LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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Reviewed: Signature on file Approved:

Signature on file

Date: 8/16/11 Date: 8/16/11

Systematic name: 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol) (ammonium salt)

Common name(s):

1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol), PG(17:0/14:1(9Z))

Chemical Molecular weight:

706.93 (proton form) 723.96 (ammonium salt)

Exact Mass Molecular Weight:

706.48 (proton form) 723.51 (ammonium salt)

Product# / Lot #:

LMGP04010007 / LM1-059A

Manufacture date:

July 19, 2011

Storage Conditions:

-16 to -24°C

Concentration: Volume: Total Amount/Ampoule:

15.46 μM (proton form) 1.0 ml 10.93 µg (proton form)

Solvent:

methanol

Structural Information:

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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TLC: Conditions: Plate A Mobile Phase: 65:25:4 CHCl3:MeOH:H2O

Plate B Mobile Phase: 65:25:4 CHCl3:MeOH:NH4OH Data:

Plate A Plate B

Lane 1: Suitability (18:1 PE + oleic acid and 18:1 LPE) Lane 2: PG(17:0/14:1(9Z))

Interpretation:

One spot in iodine and water, ninhydrin (-), phosphorus (+), char (+)

Inductively Coupled Plasma Mass Spectroscopy (Calcium): Interpretation: PG(17:0/14:1(9Z)) = 19.8 ppm Ca

1 2 1 2

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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Proton Nuclear Magnetic Resonance (1H-NMR):

Conditions: Sample conc.: 7.5 mg/ml in 8:2 CDCl3 : CD3OD Temp.: 292 K Field Strength: 400 MHz Pulse: 30° for 3.98 sec. Relaxation time: 1 sec. # of scans: 16 Line Broadening: 0.30 Hz

Data:

Interpretation: Shift (ppm) Actual Protons Theoretical Protons Assignment

Not detected 0 4 NH4 Not detected 0 2 OH

~5.34 1.999 2 olefinic protons ~5.22 1.009 1 C2 of glycero backbone ~4.40 0.985 1 C1 of glycero backbone ~4.17 1.005 1 C1 of glycero backbone ~4.05 13.868 0 water ~3.97 2.473 2 C3 of glycero backbone ~3.90 2.06 2 C3 of glycero headgroup ~3.78 1.038 1 C2 of glycero headgroup ~3.60 2.177 2 C1 of glycero headgroup ~2.32 4.018 4 α protons ~2.03 3.986 4 allylic protons ~1.60 3.991 4 β protons ~1.26 38.881 38 remaining methylenes of a yl chains ~1.18 1.672 0 IPA ~0.88 6 6 methyls on acyl chains

Note: Sum does not include water or IPA

Actual Sum Protons

Theoretical Sum Protons

69.622 74

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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Phosphorus Nuclear Magnetic Resonance (31P-NMR): Conditions: Sample conc.: 20 mg/ml in detergent solution

Temp.: 293 K Field Strength: 162.0 MHz Pulse: 30° for 0.511 sec. Relaxation time: 2 sec. # of scans: 16 Line Broadening: 1.0 Hz

Data:

Interpretation: One shift under described conditions

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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Reversed Phase HPLC with Evaporative Light Scattering Detection (HPLC/ELSD): Conditions: Mobile Phase A: 1:1 MeOH:H2O + 20 mM NH4OAC

Mobile Phase B: 100 % MeOH + 20 mM NH4OAC Gradient: Mobile Phase A Mobile Phase B

0 min 25% 75% 5 min 25% 75% 10 min 10% 90% 20 min 0% 100% 30 min 0% 100%

Column: Zorbax C8, 4.6 mm X 150 mm i.d,5µm Flow rate: 1 ml/min Detector: Sedex 85, 40 degrees C, 3.5 bar, gain of 3 Injection Volume: 10 µl @ 1 µg/µl

Data:

Minutes

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30

mV

olts

0

100

200

300

mV

olts

0

100

200

300

17

:0-1

4:1

PG

1

2.4

88

ELSD17:0-14:1 PG

NameRetention Time

Interpretation:

Purity = 99.1% AUC

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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Fatty Acid Methyl Esters by Capillary Gas Chromatography (GC-FAME): Conditions: Column: DB-225, 30M X 0.25 mm, i.d.

Temperatures: Initial Temp. = 220°C Injection = 250°C, Detector = 250°C

Carrier: Grade 5 Helium at 1 ml/min Injection: 1 µl @ 1:20 split Sample: 5 mg/ml in hexane after reaction with 0.2N sodium

methoxide in methanol at room temperature for 5 minutes.

Data:

Minutes

0 1 2 3 4 5 6 7 8 9 10

pA

0

100

200

300

400

500

600

700

800

900

1000

pA

0

100

200

300

400

500

600

700

800

900

1000

(4:0

)

(6:0

)

(8:0

)

(10:

0)

(12:

0)

(14:

0)14

:1

(15:

0)

(16:

0)(1

6:1)

17:0

(17:

1)

(18:

0)(1

8:1)

(18:

2)(1

8:3

(6-9

-12

Gam

ma)

)(1

8:3

(Alp

ha))

(20:

0)(2

0:1)

(20:

2)(2

0:3

(8-1

1-14

))(2

0:4)

(20:

3 (1

1-14

-17)

)

(20:

5)

(22:

0)(2

2:1)

(22:

2)

(22:

4)

(22:

5)(2

2:6)

(24:

0)

(24:

1)

Front Signal17:0-14:1 PG

Name

Interpretation:

Purity = 43.4% 14:1 and 56.6% 17:0 (AUC)

