低臭素化ダイオキシン類の 異性体分析 兵庫県立健康環境科学研究センター 中野 武
現状 PCDD/DF分析法は、毒性係数の示されている17異性体、4～8塩素化DD/DFのみを対象
低塩素化ダイオキシン類 の異性体分析 ● SP-2331 ● DB-DIOXIN ● DB5-MS
Column; SP-2331 and DB-DIOXIN Polarity: DB-DIOXIN < SP-2331 DB-Dioxin 44% methyl, 28% phenyl, 20% cyanopropyl 8% polyoxyethylene SP-2331 90% biscyanopropyl 10% phenylcyanopropyl
Standard: a) Authentic standards: 13, 27, 23, 36, 28, 26, 34, 46-DiCDFs Authentic standards of 136,138,146,147,149,236,238,246,267,346 -TrCDFs
b) Synthesis of PCDD by pyrolysis phenols in presence of KOH at temperature between 250C-300C c) Synthesis of PCDF by pyrolysis of respective Phenols in pyrex ampoule (370-400C, 15-30min) d) Synthesis of PCDF by pyrolysis of PCB in pyrex ampoule (400-450C, 15min)
PCDF formation from chlorophenols
Synthesis PCDF Synthesis of single isomers (in one homolog): 2-chloroPhenol : 4,6-DiCDF Synthesis of mixtures 3,4-dichloroPhenol (result in 1,2- and 2,3-fragment) 3-chloroPhenol (result in 1- and 3-fragment)
4-MCP 1,2,8-TrCDF 2,3,8-TrCDF 3,4-DCP
Synthesis of 1,4,X-T3CDF isomers from chlorophenol 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
Authentic standard NMR 1,4,7-TrCDF Cl1 H9 H8 H2 Cl7 H3 H6 Cl4 H2 ; H3 H6 H9 H8 NMR 1,4,7-TrCDF
Synthesis PCDD Synthesis of single isomers 1-MCDD, 2-MCDD, 12-DCDD, 13-DCDD, 23-DCDD, 123-TCDD, 124-TCDD, 236-TCDD, 237-T3CDD
Synthesis PCDD Most cases simple PCDD mixtures with two isomers (smiles rearangement): 2,3+2,3,4 result in 1,2,6- and 1,2,9-T3CDD Assignment in mixture of two isomers: With the same ring fragment substitution the more polar compound elutes later.
Chromatogram on Columns; Comparison Retention Behavior Chromatograms - DB-Dioxin - SP2331 column
Ambient air SP-2331
GC/MS-SIM chromatogram of PCDF DB-DIOXIN SP-2331 D2CDF 19 13 D2CDF 13 46 T3CDF T3CDF 346 137 129 137 GC/MS-SIM chromatogram of PCDF
GC/MS-SIM chromatogram of PCDD DB-DIOXIN SP-2331 D2CDD 19 13 13 D2CDD 19 T3CDD 137 129 T3CDD 129 137 GC/MS-SIM chromatogram of PCDD
Assignment for MCDD/F to T3CDD/F
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
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 15 16 T3CDF 28 8 10 23 21
R.T. Behavior : Substitution Fragments SP-2331 and DB-DIOXIN PCDF Elution order on DB-DIOXIN 2,4-, 3,7- isomers : earlier 1,2-, 1,6-, 1,9- isomers : later compare with SP-2331
GC/MS-Chromatogram of D2CDF DB-Dioxin SP-2331 24 12 SP-2331 37 19 16
GC/MS-Chromatogram of T3CDF DB-Dioxin SP-2331 SP-2331 149 129 139
R.T. Behavior : Substitution Fragments SP-2331 and DB-DIOXIN PCDD Elution order on DB-DIOXIN 1,4- isomers : earlier compare with SP-2331
GC/MS-Chromatogram of D2CDD DB-Dioxin 14 16 D CDD 2 SP-2331 DB-Dioxin 19 13 27/28 18 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
GC/MS-Chromatogram of T3CDD DB-Dioxin 147 146
低臭素化ダイオキシン類 の異性体分析 ● SP-2331 (95%cyanopropyl 5%phenyl-siloxane) 6 CN O Si C H 3 6 CN x y ● SP-2331 (95%cyanopropyl 5%phenyl-siloxane) ● BPX-35 (35%phenylpolysilphenylene-siloxane)
Standards. For the PBDF mixture all three monobromophenol isomers, Phenol and 2, 4-DBP were pyrolysed at 370ºC (15 minutes) in the presence of trace amounts of CuBr2. Authentic standards 2,7-D2BDF, 2,8-D2BDF, 2,3,8-T3BDF,
低臭素化DD/DF異性体について、市販標品のRT、標品混合物中の異性体予測生成比、異性体の溶出順序の規則性から、推定した。 使用した2-/3-/4-ブロモフェノールの量を反映し、2->1-,3->4-(4:2:2:1)であった。 DiBrDFの場合、24- 以外に28-, 18-, 27- , 17-, 19-, 26-, 37- , 16-, 36- , 46-が含まれ、 TrBrDFは、248- , 168- , 247- , 246- 、 TeBrDFの場合、2468-が主要な異性体である。 低臭素化DD/DF異性体について、市販標品のRT、標品混合物中の異性体予測生成比、異性体の溶出順序の規則性から、推定した。
M1BrDF～T3BrDFのGC/MS-SIMクロマトグラム 46 17 13 18 16 37 24 27 36 28 26 247 248 246 346 238 138/136 137 168 1 3 2 4 M1BrDF D2BrDF T3BrDF 347/236 237/149 M1BrDF～T3BrDFのGC/MS-SIMクロマトグラム SP2331 ; 60m x 0.32mmid. 0.20um; 120°C (1 min.), 20°C/min. to 200°C, 2°C /min. to 260°C (30min).; He 1.2mL/min.
