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Combination of Gas Chromatography with

Fluorescence and Phosphorescence in Analysis


of Petroleum Fractions
H. V. DRUSHEL and A. L. SOMMERS
Esso Research laboratories, Humble Oil & Refining Co., Baton Rouge, l a .

Luminescence characteristics of numerous aromatic sulfur and nitrogen


compounds were determined.
Recorded spectra were corrected for
the spectral response of the phototube
by a computer program previously
reported. Examples of the use of
luminescence spectra to characterize
selected nitrogen-rich petroleum fractions are presented. Phosphorescence,
including decay characteristics, provided more information than fluorescence in the study of the nitrogen
compounds isolated from petroleum.
The sensitivity of these techniques
made them particularly suitable for
the study of fractions trapped from
gas chromatographs.

techniques of obtaining
luminescence spectra have been
used by physical chemists for many
years, they have found little use in
analytical chemistry until recently.
One reason for this slow rate of application has been the lack of commercially
available instrumentation. However,
instruments are now available and the
photoelectric recording of fluorescence,
phosphorescence, and excitation spectra
is within the scope of most well-equipped
analytical laboratories. Fluorometry
has been used by analytical chemists
for some time but only recently has
spectrofluorometry with choice of excitation wavelengths been used for analysis
and
characterization
of
organic
materials. Parker and Rees (15) have
used the analogy that fluorometry is
related to spectrofluorometry as filter
photometry is related to absorption
spectrophotometry.
Spectrofluorometry offers a t least two
distinct advantages over absorption
spectrophotometry, uiz., greater sensitivity and several spectra (excitation
and emission) as criteria for identification. However, not all substances
which absorb radiation exhibit fluorescence.
Spectrophosphorimetry, which is
now beginning to be applied in
analytical chemistry, offers even more
information than spectrofluorometry
for the characterization and identificaLTHOUGH

10

ANALYTICAL CHEMISTRY

70827

tion of organic materials. I n addition


to the advantages cited for fluorescence,
the lifetime (7) for the time-delayed
emission (phosphorescence) of substances becomes a criterion for identification and purity.
Several different spectrometric techniques have been used for the characterization of sulfur and nitrogen
compounds isolated or concentrated
from petroleum in projects associated
with the American Petroleum Institute
(2) or in petroleum company laboratories (9). These techniques have
usually been limited to three of the
established spectrometric disciplines
(mass, infrared, and ultraviolet),
Because of the greater sensitivity and
specificity of the luminescence techniques over absorption spectrophotometry, they should find considerable use
in characterization of small amounts of
material following complex micro separation techniques such as chromatography
in its various ramifications. I n this
paper, the luminescence properties of a
variety of sulfur and nitrogen compounds are presented. In addition, the
application of fluorescence and phosphorescence to the characterization of
several nitrogen compound concentrates
from petroleum middle distillates is
demonstrated. Combination of luminescence techniques with gas chromatography is stressed.
EXPERIMENTAL

Instruments. Fluorescence, phosphorescence, and excitation spectra


were obtained with an Aminco-Kiers
spectrophosphorimeter with fluorescence attachment. Spectra were recorded with a Moseley Model 1
Autograph X-Y Recorder. Decay
curves for phosphorescence were obtained by photographing the oscilloscopic trace of emission intensity us.
time presented on a Hewlett-Packard
oscilloscope with built-in time base
sweep. Ultraviolet absorption spectra
were determined with a Cary Model
14M spectrophotometer. Gas chromatographic separations were performed
with a programmed-temperature Model
500 F&M chromatograph using a 17foot SE-30 column.
Solvents. Spectro-grsde isooctane
and carbon tetrachloride were used

in obtaining ultraviolet, fluorescence,


and infrared spectra. For phosphorescence spectra, a 50:50 (by volume)
mixture of methylcyclopentane and
methylcyclohexane
(hereafter
referred to as PH) was used which formed
a clear transparent glass a t liquid
nitrogen temperatures. Both of these
hydrocarbons were purified from Philips
pure grade materials by percolation
over silica gel. Compounds not soluble
in PH were studied in the common
solvent mixture EPA (ethyl ether,
isopentane, ethyl alcohol; 5:5:2 by
volume) which was prepared from
reagent grade ether and alcohol and
Philips pure grade isopentane.
Reference Compounds. Sulfur and
nitrogen compounds were obtained
from various sources. Most of the
compounds were purchased from commercial supply houses. Some were
made available by the American
Petroleum Institute through the
small-sample program of Research
Projects 48 and 52. The source of
each compound studied is shown in
Table I. Purities were not known.
However, in most cases, identity was
confirmed and purities were estimated
to be 98-99% by nuclear magnetic
resonance spectrometry using a Varian
-4-60 NMR spectrometer. Because the
quantum efficiencies for most of the
compounds were high, contributions
from luminescence by impurities were
assumed to be relatively minor.
Calibration.
Spectral response
characteristics of the emission monochromator with a lP28 phototube
were determined with a tungsten
lamp and a Hanovia quartz mercuryvapor arc lamp (lamp code SH).
Spectral energy distribution data for
the emission from the tungsten lamp
was calculated from Wiens law.
Relative spectral line intensities for
the mercury lamp were supplied by the
manufacturer. Values for the phototube response were compared with the
known relative intensities from these
lamps and a curve was constructed for
use in correcting recorded spectra.
An IBM 1620 computer was used to
correct recorded spectra (intensity from
photomultiplier photometer us. wavelength) to give relative intensity in
units of energy us. frequency. Since an
X-Y plotter was not available, the
spectral information was converted to
punched cards and a standard IBM
printer was used to display graphically

the resultant spectra. Details of the


correction and presentation of spectra
using the computer approach have
been presented ( 5 ) .
A mercury arc lamp was used to
calibrate the wavelength axis of the
recorded spectra. With this calibration, reported peak wavelengths are
probably reliable to =k2 mp, the estimated precision of measurement.

