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Which LC column should I use for Method 8330B explosives analysis?

12 Aug 2014

Over the years, Restek has run applications on a long list of columns for this analysis. It does require a primary and secondary column for analysis, since there is not one column that perfectly separates all compounds on the list simultaneously. Keep in mind that EPA Method 8330B allows for alternate columns versus the ones listed if you can demonstrate proficiency by presenting valid data that meets QC acceptance criteria as described in Method 8000. Our Innovations chemists have worked hard to make column selection easier for this and get you started in the right direction for optimization.

So far, we have determined optimal conditions for two pairs of columns that we recommend above all other combinations. For traditional HPLC systems with a 400 Bar pressure limit, we suggest using an Ultra C8 as the primary column and an Ultra Aromax as the secondary (confirmatory) column.

Here are chromatograms. 

Explosives by EPA 8330B on Ultra C8 with Trident Direct Guard Column

Explosives by EPA 8330B on Ultra C8 with Trident Direct Guard Column

LC_EV0526

Conditions

ColumnUltra C8 (cat.# 9103565)
Dimensions:150 mm x 4.6 mm ID
Particle Size:5 µm
Pore Size:100 Å
Temp.:30 °C
Standard/Sample
8330B Nitroaromatics and nitramine mix (cat.# 33204)
1,4-Dinitrobenzene (cat.# 33205)
Diluent:Deionized water
Conc.:20 µg/mL
Inj. Vol.:10 µL
Mobile PhaseWater:methanol (52:48)
Flow:1.2 mL/min
DetectorUV/Vis @ 210/254 nm
InstrumentShimadzu UFLCXR
NotesTrident guard cartridge holder cat.# 25083 was used to produce this chromatogram, but has since been discontinued. For assistance choosing a replacement for this application, contact Restek Technical Service or your local Restek representative.
– – – – – –
Trident direct 10 mm in-line guard cartridge holder cat.# 25083
Ultra C8 10 mm x 4 mm guard cartridge (cat.# 910350210)

This is the recommended primary column for a 2 column analysis. See LC_EV0527 for the confirmation column.

Explosives by EPA 8330B on Ultra Aromax with Trident Direct Guard

Explosives by EPA 8330B on Ultra Aromax with Trident Direct Guard

LC_EV0527

Conditions

ColumnUltra Aromax (cat.# 9127565)
Dimensions:150 mm x 4.6 mm ID
Particle Size:5 µm
Pore Size:100 Å
Temp.:35 °C
Standard/Sample
8330B Nitroaromatics and nitramine mix (cat.# 33204)
1,4-Dinitrobenzene (cat.# 33205)
Diluent:Deionized water
Conc.:20 µg/mL
Inj. Vol.:10 µL
Mobile PhaseWater:methanol (30:70)
Flow:1.2 mL/min
DetectorUV/Vis @ 210/254 nm
InstrumentShimadzu UFLCXR
NotesTrident guard cartridge holder cat.# 25083 was used to produce this chromatogram, but has since been discontinued. For assistance choosing a replacement for this application, contact Restek Technical Service or your local Restek representative.
– – – – – –
Trident direct 10 mm in-line guard cartridge holder cat.# 25083
Ultra Aromax 10 mm x 4 mm guard cartridge (cat.# 912750210)

This is the recommended confirmation column for a 2 column analysis. See LC_EV0526 for the primary column.

If you have a system that has a pressure limit of at least 600 Bar, we suggest our Raptor Biphenyl as the primary column and Raptor ARC-18 as the secondary column. Here are chromatograms.

Explosives on Raptor Biphenyl by EPA Method 8330B

Explosives on Raptor Biphenyl by EPA Method 8330B

LC_EV0531

Conditions

ColumnRaptor Biphenyl (cat.# 9309A65)
Dimensions:150 mm x 4.6 mm ID
Particle Size:2.7 µm
Temp.:35 °C
Standard/Sample
8330B Nitroaromatics and nitramine mix (cat.# 33204)
1,4-Dinitrobenzene (cat.# 33205)
Diluent:Water
Conc.:20 µg/mL
Inj. Vol.:5 µL
Mobile Phase
A:Water
B:Methanol
Time (min)Flow (mL/min)%A%B
0.001.03070
101.03070
DetectorUV/Vis @ 210/254 nm
Cell Temp:35 °C
Flow Cell Size:Semi-micro
InstrumentShimadzu Prominence
NotesThis is the recommended primary column for a two-column analysis. See LC_EV0530 for the confirmation column.

Explosives on Raptor ARC-18 by EPA Method 8330B

Explosives on Raptor ARC-18 by EPA Method 8330B

LC_EV0530

Conditions

ColumnRaptor ARC-18 (cat.# 9314A1E)
Dimensions:100 mm x 3.0 mm ID
Particle Size:2.7 µm
Temp.:35 °C
Standard/Sample
8330B Nitroaromatics and nitramine mix (cat.# 33204)
1,4-Dinitrobenzene (cat.# 33205)
Diluent:Water
Conc.:20 µg/mL
Inj. Vol.:3 µL
Mobile Phase
A:Water
B:Methanol
Time (min)Flow (mL/min)%A%B
0.001.07525
22.51.07525
DetectorUV/Vis @ 210/254 nm
Cell Temp:35 °C
Flow Cell Size:Semi-micro
InstrumentShimadzu UFLCXR
NotesThis is the recommended confirmation column for a two-column analysis. See LC_EV0531 for the primary column.

I hope this clarifies some things and gets you started in the right direction. Thank you for reading.

Products Mentioned


Colonna HPLC Ultra C8, 5 µm, 150 x 4,6 mm
Cartuccia per precolonna Ultra C8, 5 µm, 10 x 4,0 mm, 3 pz.
Colonna HPLC Ultra Aromax, 5 µm, 150 x 4,6 mm
Cartuccia per precolonna Ultra Aromax, 5 µm, 10 x 4,0 mm, 3 pz.
Colonna HPLC Raptor ARC-18, 2,7 µm, 100 x 3,0 mm
Colonna HPLC Raptor bifenilica, 2,7 µm, 150 x 4,6 mm
Miscela di esplosivi nitramminici e nitroaromatici 8330B con HPLC, 1000 µg/mL, acetonitrile, 1 mL/ampolla
Standard interno 8330 (1,4-dinitrobenzene) 2000 µg/mL, acetonitrile, 1 mL/ampolla
GNBL5554