing this protocol. Many of their constructive suggestions and criticisms
led to revisions of the draft protocol and have been incorporated into this
document.
viii
-------
INTRODUCTION
The applicability of the gas chromatography/Fourier transform infrared
(GC/FT-IR) spectrometric technique to the analysis of wastewater, soils,
sediments, hazardous wastes, and diesel participates has been demonstrated.*~8
Although this technique is currently one" to two orders of magnitude less
sensitive than gas chromatography/mass spectrometry (GC/MS), with sufficient
extract concentration, GC/FT-IR is capable of detecting about 75 percent of the
GC/MS detectable analytes.* The implementation of recently proposed improve-
ments in GC/FT-IR technology' promises to improve the sensitivity of this
technique to near that of current GC/MS technology. The feasibility of the
direct linked GC/FT-IR/MS method for multicomponent analysis promises an
economical solution to GC/FT-IR confirmation and complementation of the GC/MS
analytical method.10-12 This economy will be realized through the availabi-
lity of low cost FT-IR and MS detectors and through the development of powerful
computer software to process the large quantities of spectral data generated by
using two detectors for every analytical run.
But before the use of GC/FT-If: analysis for regulatory decisions becomes
widely accepted by the academic, industrial, and governmental communities,
validated protocols for its usage must be available. These protocols must be
validated by procedures designated by the appropriate vehicles within the
scientific community. The Association of Official Analytical Chemists (AOAC)
and American Society for Testing Materials (ASTM) are such vehicles and both
have proposed suitable validation procedures for analytical protocols.13•14
-------
CONCLUSIONS AND RECOMMENDATIONS
The analytical protocol described herein is adequate for the identifica-
tion of environmental contaminants at the mid ppb to low ppm range. The prin-
cipal problem expected in applying this protocol to environmental samples is
expected to be the concentration of solid waste extracts to 1 ml. Jn cases of
this sort, the extract may be cleaned up by gel permeation er the analyst may
settle for higher identification limits.
Computer software is required to determine real-time relative IR peak
intensities. This is required to provide relative intensity, as well as fre-
quency precision, acceptance criteria for analyte identification. In addition,
a consensus is required from the spectroscopic community on suitable chemical
candidates for vapor-phase frequency calibration of the GC/FT-IR system (one
reviewer has suggested indene for this role). This coi.sensus should bp sought
from the Coblentz Society and/or the appropriate ASTM co,Tmittee.
Finally, the minimum identifiable quantities of typical environmental
contaminants should be determined in a round-robin study. This study should
employ laboratories equipped with different model GC/FT-IR systems. This
study is necessary because there are currently 11 commercial suppliers of FT-IR
spectrometers. The round-robin study should be coordinated through the appro-
priate ASTM and/or Coblentz Society Committees.
-------
RESULTS AND DISCUSSION
The minimum Identifiable quantity (MIQ) of 54 environmentally important
compounds using on-line GC/FT-IR techniques are listed in Tables 1 and 2.
The capillary GC/FT-IR MIQ's range from 300 ng of molecules with oxygen con-
taining functional groups to 800 nanograms (ng) of the polynuclear arot . 'cs
fluorene and phenanthrene. This ccrrespcnds to a sample sensitivity of 150 to
400 parts per billion (ppb) for wastev»ater, if 1 L of water is extracted and
the extract concentrated to 1 ml v.ith 2 jiL pf.-extract analyzed. A 50 gram (gm)
solid sample undergoing workup by the Appendix A method provides samp.e sensi-
tivities ranging from 3 to 8 ppm. Note that all MIQ's listed in Tables 1 and 2
were determined using the same GC program. As a result the GC peak volumes
(elution volumes) of every analyte were not optimized. For example, Gurka et
al. report a factor of 4.5 difference in GC peak volumes between t^trachloro-
ethylene and di-n-Butyl phthalate using the some GC program.5 Although large
GC/FT-IR peak volumes mean reduced sensitivity via analyte dilution, multi-
compcnent analysis necessitates that the elution volume of each analyte cannot
be optimized with a single GC run. This resulting sacrifice In sensitivity is
made to ensure higher sample throughputs.
The minimum identifiable quantities listed in Tables 1 and 2 have been
obtained with the Environmental Monitoring Sjstems Laboratory, Las Vegas,
Nevada (EMSL-LV), GC/FT-IR system, which has been oesc^ibed elsewhere*.4.5 and
has recently been updated to include a Data General Nova 4 computer equipped
with a high speed array processor and a Lark Model, Control Data 50 megabyte
double disk drive. GC/FT-IR sensitivity values for strong infrared absorbers
ranging between 200 ng for Single beam and 50 ng for double beam systems have
been reported by Griffiths.15 However, dual-beam GC/FT-IR systems are not
commercially available at this time. Recently, Taylor has reported identifi-
able fSCC/GC/FT-IR spectra for 40 ng of some compounds.16 At this, time the
relative sensitivities of GC/FT-IR systems purchased from different manufac-
turers has not been assessed. However the MIQ's reported in this study have
been generated under routine environmental analysis conditions and should be
considered as realistically obtainable.
The parameters affecting GC/FT-IR sensitivity are listed in Table 3.
These parameters break down into the broad classes of spectrometer, interface,
chroratographic, computer, and molecular factors. In general, the a'.alyst has
control over only the chromatographic parameters and the lightpipe temperature.
In some cases the analyst may have & choice of scan rates. A factor of three
reduction in sensitivity has been reported on heating the lightpipe fron
ambient to 240°C5 but Griffiths has concluded that this effect may be entirely
eliminated by re-configuring the spectrometer collection optics.5 An average
factor t>f five improvement in sensitivity with the use of FSCC/GC-FT-IR has
been reported.1 This sensitivity gain is expected from smaller capillary
-------
TABLE 1. FUSED SILICA CAPILLARY COLUMN GAS CHROMATOGRAPHIC/FCURIER
TRANSFORM INFRARED ON-LINE AUTOMATED IDENTIFICATION LIMITS FOR
BASE-NEUTRAL EXTRACTABLES
Identification Limit, ug
No. Compound
Isophorone
Nitrobenzene
Dimethyl phthalate
Diphenyl ether
2, 4-Oinitro toluene
N-Nitroso-dimethylaaine
3-Methyl-2-butanone
1,3-Dichlorobenzene
Oi ethyl phthalate
4-Chlorojiheny!phenyl ether
Oi-n-Butyl phthalate
Di-n-Propyl phthalate
Butyl benzyl phthjlate
2-Methylnapthalene
1 ,4-Dichlorobenzene
bis-2-Chloroethyl ether
Hexachloroe thane
o-Nitrotoluene
Aniline
4-Chloroaniline
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
1 ,2 ,4-Tr ichl orofcenzene
n-Valeraldehyde
Nap thai ene
2 -Chi oronap thai ene
2, 6-Oinitro toluene
bis-2-Chloro-isopropyl ether
bis-2-Chloro-etho*y»e thane
4-Brojaophenylphenyl ether
N-Nitroso-di-propylaoine
N-Ni troso-di -phenyl aai ne
Thiophene
l^-Dichlo^benzene
Acenaptnene
Acenapthylene
1 ,3-HexachlorobuU4iene
Fl uorene
Packed*
Column
•••
1.00
1.10
1.20
2.00
2.00
2.00
2.00
2.00
2.00
2.10
...
...
...
3.00
3.00
3.00
3.30
3.30
...
...
...
...
3. SO
4.20
5.00
5.00
6.00
6.00
6.00
6.00
6.00
6.00
6.20
7.00
7.00
...
7.50
10.00
Capillary'*
Column
0.30
0.40
0.40
...