Table II. Excitation and Emission Wavelengths and Corrected Fluorescence


Intensities (Relative to Carbazole) for Some Aromatic Compounds Containing
Sulfur, Nitrogen, or Oxygen Atoms

Solvent: Spectro-Grade Iso-octane


ExcitaPredomtion
Fluorescence
inant
waveWaveCorrected
vibrational
lengths,
length,
rel.
spacing,
Compound
m/r
mP
intensity
cm.-l
Diphenylamine
267
333
0,353
...
e-Naphthoquinoline
236
346
0.472
Table 1. Source of Sulfur and Nitrogen
256
364
0.386
1400
Compounds
320
384
0,0954
1450
335
406
0.0178
1400
API Research Projects 48 & 52, Small
5,7-Dimethylbenzacridine
253
389
0.182
Sample Program
287
410
0,135
1300
1-Thiaindane
338
434
0.0346
1350
2-Thiaindane
355
460
0.0088
1300
Acridane
375
Carbazole
[2-Benzo(b)thienyl] henylthiamethane
Diphenyl ether
225
296
0.0501
...
1,2,3,4TetrahydrotRianaphtheno2,3-Benzodiphenylene
oxide
248
354
2.26
(2,3-b)thianaphthene
308
372
1.50
2,3-3',2 '-Dithianaphthene
1350
3-hIethylbenzo(b )thiophene
392
0.553
1370
Benzo ( 1,2-b,5,Pb)dithiophene
414
0.129
1380
Dithieno (2,3-b,5,4-b) thiophene
352
3,PBenzoquinoline
265
0.025
L. Light & Co., Ltd.
363sh.
0.0186
1,2-Dimethylindole
343
313
0,00226
3-Methylisoquinoline
Dimethylthiant hrene
232
350sh.
1.25
1-Methyl-2-phenylindole
5,7-Dimethylbenzacridine
291
364
1.48
...
2,2'-Dinaphthyl ether
1-Azapyrene
380
1.08
...
o-Dianisidine
231
2,2'-Dinaphthyl ether
335
0.198
2-hIethylacridine
257
349
0.158
1200
Diphenyl ether
Diphenylamine
218
302
Diphenylene oxide
0.709
a-Naphthoquinoline
237
312
0.955
...
1,2-Benzocarbazole
275
323sh.
0.480
...
1-Xethyl-2-phenylindole
3,PBenzoquinoline
275
355
0.00465
Diphenylene oxide
320
369
0.00374
1080
2,3-Benzodiphenylene oxide
335
385sh.
0.00144
1120
2-Methylindole
' Phenyl sulfide
263
310
0,00451"
...
Eastman
263
2-Methylindole
0,605
302
...
2-Amino-Pmethylpyridine
2-Amino-4methylpyridine
230
324
0,0907
Diphenyl sulfide
278
Dibenzothiophene
Di-p-tolyl sulfone
o-Dianisidine
298
363
1.16
...
h-aphthalene
Quinaldine
317
367
0.00184
...
Triphenylmethane
Thianthrene
Dimethylthianthrene
252
435
0.00180"
...
Indole
1,2-Dimethylindole
275
310
0.948
Methyl p-tolyl sulfide
Ethyl p-tolyl sulfide
Tetrahydrocarbazole
271
318
0.465
...
Di-p-tolyl disulfide
Dibenzothiophene 5-dioxide
300
360
0.0180
...
Diphenyl disulfide
330
Dibenzyl disulfide
8-Methylquinoline
Thianthrene
330
430
Not detnd.
...
Isoquinoline
1,2,3,4-TetrahydrothiaQuinoline
naphtheno-( 2,3-b)
2,6-Dimethylquinoline
thianaphthene
...
Weak fluor.
2,2 '-Bip yridine
Di-p-tolyl sulfone
[2-Benzo (b) thienyll2,4,6-Trimethylpyridine
phenylthiamethane
...
Weak fluor.
...
...
1-Thiaindane
Rutgerswerke-Aktiengesellschaft
...
Weak fluor.
...
...
3-Methylbenzo (ti)thiophene
Tetrahydrocarbazole
268
307
0.0505
...
Quinaldine
Acridane
280
338
1.86
...
7,8-Benzoquinoline
2,3-3',2'-Dithianaphthene
3-hIethylisoquinoline
255
339
0.255
...
2-Methylpyridine
300
350
0.248
...
4bIethylpyridine
320
Synthesized by N. L. Cull, Esso Research
Benzo (1,2-b,5,4-b) dithioLabs.
phene
250
332
0.143
...
287
Dibenzothiophene dioxide
I
Carbazole
235
331
1.00
...
Matheson Coleman & Bell
282
346
0.862
*..
Phenothioxin 10-dioxide
Acetophenone
a Emission increases with excitation time indicating secondary reaction.
~

