Labo farmaceutische analyse IV
Opdracht
Naam: …Smet
Yilska…………………………………………………………………………
Klasgroep: 3FBTb
Docent: M. Thierry …
Screening for drugs in saliva samples by LLE and LC-MS/MS
Purpose and procedure
The current procedure is intended for identification and quantification
of 32 different drugs and drugs of abuse in saliva samples. The
procedure is based on LLE and LC-MS/MS.
Target analytes
3-hydroxy-diazepam, 6-monoacetylmorphine (6-MAM), 7-
aminonitrazepam, 7-aminoflunitrazepam, alprazolam, amphetamine,
benzoylecgonine, bromazepam, buprenorphine, carisoprodol,
clonazepam, cocaine, codeine, diazepam, fenazepam, flunitrazepam,
lorazepam, lysergic acid diethylamide (LSD), 3,4-
methylenedioxyamphetamine (MDA), 3,4-
methylenedioxyethylamphetamine (MDEA), MDMA, meprobamat,
methamphetamine, methadone, morphine, N-desmethyldiazepam,
nitrazepam, oxazepam, THC, zolpidem and zopiclone.
Chemical structure and physicochemical properties of some of
the analytes
Amphetamine (A) Cocaine
MDMA Metamphetamine (MA)
, THC
A Cocaine MDMA MA THC
Log P 1.80 2.28 1.86 2.24 5.94
pKa 10.01 8.85 10.14 10.21 9.34
A, cocaine, MDMA and MA are basic substances, whereas THC is
slightly acidic.
Internal standard solution
Morphine-d3 (0.53 µmol/l), amphetamine-d3 (3.0 µmol/l),
metamphetamine-d11 (0.85 µmol/l), benzoylecgonine-d8 (0.31 µmol/l),
MDMA-d5 (0.77 µmol/l), 7-aminoflunitrazepam-d7 (0.018 µmol/l), N-
desmethyldiazepam-d5 (0.045 µmol/l and methadone-d9 (0.27 µmol/l)
Sample collection and pretreatment
A special oral fluid collection kit is used. The device consists of a
collector pad on a plastic handle and a collection vial that contains
0,8 ml of stabilizing buffer solution. The collector pad is treated with
NaCl, citric acid, sodium benzoate, potassium sorbate, gelatin, sodium
hydroxide, and deionized water. The collection vial contains
chlorhexidine digluconate, a blue dye, Tween 20 (nonionic
surfactant), and deionized water. The collector plate is whipped
between gum and cheek to stimulate the saliva production, and
sampling is performed for 2 minutes. After sampling, the collector pad
is placed in the collection vial for storage at +4°C. The volume of the
saliva collected by the pad is determined by weighing the pad before
and after sampling. The content of the pad is centrifuged and 0,5 ml
of the supernatant is transferred to a 5 ml polypropylene tube.
Sample preparation
Opdracht
Naam: …Smet
Yilska…………………………………………………………………………
Klasgroep: 3FBTb
Docent: M. Thierry …
Screening for drugs in saliva samples by LLE and LC-MS/MS
Purpose and procedure
The current procedure is intended for identification and quantification
of 32 different drugs and drugs of abuse in saliva samples. The
procedure is based on LLE and LC-MS/MS.
Target analytes
3-hydroxy-diazepam, 6-monoacetylmorphine (6-MAM), 7-
aminonitrazepam, 7-aminoflunitrazepam, alprazolam, amphetamine,
benzoylecgonine, bromazepam, buprenorphine, carisoprodol,
clonazepam, cocaine, codeine, diazepam, fenazepam, flunitrazepam,
lorazepam, lysergic acid diethylamide (LSD), 3,4-
methylenedioxyamphetamine (MDA), 3,4-
methylenedioxyethylamphetamine (MDEA), MDMA, meprobamat,
methamphetamine, methadone, morphine, N-desmethyldiazepam,
nitrazepam, oxazepam, THC, zolpidem and zopiclone.
Chemical structure and physicochemical properties of some of
the analytes
Amphetamine (A) Cocaine
MDMA Metamphetamine (MA)
, THC
A Cocaine MDMA MA THC
Log P 1.80 2.28 1.86 2.24 5.94
pKa 10.01 8.85 10.14 10.21 9.34
A, cocaine, MDMA and MA are basic substances, whereas THC is
slightly acidic.
Internal standard solution
Morphine-d3 (0.53 µmol/l), amphetamine-d3 (3.0 µmol/l),
metamphetamine-d11 (0.85 µmol/l), benzoylecgonine-d8 (0.31 µmol/l),
MDMA-d5 (0.77 µmol/l), 7-aminoflunitrazepam-d7 (0.018 µmol/l), N-
desmethyldiazepam-d5 (0.045 µmol/l and methadone-d9 (0.27 µmol/l)
Sample collection and pretreatment
A special oral fluid collection kit is used. The device consists of a
collector pad on a plastic handle and a collection vial that contains
0,8 ml of stabilizing buffer solution. The collector pad is treated with
NaCl, citric acid, sodium benzoate, potassium sorbate, gelatin, sodium
hydroxide, and deionized water. The collection vial contains
chlorhexidine digluconate, a blue dye, Tween 20 (nonionic
surfactant), and deionized water. The collector plate is whipped
between gum and cheek to stimulate the saliva production, and
sampling is performed for 2 minutes. After sampling, the collector pad
is placed in the collection vial for storage at +4°C. The volume of the
saliva collected by the pad is determined by weighing the pad before
and after sampling. The content of the pad is centrifuged and 0,5 ml
of the supernatant is transferred to a 5 ml polypropylene tube.
Sample preparation