0.30
0.30
...
0.40
0.40
0.60
0.80
0.40
0.40
0.80
0.30
1.00
0.40
...
tf.40
0.40
0.40
0.40
0.5S
0.40
...
0.40
0.80
0.30
0.40
0.40
0.80
0.30
0.80
...
0.50
...
0.80
0.80
0.80
(continued)
-------
TABLE 1. (Continued)
Identification Limit, ug
NO.
Compound
Packed4
Column
Capillary^
Coluan
Hexachlorocycl opentadtene 10.00
Phenanthrene — 0.80
Benzyl alcohol — 1.0.
•••••••>•••«***•••••*•••BUS•*>•«•*••«»«a««««»»»«o««ma«anooo»»o»«»»«»»*«»«ti .mm*
4 Determined on-the-fly using on-coluan injection and with a 6 ft A 1/8 inch
i.d. glass column packed with 1.51 OV-1? and 1.951 QF-1 on 80/100 oesii Fas
Chroa Q. A heliua flow of 30 oL/iain was used and the GC was programmed froo
70* to 225* at 10Vain. The Interferometer scan rats was 0.3 en/sec and 3
scans/2 sec were collected. The lightpipe was gold-coated with dimensions
of 60 ca x 2.4 on i.d. and was maintained at 240*C. A narrow band HgCdTe
detector (3800-700ca~M with a 2 iwf focal chip was used.5
0 Deterained on-the-fly using splitless injection and a J ft W DB-5 30 H x 0.32
on fused silica capillary colusm (1.0 urn film thickness) at a helium flow of
I oL/nin and no makeup gas. The GC was progranoed front 40* to 260*C at
10*/Bln. The interferooeter scan rate was 1.2 era/sec and '£ scans per second
were collected onto aagnetic disk. The lightpipe was gold-coated with dimen-
sions of 12 ca x 1.5 (or 2.0) ma I.d. and was maintained at 230*C. A nedlun
band HgCdTe detector f3800-700ca-1; Devalue Upeak 1000 Hz, 1) > 1.0 x 1010cm
Hzl/2y-l] with a 1 00* fo^ chip was used. These identification Units are
a factor of 2.75 better than those reported in reference 1 and result froa
updating the systea coaputer froa 32K to 6*K of napped memory and collecting
scans to disk rather than aagnetic tape.
coluam, compared to packed column, elution volumes.*' Clearly the relative
sensitivities of &C/FT-1R systeas, fro» different manufacturers, Snould be
assessed via a round-robin study nith standard solutions.
To establish acceptance criteria for analyte identification, the precision
of real-tine spectral frequencies were determined using the Digi'ab GC/S
software. This software determines the nominal frequency in car1 at the peak
top of the roost intense IR bands within each spectrum. These,frequency preci-
sions are listed in Table 4 and for sharp IR peaks are ±1 cm"* or less. Thus
to report an analyte as "identified," the frequencies of the major IR bands in
the analyte and library spectra should agree to at least ±1 cu~* and the
noninal spectral frequencies for the analyte and library spectra should be
determined with the same coaputer software. Acceptance criteria for the rela-
tive Intensities of the oajor IR bands in the analyte and library spectra
should also be established, but at present the necessary computer software is
not available.
-------
TABLE 2. FUSED SILICA CAPILLARY COLUMN GAS CHROMATOGRAPHIC/FOURIER TRANSFORM
INFRARED ON-LINE AUTOMATED IDENTIFICATION LIMITS FOR ACIDIC EXTRACTABLES
««Caaa3a33aaa33aaa333a33333a3333S3a3a===3333::3333S33a3333333aaa33aa3a333333=333
NO.
Compound
Identification Limit, nga
1
2
3
4
5
6
7
8
9
10
11
12
88883388
Phenol
2-Chlorophenol
2-Cresol
4-Cresol
2-Nitrophenol
Qenzcic acid
2,4-Dichlorophenol
4-Chlorophenol
2,4,6-Trichlorophenol
2,4, 5«Tri chl orophenol
2,4-Dinitrophenol
4,6-Dinitro-2-cresol
0.55
0.55
0.55
0.55
0.40
0.55
0.55
0.95
0.9'J
0.95
0.55
0.55
a Operating conditions are the same as those cited in footnote b of Table 1.
Location
Interface
Molecule
TABLE 3. GC/FT-IR SENSITIVITY PARAMETERS
Nature Effect
Spectrometer
Scan Rate
Source Output
Computer
A/D Convertor
Spectral S/N (S/N o /Nscans)
Signal Intensity
Data Treatment Capacity
Data Transmission Capacity
GC/Column Type
(packed or capillary)
Analyte Elution Volume
Lightpipe temperature
Detector D*
Detector Element Area
(AD)
Makeup Gas
Hater Vapor
Flow Rate
Structure
Molecular Weight
Boiling Point
Analyte Concentration
Analyte Concentration
Spectral S/N (S/N I temperature)
a
Intrinsic Detector Sensitivity
Signal Density at Detector
{Noise-Equivalent-Power (NEP) =
Analyte Dilution
Reduces S/N
Analyte Concentration
Intensity of Absorption
Number of Molecules
Analyte Concentration
============
-------
TABLE 4. REPRODUCIBILITY OF SOFTUARE-DETERMINED GC/FT-IR SPECTRAL FREQUENCIES
==================================
Compound
n-N1 troso-dlmethyl ami ne
1 ,3-D1ch1 oro!".'r;zene
1,4-Dlchlorobenzene
1,2-Dlchlorobenzene
bls-2-Chloroethyl ether
n-N1troso-d1-propyl ether
Nitrobenzene
Isophorone
2958.1
1489.0
1476.4
1284.1
1007.9
1577.4
1454.9
1078.0
783.5
1476.6
1092.0
1013.5
818.6
1458.0
1126.3
1036.7
746.5
2991.2
1132.6
1088.8
755.3
2974.9
2939.9
2888.8
1484.1
1042.5
1540.0
1353.7
852.5
2963.0
2904.6
2884.7
1693.6
1372.0
No.
Runs
14
14
10
13
14
13
13
10
13
14
14
13
13
13
11
12
12
9
11
10
6
14
14
9
-J3
13
12
12
4
11
10
7
11
4
No.
Days
8
8
7
7
8
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
6
8
8
7
8
8
8
8
4
7
7
6
7
3
============3
S.D.C
1.70
0.00
1.26
2.40
1.70
1.50
2.08
^.11
2.33
1.09
0.00
1.92
1.50
0.00
2.72
3.55
1.57
8.39
2.02
1.69
3.01
1.70
1.38
3.35
1.75
1.13
2.49
1.15
5.20
0.00
1.26
5.62
1.29
2.00
(continued)
-------
TABLE 4. (Continued)
Compound
b1s-2-Chloro-ethoxyraethane
Hexachl orobutadi ene
2,6-Dinitrotoluene
2,4-Dinitrotoluene
Fluorene
Di-Phen.ylamine
Phtfnanthrene
*a8SS3333S33S3383833SS3a38B33s:
i/.CH-la.b
2959.0
2892. 6
1159.8
1119.6
1033.6
1559.1
982.7
853.0
798.9
1551.4
1357.3
1605.0
1548.0
1350.0
3071.0
1451.3
737.0
159.,. 2
1501.2
1300.0
745.4
3067.0
605.6
730.5
:S333383 3388833 338
No.
Runs
6
7
10
7
11
7
9
9
9
11
11
10
13
12
10
6
8
13
13
13
7
10
10
10
83833331
No.