VOL. 38, NO. 1, JANUARY 1966

11

Figure 1 , Typical recorded excitation, fluorescence, and


phosphorescence spectra
Compound: 1, 2-dimethylindole

Fluorescence

Phosphorescence

1 .O pg./ml.
Isooctane
Curve 4
Curve 2

Concn.
Solvent
Excitation
Emission

0.58 mg./ml.
EPA
Curve 3
Curve 1

carbazole. These relative intensity


data are not very precise because of the
long period of time over which data were
accumulated and the wide variety of slit
combinations used. The main purpose
in accumulating the luminescence data
for reference compounds was to provide
a basis for the qualitative identification
of compound types in trapped gas
chromatographic fractions. For this
reason the vibrational fine structure and
the corrected relative intensities within
a given spectrum are important. The
~~

~~~

~~

RESULTS AND DISCUSSION

Luminescence Characteristics of
Reference Compounds. Typical ex-

citation, fluorescence, and phosphorescence spectra for one of the compounds


as recorded are reproduced in Figure 1.
Data on relative intensities, which are
affected by the choice of slit combination and other instrumental parameters, will be presented below.
Data from the recorded spectra were
corrected and converted to relative
intensity us. frequency (micron-') by
means of the developed computer program. The luminescence properties of
many of the compounds studied have
not been reported previously in the
literature. Several oxygen-containing
analogs were included to determine the
effects of other heteroatoms.
Corrected relative fluorescence intensities are listed in Table 11. Intensities
were converted to values equivalent to
a single arbitrary set of experimental
conditions (photomultiplier setting, sensitivity, slit combination, and concentration). Solutions were degassed
and blanketed with high-purity dry
nitrogen to eliminate the possibility of
oxygen quenching. Most compounds
were studied a t concentrations of 1 to
10 pg./ml. to reduce effects of concentration quenching or other phenomena which normally give rise to nonlinearity with concentration. All relative peak heights were corrected for
spectral response of the phototube by
use of the technique described earlier
(5)*
Although different wavelengths were
used for excitation, no correction was
made for differences in excitation energy
a t different wavelengths. Rather than
use arbitrary units, intensities were
calculated relative to the intensity of

12

ANALYTICAL CHEMISTRY

Table 111. Excitation and Emission Wavelengths and Phosphorescence Intensities


(Relative to Carbazole) for Some Aromatic ComPounds Containina Sulfur, Nitrogen,
or Oxygen Atoms

Compound
Tetrahydrocarbazole

Solvent
EPA

1,2-Dimethylindole

EPA

Dimethylthianthrene

EPA

Quinaldine

EPA

Excitation
wavePhosphorescence
lengths. Wavelength. Corrected
mp
mp
rel. intensity
292
411
0.36
224
439
0.53
0.44
460sh.
0.28
500b.sh.
414
288
0.31
441
0.44
454sh.
0.34
464sh.
0.33
285
422?
...
482
0.14
46 1
300
0.086
471sh.
0.059
0.13
493
0.11
529
0,042
568
0.0080
332
590
Y

5,7-Dimethylbenzacridine

EPA

2,2'-Dinaphthyl ether

EPA

279
315

1-Asapyrene

EPA

368

370

485

515
-

542sh.
524

561
-

o-Dianisidine

EPA

305

2-Amino4methylpyridine
2-Methylacridine

EPA
EPA

302
292

Diphenyl sulfide
Diphenyl ether

EPA
EPA

Diphenylamine

EPA

285
234
264
305

597
498
524
439
437
451sh.
464
416
404
40 1
417
427

0.16
0.27
0.20
0.00053
0,00055
0.00041
0.053
0.078
1.02
0.0012
0,0013
0.0014

Predominant
vibrational
spacing,
cm.-l

...
1580

...
...
...
1450
...
...

...
...
...

1400
1400
1500

...

...
...

1210

iib
1090

...
...
...
...
...

1300

1.4

...

0.44

...

3.6
2.8
2.5

...
...
...
(Continued)

Table 111. Excitation and Emission Wavelengths and Phosphorescence Intensities


(Relative to Carbazole) for Some Aromatic Compounds Containing Sulfur, Nitrogen,
or Oxygen Atoms (Confinued)

Compound
Dithieiio (2,3-b,5,4b)thiophene
B ~ ~.( lJ2-b,5,4-b)z o
dithiophene

Solvent
PH
PH

Excitation
wavePhosphorescence
lengths, Wavelength. Corrected
rel. intensity
mw
mp
272
287

3-Methylbenzo ( b )
thiophene

PH

280

1-Thiaindane
2-Thiaindane
[2-Benzo ( b ) thienyllphenylthiamethane

PH
PH
PH

293
280
300

1,2,3,PTetrahydrothianaphtheno-(2,3-b)thianaphthene
Acridane

PH

297

EPA

308

465
459
466
476
494
ash.
511sh.
527sh.
440sh.
450sh.
466

497

PH

295

318

Carbazole

a-Naphthoquinoline

7,8-Benzoquinoline

1,2-Benzocarbazole

2-Methylindole

EPA

EPA

EPA

EPA

EPA

1-Methyl-2-phenylindole

EPA

314-Benzoquinoline

EPA

Diphenylene oxide

EPA

285
325
315

332
-

282
317
329
317

338
285

315
315

330
-

284

440
435
460sh.
485
515sh.
474
414
__
425
437
479
492
501
518
531
54lsh.
559
574
410

426
439
456
465sh.
466
475
501
538
575sh.
468
477
504
514sh.
540
580sh.
483
497
519
553sh.
408
420sh.
434
446sh.
458
500
531
568sh.
455
460sh.
488
523
560sh.
414
435sh.
444

461
469
479

0.0031

Predominant
vibrational
spacing,
cm.-l

...

1.1

1.1
1.0
1.1
1.0
0.97
0.88
0.145
0.16
0.28
0.27
0.069

300
430
1530

0.0001

...

...
...
...
...
...
...

...

...

1.6
2.8
2.8
0.62

...
...
...
...

1.5
1.3

...

1.2

0.31
0.28
0.20
0.44
0.38
0.27
0.27
0.19
1.0
0.55
0.99
0.55

0.52
0.047
0.036
0.064
0,045
0.045
0.19
0,090
0.23

...

0.17

0.034

0.031
0.063
0.042
0.38
0.27
0.44
0.30
0.28
0.011
0.019
0.012
0.12
0.11
0.20
0.17
0.078
0.75
0.50

0.81
0.45
0.44
0.42

600
680

...
500
...

1550
480

...

...
...
...

...
1600

...
...
...
...

1500

...
...
...

1600

...
...
...
...
...

1500

...
...

1500

...

...
...
...
...
