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Acetaminophen in Calf Serum
Using 60mg 3mL HyperSep Retain PEP Extraction Column
(Part Number: 60107-203)
Sample Preparation
Dilute 10mg of acetaminophen in 100mL of water to give
a 100ppm solution
Mix 25mL of acetaminophen (100ppm) solution in a
50mL flask and dilute to volume with water
(standard solution 50ppm)
Mix 25mL of acetaminophen (100ppm) solution in a
50mL flask and dilute to volume with serum and 1%
H
3
PO
4
(sample 50% serum and 1% H
3
PO
4
)
Condition Retain PEP Extraction Column
2mL of methanol
2mL of DI H
2
O
Apply Sample
Load 2mL sample solutions
Wash Column
2mL of methanol/DI H
2
O (5/95, v/v)
Dry column (5 to 10 minutes at >10"Hg/full flow for
positive pressure manifold)
Elute Acetaminophen
2mL of methanol
Dry Eluate and Reconstitute
Evaporate to dryness at <50°C using nitrogen
Reconstitute sample using 1mL of mobile phase
Acetaminophen in Calf Serum
Using 60mg 3mL HyperSep Retain-CX Extraction Column
(Part Number: 60107-303)
Sample Preparation
Dilute 10mg of acetaminophen in 100mL of water to give
a 100ppm solution
Mix 25mL of acetaminophen (100ppm) solution in a
50mL flask and dilute to volume with water
(standard solution 50ppm)
Mix 25mL of acetaminophen (100ppm) solution in a
50mL flask and dilute to volume with serum and 1%
H
3
PO
4
(sample 50% serum and 1% H
3
PO
4
)
Condition Retain-CX Extraction Column
2mL of methanol
2mL of DI H
2
O
Apply Sample
Load 2mL sample solutions
Wash Column
2mL of methanol/DI H
2
O (5/95, v/v)
Dry column (5-10 minutes at >10"Hg/full flow for
positive pressure manifold)
Elute Acetaminophen
2mL of methanol
Dry Eluate and Reconstitute
Evaporate to dryness at <50°C using nitrogen
Reconstitute sample using 1mL of mobile phase
Pharmaceutical/Biotech
Applications
Acetaminophen Extraction and Cleanup from Hyclone
Using 60mg 6mL HyperSep Retain PEP and -CX Extraction Columns
(Part Number: 60107-203 and 60107-308)
Prepare Sample
Dissolve 10mg of Ethylamino-phenol in 100mL of
water (100ppm)
Dilute 25mL of Ethylamino-phenol solution to 50mL
with water
Dilute 25mL of Ethylamino-phenol solution to 50mL
with serum
Condition Column
1 x 2mL CH
3
OH
1 x 2mL H
2
O water 2mL
Apply Sample
Load 2mL of sample
Wash Column
1 x 2mL 5% CH
3
OH
Elute Sample
1 x 2mL CH
3
OH
Results
SPE Column
Recovery (%)
Pretreatment 1
Pretreatment 2
HyperSep Retain PEP + 50% Serum
58.5
40.3
HyperSep Retain-CX + 50% Serum
94.6
94.6