cis-DCCA
(nd; 0.1);
DF, 8%
trans-DCCA
(nd;
0.1); DF, 7%
DBCA (nd; 0.1); DF,
11%
3PBA (nd; 0.1); DF,
80%
V: 3 mL
H: acidic (0.6 mL HCl;
90 min, 95
C)
E: LLE
C: LLE
I: GC-MS (ion trap, SIS)
LLE
• Extraction: 2
 4 mL hexane
• Re-extraction to 0.5 mL
0.1 M NaOH
• Re-extraction (0.1 mL
HCl + 2 mL hexane)
• Evaporation
– under stream
of nitrogen
• 10
μL HFIP, 15
μL DIC,
250
μL hexane
– 10 min, RT
• Wash: 1 mL 5% K
2 CO
3
• External quality
control: ClinCal
Urine Calibrator
[24]
HOCH2-FB-Al (0.01;
0.1); DF, 82%
MTFBL (0.05; 0.31);
DF, 54%
MMTFBL (0.05;
0.31); DF, 56%
FB-Al (0.02, 0.31);
DF, 84%
cis-DCCA
(0.03;
0.10); DF, 88%
trans-DCCA
(0.02;
0.10); DF, 100%
3PBA (0.01; 0.05);
DF, 100%
trans-CDCA
(0.06,
0.31); DF, 68%
CH3-FB-Ac (0.01;
0.31); DF, 38%
V: 2.5 mL
H: acidic (0.6 mL, 6 mol/L
HCl; 2 h, 106
C)
E: LLE (24-well plate;
Extrahera)
C: No
I: GC-MS/MS (triple
quadrupole, EI)
LLE
• Extraction: 3 mL and 2.5 mL
MTBE
• Organic phase divided into
two parts
• Evaporation under vacuum
(4
C in ice block)
Part I:
100
μL acetone + 50
μL BSTFA:
TMCS (99:1) 30 min, room temperature
• Wash with 1 mL NaHCO
3
(1 mol/L) + 100
μL iso-octane
Part II:
100
μL of ACN + 15
μL
HFIP + 10
μL DIC 30 min, room
temperature
• Wash with 0.5 mL NaHCO
3
(1 mol/L) + 100
μL iso-octane
• Population:
3-year-old children
(n
¼ 50), Japan
[8]
MTFBL (0.01; 0.04);
DF, nd
MMTFBL (0.02;
0.07); DF, nd
TMCA (0.04; 0.12);
DF, 0%
V: 25 mL
H: enzymatic
E: LLE
C: No
I: GC-MS (EI, SIM)
LLE
• Extraction: 2
 2 mL toluene
• Dehydration: 1 g Na
2 SO
4
• +1 mL toluene (washing
Na
2 SO
4 )
• Alcoholic group (MTFBL,
MMTFBL): 50
μL TMSI
(30 min, 70
C)
• Carboxylic acid: 30
μL
MTBSTFA (30 min, 70
C)
• Population: six
subjects without
occupational exposure, Japan
[20]
(continued)
Analytical Methods for Determination Urinary Metabolites of Synthetic. . .
65
(nd; 0.1);
DF, 8%
trans-DCCA
(nd;
0.1); DF, 7%
DBCA (nd; 0.1); DF,
11%
3PBA (nd; 0.1); DF,
80%
V: 3 mL
H: acidic (0.6 mL HCl;
90 min, 95
C)
E: LLE
C: LLE
I: GC-MS (ion trap, SIS)
LLE
• Extraction: 2
 4 mL hexane
• Re-extraction to 0.5 mL
0.1 M NaOH
• Re-extraction (0.1 mL
HCl + 2 mL hexane)
• Evaporation
– under stream
of nitrogen
• 10
μL HFIP, 15
μL DIC,
250
μL hexane
– 10 min, RT
• Wash: 1 mL 5% K
2 CO
3
• External quality
control: ClinCal
Urine Calibrator
[24]
HOCH2-FB-Al (0.01;
0.1); DF, 82%
MTFBL (0.05; 0.31);
DF, 54%
MMTFBL (0.05;
0.31); DF, 56%
FB-Al (0.02, 0.31);
DF, 84%
cis-DCCA
(0.03;
0.10); DF, 88%
trans-DCCA
(0.02;
0.10); DF, 100%
3PBA (0.01; 0.05);
DF, 100%
trans-CDCA
(0.06,
0.31); DF, 68%
CH3-FB-Ac (0.01;
0.31); DF, 38%
V: 2.5 mL
H: acidic (0.6 mL, 6 mol/L
HCl; 2 h, 106
C)
E: LLE (24-well plate;
Extrahera)
C: No
I: GC-MS/MS (triple
quadrupole, EI)
LLE
• Extraction: 3 mL and 2.5 mL
MTBE
• Organic phase divided into
two parts
• Evaporation under vacuum
(4
C in ice block)
Part I:
100
μL acetone + 50
μL BSTFA:
TMCS (99:1) 30 min, room temperature
• Wash with 1 mL NaHCO
3
(1 mol/L) + 100
μL iso-octane
Part II:
100
μL of ACN + 15
μL
HFIP + 10
μL DIC 30 min, room
temperature
• Wash with 0.5 mL NaHCO
3
(1 mol/L) + 100
μL iso-octane
• Population:
3-year-old children
(n
¼ 50), Japan
[8]
MTFBL (0.01; 0.04);
DF, nd
MMTFBL (0.02;
0.07); DF, nd
TMCA (0.04; 0.12);
DF, 0%
V: 25 mL
H: enzymatic
E: LLE
C: No
I: GC-MS (EI, SIM)
LLE
• Extraction: 2
 2 mL toluene
• Dehydration: 1 g Na
2 SO
4
• +1 mL toluene (washing
Na
2 SO
4 )
• Alcoholic group (MTFBL,
MMTFBL): 50
μL TMSI
(30 min, 70
C)
• Carboxylic acid: 30
μL
MTBSTFA (30 min, 70
C)
• Population: six
subjects without
occupational exposure, Japan
[20]
(continued)
Analytical Methods for Determination Urinary Metabolites of Synthetic. . .
65