...
...

1400

...
...
...
...

1650

...

...
(Cmkked)

spectral sensitivity of the phototube


is thus taken into account. The evaluation of absolute intensities or of
quantum efficiencies ( 5 ) is beyond the
scope of the present application. The
listing of the corrected intensities relative to carbazole provides some idea
of the sensitivity of the technique in
qualitative identification. Because of
the many variables involved (slit combinations] photomultiplier settings,
excitation wavelengths, etc.) the listed
data should not be used for quantitative
calculations.
Relative phosphorescence intensities
were more difficult to evaluate because
of slight variations related to positioning
of the small sample tube in the quartz
Dewar flask.
;ilso, since phosphorescence intensities are much lower
than fluorescence intensities] spectra
were obtained a t high concentrations
which may have produced concentration
quenching effects.
However, some
approximate relative phosphorescence
intensities were calculated as shown in
Table 111. Again, as with fluorescence,
the intensity data correspond to a
single set of experimental conditions
(at 1 mg./ml.) relative to the intensity
for carbazole. Band positions for
excitation are also included. The same
limiting factors as discussed under
fluorescence intensities apply to the
phosphorescence data in Table 111.
Values of the half life for phosphorescence are presented in Table IV. Most
values were determined by manual
rotation of the shutter. For very short
half lives the shutter was rotated a t an
appropriate speed by the shutter motor.
Some information on the luminescence
of complex compounds of carbon,
hydrogen, and sulfur or nitrogen has
been published (these data are collected
in Table V for convenience). The
fluorescence characteristics of aniline,
methylaniline, dimethylaniline, indole,
skatole, and 2-methylindole were compiled by the American Instrument Co.
(20). Several nitrogen heterocyclic compounds were included in the report of
the fluorescence spectra of some aromatic compounds by the Chicago Medical
School (13). A bibliography of phosphorescent molecules compiled by Chen
for the American Instrument Co. lists
several nitrogen compounds (4). Russian workers recently reported on the
luminescence of a series of nitrogen
compounds (6) polarization of the
fluorescence of porphyrins (8),and the
luminescence of acridine (17) and quinoline ( 7 ) . Van Duuren studied the
fluorescence of various heterocyclic
aromatic compounds ($1). Two dibenaacridines were also studied by Schoental and Scott (16). The fluorescence of
a series of unusual azaindoles was presented by Adler (1). Heckman (10)
studied the phosphorescence spectra of
indole, thianaphthene (benzothiophene),
VOL. 38, NO. 1, JANUARY 1966

13

Table Ill. Excitation and Emission Wavelengths and Phosphorescence Intensities


(Relative to Carbazole) for Some Aromatic Compounds Containing Sulfur, Nitrogen,
or Oxygen Atoms (Continued)

PredomExcitainant
tion
vibrawavePhosphorescence
tional
lengths, Wavelength, Corrected spacing,
Compound
Solvent
mfi
mr
rel. intensity cm. -1
2,3-Benzodiphenylene oxide
EPA
318
520
0.094
...
337
528sh.
0.058
...
542
0,063
...
563
0.099
1460
588sh.
0.060
...
3-Methylisoquinoline
EPA
282
495sh.
...
310
515
o.ooi2
...
Wsh.
...
...
Older data (different 1P28 phototube)
Dibenzothiophene 5-dioxide
EPA
285
445
0.30
...
315
478
0.45
...
510
0.39
...
EPA
312
425
Dibenzothiophene
1.5
...
439
1.3
...
456
1.5
...
EPA
270
370sh.
Not
detnd.
p-Tolyl sulfone
.
..
385
Not detnd.
...
EPA
285
364
0.99
Phenothioxin-10-dioxide
382
1.2
iioo
400
0.87
...
475
PH
290
0,019
Naphthalene
...
487
0.012
. .
510
0,027
1450
549
0,019
...
PH
280
...
354sh.
Triphenylmethane
...
373
0.27
...
386
0.28
...
405sh.
0.19
...
EPA
285
480
0.13
Thianthrene
...
410
EPA
283
0.070
Indole
...
437
0.097
1550
450sh.
0,052
...
414
EPA
285
0.27
Methyl p-tolyl sulfide
*..
285
420
0.34
Ethyl p-tolyl sulfide
...
Very weak phosphorescence
EPA
Di-p-tolyl disulfide
...
Very weak phosphorescence
EPA
Diphenyl disulfide
...
Very weak phosphorescence
EPA
Dibenzyl disulfide
...
303
EPA
8-Meth ylquinoline
474
0.0069
,..
486
0.0045
...
0.0097
1400
508
546
0.0069
...
EPA
308
472
0.0014
Isoquinoline
550
484
0.0016
350
493
0.0025
500
506
0.0031
519
550
0.0030
0,0039
529
350
543
0.0030
500
458
0.028
EPA
Quinoline
298
...
470
0.020
550
490
0.036
1450
502sh.
0.025
...
526
0.027
...
0.025
EPA
310
473
2,6-Dimethylquinoline
...
0.022
482sh.
...
508
0.044
1450
544
0,033
...
429
2,2 -Bipyridine
EPA
295
0.025
...
449
...
0.025
1600
460
0.036
0.031
480
...
0.063
PH
240
389
Acetophenone
..*
1650
416
0.095
278
1750
320
448
0.070
0.041
1600
483

14

ANALYTICAL CHEMISTRY

carbazole, dibenzofuran, and dibenzothiophene. He found that there was no


phosphorescence for pyrrole, furan,
and thiophene. Yoshida (23) obtained
the fluorescence spectra of a variety of
aromatic compounds with -NHz
groups. Van Duuren (82) has also
studied the fluorescence of various substituted indoles. Shimada (18, 19)
carried out a vibrational analysis of the
phosphorescence spectra of pyrazine
and pyrimidine.

Table IV. Lifetime for Decay of


Phosphorescence Emission for Some
Selected Compounds

Compound
Aromatic hydrocarbons
Acenaphthene
Indene
1-Methylnaphthalene
2,7-Dimethvlnaphthalkne
Oxygen compounds
Diphenyl ether
Dibenzofuran(diphenylene oxide)
2,3-Benxodiphenylene
oxide
Nitrogen compounds
Diphenylamine
Quinoline
Quinaldine
2-Methylacridine
Tetrahydrocarbazole
1,2-Dimethylindole