Days
6
7
8
7
8
5
7
7
7
8
8
8
8
8
8
5
6
8
8
8
7
8
7
7
[33833838381
S.D.c
3.58
5.53
1.93
4.28
2.16
1.95
2.00
0.00
1.76
1.26
2.41
0.00
3.00
0.00
0.00
2.07
0.00
2.08
0.83
0.00
1.13
1.89
1.26
2.17
t88r.88888B838
a Mean value for the indicated number of runs.
b Determined from the peak maximum using Digilab GC/S* software.
c Standard deviation for the indicated number of runs.
The wastewater*8 and solid sample19"2! workup methods used in this pro-
tocol have been described in detail elsewhere. The wastewater work up is based
on Method 625 and has been modified to allow the Fused Silica Capillary (FSCC)
extract analysis method of Sauter and Betowski."»" The solid sample workup
technique has been developed by the Battelle. Midwest end Southern Institutes.
This procedure has been validated for the largest solid sample size of any
currently available method. A gel permeation option has been included for
those extracts which cannot be sufficiently concentrated to attain the desired
detection limit.
8
-------
In addition to the earlier described frequency precisions, many other
Quality Assurance/Quality Control (QA/QC) procedures are Incorporated within
the protocol. These include criteria for the instrumental centerburst inten-
sity as a function of temperature and spectrometer day-to-day stability. QA/QC
procedures are roughly divided into classes of Daily, Periodic, and Initial
Setup checks. Dally QA/QC includes a 100 percent Line Test, Single Beam Test,
Spectrometer and Mirror Align Tests, and Lightpipe and Beam Splitter protective
procedures. Periodic QA/QC procedures include a detector check, frequency
calibration, and capillary and packed column sensitivity tests. Initial setup
QA/QC includes interferometer and detector checks, frequency calibration, an
Interface sensitivity check, the determination of minimum identifiable quanti-
ties of target compounds, and the preparation of a calibration plot of detector
centerburst intensity versus lightpipe temperature.
-------
REFERENCES
1. D. F. Gurka and M. H. Hiatt. Anal. Chem. 56, 1102, 1984.
2. K. H. Shafer, T. L. Hayes, J. W. Brasch, and R. J. Jakobsen. Anal. Chem.
56, 237, 1984.
3. S. L. Smith, S. E. Garlock, and G. E. Adams. Appl. Spectrosc. 37, 192,
1984. ~~
4. D. F. Gurka and L. Betowski. Anal. Chem. 54, 1819, 1982.
5. D. F. Gurka, P. R. Laska, and R. Titus. J. Chromatogr. Sci. 20, 145,
1982. ~~
6. L. V. Azarrage and C. A. Potter. J_. High Resolut. Chromatogr. Chromatogr.
Commun. 4, 60, 1981.
7. K. H. Shafer, A. Bjorseth, J. Tabor, and R. J. Jakobsen. J_. High Resolut.
Chromatogr. Commun. 3_, 87, 1980.
8. D. L. Newton, M. 0. Erickson, K. B. Tomer, E. D. Pellizari, P. Gentry, and
R. B. Zweidinger. Environ. Sci. Techno!. 16, 206, 1982.
9. P. R. Griffiths. Eastern Analytical Symposium, New York, New York, 1983.
10. C. L. Wilkins, G. N. Giss, G. M. Brissey, and S. Steiner. Anal. Chem. 53,
113, 1980. ~
11. C. L. Wilkins, G. N. Giss, R. L. White, G. M. Brissey, and E. C. Onyiriuka.
Anal. Chem. 54, 2260, 1982.
12. R. W. Crawford, T. Hirshfeld, R. H. Sanborn, and C. M. Hong. Anal. Chem.
54, 817, 1982.
13. H. Horwitz. JAOAC, 66, 455, 1983.
14. ASTM, 41_, E691, 959, 1930.
15. H. M. Gomez-Taylor and P. R. Griffiths. Anal. Chem. 50, 422, 1978.
16. J. R. Cooper and L. T. Taylor. Appl. Spectrosc. 38, 366, 1984.
17. P. R. Griffiths. Appl. Spectrosc. 31_, 284, 1977.
10
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18. J. E. Longbottom and J. J. Lichtenberg. "Methods for Organic Chemical
Analysis of Municipal and Industrial Waste Water," EPA-600/4-82-057 , July
19. U.S. EPA Contract No. 68-03-2624 to the Battelle Memorial Institute,
1978.
20. U.S. EPA Contract No. 68-03-2695 to the Midwest Research Institute,
1979.
21. U.S. EPA Contract No. 68-02-2685 to the Southern Research Institute,
1980.
22. A. D. Sauter and V. Lopez-Avila. Quality Control Protocol for the Fused
Silica Capillary Column GC/MS Determination of Semi volatile Priority
Pollutants. EPA Report (in preparation), U.S. Environmental Protection
Agency, Las Vegas, Nevada.
23. A. D. Sauter, L. D. Betowski, T. R. Smith, V. A. Strickler, R. G. Beimer,
B. N. Colby, and J. E. Wilkinson. J. High Resolut. Chroma togr. Commun. 4,
366, 1981. ~
11
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APPENDIX A-l
Interim Protocol for the Automated Analysis of Semlvolatile Organic Compounds
by Gas Chromatography/Fourier Transform Infrared (GC/FT-IR) Spectrometry
A-l
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CONTENTS
Page
1.0 Scope and Application A-3
2.0 Summary of Method for GC/FT-IR Analysis A-3
3.0 Safety A-4
4.0 Interferences. ........................... A-5
5.0 Apparatus and Material A-5
6.0 Reagents A-7
7.0 Calibration Standards and Internal Standards A-7
8.0 Extraction of Solid Samples (Base/Neutrals) A-8
9.0 Sample Extraction (Acids) A-8
10.0 Extract Cleanup and Calibration (Optional) A-9
11.0 GPC Cleaned Extract Concentration A-10
12.0 Extract Drying A-10
13.0 Extract Concentration A-10
14.0 Daily FT-IR Quality Assurance/Quality Control A-ll
15.0 Periodic FT-IR Quality Assurance/Quality Control A-12
16.0 Initial FT-IR Quality Assurance/Quality Control A-12
17.0 GC/FT-IR Extract Analysis A-13
18.0 Quantitation A-14
A-2
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EMSL-LV, JULY 1984
INTERIM PROTOCOL FOR THE AUTOMATED ANALYSIS OF SEMIVOLATILE ORGANIC
COMPOUNDS BY GAS CHROMATOGRAPHY/FOURIER TRANSFORM
INFRARED (GC/FT-IR) SPECTROMETRY
1.0 Scope and Application
1.1 This method covers the automated identification of solvent extract-
able semivolatile organic compounds, which are amenable to gas
chromatography, by GC/FT-IR.
1.2 This method is applicable to the determination of extractable semi-
volatile organic compounds in wastewater, soils and sediments, and
solid wastes. For example, benzidine can be subject to oxidation
losses during solvent concentration; a-BHC 6-BHC, endosulfan I and
II, and endrin are subject to decomposition under the alkaline
conditions of the extraction step; and hexachlorocyclopentadiene and
N-nitrosodiphenylamine decompose at higher temperatures. Other
extraction and/or instrumentation procedures should be considered
for unstable chemicals.
1.3 The identification limit of this method may depend strongly ttpon the
level and type of gas chromatographicable (GO volatile extractants.
In addition, packed column GC/FT-IR identification limits are about a
factor of five higher than the corresponding capillary column values.