BPX-35 ; 30m x 0.32mm id. 0.25um; 120°C (2 min.), 20°C/min. to 180°C, 10 11 Retention Time (min) 2726152 Intensity M1BDF / Average 13 14 15 503 14887 BrClDF / Average 16 17 18 19 792528 D2BDF / Average 1 3 2 4 13 46 24 27 28 26 17 18 BPX-35 ; 30m x 0.32mm id. 0.25um; 120°C (2 min.), 20°C/min. to 180°C, 2°C /min. to 220°C, 10°C /min. to 300°C (10min).; He 1.0mL/min.
BPX-35 ; 30m x 0.32mm id. 0.25um; 120°C (2 min.), 20°C/min. to 180°C, 26 27 28 29 Retention Time (min) 312320 Intensity T3BDF / Average 31 32 33 87023 T4BDF / Average BPX-35 ; 30m x 0.32mm id. 0.25um; 120°C (2 min.), 20°C/min. to 180°C, 2°C /min. to 220°C, 10°C /min. to 300°C (10min).; He 1.0mL/min.
Standards. The PBDD standards were synthesized by pyrolysis of bromophenols in Pyrex ampoules. The PBDD was synthesized by condensation of 2-monobromo phenol, 2, 4- and 2, 6- dibromophenol (DBP) and 2, 4, 6-tribromophenol (TBP) at 350ºC (30 min). Authentic standards: 2,7/2,8-D2BDD, 2,3,7-T3BDD.
合成した臭素化DD標品混合物は、 DiBrDDは、13-、27/28- 、 TrBrDDは、137-, 138- 、 136-, 139- TeBrDDは、1368-, 1379- が含まれていた。
M1BrDD～T3BrDDのGC/MS-SIMクロマトグラム 138 124 237 129 136 137 2 1 13 19 12 27/28/23 18 16 14/17 M1BrDD D2BrDD T3BrDD M1BrDD～T3BrDDのGC/MS-SIMクロマトグラム SP2331 ; 60m x 0.32mmid. 0.20um; 120°C (1 min.), 20°C/min. to 200°C, 2°C /min. to 260°C (30min).; He 1.2mL/min.
BPX-35 ; 30m x 0.32mm id. 0.25um; 120°C (2 min.), 20°C/min. to 180°C, 10.4 10.8 Retention Time (min) 4457267 Intensity M1BDD / Average 14 15 16 524 9893 BrClDD / Average 17 18 19 2389477 D2BDD / Average 2 1 13 27/28 BPX-35 ; 30m x 0.32mm id. 0.25um; 120°C (2 min.), 20°C/min. to 180°C, 2°C /min. to 220°C, 10°C /min. to 300°C (10min).; He 1.0mL/min.
BPX-35 ; 30m x 0.32mm id. 0.25um; 120°C (2 min.), 20°C/min. to 180°C, 27 28 29 Retention Time (min) 1307231 Intensity T3BDD / Average 31.0 31.5 32.0 32.5 191064 T4BDD / Average 1368 1379 BPX-35 ; 30m x 0.32mm id. 0.25um; 120°C (2 min.), 20°C/min. to 180°C, 2°C /min. to 220°C, 10°C /min. to 300°C (10min).; He 1.0mL/min.
BrDF>>BrClDF＞BrCl2DF、 BrDD>>BrClDD＞BrCl2DD 排ガス試料 PCB濃度が高い場合、 DiBrDF(323.8785, 325.8765) は PeCB(323.8834,325.8805)の妨害を受ける。 臭素系DD/DF は BrDF>>BrClDF＞BrCl2DF、 BrDD>>BrClDD＞BrCl2DD
PCB濃度が高い場合、DiBrDF(323.8785, 325.8765) はPeCB(323.8834,325.8805)の妨害を受ける。 16 20 24 Retention Time (min) 649323 568627 D2BrDF STD mix Sample 27 28 #118 #105 #114 GC/MS-SIMクロマトグラム(D2BrDF) SP2331 ; 60m x 0.32mmid. 0.20um; 120°C (1 min.), 20°C/min. to 200°C, 2°C /min. to 260°C (30min).; He 1.2mL/min.
臭素系DD/DF はBrDF>>BrClDF＞BrCl2DF、BrDD>>BrClDD＞BrCl2DD 14 16 18 20 Retention Time (min) 171522 86858 BrClDF Sample-B Sample-A GC/MS-SIMクロマトグラム(BrClDF) SP2331 ; 60m x 0.32mmid. 0.20um; 120°C (1 min.), 20°C/min. to 200°C, 2°C /min. to 260°C (30min).; He 1.2mL/min.