5,7-Dimethylbenzacridine
3,CBenzoquinoline
7,8-Benzoquinoline
2-Methylpyridine
CMethylpyridine
2,4,6-Trimethylpyridine
2,2-Bipyridine
Acridane
Carbazole

Mean
lifetime,
Sol- secvent onds
PH
PH
PH

2.8
0.5
2.0

PH

1.8

EPA 0.45
EPA 6 . 1
EPA 1 . 5

EPA
PH
EPA
EPA
EPA
EPA

1.9
0.8
1.7
3.8
4.8
5.2

EPA
EPA
EPA
PH
PH
PH
PH
EPA
EPA

0.26
1.2
2.1
1.3
1.7
1.8

1.0
3.0

8.1

Sulfur compounds
Di-p-tolyl sulfone
EPA 1 . 4
Phenothioxin 10dioxide
EPA 0.75
Diphenyl sulfide
EPA 0.034
Dimethylthianthrene
EPA 0.072
EPA 0.063
Methyl p-tolyl sulfide
Thianthrene
EPA 0.040
[2-Benzo ( b ) thienyl]
phenylthiamethane
PH 0.055
1-Thiaindane
PH 0.045
3-Methylbenzo ( b )
thiophene
PH 0.22
2,3-3,2-Dithianaphthene
P H 0.034
Benzo ( 1,2-b15,4b)dithiophene
PH 0.26
1,2,3,PTetrahydrothianaphthenoP H 0.074
(2,3-b) thianaphthene PH 0.074
a Time required to reach 36.8y0 of
initial intensity.

Theory, Interpretation, and Discussion of Spectra. I n the fluorescence process, the molecule is raised
from a singlet ground state ( 8 ) to an
or vibrational
excited singlet state (S*)
level thereof, followed by vibrational
deactivation to the zero vibrational
level of S*, thence via fluorescent emission to S or vibrational level thereof.
Since the vibrational levels of S and S*
are similar, the emission spectrum is
usually an approximate mirror image of
the absorption spectrum (12). Thus,
where fine structure may be observed,
something of the vibrational frequencies
in the molecules may be determined
and used as a means of characterization
or identification.
Phosphorescence involves a metahaving a longer
stable triplet state (T*)
lifetime and a lower energy than the
singlet excited state. Electronic transitions between two states involving a
change in multiplicity, as in the T* +
S phosphorescent emission, are normally
forbidden. The probability of transitions between these states, however, is
governed by the degree of spin-orbital
interaction (magnetic interaction between the spin and orbital motions of an
electron). I n atoms of low atomic
number the spin-orbital coupling is
small, while for heavier atoms it is
larger, The constant in the expression
describing this magnetic interaction is
proportional to the fourth power of the
effective nuclear charge. McClure (14)
found that substituting heavy atoms in
organic molecules produced the same
effect as observed in atoms. Although
this substitution causes little change in
the wavelength and intensity of the
absorption spectra, increased probability of T*-+ S transitions decreases
the lifetime in the T* state. For the
halogenated naphthalenes substituted
in the a-position, McClure (14) found
the following lifetimes for phosphorescence: F = 1.5, C1 = 0.30, Br =
0.018, and I = 0.0025 second.
Nitrogen and oxygen analogs of
aromatic hydrocarbons should exhibit
phosphorescence lifetimes equivalent to
their hydrocarbon analogs (Tables IV
and V). Sulfur compounds show much
shorter lifetimes and higher intensities
than the hydrocarbon or oxygen analogs
because of increased spin-orbital interaction (Table IV).
The phosphorescence intensities of
many of the oxygen, nitrogen, and
sulfur compounds shown in Table I11
are high compared to their fluorescence intensities (Table 11). This is
contrasted to the behavior of the hydrocarbon analogs which show weak
phosphorescence. It is for this reason
that spectrophosphorimetry becomes
useful in the characterization of sulfur
and nitrogen compounds, particularly
when micro separation techniques are
used and sensitivity is needed.

Table V.

Some Luminescence Characteristics of Pertinent Nitrogen Compounds


Compiled from the Literature

Excitation
max ,, Emission max., mp
Phos.
Compound
mr Fluor.
280
340
...
Aniline
...
290
360
Methylaniline
...
255
370
Dimethylaniline
...
280
350
Indole
...
290
370
Skatole
...
355
280
2-Methylindole
...
. . . 404
Benz(c)acridine
. . . 406
...
11H-Benzo(a)carbazole
...
11H-Benzo(b)carbazole
...
408
396
...
7H-Benzo(c)carbazole
. . . . . . 372' *
Aniline
...
. . . 408, 438
Carbazole
...
. . . 865
Chlorophyll-b
...
. . . 493
Dimethyl-a-naDhthvl- amine
2,4-Dimethylquinoline
...
. . . 458
. . . . . . 420
p-Dimethyltoluidine
...
. . . 395, 412,
Diphenylamine
423
...
. . . 680-870
Metal-etioporphyrin11
complexes
400-600
...
...
Indole
462
...
Quinoline
400-600
323
...
Isoquinoline
660-760
,..
hletal-mesoporphyrinIX . . .
complexes
470
...
...
8-Meth ylquinoline
a-Naphthylamine
...
. . . 498
. . . . . . 498
p-Naphthylamine
502
...
...
Di-@-naphthylamine
. . . . . . 494, 540,
a-Naphthonitrile
587
... . . . 484
p-Naphthonitrile
. . . 495
Phenyl-p-naphthylamine . . .
335
350
...
Phenanthridine
367
385
400
300
...
395
Dibenz(a,h)acridine
403
418
445
470
300
...
395
Dibenx(a,j)acridine
400
416
440
465
280
348
...
Indole
256
322
...
Isoindazole
290
250
...
310
Benzimidazole
275
-

Hal flife
of phos.,
seconds

...
...
...
...
...
...
...
*..

...
...

Long
Long
10-4
Long
Long

...

Long
10-4

...

Long

5 X'io-4

4-Axaindole
5-Azaindole
6Axaindole
7-Axaindole
Benztriazole
3,4-Diaxindole
3,5-Diazaindole
Purine
Aniline
N,N-Dimethylaniline
N,N-Diethylaniline
Diphenylamine
Triphenylamine

Long
Long
Long
Long
Long
Long
Long

...

...

...

...
...

2.5gb
3.80b

...

4.45b

417
404

...

*..

...
...

1.096
0.276

392

...

...

2.16b

391
350
320

...

...

*..

...
...
...

0.026
4,466

...
...
...
...

355
370
33OC
336O
336c
345c

...
...

...
...

0.286
0. 14b

...

35OC

299
270
282
266
294
293
270
255
285
280
281

...

...

408
417C
412c
398"
415"
422c
410~
