
ACE UHPLC and HPLC Columns
Benzodiazepines from Drugs of Abuse Screen
(#AN2190)
Conditions
Column: ACE Excel 1.7 C18
Dimensions: 100 x 2.1 mm
Part Number: EXL -171-1002U
Mobile Phase: A: 5 mM ammonium
acetate in H2O
B: 5 mM ammonium
acetate in MeOH
Gradient: Time (mins) %B
0.0 10
10.0 90
11.9 90
13.4 10
15.5 10
Flow Rate: 0.3 mL/min
Injection: 10 μL
Temperature: 40 °C
Detection: MS Quattro Premier XE
triple quad
MRM, positive and
negative ESI mode
Desolvation
temperature: 450 °C
IonSource temperature:
150 °C
Collision gas pressure:
3.5 x10-3 mbar
18
20
Analytes
18. 7-Amino-clonazepam
(m/z 286.2 121.0)
20. 7-Amino-flunitrazepam
(m/z 284.2 135.0)
27. Bromazepam
(m/z 316.1 182.1)
29. Clonazepam
(m/z 316.1 270.1)
30. Nitrazepam
(m/z 282.2 236.1)
31. a-Hydroxytriazolam
(m/z 359.1 331.1)
32. Flunitrazepam
(m/z 314.2 268.2)
33. a-Hydroxyalprazolam
(m/z 325.2 297.1)
34. Estazolam
(m/z 295.2 267.2)
36. Triazolam
(m/z 343.0 308.1)
37. 2-Hydroxyethylflurazapam
(m/z 333.2 109.0)
38. Lorazepam
(m/z 321.1 275.1)
39. Oxazepam
(m/z 287.2 241.0)
40. Alprazolam
(m/z 309.2 281.2)
42. Temazepam
(m/z 301.1 255.1)
43. Nordiazepam
(m/z 271.1 139.9)
44. Midazolam
(m/z 326.2 291.2)
45. Diazepam-d5
(m/z 290.2 154.0)
46. Diazepam
(m/z 285.2 154.0)
47. Flurazepam
(m/z 388.2 315.1)
33, 34
31, 32
29, 30
27
Application #AN2370
36 - 40
44 - 47
4.4 4.6 4.8 5.0 5.2 5.4 5.6 5.8 6.0 6.2 6.4 6.6 6.8 7.0 7.2 7.4 7.6 7.8 8.0 8.2 8.4 8.6 8.8 9.0 9.2
Time - Minutes
100
%
0
Reproduced with permission of Biotage GB Ltd, UK
42
43
Biomarker Analysis for Gaucher Disease
by LC-MS/MS
Conditions
Glucosylsphingosine
0.2
0.4
0.6
0.8
1.0
1.2
Gaucher Disease patient
0.2
0.4
0.6
0.8
1.0
1.2
Gaucher Disease patient
1.2
1.4
1.4
1.4
1.6
1.6
1.6
1.8
1.8
1.8
Time - Minutes
Application #AN3490
Analytes
1. Glucosylsphingosine
(m/z 462 282)
2. Lyso-Gb2 (IS)
(m/z 624 282)
Column: ACE 3 C8
Dimensions: 50 x 2.1 mm
Part Number: ACE-112-0502
Mobile Phase: A: 50 mM formic acid in H2O
B: 50 mM formic acid in
MeCN/acetone (1:1 v/v)
Gradient: Time (mins) %B
0.0 5
4.0 66
4.1 100
5.1 100
5.9 5
Flow Rate: 0.9 mL/min
Injection: 5 μL
Temperature: 60 °C
Detection: API 4000 triple quad MS
ESI in positive ion mode
Temperature: 500 °C
Healthy control
1
2.6
1
2
2.6
1
2
2.8
2.8
2
0.2
1.6e4
1.4e4
1.2e4
1.0e4
8000.0
6000.0
4000.0
2000.0
0.0
1.4e4
1.2e4
1.0e4
8000.0
6000.0
4000.0
2000.0
0.0
0.4
0.6
0.8
1.0
2.0
2.0
2.0
2.2
2.2
2.2
2.4
2.4
2.4
2.6
2.8
3.0
3.0
3.0
3.0e5
2.8e5
2.6e5
2.4e5
2.2e5
2.0e5
1.8e5
1.6e5
1.4e5
1.2e5
1.0e5
8.0e4
6.0e4
4.0e4
2.0e4
0.0
Rolfs A, Giese AK, Grittner U, Mascher D, Elstein D, et al. (2013)
Glucosylsphingosine Is a Highly Sensitive and Specific Biomarker for Primary
Diagnostic and Follow-Up Monitoring in Gaucher Disease in a Non-Jewish,
Caucasian Cohort of Gaucher Disease Patients. PLoS ONE 8(11): e79732.
doi:10.1371/journal.pone.0079732
Intensity, cps Intensity, cps Intensity, cps
46
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