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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Mass Spectroscopy: Qualitative Conditions:

Period 1: -------------- Scans in Period: 75 Relative Start Time: 0.00 msec Experiments in Period: 1 Period 1 Experiment 1: ---------------------------- Scan Type: Enhanced MS (EMS) Polarity: Negative Scan Mode: Profile Ion Source: Turbo Spray # Scans to Sum: 1 Scan Rate: 1000 amu/s Intensity Thres.: 0.00 cps Settling Time: 0.0000 msec MR Pause: 5.0070 msec Q0 trapping: No MCA: Yes Center/Width: No LIT fill time: 5.00 msec Q3 Entry Barrier: 8.00 V Step Size: 0.08 amu Start (amu) Stop (amu) Time (sec) Param Start Stop 510.50 910.50 0.4001 AF3 0.61 0.83 EXB 149.46 145.47 Parameter Table(Period 1 Experiment 1) CUR: 10.00 IS: -4500.00 TEM: 100.00 GS1: 50.00 GS2: 0.00 ihe: ON CAD: High DP 0.00 CES 0.00 CE -5.00

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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Data:

Interpretation: -EMS: [M-H]- = 705.76 u

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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Qualitative Conditions:

Period 1: -------------- Scans in Period: 222 Relative Start Time: 0.00 msec Experiments in Period: 1 Period 1 Experiment 1: ---------------------------- Scan Type: Enhanced Resolution (ER) Polarity: Negative Scan Mode: Profile Ion Source: Turbo Spray # Scans to Sum: 1 Resolution Q1: Open Scan Rate: 250 amu/s Intensity Thres.: 0.00 cps Settling Time: 0.0000 msec MR Pause: 5.0070 msec Q0 trapping: No MCA: Yes Center/Width: Yes LIT fill time: 20.00 msec Dynamic Fill Time: On TIC Target EMS Scan: 1.00 x1e7 cps. TIC Target ER Scan: 0.50 x1e7 cps. TIC Target EPI Scan: 2.00 x1e7 cps. Max Fill EMS Scan: 150 msec Max Fill ER Scan: 250 msec Max Fill EPI Scan: 250 msec Min Fill EMS Scan: 2 msec Min Fill ER Scan: 2 msec Min Fill EPI Scan: 2 msec Default Fill EMS Scan: 20 msec Default Fill ER Scan: 20 msec Default Fill EPI Scan: 20 msec Q3 Entry Barrier: 8.00 V Step Size: 0.03 amu Start (amu) Stop (amu) Time (sec) Param Start Stop 690.68 720.68 0.1201 AF3 0.27 0.28 EXB 151.00 150.84 Parameter Table(Period 1 Experiment 1) CUR: 10.00 IS: -4500.00

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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TEM: 200.00 GS1: 50.00 GS2: 50.00 ihe: ON CAD: High DP 0.00 CES 0.00 CE -5.00

Data:

Interpretation:

-ER: [M-H]- = 705.60 u

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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Qualitative Conditions:

Period 1: -------------- Scans in Period: 33 Relative Start Time: 0.00 msec Experiments in Period: 1 Period 1 Experiment 1: ---------------------------- Scan Type: Enhanced Product Ion (EPI) Polarity: Negative Scan Mode: Profile Ion Source: Turbo Spray # Scans to Sum: 2 Product Of: 705.68 amu Resolution Q1: Unit Scan Rate: 1000 amu/s Intensity Thres.: 0.00 cps Settling Time: 0.0000 msec MR Pause: 5.0070 msec Q0 trapping: Yes MCA: Yes Center/Width: No LIT fill time: 20.00 msec Q3 Entry Barrier: 8.00 V Step Size: 0.08 amu Start (amu) Stop (amu) Time (sec) Param Start Stop 50.00 70.00 0.0201 AF3 0.35 0.36 EXB 154.17 153.93 Start (amu) Stop (amu) Time (sec) Param Start Stop 65.00 137.20 0.0723 AF3 0.36 0.40 EXB 153.99 153.14 Start (amu) Stop (amu) Time (sec) Param Start Stop 132.00 492.59 0.3607 AF3 0.40 0.60 EXB 153.20 148.91 Start (amu) Stop (amu) Time (sec) Param Start Stop 487.00 750.00 0.2631 AF3 0.60 0.74 EXB 149.69 147.07 Parameter Table(Period 1 Experiment 1) CUR: 10.00 IS: -4500.00

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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TEM: 200.00 GS1: 50.00 GS2: 50.00 ihe: ON CAD: Medium DP -30.00 CES 0.00 CE -45.00

Data:

Interpretation:

Consistent with theoretical structure

LIPID MAPS Certificate of Analysis Product Number / Lot Number: LMGP04010007 / LM1-059A

Mass Spectrometry Reference Standard 1-heptadecanoyl-2-(9Z-tetradecenoyl)-sn-glycero-3-phospho-(1'-rac-glycerol)

(ammonium salt)

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Quantitative Conditions:

HPLC/MS/MS Comment: Mercury 20 x 2 mm, C18, 3µ Mobile Phase A: 1:9 MeOH:H2O + 10 mM NH4OAc Mobile Phase B: MeOH + 10 mM NH4OAc

MRM Transition Q1 Mass (amu) Q3 Mass (amu) Dwell(msec)

705.49 225.00 1000.00 Step Table

Step Total Time (min)

Flow Rate (µl/min)

A (%) B (%)

0 0.00 1000 8.0 92.0 1 2.00 1000 8.0 92.0

Data:

Interpretation: Total PG(17:0/14:1(9Z)) = 15.46 µM