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2007.01
Clinical/Forensics
Allure® Biphenyl Columns (USP L11)
+MRM transitions per compound to verify com-
pound identity at approximately 1ng on-column.
Table 1 shows the +MRM transitions and the
source conditions for approximately 1ng each of
several cannabinoid metabolites.
Based on this work, we conclude an Allure®
Biphenyl column, coupled with an API MS/MS
3200 detector and a Shimadzu LC20AD
Prominence, can be used to quantify low levels of
cannabinoid analytes from underivatized sample,
and can achieve baseline separation of
Δ
9
-THC and
cannabidiol, in less than 5 minutes.
References:
1 Abbara, C., R. Galy, A. Benyamina, M. Reynaud and L. Bonhomme-
Faivre,
Development and validation of a method for quantitation of
Δ
9
-tetrahydrocanabinol in human plasma by high performance liquid
chromatography after solid phase extraction
J. Pharma. Biomed. Anal.
41 (2006) 1011-1016.
2 Sellers, K.
Reliably Confirm Cannabinoids by GC/MS
Restek
Advantage 2006.04 (2006) 16-17.
3 Weinmann, W., S. Vogt, R. Goerke, C. Muller and A. Bromberger,
Simultaneous determination of THC-COOH and THC-COOH-glucuronide
in urine samples by LS/MS/MS
Forens. Sci. Intl. 113 (2000) 381-387.
Reference 2 available from Restek – request lit. cat.# 580120.
Figure 2
Fast, selective separation of
Δ
9
-THC and its metabolites, using
an Allure® Biphenyl HPLC column (extracted ion chromatography).
LC_PH0424
Table 1
MRM transitions for THC and metabolites: multiple transitions
are monitored for each compound for definitive identifications.
Analyte
Q1 Mass Q3 Mass Time (ms)
DP (V)
EP (V)
CE (V) CXP (V)
Hydroxy-THC (MRM1)
331.2
313.1
100
36
5
21
10
Hydroxy-THC (MRM2)
331.2
193.1
100
36
5
35
6
Carboxy-THC (MRM1)
345.2
327.0
100
41
4.5
21
10
Carboxy-THC (MRM2)
345.2
299.3
100
41
4.5
25
6
Cannabidiol (MRM1)* 315.2
193.2
100
36
4.5
31
6
Cannabidiol (MRM2)* 315.2
123.2
100
36
4.5
43
6
Cannabinol (MRM1)
311.2
223.0
100
46
8.5
27
8
Cannabinol (MRM2)
311.2
222.5
100
46
8.5
37
10
Δ
9
-THC (MRM1)*
315.2
193.2
100
41
4.5
33
6
Δ
9
-THC (MRM2)*
315.2
123.1
100
41
4.5
43
6
Δ
9
-THC-d3 (MRM1)
318.3
196.3
100
36
4.5
31
6
Δ
9
-THC-d3 (MRM2)
318.3
123.2
100
36
4.5
43
6
*Note, cannabidiol and
Δ
9
-THC share the same transitions, but are separated chromatographically.
DP – declustering potential, EP – entrance potential, CE – collision energy, CXP – collision cell exit potential
Sample:
Inj.:
10µL
Conc.:
100ng/mL each component
Sample diluent: mobile phase
Column:
Allure
®
Biphenyl
Cat. #:
9166332
Dimensions:
30 x 2.1 mm
Particle size:
3µm
Pore size:
60Å
Conditions:
Mobile phase:
A: 0.1% formic acid in water
B: 0.1% formic acid in methanol
Time (min.) %B
0.5
50
2.5
90
5.0
90
6.0
50
6.1
—
Flow:
0.5mL/min.
Temp.:
ambient
Det.:
Spectrum Model
(Sciex API 3200 Triple Quadrupole)
Interface:
ESI
Ion mode:
positive
Temp.:
500°C
Ion source:
5500V
1. 11-hydroxy-
Δ
9
-tetrahydrocannabinol
2. cannabidiol
3. 11-nor-9-carboxy-
Δ
9
-tetrahydrocannabinol
4. cannabinol
5.
Δ
9
-tetrahydrocannabinol
6.
Δ
9
-tetrahydrocannabinol-d
3
Individual
Compound
CAS# cat.#
price
Cannabinoid & Metabolites
cannabidiol
13956-24-1 34011
$23
cannabinol
521-35-7
34010
$23
Δ
9
-THC
1972-08-3 34067
$33.00
±11-nor-9-carboxy-
Δ
9
-THC 104874-50-2 34068
$35.00
Exempted Drug of Abuse
Reference Materials
1,000µg/mL in P&T methanol, 1mL/ampul
No datapacks available.
3µm Column, 2.1mm
cat. #
price
30mm
9166332
$379
50mm
9166352
$379
100mm
9166312
$405
30mm (with Trident
™
Inlet Fitting)
9166332-700
$394
50mm (with Trident
™
Inlet Fitting)
9166352-700
$394
100mm (with Trident
™
Inlet Fitting)
9166312-700
$420
ordering
note
For guard cartridges for these columns, visit our website at
www.restek.com.
thank
you
Instrument provided courtesy
of Applied Biosystems
www.appliedbiosystems.com