The values listed in Tables A-l and A-2 represent the minimum quanti-
ties of semivolatile organic compounds which have been identified by
the specified GC/FT-IR system, using this method and under routine
environmental automated-analysis conditions. The corresponding
minimum visually identifiable quantities are about a factor of two
lower than the Table A-l values. Capillary GC/FT-IR wastewater
identification limits of 400 ppb may be achieved for weak infrared
absorbers (polynuclear aromatics) with this method while the
corresponding identification limits for solid samples are about 8 ppm.
GC/FT-IR response factor studies indicate that the identification
limits of the strongest infrared absorbers are at least a factor of 10
lower than the corresponding limits for weak infrared absorbers.
2.0 Summary of Method for GC/FT-IR Analysis
If this method is used to confirm gas chromatography/mass spectrom-
etry (GC/MS) the GC/MS analyzed extract should be concentrated, at
least 10-fold, using Kuderna-Danish (K-D) techniques, prior to
A-3
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6C/FT-IR analysis. Otherwise, the following preparation techniques
may be used.
2.1 Wastewater Analysis
A 1 to 2-liter sample of wastewater is extracted and worked up by EPA
Method 625 (1). If fused silica capillary column GC/FT-IR analysis
is employed the 625 base neutral (B/N) and acid extracts should be
concentrated to 1-mL by K-D techniques. Just prior to analysis the
two 1-mL extracts should be combined and concentrated to 1-mL by K-D.
For packed column GC/rT-IR analysis, 1-mL B/N and acid extracts
should be separately analyzed. At leest two nL of extract should be
injected into the gas chromatograph for both packed and capillary
analyses.
Z.t Solid Waste Analysis
The isolation and cleanup procedures as originally developed for the
analysis of municipal sludges for priority pollutants are the basis
of this analysis (2-4). If fused silica capillary column GC/FT-IR
analysis is employed the 625 base -.eutral (B/N) and acio extracts
should be concentrated to 1-rt by K-D techniques. Subsequently, the
two 1-mL extracts should be combined and concentrated to 1-mL by K-D.
For packed column GC/FT-IR analysis, 1-mL B/IJ and acid extracts
should be separately analyzed. At least two uL of extract should be
injected into the gas chromatograph for both packed and capillary
analyses.
2.3 A 50-g sample of chilled residual waste is extracted with methylene
chloride using wet residual waste/solvent techniques aided by a
high-speed homogenizer. Samples are extracted at pH 11 and again at
pH 2 to extract base/neutral and acidic compounds, respectively.
3.1 The toxicity or carcinogeniclty of each reagent used in this method
has not been precisely defined; however, each chemical compound
should be treated as a potential health hazard. From this viewpoint,
exposure to these chemicals must be itiinimized by whateve" means
available. The laboratory is responsible for maintaining a current
awareness file of OSHA regulations regarding the safe handling of the
chemicals specified in this method. A reference file of material
data handling sheets should be made available to all personnel
involved in the chemical analysis.
3.2 All operations involving the use of methylene chloride, including the
extraction of the waste sample, filiations of the extract, and con-
centration of the extract, must be performed in a fume hood. Care
should be taken to avoid skin contact with methy!ene chloride.
A-4
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4.0 Interferences
4.1 Glassware and .other sample processing hardware must be thoroughly
cleaned to prevent contamination and misinterpretation. All of
these materials must be demonstrated to be free from interferences
under the conditions of the analysis by running method blanks.
Specific selection of reagents or purification of solvents by dis-
tillation in all-glass systems may be required.
4.2 Matrix interference will vary considerably from source to source,
depending upon the diversity of the residual waste being sampled.
While general cleanup techniques are provided as part of this method,
unique samples may require additional cleanup to isolate the analytes
of Interest from interference;. In order to achieve maximum sensiti-
vity.
4.3 Clean all glassware as soon as possible after use by rinsing with the
last solvent used. Glassware should be sealed/stored in a clean
environment immediately after drying to prevent any accumulation of
dust or other contaminants. See reference 5 for further guidelines
on glassware cleaning.
5.0 Apparatus and Material
5.1 Extracting equipment.
5.1.1 ADT tissumizer (Tekmar SOT 182EN or equivalent).
5.1.2 Centrifuge (IEC CU-5000 or equivalent).
5.1.3 Screw-capped centrifuge bottles, 200 ml (Scientific Products
C4144) with TFE-lined screw caps.
5.1.4 Fleakers®- 300 ml or equivalent.
5.1.5 Glass Syringe - 50 mi. equipped with a 150 mm x 5 mm ID TFE
tube.
5.2 Gel permeation chromatography cleanup.
5.2.1 Chromatography column - 500 mm x 19 mm ID (Scientific Products
C-4670-106 or equivalent).
5.2.2 Bio-Beads SX-3, 200/400 mesh (Blo-Rad Laboratories 152-2750).
5.2.3 Glass wool.
5.2.4 Graduate cylinders - 100 ml.
5.2.5 GPC Autoprep (Analytical Biochemistry Labs, Inc., 1002 or
equivalent with 25 mm ID column containing 50 to 60 g of
Bio-Beads SX-3). (Optional)
A-5
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5.3 Gas Chromatographic/Fourier Transform Infrared Spectrometric
Equipment
5.3.1 Fourier Transform Infrared Spectrometer - A spectrometer
capable of collecting at least one scan set per second at
8cm'1 resolution is required. A state-of-the-art A/D conver-
ter is required since it has been shown that the throughput of
single beam GC/FT-IR systems is A/0 converter limited (6).
5.3.2 GC/FT-IR Interface - The interface should be ligntpipe volume-
optimized for the selectecS cliromatographic conditions (Light-
pipe volume about 5 ml for packed columns; lightpipe volume
200-400 uL for capillary columns). The shortest possible
inert transfer line (preferably fused silica) should be used
to interface the end of the chromatographic coluui to the
light-pipe. If fused silica cafillary columns are employed,
the end of the GC column can serve as the transfer line.
Griffiths has demonstrated that the optimum lightpipe volume
is equal to the full width at half height of the GC eluate
peak (see general reference number 5).
5.3.3 Packed Column 1 - For base/neutral compounds and pesticides a
6-foot glass column (1/4 in OD x 2 mm 10) packed with 3% SP-
2250 coated on 100/120 Supelcoport (or equivalent).
5.3.4 Packed Column 2 - For acids, a 6-foot glass column (1/4 in 00
x 2 mm 10) packed with 1% SP-1240 OA coated on 100/120 mesh
Supelcoport (or equivalent).
5.3.5 Capillary Column - A fused silica DB-5 30 M x 0.32 mm capillary
column with 1.0 taa film thickness (or equivalent).
5.3.6 Data Acquisition - A computer system dedicated to the GC/FT-IR
system to allow the continuous acquisition, of scan sets for a
full chromatographic run. Peripheral data storage systems
should be available (magnetic tape and/or disk) for the stor-
age of all acquired data. Software should be available to
allow the acquisition and storage of every scan set, to locate
the file numbers and transform high S/N scan sets, and to pro-
vide a real time reconstructed chromatogram.
5.3.7 Detector - A cryoscopic, medium-band HgCdTe (MCT) detector
with the smallest practical focal area. Typical narrow-band
MCT detectors operate from 3800-800 cm-1 but medium-band MCT
detectors can reach 650 cm"*. A 650 cnr* cutoff (or lower) is
desirable since it allows the detection of typical carbon-
chlorine stretch and aromatic out-of-plane carbon-hydrogen
vibrations of environmentally important organo-chlorine and
polynuclear aromatic compounds. The MCT D* should be > 1 x
lD
A-6
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5.3.8 lightpipe - Con? true ted of inert natertals. gold coated, and
voluoe-optiaized for the desired chrooatographic conditions (see
GC/FT-IR Interface. Section 5.3.2).
