LIPID MAPS Certificate Of Analysis
Reagent Number / Lot Number:
RF0000000780 / LM39-095A
Mass Spectrometry Reference Standard
d5-TG Internal Standard Mixture I
4 μM each of compounds LMGL03010008 – LMGL03010016
in 1:1 toluene:methanol
Systematic
name(s):
1,3-di-(5Z,8Z,11Z,14Z,17Z-eicosapentaenoyl)-2-(4Z,7Z,10Z,13Z,16Z,19Z-docosahexaenoyl)-snglycerol-d5
1,3-ditetradecanoyl-2-(9Z-hexadecenoyl)-sn-glycerol-d5
1,3-dipentadecanoyl-2-(9Z-octadecenoyl)-sn-glycerol-d5
1,3-dihexadecanoyl-2-octadecanoyl-sn-glycerol-d5
1,3-diheptadecanoyl-2-(10Z-heptadecenoyl)-sn-glycerol-d5
1,3-dinonadecanoyl-2-dodecanoyl-sn-glycerol-d5
1,3-dieicosanoyl-2-(11Z-eicosenoyl)-sn-glycerol-d5
1,3-di-(11Z,14Z-eicosadienoyl)-2-(6Z,9Z,12Z-octadecatrienoyl)-sn-glycerol-d5
1,3-di-(5Z,8Z,11Z,14Z-eicosatetraenoyl)-2-(9Z,12Z-octadecadienoyl)-sn-glycerol-d5
Common
name(s):
1,3-eicosapentaenoyl-2-docosahexaenoyl-sn-glycerol-d5, (20:5-22:6-20:5 TG-d5)
1,3-dimyristoyl-2-palmitoleoyl-sn-glycerol-d5, (14:0-16:1-14:0 TG-d5)
1,3-dipentadecanoyl-2-oleoyl-sn-glycerol-d5, (15:0-18:1-15:0 TG-d5)
1,3-dipalmitoyl-2-stearoyl-sn-glycerol-d5, (16:0-18:0-16:0 TG-d5)
1,3-diheptadecanoyl-2-heptadecenoyl-sn-glycerol-d5, (17:0-17:1-17:0 TG-d5)
1,3-dinonadecanoyl-2-lauroyl-sn-glycerol-d5, (19:0-12:0-19:0 TG-d5)
1,3-diarachidoyl-2-eicosenoyl-sn-glycerol-d5, (20:0-20:1-20:0 TG-d5)
1,3-dieicosadienoyl-2-linolenoyl-sn-glycerol-d5, (20:2-18:3-20:2 TG-d5)
1,3-arachidonoyl-2-linoleoyl-sn-glycerol-d5, (20:4-18:2-20:4 TG-d5)
Molecular
weight(s):
20:5-22:6-20:5 TG-d5 = 976.47
14:0-16:1-14:0 TG-d5 = 754.23
15:0-18:1-15:0 TG-d5 = 810.34
16:0-18:0-16:0 TG-d5 = 840.40
17:0-17:1-17:0 TG-d5 = 852.41
19:0-12:0-19:0 TG-d5 = 840.40
20:0-20:1-20:0 TG-d5 = 978.65
20:2-18:3-20:2 TG-d5 = 938.51
20:4-18:2-20:4 TG-d5 = 932.46
Exact
Mass(es):
20:5-22:6-20:5 TG-d5 = 975.74
14:0-16:1-14:0 TG-d5 = 753.69
15:0-18:1-15:0 TG-d5 = 809.75
16:0-18:0-16:0 TG-d5 = 839.80
17:0-17:1-17:0 TG-d5 = 851.80
19:0-12:0-19:0 TG-d5 = 839.80
20:0-20:1-20:0 TG-d5 = 977.94
20:2-18:3-20:2 TG-d5 = 937.81
20:4-18:2-20:4 TG-d5 = 931.77
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Page 1 of 5
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LIPID MAPS Certificate Of Analysis
Reagent Number / Lot Number:
RF0000000780 / LM39-095A
Mass Spectrometry Reference Standard
d5-TG Internal Standard Mixture I
4 μM each of compounds LMGL03010008 – LMGL03010016
in 1:1 toluene:methanol
Product# /
Lot #:
LM-6000 / LM39-095A
Manufacture
date:
April 25, 2012
Storage
Conditions:
-16 to -24° C
Concentration
/ Amount(s):
Solvent:
20:5-22:6-20:5 TG-d5 = 3.92 µM / 3.82 µg per vial
14:0-16:1-14:0 TG-d5 = 3.97 µM / 3.00 µg per vial
15:0-18:1-15:0 TG-d5 = 3.98 µM / 3.23 µg per vial
16:0-18:0-16:0 TG-d5 = 3.91 µM / 3.29 µg per vial
17:0-17:1-17:0 TG-d5 = 3.94 µM / 3.36 µg per vial
19:0-12:0-19:0 TG-d5 = 3.98 µM / 3.35 µg per vial
20:0-20:1-20:0 TG-d5 = 3.97 µM / 3.88 µg per vial
20:2-18:3-20:2 TG-d5 = 3.97 µM / 3.72 µg per vial
20:4-18:2-20:4 TG-d5 = 3.99 µM / 3.72 µg per vial
1:1 (v/v) Toluene : Methanol
D
R1
D
Structural
Information:
D
R2
D
R3
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* R1, R2 and R3 are fatty acids
in described positions.
