Page%201%20Analysis%20of%20Low%20Chlorinated%20PCDD/F%20-%20Isomer%20Specific%20Analysis%20of%20%20MCDD/MCDF%20to%20T3CDD/T3CDF%20on%20DB-Dioxin-Column%20

1

• Analysis of Low Chlorinated PCDD/F
• Isomer Specific Analysis of
• MCDD/MCDF to T3CDD/T3CDF
• on DB-Dioxin-Column

T.NAKANO and R.WEBER
Hyogo Pref. Inst. of Public Health Environ.
Sci. Inst. of Organic Chem., Univ. of Tuebingen
2
Errata
I found my fixed abstract was not received.
Vol.55, p.123-126
Table 1 and Figure 2 ( 2,3-D
2,8-D )
DCDD 13 28.51 27 29.57 23 29.57 28 29.73
DCDD 13 28.51 27 29.57 28 29.57 23 29.73
Roland Weber pointed out before, that the
2,7-D is not single authentic standard. (2,7-D
contains 2,8-D as impurity, and can not separate)
I am hating computer virus (Klez etc)
3
Introduction Relevance of isomer specific
analysis of T4CDD/F-OCDD/F
 Due to the high toxicity of PCDD/F substituted
in 2,3,7,8-position, a lot of attention was paid
to the isomer specific analysis of tetra- to
octachlorinated DD/DF in the last two decades.
Since, exclusively, the congeners substituted in
all four 2,3,7,8-position exhibit the dioxin
related toxicity, lower chlorinated congeners
were not assigned with TEQ values and, therefore,
not a lot of attention was paid to them.
4
Introduction Isomer specific analysis of low
chlorinated DD/F 
The analysis of low chlorinated isomers seems
interesting from several points of view
     For the investigation of the mechanisms of
PCDD/F formation/degradation, the low chlorinated
compounds offer valuable additional information.
5
For on-line measurement of PCDD/F in waste
incineration, some recent research projects
focused on estimating TEQ values by measurement
of low chlorinated PCDD/F as indicator compound.
To establish this correlation, it seems
important to measure not only the total amount of
the low chlorinated homologues but to calculate
and correlate specific isomers.
6
Furthermore, the low chlorinated DD/F may
give valuable information for environmental
samples. The analysis of isomer
distributions in environmental samples is the key
to estimating the origin of dioxins and related
compounds.
(Also in this field, many data are published for
T4CDD/F-OCDD/F, however, insufficient data are
available for mono- to tri-chlorinated
dioxin/furan.)
7
GC/MS Analysis. The analysis was carried out
using an HP 5890 II gas chromatograph connected
to a JMS-700 mass spectrometer (JEOL Ltd. Japan)
(operating at a resolution gt10 000).
8
SP-2331 and DB-DIOXIN
Column

• Polarity DB-DIOXIN lt SP-2331
• DB-Dioxin
• 44 methyl, 28 phenyl,
• 20 cyanopropyl
• 8 polyoxyethylene
• SP-2331
• 90 biscyanopropyl
• 10 phenylcyanopropyl

9
DB-Dioxin (60m, 0.25mm i.d., 0.15um, JW)
Temperature program used for isomer specific
separation of the MCDD/F-T3CDD/F on DB-Dioxin
column 50?C, 1 min. isothermal 20?C/min. to
150?C, 3?C /min. to 270?C, 14min. isothermal.
Carrier gas flow rate He 1.0mL/min.
Temperature program
10
SP-2331 (60m, 0.25mm i.d., 0.25um, SUPELCO)
Temperature program used for isomer specific
separation of the MCDD/F-T3CDD/F on SP-2331
column 120?C, 2 min. isothermal 30?C/min. to
200?C, 2?C/min. to 260?C, 6 min. isothermal.
Carrier gas flow rate He 1.0mL/min.
Temperature program
11
Standard a) Authentic standards commercially
available13, 27, 28, 23, 36, 26, 34, 46-DiCDFs
(8 isomers / 16 isomers)Authentic standards of
136,138,146,147,149,234,236,238,246,247,267,346
-TrCDFs(12 isomers / 28 isomers)
12
b) Synthesis of PCDD by pyrolysis of
chlorophenols in presence of KOH (250?C-300?C,
60min)
c) Synthesis of PCDF by pyrolysis of
chlorophenols (370-400?C, 15-30min)
d) Synthesis of PCDF by pyrolysis of PCB
(400-450?C, 15min)
13
Synthesis PCDFSynthesis of single isomers (in
one homolog) 2-chloroPhenol
4,6-DiCDFSynthesis of mixtures
3,4-dichloroPhenol (result in 1,2- and
2,3-fragment) 3-chloroPhenol (result in
1- and 3-fragment)
14
2-MCP
3-MCP
1,4,9-TrCDF
1,4,6-TrCDF
2,5-DCP
1,4,7-TrCDF
4-MCP
1,4,8-TrCDF
Authentic standard (1,4,7- NMR)
Synthesis of 1,4,X-T3CDF isomers from
chlorophenol
15
Synthesis PCDDSynthesis of single isomers
1-MCDD, 2-MCDD, 12-DCDD, 13-DCDD, 23-DCDD,
123-TCDD, 124-TCDD, 236-TCDD, 237-T3CDD
16
Most cases simple PCDD mixtures with two isomers
(smiles rearangement) 2,32,3,4 result in
1,2,6- and 1,2,9-T3CDD
Synthesis PCDD
1,2,6-D
1,2,9-D
2,3-CP2,3,4-DCP
Assignment in mixture of two isomers With the
same ring fragment substitutionthe more polar
compound elutes later.
17
DB-DIOXIN
SP-2331
D2CDD
19
13
13
D2CDD
19
T3CDD
137
129
T3CDD
129
137
GC/MS-SIM chromatogram of PCDD
18
Elution order for D2CDD to T3CDD
elutes earlier
elutes later
19
DB-DIOXIN
SP-2331
D2CDF
19
13
D2CDF
13
46
T3CDF
T3CDF
346
137
129
137
GC/MS-SIM chromatogram of PCDF
20
Elution order for D2CDF to T3CDF
elutes earlier
elutes later
21
SP-2331 and DB-DIOXIN
R.T. Behavior Substitution Fragments
PCDD