5.3.9 Gas Chromatograph - The FT-IR spectrometer should be inter-
faced to a teatperature programmable gas chromatograph equipped
with a Grob-type (or equivalent) purged spHrless injection
systea (7-9) suitable for capillary glass columns or an
on-coluon injector systea suitable for packed-glass coluons.
A short, inert, transfer line should interface the gas chroaa-
tograph to the FT-IR lightpipe and. if applicable to the GC
detector. Fused silica GC columns eay be directly interfaced
to the light pipe inlet.
5.4.0 Dry Purge Gas - If the spectrometer is the purge-type, provi-
sions should be made to provide a suitable continuous source
of dry purge-gas to the FT-IR spectroaeter.
6.0 Reagents
6.1 Sodiua hydroxide - (ACS) 10 N In distilled Mater.
6.2 Hydrochloric acid - (ACS) concentrated, 12 K.
6.3 Sodiua sulfate - (ACS) granular anydrous; conditioned at 40J*C for 4
hours and rinsed with dry nethylene chloride (20 ol/g).
6.4 Nethylene chloride - Pesticide quality (Burdick and Jackson or
equivalent).
6.5 Stock standards - Prepare rtock standards fro» EPA Priority Pollu-
tants Kit containing the pro-analysed neat compounds (Chea Service or
equivalent).
6.6 GPC calibration solutions:
a. Corn oil - 200 og/oL in dichloroaethane.
b. bis(2-ethylhexylphthalate) and penUchlorophenol - 4.0 ng/nL in
dTchloroaethane.
7.0 Calibration Standards and Internal Standards
7.1 If quantitation is desired, prepare calibration standards that con-
tain the compounds of interest, either singly or nixed together. The
standards should be prepared at concentrations that Hill completely
bracket the working range of the chromatographic system (t*o or more
orders of oagnitude are suggested).
7.2 If GC/FT-IR will not be used to confirm GC/HS results then prepare
internal standard solutions. Suggested internal standards are 1-
fluoronapthalene, anthracene, terphenyl, 2-chlorophenol, pnenol, bis
(2-chloroethoxy) methane, 2,4*dichlorophenol. phenanthrene,
A-7
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anthracene, and butyl benzylphthaiate. Determine the Internal stan-
dard concentration levels from the minimum identifiable quantities
(see Section 16.4).
8.0 Extraction of Solid Samples (Base/Neutrals)
8.1 The sample which should have been received and maintained at ice
teaperature is thoroughly mixed by homogenization in the sample
bottle. Weigh a 50-g aliquot, or an appropriate weight as pre-
determined by screening analyses, into a 250 ml centrifuge tube.
8.2 Adjust the pH of the sample with 10 N sodium hydroxide to a pH of 11
or greater. Mix briefly with the homogenizer to ensure uniform
sample pH.
8.3 Add 60 ml of roethylene chloride to the sample tube and homogenize for
2 minutes at high speed. Rinse homogenizer off with a minimum
quantity of reagent water, tnen with about 5-10 ml of methylene
chloride. Additional amounts of methylene chloride may be added
until total liquid level is near the top of the centrifuge tube. It
nay be necessary to extract abrasive samples using a sonication
apparatus.
8.4 Centrifuge the samples at 1400 R.C.F. for 15 minutes. The mixture
will separate into an squeous layer over the methylene chloride
extract. A solid cake or emulsion may form at the water-methylene
chloride Interface. If the emulsion interface between layers is more
than on»-half the size of the solvent layer, the analyst must employ
a smaller sample to complete the phase separation. The optimum
technique will depend upon the total solid content of the sample.
Withdraw the organic extract from the centrifuge tube with a 50-mL
glass syringe that has been equipped with a 150 mm x 5 mm 10 TFE
tube. Discharge the extract Into a 300-mL fleaker*.
8.S Add a second 60-nL volume of methylene chloride to tie sample tube
and complete the extraction procedure a second time. Combine the
extracts in the fleaker*.
8.6 Perform a third extraction 1n the same manner, and then dry the
combined extracts as stipulated in the Extract Drying Section.
9.0 Sample Extraction (Acids)
9.1 Adjust the pH of the sample, previously extracted for base/neutrals,
w'th hydrochloric acid to a pH of 2 or less. The acid must be added
slowly and with instant mixing to minimize foaming"oT the sample.
9.2 Ext.'act the sample again using the procedures described in Sections
8.3 to 8.6. Discard the extracted residual water aliquots.
A-8
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10.0 Extract Cleanup and Calibration (Optional)*
10.1 Place 20-25 g of Bio-Beads SX-3 into 200 ml beaker. Cover the beads
with methylene chloride and allow the beads to swell overnight before
packing the column. Put a glass woo! plug in the bottom of a glass
chromatographic column. Transfer the swelled beads to the column and
continue to rinse the packed column with methylene chloride. Add to
the top of the packed column a glass wool plug followed by a layer of
glass beads which will prevent the Bio-Beads from floating to the top
of the elution solvent. Wash the column with about 200 ml of methyl-
ene chloride. Just prior to exposure of the GPC packing, stop the
elution by closing the stopcock on the chromatography column. Dis-
card the eluate.
10.2 Transfer 5 nl of the GPC calibration solution (Section 6.6) to the
Bio-Beads SX-3 column. Drain the column into a 100-mL graduated cen-
trifuge tube until the liquid is just above the surtace of the GPC
packing. Wash the calibration solution onto the column with several
1-mL aliquots of methylene chloride. Next, elute the column with 200
ml of methylene chloride and collect 10-mL fractions. Analyze the
fractions for bis-(2-ethylhexyl)phthalate and pentachlorophenol by
GC/FID on a 1% SP-1240 DA column. Determine the corn oil elution
pattern by evaporation of each fraction to dryness followed by gravi-
metric determination of the residue. Plot the concentration of each
component in each fraction versus the total eluant volume. Discard
the first fractions that elute up to a retention volume represented
by ±851 recovery of the bis(2-ethylhexy!)phthalate. This corresponds
to removal of most of the corn oil. Collect the fractions that elute
up to a retention volume represented by 50 ml after the elution of
pentachlorophenol. A typical procedure is to discard the first 60
ml, to collect the next 110 ml, and to wash the column with 250 mL of
methylene chloride between samples.
10.3 Apply the above GPC separation procedure to an aliquot (1-4 ml) of
the base/neutral or acid concentrate. The volume of concentrate sub-
mitted to GPC is determined by the amount of residue in the concen-
trate. Determine a residue weight of the concentrate by placing a
1-mL aliquot on tared aluminum foil pan, allowing the solvent to
evaporate, and rewelghing the pan. The volume of extract submitted
to GPC should not exceed the capacity of t>>e column or approximately
200 mg.
10.4 Collect the first 60 ml of eluant in a 100 ml graduate cylinder and
pass the next 110 ml of eluant through a drying column containing
anhydrous sodium sulfate and collect in a 500 nl K-D flask equipped
* A test sample should be extracted as described above and tested as delineated
in Section 10.3. If thi» residue weight is of the order of 1-5 mg the GPC
workup may be disregarded. GPC cleanup 1s usually necessary If the sample
extracts cannot be concentrated to the desired volume.
A-9
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with a 10 ml concentrator tube. The drying column should be packed
to a height of 60 mm with anhydrous sodium sulfate. Rinse the drying
column with three 25-mL portions of methylene chloride.
11.0 GPC Cleaned Extract Concentration
11.1 Concentrate the GPC cleaned extract as described in Extract Concentra-
tion Procedure (Section 13.0).
11.2 Transfer the cleaned, concentrated extract to a 6-mL serum TFE capped
bottle and store at 4°C for GC/FT-IR analysis.
