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

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

LC_EV0526

Peaks

	Peaks
1.	HMX
2.	RDX
3.	1,3,5-Trinitrobenzene
4.	1,4-Dinitrobenzene (SS)
5.	1,3-Dinitrobenzene
6.	Nitrobenzene
7.	3,5-Dinitroaniline
8.	Tetryl
9.	2,4,6-Trinitrotoluene

	Peaks
10.	Nitroglycerin
11.	2,4-Dinitrotoluene
12.	4-Amino-2,6-dinitrotoluene
13.	2,6-Dinitrotoluene
14.	2-Amino-4,6-dinitrotoluene
15.	2-Nitrotoluene
16.	4-Nitrotoluene
17.	3-Nitrotoluene
18.	PETN

Conditions

Column	Ultra 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 Phase	Water:methanol (52:48)
Flow:	1.2 mL/min

Detector	UV/Vis @ 210/254 nm
Instrument	Shimadzu UFLCXR
Notes	Trident 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.

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

LC_EV0527

Peaks

	Peaks
1.	HMX
2.	RDX
3.	1,3,5-Trinitrobenzene
4.	1,4-Dinitrobenzene (SS)
5.	1,3-Dinitrobenzene
6.	Nitrobenzene
7.	3,5-Dinitroaniline
8.	Tetryl
9.	2,4,6-Trinitrotoluene

	Peaks
10.	Nitroglycerin
11.	2,4-Dinitrotoluene
12.	4-Amino-2,6-dinitrotoluene
13.	2,6-Dinitrotoluene
14.	2-Amino-4,6-dinitrotoluene
15.	2-Nitrotoluene
16.	4-Nitrotoluene
17.	3-Nitrotoluene
18.	PETN

Conditions

Column	Ultra 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 Phase	Water:methanol (30:70)
Flow:	1.2 mL/min

Detector	UV/Vis @ 210/254 nm
Instrument	Shimadzu UFLCXR
Notes	Trident 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.

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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

LC_EV0531

Peaks

	Peaks
1.	HMX
2.	RDX
3.	1,3,5-Trinitrobenzene
4.	1,4-Dinitrobenzene
5.	1,3-Dinitrobenzene
6.	3,5-Dinitroaniline
7.	Nitrobenzene
8.	Nitroglycerin
9.	Tetryl

	Peaks
10.	2,4,6-Trinitrotoluene
11.	2-Amino-4,6-dinitrotoluene
12.	4-Amino-2,6-dinitrotoluene
13.	2,4-Dinitrotoluene
14.	2,6-Dinitrotoluene
15.	2-Nitrotoluene
16.	4-Nitrotoluene
17.	3-Nitrotoluene
18.	PETN

Conditions

Column	Raptor 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 0.00 1.0 30 70 10 1.0 30 70

Detector	UV/Vis @ 210/254 nm
Cell Temp:	35 °C
Flow Cell Size:	Semi-micro
Instrument	Shimadzu Prominence
Notes	This is the recommended primary column for a two-column analysis. See LC_EV0530 for the confirmation column.

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Explosives on Raptor ARC-18 by EPA Method 8330B

LC_EV0530

Peaks

	Peaks
1.	HMX
2.	RDX
3.	1,3,5-Trinitrobenzene
4.	1,4-Dinitrobenzene
5.	1,3-Dinitrobenzene
6.	3,5-Dinitroaniline
7.	Nitrobenzene
8.	Nitroglycerin
9.	Tetryl

	Peaks
10.	2,4,6-Trinitrotoluene
11.	2-Amino-4,6-dinitrotoluene
12.	4-Amino-2,6-dinitrotoluene
13.	2,4-Dinitrotoluene
14.	2,6-Dinitrotoluene
15.	2-Nitrotoluene
16.	4-Nitrotoluene
17.	3-Nitrotoluene
18.	PETN

Conditions

Column	Raptor 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 0.00 1.0 75 25 22.5 1.0 75 25

Detector	UV/Vis @ 210/254 nm
Cell Temp:	35 °C
Flow Cell Size:	Semi-micro
Instrument	Shimadzu UFLCXR
Notes	This is the recommended confirmation column for a two-column analysis. See LC_EV0531 for the primary column.

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