LC and LC/MS Applications

Environmental

Anions with Agilent HPLC Ion-Kit Catechol and Various Metabolites Separated on a
Wide-Pore Column

1. Catechol
2. Anthranilic acid
3. 2,3 Dihydroxybiphenyl diol
4. Carbazole

Sample: Anions
Column: Asahipak ODP- 50, 4 x 125 mm, 5 µm, (P/N: 79923OP-564)
Mobile phase: Water/acetonitrile, 86/14% + modifier (P/N: 5062-2480)
adjusted to pH 8.6 with carbonate free NaOH Sample: 15 µL, 250 - 500 µg/mL each compound
Flow rate: 1.5 mL/min Column: ZORBAX 300SB- C18, 4.6 x 150 mm, 5 µm, (P/N: 883995- 902)
Temperature: 40°C Mobile phase: A = H2O with 0.1% TFA to pH 3 with TEA
Detection: 266 nm B = ACN with 0.085% TF A
Gradient: 5- 60% in 20 min., Wash, 95%B for 2 min.
Flow rate: 1.5 mL/min
Temperature: 60°C
Detection: UV 233 nm

LC and LC/MS
Explosives Explosives (EPA 8330) using Hypersil BDS
1. Octogen 10. 2- Amino4,6 - Dinitrotoluene
5 ng 2,4-DNB

2. Nitroglycol 11. 4- Amino2,6 - Dinitrotoluene

3. Hexogen 12. 2,4- Dinitrotoluene
NB

12 ng Nitropenta

4. Nitrodiglycol 13. 2,6- Dinitrotoluene

5 ng 1-A-6-NT 1,2-DNB

10 ng TNT

5. 1,3,5- Trinitrobenzene 14. 2- Nitrotoluene

2,6-DNB

4 ng 3-NT
11 ng

6 ng 2-NT

6. 1,3- Dinitrobenzene 15. 4- Nitrotoluene

5 ng 4-NT
2-A-4,6-NT
4 ng 2-A-4,6-NT
5 ng Hexogen

6 ng Tetryl

214,1 nm
5 ng 1,3-DNB

7. Nitroglycerine 16. 3- Nitrotoluene

5 ng
1,3,5-TNB

8. Tetryl 17. Nitropenta

10 ng Octogen

9. 2,4,6 Trinitrotoluene 18. Diphenylamine

3 ng
6 ng 2,4,6-TNP

1,2-DNB

235,4 nm
18
5 15
91011 12 16
3 6 14
0 5 10 15 20 1 13 17 DAD1 B, Sig. = 215, 20 nm
Time (min) 2 4 78 Ref = 350, 50 nm

DAD1 A, Sig = 235, 20 nm

Sample: Explosive Compounds
Ref = 400, 80 nm
Column: Hypersil BDS- C18, 4 x 100 mm, 3 µm, (P/N: 79926OB-354)
Mobile phase: A = Water, B = Methanol (MeOH)
0 5 10 15 20 25 30 35 40 min.
Gradient: at 0.0 min 26% B
at 10 min 40% B
at 20 min 55% B Sample: Trace Level Explosives
at 30 min 70% B Column: Hypersil BDS - C18, 2 x 125 mm, 3 µm, (P/N: 79926BD - 362)
at 31 min 26% B Mobile phase: A = Water, B = Methanol
Flow rate: 0.72 mL/min Flow rate: 0.5 mL/min
Temperature: 38°C Temperature: 38°C
Detection: 214 nm and 235 nm Detection: UV 214+235 nm, see above

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