D
Page 2 of 5
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LIPID MAPS Certificate Of Analysis
Reagent Number / Lot Number:
RF0000000780 / LM39-095A
Mass Spectrometry Reference Standard
d5-TG Internal Standard Mixture I
4 μM each of compounds LMGL03010008 – LMGL03010016
in 1:1 toluene:methanol
Formulation of Mixture to contain 4 µM of each d5-TG
Working stocks of each compound were formulated and the concentration confirmed by
HPLC/MS/MS. A concentrated stock mixture was formulated at 40 µM and then
subsequently diluted to 4 µM(1:10 dilution) using volumetric glassware. Solvent is
filtered and degassed 1:1 (v/v) toluene : methanol. The 4 µM mixture was then packaged
as 1 ml / ampoule units.
Lot #
Stock conc
(μM)
Target
conc (μM)
Vol added
to 100 mL
LM-3208
LM5-059
347.7
40
11.51
LM-3207
LM26-125
419.0
40
9.55
LMGL03010010
LM-3209
LM5-060
304.0
40
13.16
840.4
LMGL03010011
LM-3210
LM5-066
333.3
40
12.00
852.4
LMGL03010012
LM-3211
LM5-062
337.7
40
11.85
19:0(2)-12:0 d5-TG
840.4
LMGL03010013
LM-3212
LM5-064
325.3
40
12.30
20:0(2)-20:1 d5-TG
978.7
LMGL03010014
LM-3213
LM5-063
336.7
40
11.88
20:2(2)-18:3 d5-TG
938.5
LMGL03010015
LM-3214
LM26-120
387.7
40
10.32
20:4(2)-18:2 d5-TG
932.5
LMGL03010016
LM-3215
LM26-122
403.3
40
9.92
Compound
M.W.
LM #
Product #
14:0(2)-16:1 d5-TG
754.2
LMGL03010009
20:5(2)-22:6 d5-TG
976.5
LMGL03010008
15:0(2)--18:1 d5-TG
810.3
16:0(2)-18:0 d5-TG
17:0(2)-17:1 d5-TG
HPLC/MS/MS was used to quantify the final concentration of each d5-TG in solution.
Three (3) ampoules were assayed in triplicate (n=9) against a dilution of a 40 μM
working stock mixture as calibrator.
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Page 3 of 5
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LIPID MAPS Certificate Of Analysis
Reagent Number / Lot Number:
RF0000000780 / LM39-095A
Mass Spectrometry Reference Standard
d5-TG Internal Standard Mixture I
4 μM each of compounds LMGL03010008 – LMGL03010016
in 1:1 toluene:methanol
Quantitative
Conditions:
HPLC/MS/MS
Column: Mercury C8, 20 X 2 mm, 3µ
Mobile phase A: 1:9 (v/v) Methanol : H2O + 10 mM NH4OAC
Mobile phase B: Methanol + 10 mM NH4OAC
Total Time
(min)
3.00
2.00
4.00
4.01
7.00
Parameter Table
CUR:
IS:
TEM:
GS1:
GS2:
ihe:
CAD:
EP
d5-TG
20:5(2)-22:6
14:0(2)-16:1
15:0(2)-18:1
19:0(2)-12:0
17:0(2)-17:1
16:0(2)-18:0
20:0(2)-20:1
20:2(2)-18:3
20:4(2)18:2
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Flow Rate
(µl/min)
600
600
600
1000
1000
A (%)
3.0
0.0
0.0
0.0
0.0
B (%)
97.0
100.0
100.0
100.0
100.0
(Period 1 Experiment 1)
10.00
5000.00
500.00
50.00
50.00
ON
Medium
10.00
Q1 Mass
(amu)
993.88
771.75
827.79
857.83
869.83
857.83
996.00
955.84
949.94
Q3 Mass
(amu)
674.60
526.30
568.30
584.50
582.40
542.40
666.50
630.40
628.60
Page 4 of 5
Dwell (msec)
150.00
150.00
150.00
150.00
150.00
150.00
150.00
150.00
150.00
Param
CE
CE
CE
CE
CE
CE
CE
CE
CE
37.00
33.00
33.00
35.00
41.00
35.00
41.00
43.00
37.00
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LIPID MAPS Certificate Of Analysis
Reagent Number / Lot Number:
RF0000000780 / LM39-095A
Mass Spectrometry Reference Standard
d5-TG Internal Standard Mixture I
4 μM each of compounds LMGL03010008 – LMGL03010016
in 1:1 toluene:methanol
Data:
X
ICof +M
R
M(9pairs): 993.9/674.6am
ufromSam
ple3(Am
poule1, Inj. 1)of TGd5M
ixLM
-6000LM
39-095AStability042512.w
iff (Tu...
M
ax. 1.1e5cps.
6.4e5
6.0e5
5.5e5
5.0e5
4.5e5
4.0e5
3.5e5
3.0e5
2.5e5
2.0e5
1.5e5
1.0e5
5.0e4
0.0
Interpretation:
Reviewed:
Date:
0.5
1.0
1.5
d5-TG
20:5(2)-22:6
14:0(2)-16:1
15:0(2)-18:1
19:0(2)-12:0
17:0(2)-17:1
16:0(2)-18:0
20:0(2)-20:1
20:2(2)-18:3
20:4(2)18:2
Signature on file
2.0
2.5
(µM)
3.92
3.97
3.98
3.98
3.94
3.91
3.97
3.97
3.99
3.0
3.5
Tim
e, m
in
4.0
s.d.
0.06
0.14
0.07
0.07
0.06
0.04
0.04
0.05
0.03
5.0
Date:
Page 5 of 5
5.5
µg / vial
3.82
3.00
3.23
3.35
3.36
3.29
3.88
3.72
3.72
Approved:
April 27, 2012
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4.5
6.0
6.5
s.d.
0.06
0.11
0.05
0.06
0.05
0.04
0.04
0.05
0.03
Signature on file
April 27, 2012
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