12.0 Extract Drying
12.1 When the extract has not been subjected to a drying step (Na2S04,
prior to cleanup for example), the following extract drying procedure
will be employed.
12.2 Pour the combined extracts, for each fraction resulting from the
isolation procedure, through a drying column containing 3 to 4 inches
of organics-free anhydrous sodium sulfate. The dried extract should
be collected in a 500 ml K-D flask equipped with a 10-mL concentrator
tube. The flask which originally contained the extract and the
drying tube should be washed three times with 30-mL aliquots of the
extraction solvent. These washes should be collected in a chilled
K-D flask.
13.0 Extract Concentration
The dried extract will be concentrated as close as possible to 1-mL.
13.1 Add 1 to 2 clean boiling chips to the flask and attach a three-ball
macro-Snyder column. Prewet the column by adding approximately 1 ml
of the extract?ng solvent through the top to the column. Place the
apparatus in a 60 to 65°C water bath with the concentrator tube
partially immersed in the water and the lower rounded surface of the
flask bathed with water vapor. Adjust the apparatus to complete
concentration to approximately 10 ml in 15 minutes. At the proper
rate of distillation, the balls of the column will chatter but the
chambers will not flood. When the liquid has reached an approximate
volume of 4 tri, remove the apparatus from the water bath and allow
the solvent to drain for at least 10 minutes while cooling. Remove
the Snyder column, and rinse the contents of the flask and its lower
joint into the concentrator tube with 1 to 2 ni of the solvent
employed in the extraction.
13.2 Remove the concentrator tube from the water bath, then fit with a
mlcro-Snyder column. Organic-free nitrogen is employed to reduce
the volume of the extract to approximately 1-mL. A Pasteur pipette
is inserted into the top of the Snyder column to accomplish the final
concentration. The concentrator tube and the pipette are washed with
A-10
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approximately two 0.2-tnL volumes of the extracting solvent. Dilute
the final extract to 1 ml for GC/FT-IR analysis.
13.3 If the extract is to be stored before GC/FT-IR analysis, the extract
can be transferred to an appropriately sized serum vial equipped with
a Teflon-lined rubber septum and crimp cap. The extracts' volume
should be scored on this vial, and appropriate sample identification
consistent with the quality control and chain-of-custody requirements
must be affixed to the vial. The extracts can then be stored in the
dark at 4*C.
13.4 It is possible that samples which contain high concentrations of
extractable organic compounds will not concentrate to 1.0 ml. For
extracts of this type, the GPC Cleanup procedure may be used or the
final volume after concentration should be adjusted to a minimal
volume that affords an extract of suitable viscosity for micro
syringe sampling.
14.0 Daily FT-IR Quality Assurance/Quality Control
14.1 One Hundred Percent Line Test - Set the GC/FT-IR operating conditions
to those employed for the Sensitivity Test (see Section 14.2 or 14.3).
Collect 16 scans over the entire detector spectral range. Plot the
tsst and measure the peak-to-peak noise between 1800 and 2000 era*1.
This noise should be £ 0.15%. Store this plot for future reference.
14.2 Single Beam Test - With the GC/FT-IR at analysis conditions, collect
16 scans in the single beam mode. Plot the co-added file and compare
with a subsequent file acquired in the same fashion several minutes
later. Note if the spectrometer is at purge equilibrium. Also check
the plot for signs of deterioration of the light-pipe potassium bro-
mide windows. Store this plot for future reference.
14.3 Align Test - With the light pipe and HCT detector at thermal equili-
brium, check the intensity of the centerburst versus the signal-
temperature calibration curve (see Section 16.6 and Figure 1).
Signal intensity deviation from the predicted intensity may mean
thermal equilibrium has not yet been achieved, loss of detector
coolant, decrease in source output, or a loss in signal throughput
resulting from lightpipe deterioration.
14.4 Mirror Alignment - Adjust the interferoiroter mirrors to attain the
roost intense align interferogram. Data collection should not be
initiated until the align interferogram is stable. If necessary,
align the mirrors prior to each GC/FT-IR run.
14.5 Lightpipe - "the lightpipe and lightpipe windows should be protected
from moisture and other corrosive substances at all times. For this
purpose, maintain the lightpipe temperature above the maximum GC
program temperature but below its thermal degradation limit. When
not in use, maintain the lightpipe temperature slightly above ambient.
A-ll
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At all times maintain a flow of dry, Inert, carrier gas through the
lightplpe.
14.6 Beamsplitter - If the spectrometer Is thermostated, maintain the
beamsplitter at a temperature slightly above ambient at all times.
If the spectrometer is not thermostated minimize exposure of the
beamsplitter to atmospheric water vapor,
15.0 Periodic FT-IR Quality Assurance/Quality Control
15.1 With an oscilloscope, check the detector centerburst intensity
versus the manufacturers specifications. Increase the source vol-
tage, If necessary, to meet these specifications. For reference
purposes, prepare a plot of time versus detector voltage over at
least a 5-day period (see Figure 2).
15.2 frequency Calibration - At the present time, no consensus exists
within the spectroscopic community on a suitable frequency reference
standard for vapor-phase FT-IR. One reviewer has suggested the use
of indene as an on-the-fly standard. See reference 10 for other
reference standards.
15.3 Capillary Column Interface Sensitivity Test - Install a 30 H x 0.32
mm fused silica capillary column coated with 1.0 uM of OB-5 (or
equivalent). Set the lightpipe and transfer lines at 2SO'C, the
Injector at 225*C and the GC detector at 280*C (if used). Under
splitless Grob-type injection conditions, inject 400 ng of nitro-
benzene, dissolved in methylene chloride, while programming the GC
from 40* to 280*C at I0*/min with a carrier gas flow of 1 nt/roin.
The nitrobenzene should be Identified by the on-line library software
search within the first five hits (nitrobenzene should be contained
within the search library).
15.4 Packed Column Interface Sensitivity Test - Install a 6 ft x 1/8 inch
glass column packed with 1.5% OV-17 and 1.951 QF-1 on 80/160 mesh Gas
Chrom Q (or equivalent). Set the lightplpe at 240*C, transfer lines
at 220'C, the injector at 225*C, and the GC detector at 280*C (if
used). Inject on-column 1 microgram of nitrobenzene, dissolved in at
least 2 pi of methylene chloride, while programming the GC frota 70"
to 220'C at lO'/rain with a carrier gas flow at 30 .-nL/min. The nitro-
benzene should be identified by the on-line library software search
within the first five hits (nitrobenzene should be contained within
the search library).
15.5 Checkup By Manufacturers' Servicemen - The spectrometer and interface
should be checked by an authorized repairman for conformance to
factory specifications.
A-12
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16.0 Initial FT-IR Quality Assurance/Quality Control*
16.1 Interferometer - If the interferometer 1s a1r-dr1ven, adjust the
Interferometer drive air pressure to manufacturers specifications.
16.2 MCT Detector Check - If necessary, install a new source and check the
MCT centerburst with an oscilloscope versus the manufacturers speci-
fications (if available). Allow at least five hours of new source
operation before data acquisition.
16.3 Frequency Calibration - At the present time, no consensus exists
within the spectroscopic community on a suitable frequency reference
standard for vapor-phase FT-IR. One reviewer has suggested the use
of indene as an c.i-the-fly standard. See reference 10 for other
reference standards.
16.4 Minimum Identifiable Quantities - Using the GC/FT-IR operating para-
meters specified in Tables A-l and A-2 (or equivalent) determine the
minimum identifiable quantities for the compounds of interest.
16.5 Sensitivity Check - Determine the sensitivity of the GC/FT-IR inter-
face (see Section 15.3 and 15.4).