427"
435c

0.3gb

4.2
2.4
2.0
1.9

0.70

(6)

(Continued)
VOL. 38, NO. 1, JANUARY 1966

15

To ble V.

Some Luminescence Characteristics of Pertinent Nitrogen Compounds


Compiled from the Literature (Continued)

Compound
9,10-Dihydroacridine
Indole
Carbazole
Indole
Carbazole
5H-Benzo(b)carbazole
7H-Dibenzo(a,g)carbazole
7H-Dibenzo(c,g)
carbazole

Excitation
max., Emission max., mru
mp Fluor. Phos.
, , .
347~ 412O
. ..
439c
297c 403c
. . . 43lC
455c
. . . 342< 406c
357c 41SC
. . . 435c
280
297
...
32 1
335
...
348
330
387
...
411
440
340
367
386
408
430
350
363
...
382
400
424
330
...
317
400
360
...
297
285
306
280
315
280
291
315
...
282
320
...
290
337
306

Half life

of phos.,

seconds
1.6

Ref.

0.36

(6)

0.55

(6)

Remarks

(6)

. ..
...

(21)
(21)

...

(21)

IN A

LATE.

...

(21)

* ..
(21)
Quino1ine
...
(21)
Acridine
Indole
2-Methylindole
3-Methylindole
1,2-Dimethylindole
2,3-Dimethylindole
...
(82)
2-( 3-Indolyl)-2,3dihydroindole
(indole dimer)
360
..
(22)
1-Methyl-2-phenylindole 310
..
(22)
332
290
Carbazole
348
318
330
Ap roximate limit of detection in pg./ml.
b RePative intensity in water at p~ of maximum emission.
c Values read from reproduced spectra (relative intensity us. frequency).
d Relative intensity in cyclohexane (uncorrected).

55

50

45

35

40

30

6.0d
8.2d
64. Od
5.4d

25

TIME (MINUTES)

Figure 2. Gas chromatogram of a basic nitrogen extract from a


petroleum fraction
Column
Programming rate
Initial temp.
Sample size
(Fraction cut-points as shown)

16

ANALYTICAL CHEMISTRY

The vibrational fine structure corresponds to vibrational frequencies (as


observed in infrared and Raman
spectra) representing fundamental skeletal or C-C,
C-0, C-N, or C-S
vibrations. Some approximate vibrational frequencies are calculated for
several of the compounds showing fine
structure and the values are included in
Tables I1 and 111. In most cases, the
values seem to correspond to the heavy
atom-carbon stretching frequencies or
the skeletal vibrations in aromatic
hydrocarbons. It is this vibrational
fine structure which becomes useful in
the identification of certain molecular
types by luminescence.
Application of Luminescence to
Characterization of Petroleum Fractions. BASICKITROGEXCOMPOUXDS

430-650" F.

17-Foot SE-30 on Chromasorb W


C./minute

4'

150' C.
30 pl.

STRAIGHT-RUN
MIDDLEDISTILX 430-650" F. distillation

cut of a crude petroleum was extracted with dilute (1:1) hydrochloric


acid to concentrate the basic nitrogen
compounds. The acid extract was
neutralized with sodium hydroxide and
the released nitrogen compounds were extracted into hexane. After thoroughly
washing the hexane with water, the
solution was evaporated in a nitrogen
atmosphere to yield the basic nitrogen
concentrate.
Components of the basic nitrogen
extract were partially separated by gas
chromatography on an SE30 column
(Figure 2). The small amount of material represented by each trapped fraction
did not permit the use of infrared
spectrophotometry for characterization.
However, even upon diluting to several
milliliters, there was sufficient material
in each fraction to obtain ultraviolet
spectra (Figure 3). Although evidence
for quinolines is apparent, it would be
difficult to detect the presence of substituted pyridines or tetrahydroquinolines in this fraction on the basis of the
ultraviolet absorption bands.
Phosphorescence spectra provided information not readily obtained from the
ultraviolet spectra. By changing the
excitation wavelength, it was possible to
observe both pyridine (or tetrahydroquinoline) and quinoline species in the
mixture of compounds represented in
each trapped fraction (Figure 4).
Pyridines and tetrahydroquinolines were
excited a t 260 mp while quinolines
were excited at 295 mp with several
characteristic emission bands between
460 and 550 mp. Further evidence
for the identity of the molecular
types present was provided by the
half life for decay of phosphorescence.
The decay characteristics for the compounds in the gas chromatographic
fractions are listed in Table VI. A
gradual increase in the half life was
observed with increase in cut number
(increase in boiling point or molecular
weight). This was expected on the

1.0

0.8

0.6

Y
m
<
0.4

IIAF"WE

(Mp)

Figure 4. Phosphorescence and excitation spectra of gas chromatographic fraction No. 4 of a basic nitrogen extract of a 430650" F. petroleum fraction
Curve
Curve
Curve
Curve

0.2

1.
2.
3.
4.

Excitation spectrum (emission set at 4 0 5 mp)


Excitation spectrum (emission set at 4 9 5 mp)
Phosphorescence spectrum (excitation set at 260 m p )
Phosphorescence spectrum (excitation set at 2 9 5 mp)

Figure 3. Ultraviolet spectrum of gas chromatographic fraction No. 4 of a basic nitrogen extract of
a 430-650" F. petroleum fraction

basis of the half life values determined


for the reference compounds (Table IV).
Although the object of this study was
not a quantitative analysis of this
fraction, some idea of the relative
amounts of pyridines or tetrahydroquinolines and quinolines present in
each fraction was obtained from the
ratio of emission intensities a t 400 and
500 mp with excitation at 265 and 300
mp, respectively (Table VII). These
data showed that the pyridines and
tetrahydroquinolines are concentrated
in the first three fractions and that as the
molecular weight increases, the quinoline types predominate. The high
intensities at 400 mp for the first three
fractions may also be derived from the
presence of some anilines or other related aromatic amines.