• Elution order on DB-DIOXIN
• 1,4- fragment elutes earlier
• compare with SP-2331

22
GC/MS-Chromatogram of D2CDD
DB-Dioxin

16

14
D
CDD

2
SP-2331

DB-Dioxin
19




13
18
27/28
12

17


23
14











28.0
28.4
28.8
29.2
29.6
30.0
30.4
30.8
31.2
31.6
32.0
23
GC/MS-Chromatogram of T3CDD
DB-Dioxin
147
146
24
SP-2331 and DB-DIOXIN
R.T. Behavior Substitution Fragments
PCDF

• Elution order on DB-DIOXIN
• 2,4-, 3,7- elutes earlier
• 1,2-, 1,6-, 1,9- elutes later
• compare with SP-2331

25
GC/MS-Chromatogram of D2CDF
DB-Dioxin
SP-2331
24
12
SP-2331
37
19
16
26
GC/MS-Chromatogram of T3CDF
DB-Dioxin
SP-2331
SP-2331
149
129
139
27
Separation of low chlorinated DD
The No. of separation peaks
isomer ULTRA-2 DB-5MS SP-2331 DB-DIOXIN
MCDD 2 2 2 2 2
D2CDD 10 5 4 8 9
T3CDD 14 6 5 12 12
28
Separation of low chlorinated DF
The No. of separation peaks
isomer ULTRA-2 DB-5MS
SP-2331 DB-DIOXIN MCDF 4 4 4
4 4 D2CDF 16 8 10
14 15 T3CDF 28 8 10 23
21
29
Conclusions a) The isomer specific analysis of
MCDD/MCDF to T3CDD/T3CDF was established on three
standard columns     SP-2331      DB
Dioxin     DB 5MS
30
Conclusionsb) The low chlorinated PCDD/DF can
be analysed together with T4CDD/F to OCDD/F ( one
GC/MS injection).c) The DB-Dioxin and the
SP-2331 column is effective for an isomer
specific analysis of low chlorinated PCDF as it
is requested for detailed mechanistic studies.
31
Acknowledgement The authors would like to
express their thanks to Dr. Brian Gullett and Dr.
Dennis Tabor(US-EPA) , Dr. Junji Ogakifor
donating some standards, to Terry Humphries for
critical reading of the manuscript.
32
See you next!Thank you for your attention.
33
Homologue distribution of PCDF in Ambient air
34
4-MCP
1,2,8-TrCDF
2,3,8-TrCDF
3,4-DCP
35
Authentic standard
Cl1
H9
H8
H2
Cl7
H3
H6
Cl4
H2 H3
H6
H9
H8
NMR 1,4,7-TrCDF
36
PCDF formation from chlorophenols
We can control the ratio of isomers by changing
the ratio of chlorophenol.
Case of 3-MCP 2-MCP if 2-MCP gt gt 3-MCP
46- gt 16- gt 36- gt 17- gt 37- gt 19-F if 3-MCP gtgt
2-MCP 17- gt 37- gt 19- gt 16- gt 36- gt 46-F
37
Ambient air
SP-2331
38
2,5-dichloroPhenol 3-chloroPhenol 1,4,9-
and 1,4,7- (1,4,9- , 1,4,7- authentic
standard)3-chloroPhenol and 3,4-dichlorophenol
1,2,7-, 1,2,9-, 1,7,8-, 2,3,7-
Assignment additional synthesis via PCB.
2,3,4' T3CB result in 1,2,7-T3CDF
39
Roland Weber pointed out before, that the
2,7-D is not single authentic standard. (2,7-D
contains 2,8-D as impurity, and can not
separate) As for 2,7-D2CDD, Basically the
position is 2,7-D should be presented as an
unknown ratio mixture of the two compounds
2,7-/2,8-D .
40
D2CDD
2,7-/2,8-D
13C-D2CDD
2,7-/2,8D
2,3-D
Dennis Tabor (EPA)
41
But, I believe the poor peak shape of my standard
for 2,3-DCDD has lead to an poor assignment. I
have 2,7 and 2,3 in my 13C standards and have
not gotten the poor peak shape from them. I
believe that the toluene or other solvent must
be causing you a problem. In my standard which
do not have toluene 2,7 comes out before 2,3. The
first two traces are the 252/254 of the native
DCDD. The second two traces are the 264/266 of
the 13C DCDD. The current GC/MS that I am using
is Low Res. Therefore the 13C 2,8-DCDF causes
me not to be able to see 1,4-DCDD.
42
Basically the low chlorinated PCDD/PCDF can be
analysed together with T4CDDF-OCDD/F. Behavior
on Alumina column. Low chlorinated isomers may
be oxidized during extensive Sulfuric acid
treatment.  
43
Assignment for MCDD/F to T3CDD/F
44
Elution order for D2CDD to T3CDD
45
Elution order for D2CDD to T3CDD
46
Elution order for D2CDF to T3CDF