16.6 Prepare a plot of lightpipe temperature versus MCT centerburst inten-
sity (in volts or other vertical height units). This plot should
span the temperature range between ambient and the Hghtpipe thermal
limit in increments of about 20°C. Use this plot for daily QA/QC
(see Section 14.3 and Figure 1).
17.0 GC/FT-IR Extract Analysis
17.1 Analysis - Analyze the dried methylene chloride extract using the
chromatographic conditions specified in Table A-l for packed or
capillary column interfaces.
17.2 GC/MS Confirmation - Visually compare tho analyte infrared (IR)
spectrum versus the search library spectrum of the most promising
on-line library search hits. Report, as identified, those analytes
with IR frequencies for the five (maximum number) most intense IR
bands (S/N >^ 5) which are within ±1.0 cn~^ of the corresponding bands
in the library spectrum. Choose IR bands which are sharp and well-
resolved. If the analyte cainot be unequivocally identified, report
its chemical functionality. See Table A-3 for nominal frequency
precisions obtained for typical environmental contaminants using the
EMSL-LV GC/FT-IR system with Digilab GC/S® software, and the operat-
ing conditions listed in footnote b of Table A-l. The software used
*•«» locate spectral peaks should employ the peak "center of gravity"
* See Reference 11 for detailed criteria.
A-13
-------
technique (12). In addition the IR frequencies of the analyte and
library spectra should be determined with the same computer software.
17.3 6C/FT-IR Confirmation - After visual comparison of the analyte and
library spectrum as described in Section 17.2 compare the relative
retention times (RRT) of the analyte and an authentic standard of the
most promising library search hit. The standard and analyte RRT
should agree within ± 0.01 RRT units when both are determined at the
same chromatographic conditions.
18.0 Quantitation
Although this protocol is primarily designed to aid GC/MS identifica-
tion via confirmation, some quantitation guidelines are provided.
Although liquid-phase IR quantitation is well-documented, little has
been reported concerning on-line vapor-phase quantitation techniques
(13).
18.1 Infrared Reconstructed Chromatogram (IRC) Technique - After analyte
identification, construct a standard calibration curve of concentra-
tion versus infrared reconstructed Chromatogram peak ar.?a spanning at
least two orders of concentration magnitude. Choose the working
range to bracket the analyte concentration. This method is most
practical for repetitive, target compound analyses.
18.2 Infrared Band Technique - After analyte identification construct a
standard calibration curve of concentration versus infrared band
intensity. For this purpose choose an intense, symmetrical and well-
resolved IR band. The calibration curve should span at least two
orders of magnitude and the working range should bracket the analyte
concentration. This method is most practical for repetitive, target
compound analyses.
18.3 Supplemental GC Detector Technique - If a GC detector is used in
tandem with the FT-IR detector, the following technique may be used;
after analyte identification construct a standard calibration curve
of concentration versus integrated peak a^ea. The calibration curve
should span at least two orders of magnitude and the working range
should bracket the analyte concentration. This method is most practi-
cal for repetitive, target compound analyses.
A-14
-------
TABLE A-l. FUSED SILICA CAPILLARY COLUMN GAS CHROMATOGRAPHIC/rOURIER
TRANSFORM INFRARED ON-LINE AUTOMATED IDENTIFICATION LIMITS FOR
BASE-NEUTRAL EXTRACTABLES
a===========s=s===i=^============== =============:
No. Compound
Isophorone .
Nitrobenzene
Dimethyl phthalate
Diphenyl ether
2,4-Dinitrotoluene
N-Ni troso-dimethyl ami ne
3-Methyl-2-butanone
1,3-Dichlorobenzene
Di ethyl phthalate
4-Chlorophenylphenyl ether
Di-n-Butyl phthalate
Di-n-Propyl phthalate
Butyl benzyl phthalate
2-Methylnapthalene
1,4-Dichlorobenzene
bis-2-Cnloroethyl ether
Hexachlorcethane
o-Nitrotoluene
Aniline
4-Chloroaniline
, 2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
1,2,4-Trichlorobenzene
n-Valeraldehyde
Napthalene
2-Chloronapthalene
2,6-Dinitrotoluene
bis-2-Chloro-isopropyl ether
bis-2-Chluro-ethoxymethane
4-Bromophenylphenyl ether
N-Ni troso-di -propy 1 ami ne
N-Ni troso-di-phenyl ami ne
Thiophene
1,2-Dichlorobenzene
Acenapthene
Acenapthylene
1 ,3-Hexachlorobutadiene
Fl uorene
Identification Limit, ug
Packed3
Col umn
...
1.00
1.10
1.20
2.00
2.00
2.00
2.00
2.00
2.00
2.10
...
—
—
3.00
3.00
3.00
3.30
3.30
...
«• ••»
»•
• •»•»
3.50
4.20
5.00
5.00
6.00
6.00
6.00
6.00
6.00
6.00
6.20
7.00
7.00
7.50
10.00
Cap1llaryb
Column
0.30
0.40
0.40
...
0.30
0.30
...
0.40
0.40
0.60
0.80
0.400
0.400
0.800
0.3
1.00
0.40
...
0.40
0.40
0.40
0.40
0.55
0.40
—
0.40
0.80
0.30
0.40
0.40
0.80
0.30
0.80
0.50
—
0.80
0.8C
0.80
A-15
(continued)
-------
TABLE A-l. (Continued)
OM3a========33=======.:=========================================================
Identification Limit, uq
Packed^ Capillary0
No. Compound Column Column
Hexachlorocyclopentadiene 10.00 —
Phenanthrene — 0.80
Benzyl alcohol — 1.00
===============================================================================
a Determined on-the-fly using on-column Injection and with a 6 ft x 1/8 Inch
1.d. glass column packed with 1.5% QV-17 and 1.95% QF-1 on 80/100 mesh Gas
Chrom Q. A helium flow of 30 mL/min was used and the GC was programmed from
70* to 225" at 10°/min. The interferometer scan rate was 0.3 cm/sec and 3
scans/2 sec were collected. The lightpipe was gold-coated with dimensions
of 60 cm x 2.4 nsr» i.d. and was maintained at 240°C. A narrow band HgCdTe
detector (3800-700cm-1) with a 2 mm2 focal chip was used (14).
b Determined on-the-fly using splitless injection and a J & U DB-3 30 M x 0.32
mm fused silica capillary column (1.0 um film thickness) at a helium flow of
1 mL/min and no makeup gas. The GC was programmed from 40° to 280°C at
lO'/min. The interferometer scan rate was 1.2 cm/sec and 2 scans per second
were collected onto magnetic disk. The lightpipe was gold-coated with dimen-
sions of 12 cm x 1.5 (or 2.0) mm i.d. and was maintained at 280°C. A medium
band HgCdTe detector T3800-700cm-1; 0*value (Xpeak 1000 Hz, 1) > 1.0 x IQlOcm
HZl/2n-l] with a 1 mm2 focal chip was used. These identification limits are
a factor of 2.75 better than those reported in reference 15 and result from
updating the system computer from 32K to 64K of mapped memory and collecting
scans to disk rather than magnetic tape.
A-16
-------
TABLE A-2. FUSED SILICA CAPILLARY COLUMN GAS CHROMATOGRAPHIC/FOURIER TRANSFORM
INFRARED ON-LINE AUTOMATED IDENTICICATION LIMITS FOR ACIDIC EXTRACTABLES
===5===========================================================================
No. Compound Identification Limit, uga
1
2
3
4
5
6
7
8
9
10
11
12
Phenol
2-Chlorophenol
2-Cresol
4-Cresol
2-Nitrophenol
Benzole acid
2,4-Dichlorophanol
4-Chlorophenol
2,4,6-Trichlorophenol
2,4,5-Trichlorophenol
2,4-Dinitrophenol
4,6-Dinitro-2-cresol
0.55
0.55
0.55
0.55
0.40
0.55
0.55
0.95
0.95
0.95
0.55
0.55
===============================================================================
a Operating conditions are the same as those cited in footnote b of Table A-l.