The fluorescence of pyridines and
quinolines is weak as in the case of their
hydrocarbon analogs. However, acridines and benzacridines fluoresce
strongly with well-resolved vibrational
fine structure. Examination of the gas
chromatographic fractions showed an
emission very characteristic of acridine
or benzacridine (Figure 5 ) in all

\
\

'-v

400

300

:\,

2(

WAVELENGTH (H?)

Figure 5. Fluorescence and excitation spectra of


gas chromatographic fraction No. 4 of a basic nitrogen
extract of a 430-650" F. petroleum fraction

fractions. These compounds were expected in only the last several fractions.
Because this fluorescence occurred in all
fractions at about the same intensity, it
was concluded that the acridines were
bleeding from the column as a result of
the sample having been run several
times before trapping was done. These
data emphasize the fact that high-boiling

polar compounds tend to tail or be retained in a gas chromatographic column


causing contamination of subsequent
trapped fractions. The retention and
subsequent bleeding by high-boiling
polar compounds may not be as serious
a problem when using less sensitive
techniques such as infrared spectrophotometry.
VOL. 38, NO. 1, JANUARY 1966

17

OC PEAK 2

GC PEAK 7

----wc.
-EXC.

a10ql

OC PEAKS 9 & 10

A -EXC.

---EXC. 250
288

298W

WAVELENWCK @)

Figure 7. Phosphorescence spectra of trapped gas chrornatographic fractions of product from catalytic hydrogenation of
quinoline

TIME (MIN.)
Figure 6. Gas chromatogram of nitrogen compounds
from catalytic hydrogenationof quinoline
Trapped fractions are indicated by number

Infrared and NMR spectra of this


basic nitrogen fraction showed a
relatively high methyl rather than
methylene group content, indicating a
high degree of short cha.in or methyl
group substitution. The infrared spectrum, in the hydrogen out-of-plane bending region, suggested very little substitution on the nonheterocyclic rings.
The XMR spectrum indicated almost
complete substitution of the a-hydrogen
by alkyl groups (or fused naphthene
rings).
NITROGENCOMPOUNDS
FROM CATALYTIC HYDROGENATION
OF QUINOLINE.
Phosphorescence proved invaluable in

Table VI.
Lifetime for Phosphorescence of Trapped Fractions

Basic Nitrogen Extra.& of 430-650' F.


Fraction
Mean lifetime for
GC
phosphorescence, seconds
fraction Excitation: 265 mp 290 mp
No. Emission:
4 0 5 m ~ 500mp
0.8

1.2
1.0
1.1
1.4

1
2
3
4

0.9

0.8
1.1

1.3
1.3
1.4

5
6
7
8
9

1.2
1.2

1.5
1.2
1.4

13
14
15

1.2

12

Total sample

1.2

1.4

1.3

1.3

Reference compounds
Quinoline
No.emis2-Methylpyridine

sion
1. 3

PMethylpyridine

1.7

2,4,6-Trimethylpyridine

1.8

18

ANALYTICAL CHEMISTRY

n.6

0.3

13

0.6
1.0
15
0.7
Intensity of emission from pyridines
and tetrahydroquinolines.
b Intensity of emission from quinolines,
14

gen compounds is presented in Table


From this information, the
reaction paths below were deduced,

VIII.

catalytic
alkyl chain
transfer

catalytic

J.
Zmethylquinoline
2-ethylquinoline
Zisopropylquinoline
I

catalyst

0.8

No. emission
No.emission
No.emission
~

11
12

hydrogen
(by transfer)

1.2

1.3
1.2
1.1
1.2

Basic Nitrogen Extract of 430-650' F.


Petroleum Fraction
Corrected rel.
intensity ratio
GC
405 m p (Exc. at 265 mpc)"
fraction
No.
500 mp (Exc. at 302 mp)*
1
31.8
2
17.6
3
19.9
4
3.2
5
1.2
6
1.0
7
0.9
8
0.9
9
1.1
10
0.8

uinoline

1.2
1.2
1.0
1.1

1.4
1.2

10
11

characterizing a nitrogen compound


concentrate from catalytic hydrogenation of quinoline. This mixture was
complex as shown by the chromatogram
(Figure 6) obtained on a '/cinch S.E. 30
column using a small sample (injected
1 pl.). Larger samples (ca. 25 pl.) and
larger columns (0.25-inch) were required for infrared spectral study of
trapped fractions. Under these conditions, the polar nitrogen compounds
were poorly resolved, particularly the
smaller components appearing near the
larger ones. The greater sensitivity of
the phosphorescence technique permitted the use of small samples (5 pl.)
for improved resolution during trapping
from the 0.25inch column.
Compound type identification was
easily accomplished from the phosphorescence spectra. Choice of excitation wavelengths permitted identification of several compound types even
when they appeared in the same trapped
fraction. Certain components too small
to be detected by infrared spectrophotometry were observed by phosphorescence. By combining the phosphorescence and infrared spectral data
with relative retention data (expressed

Table VII. Intensities for the Tetra.


hydroquinolines or Pyridines Relative
to the Quinolines in Trapped Fractions

as boiling points), the majority of the


compounds were identified.
Some
typical
phosphorescence
spectra
exemplifying the compound types found
are combined in Figure 7. The auantitative distribution o?. the identified nitro-

ACKNOWLEDGMENT

The authors thank


Research
Laboratories, hmble oil &z Refining
CO., for permission to publish this
material.

The assistance of R. C. Cox in writing


the computer program, W. L. Senn, Jr.,
in obtaining the NMR spectra, and J. S.
Table VIII. Nitrogen Compounds Identified in Product from Catalytic Hydrogenation of Quinoline by Hydrogen
Transfer from Diluent

GC
peak
No.
1

2
3
4
5
6
7

8
9
10
11
12

Compound identified Wt., yo


Aniline
12.3
o-Toluidine
5.3
&Ethylaniline
1.2
N-Ethyl-o-toluidine
0.3
o-Propylaniline (and other
4.2
compounds)
Quinoline
47.8
2-Methylquinoline and
14.1
1,2,3,4tetrahydroquinoline
3-Methylquinoline
