MOOC: Validation of liquid chromatography mass spectrometry (LC-MS) methods (analytical chemistry) course

Self test 2.7

HPLC analysis of one analyte in one sample and 5 calibration solutions was carried out.

Assess if the analyte presence in the sample can be confirmed by the retention time criterion set by SANTE.

Analysis	Ret. Time of analyte, min
Calibrant 1	4.298
Calibrant 2	4.413
Calibrant 3	4.211
Calibrant 4	4.368
Calibrant 5	4.183
Sample	4.122

Confirmed

Not confirmed

HPLC analysis of one analyte in one sample and 5 calibration solutions was carried out.

Assess if the analyte presence in the sample can be confirmed by the retention time criterion set by 2002/675/EC.

Analysis	Ret. Time of analyte, min	Ret. Time of ret.time standard compound, min
Calibrant 1	4.298	11.191
Calibrant 2	4.413	11.24
Calibrant 3	4.211	11.144
Calibrant 4	4.368	11.309
Calibrant 5	4.183	11.195
Sample	4.122	11.277

Not confirmed

Confirmed

LC with MS detection is perhaps the most reliable means for confirmation of analyte‘s identity. Please rank the following identity confirmation situations according to reliability (1 is the most reliable). The directive 2002/657/EC can be used as guidance.

LC retention time (k > 1) and a single MS/MS transition	4 Unselect 1 Unselect 2 Unselect 3 Unselect
LC retention time on two different columns (both have k > 1) and a single MS/MS transition (same transitions for both columns).	4 Unselect 1 Unselect 2 Unselect 3 Unselect
LC retention time (k > 1) and MS/MS with a single precursor ion with two product ions	4 Unselect 1 Unselect 2 Unselect 3 Unselect
LC retention time (k > 1), MS/MS with two transitions, of which one of the product ions is detected with HRMS	4 Unselect 1 Unselect 2 Unselect 3 Unselect