A-17
-------
TASLE A-3. REPRODUCIBILITY OF SOFTWARE-DETERMINED
GC/FT-IR SPECTRAL FREQUENCIES
53=5S=====3S:=5£ 5===S5 = = S = SSS = = S
Cot .pound
n-N i troso-dimethyl ami ne
1 ,3-Dichl orobenzene
1 ,4-D1chl orobenzene
1 ,2-Di chl orobenzene
bls-2-Chloroethyl ether
n-N1troso-d1-propyl ether
Nitrobenzene
Isophorone
v,CM-la,b
2958.1
1489.0
1476.4
1284.1
1007.9
1577.4
1454.9
1078.0
783.5
1476.6
1092.0
1013.5
818.6
1458.0
1126.3
1036.7
746.5
2991.2
1132.6
1088.8
755.3
2974.9
2939.9
2883.8
1484.1
1042.5
1540.0
1353.7
852.5
2963.0
2904.6
2884.7
1693.6
1372.0
No.
1 Runs
14
14
10
13
14
13
13
10
13
14
14
13
13
13
11
12
12
9
11
10
6
» 14
14
9
13
13
12
12
4
11
10
7
11
4
No.
Days
8
8
7
7
8
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
6
8
8
7
8
8
8
8
4
7
7
6
7
3
=33 ========= 3
S.D.c
1.70
0.00
1.26
2.40
1.70
1.50
2.08
2.11
2.33
1.09
0.00
1.92
1.50
0.00
2.72
3.55
1.57
8.39
2.02
1.69
3.01
1.70
1.38
3.35
1.75
1.13
2.49
1.15
5.20
0.00
1.26
5.62
1.29
2.00
(continued)
A-18
-------
vi*VHibi HUCU/
Compound
bi s-2-Chl oro-ethoxymethane
Hexachl orobutadi ene
2,6-Dinitrotoluene
2.4-Dinitrotoluene
FT uorene
Di-Phenylamine
Phenanthrene
i>,CM-la,b
.2959.0
2892.6
1159.8
1119.6
, 1033.6
1559.1
982.7
853.0
798.9
1551.4
1357.3
160^.0
1546.0
1350.0
3071.0
1451.3
737.0
1595.2
1501.2
1300.0
745.4
3067.0
805.6
730.5
==========:
NO.
Runs
6
7
10
7
11
7
9
9
9
11
11
10
13
12
10
5
8
13
13
13
7
10
10
10
No.
Days
6
7
8
7
8
5
7
7
7
8
8
8
8
8
8
5
6
8
8
8
7
8
7
7
S.D.c
3.58
5.53
1.93
4.28
2.16
1.95
2.00
0.00
1.76
1.26
2.41
0.00
3.00
0.00
0.00
2.07
0.00
2.08
0.83
0.00
1.13
1.89
1.26
2.17
3==============i=====================================================3========3
3 Mean value for the indicated number of runs.
b Determined from the peak maximum using Digilab GC/S® software.
c Standard deviation for the indicated number of runs.
A-19
-------
20.0i
O)
15.0
e
U
10.0
I
5.0-
100 200 300
Light Pipe Temperature Degrees Centigrade
Figure 1. Lightpipe temperature effect on GC/FT-IR signal Intensity (14),
A-20
-------
IN)
s?
S
e>
o
CT>
I
(0
o
> 4.
§3
O)
2-
i
a
a?
03 0
O i
6 8 10 12 14 16 18 20
Experiment Day
-------
REFERENCES
1. J. E. Longbottjra and J. J. Lichtenbsrg. "Methods for Organic Chemical
Analysis of 'lunicipal and Industrial Waste Water," EPA-600/4-82-057,
July 198c.
2. U.S. EPA Contract No. 68-03-2624 to the Battelle Memorial Institute (1978).
3. U.S. EPA Contract No. 68-03-2695 to the Midwest Research Institute (1979).
4. U.S. EPA Contract No. C8-02-2685 to the Southern Research Institute (1980).
5. Handbook for Analytical Quality Control in Water and Wastewater
Laboratories. EPA-600/4-79-019, U.S. EPA Environmental Monitoring and
Support Laboratory, Cincinnati, Ohio, March 1979, Section 4.
6. M. M. Gomez-Taylor and P. R. Griffiths. "On-Llne Identification of Gas
Chromatographic Effluents by Dual-Bean Fourier Transform Infrared
Speciroraetry," Anal. Chera., 50 422(1978).
7. R. R. Freeman. Hewlett Packard Application Note "Quantitative Analysis
Using a Purged Splitless Injection Technique." ANGC 7-76.
8. K. Grob and A. Romann. J. Chrom., 214, 118(1981).
9. K. Grob and G. Grob. J. Chrom. Sci., 587(1969).
10. R. H. Cole. "Tables of Wavenumbers for the Calibration of Infrared
Spectrometers," Pergamon Press, New York, 1977.
11. J. G. Grasselli, P. R. Griffiths and R. W. Hannah. "Criteria for Pre-
sentation of Spectra from Computerized IR Instruments," Appl. Spectrosc.,
36 87(1982).
12. 0. G. Cameron, J. K. Kauppinen, D. J. Moffat, and H. H. Mantsch. Appl.
Spectroso., 36, 245(1982).
13. 0. T. Sparks, R. B. Lam, and T. L. Isenhour. "Quantitative Gas Chromato-
graphy/Fourier Transform Infrared Spectrometry with Integrated Gram-
Schmidt Reconstruction Intensities," Anal. Chem., 54 1922(1982).
14. D. F. Gurka, and P. R. Laska. "The Capability of GC/FT-'.R to Identify
Toxic Substances in Environmental Sample Extracts," J. Chromatogr. Sci.,
20, 145(1982).
A-22
-------
15. 0. F. Gurka, M. Hlatt, and R. Titus. "Analysis of Hazardous Waste and
Environmental Extracts by Capillary Gas Chromatography/Fourier Transform
Infrared Spectrometry and Capillary Gas Chroraatography/Mass Spectrometry,1
Anal. Chem., 56, 1102(1984).
16. Final Report on U.S. EPA Contract Ho. 68-03-3122, "Measurement of FT-IR
Spectra for Identification of Potentially Hazardous and Toxic Chemicals."
Research Triangle Institute, March 1984. Submitted to Applied Spectro-
scopy, July 1984.
A-23
-------
GENERAL REFERENCES
1. P. R. Griffiths. "Chemical Infrared Fourier Transform Spectroscopy,"
Wiley-Interscience, New York, 1975.
2. P. R. Griffiths. "Fourier Transfoi-rn Infrared Spectrometry," Science, 222,
297(1984).
3. P. R. Griffiths, J. A. de Haseth, and L. V. Azarraga. "Capillary GC/FT-
IR," Anal. Chem., 55, 1361A(1983).
4. M. D. Erickson. "Gas Chromatography/Fourier Transform Infrared Spectroscopy
Applications," Appl. Spectrosc. Rev., :15, 261(1979).
5. P. R. Griffiths. "Fourier Transform Infrared Spectroscopy Applications
to Chemical Systems," Academic Press, New York, 143(1978). Edited by J. R.
Ferraro and L. J. Basile.
A-24
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