1.3
Indole
4.4
2-Ethylquinoline
4.4
2-Isopropylquinoline
2.6
Dimers and codimers of
2.1
partially hydrogenated
quinoline, alkylquinolines and indoles
100.0

Ellerbe and J. A. Hodgeson in obtaining


some of the luminescence spectra is
gratefully acknowledged. We also
thank J. B. Zachry for supplying the
sample from denitrogenation studies
and T. P. Hawes for assistance in obtaining some gas chromatograms.
LITERATURE CITED

(1) Adler, T. K., ANAL. CHEM.34, 685


(1962).
(2) Ball, J. S., Rall, H. T., Proc. A m .
Petrol. Inst. Sect. 111 42, 128 (1962).
(3) Becker, R. S., Ph.D. thesis, Florida
State University, Tallahassee, Fla.,
1955.
(4) Chen, M., Bibliography of Phosphorescent Molecules, American Instrument Co., Inc., 1960.
(5) Drushel, H. V., Sommers, A. L.,
Cox, R. C., ANAL. CHEM.35, 2166
(1963).
(6) Ermolaev,V. L., Opt. Spectry. (USSR)
English Transl.) 11, 266 (1961).
(7) Ermolaev, V. L., Kotlyar, I. P., Ibid.,
9, 183 (1960).
(8) Gurinovich, G. P., Sevchenko, A. N.,
Solovev, K. N., Ibid., 10, 396 (1961).
(9) Hartung, G. K., Jewell, D. M., Anal.
Chim. Acta 26, 514 (1962).
(10) Heckman, R. C., J . Mol. Spectry.
2, 27 (1958).
\ - - - - I

(11) Lewis, G. N., Kasha, M., J . A m .


Chem. SOC.66, 2100 (1944).
(12) Lewschin, W. L., 2. Physik 27, 368,
382 (1931).
(13) Lijinsky, W., Chestnut, A., Raha,
C. R., Chicago Med. School Quart. 21,
49 (1960).
(14) McClure, D. S., J . Chem. Phys. 17,
905 (1949).
(15) Parker, C. A., Rees, W. T., Analyst
87, 83 (1962).
(16) Schoental, R., Scott, E. J. Y., J.
Chem. Soc. (London)1949,1683.
(17) Shablya, A. V., Terenin, A. N., Opt.
Spectry. 10,324 (1961).
(18) Shimada, R., Spectrochzm. Acta 17,
14 (1961).
\----,.

(lG)Ibid., p. 30.
(20) Supplemental DBta Sheet to Aminco
Bulletin No. 2278, American Instrument Co., Inc., Silver Spring, Md.,
Atxi1 1959.
(21)Van Duuren, B. L., ANAL. CHEM.
32, 1436 (1960).
(22) Van Duuren, B. L., J . Org. Chem. 26,
2954 (1961).
(23) Yoshida, T., Igaku Kenkyu 27,
443 (1957); C . A . 52, 10721i (1958).

RECEIVED
for review February 18, 1965.
Accepted October 7, 1965. Presented at
the Symposium on Sulfur, Nitrogen, and
Oxygen
Compounds, Division of
Petroleum Chemistry, 149th Meeting,
ACS, Detroit, April 1965.

Isolation and Identification of Nitrogen


Compounds in Petroleum
H. V. DRUSHEL and A.

L. SOMMERS

Esso Research laboratories, Humble Oil & Refining Co., Baton Rouge, l a .

b A separation scheme for the isolation of nitrogen compounds from


petroleum was devised which provides
fractions of specific chemical classes.
Separation was achieved by use of
solid-liquid chromatography, chemical
extraction, and gas chromatography,
Included among the extractants were
sodium aminoethoxide in ethanolamine
and 7270 perchloric acid which were
used to isolate a variety of weakly
acidic compounds. Characterization
of the fractions from gas chromatography relied upon use of the usual
infrared, ultraviolet, and mass spectral
methods as well as sensitive fluorescence and phosphorescence techniques.
Application of this scheme to a light,
catalytic cycle oil resulted in identification of pyridines, quinolines, pyrindines, cyclopentaquinolines, indoles,
carbazoles, pyrroloquinones, phenols,
and other hydroxy compounds. Many
of these compounds are believed to be
responsible for the color, odor, and
gum-forming characteristics of heating
oils and other related petroleum
fractions.

70821

ITROGEN COMPOUNDS in petroleum

adversely affect many of the


important refining processes. They are
believed to reduce the activity of
cracking or hydrocracking catalysts because of their polarity and basicity.
It has also been suspected that nitrogen
compounds are to a great extent involved in gum formation, color formation, odor, and the poor storage properties of fuels. To overcome the deleterious effects of the nitrogen compounds
it is helpful to know the various types of
compounds that exist in petroleum
throughout different phases of refining.
Numerous studies have been made on
the nature of nitrogen compounds in
petroleum and several separation and
identification schemes have been reported. Hartung and Jewell (6) used
alumina adsorption, perchloric acid
extraction, and spectrophotometric examination to identify indoles, carbazoles, phenazines, and dibenzofuran
in a hydrogenated, catalytically-cracked
furnace oil. The same authors also
reported (7) the use of zinc chloride and
ferric chloride as complexing agents for

the isolation of nitriles from a hydrogenated furnace oil. Carbazole was


identified in Wilmington, Calif., petroleum by Helm and coworkers (9) using
distillation, adsorption, chemical treatment, gas chromatography, and spectrometry. As a means of identifying
nitrogen compounds in gas chromatographic effluents Thompson and coworkers (18) developed a catalytic
hydrogenation micromethod in which
the nitrogen free products are identified.
Snyder and Buell (16, 17) have used
linear elution adsorption chromatography and ion exchange resins to isolate
basic nitrogen compounds, carbazoles,
and other nitrogen compounds from
gasolines through cracked gas oils.
They used ultraviolet spectrophotometry to estimate indoles, carbazoles, and
benzcarbazoles in the isolated fractions.
Sauer and coworkers (15) applied
mass spectrometry to estimate carbazoles, indoles, pyrroles, pyridines, and
quinolines in concentrates isolated from
heating oils. La Lau (12) used mass
spectrometry to identify and estimate
the quantity of pyridines, quinolines,
VOL. 38, NO. 1, JANUARY 1966

19