MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

A) All chromatograms were run using the HPLC method described on the Product Data Sheet.

Concentrations and volumes:

Standard solution concentration was 5.0 mg/ml.

Citric acid, [1,5-14C]- concentration was 50.0 µCi/ml. Volume of standard alone injection was 10.0 µl.

Volume of Citric acid, [1,5-14C]- alone injection was 1.0 µl.

Co-injection solution consisted of 1.0 µl Citric acid, [1,5-14C]- + 10.0 µl standard.

Volume of co-injection was 11.0 µl.

Volume of blank injection was 10.0 µl.

B) Mass spectrometry – Negative mode C) NMR

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Chromatogram Info:

File Name : L:\INTB8259.PRM File Created : 10/8/2013 3:05:40 PM

Origin : Acquired, Acquisition started 10/8/2013 2:04:46 PM Acquired Date : 10/8/2013 2:44:45 PM

Project : c:\Clarity\Projects\Test.PRJ By : Administrator

Method : Unit11_40_min_run By : Administrator

Description : Radiochemical trace of 14C material alone

Created : 11/6/2007 2:08 PM Modified : 10/9/2013 2:31 PM

Column : Detection : Radiochemical

Mobile Phase : Temperature :

Flow Rate : Pressure :

Note :

[min.]Time

0 10 20 30 40

[V]

Volta

ge

0.0

0.2

0.4

0.6

0.8

1.0

9.7

1

10.9

2

11.2

3

11.7

4

L:\ I NTB8259 - Unit 11 Radio

10/9/2013 2:31 PM Page 1 of 2Chromatogram L:\INTB8259.PRM

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Result Table (Uncal - L:\INTB8259 - Unit 11 Radio)

Reten. Time[min]

Area[mV.s]

Height[mV]

Area[%]

Height[%]

W05[min]

1 9.670 21112.788 944.856 99.45 99.1 0.30

2 10.883 22.661 2.614 0.11 0.3 0.08

3 11.187 62.907 2.478 0.30 0.3 0.14

4 11.737 30.578 3.546 0.14 0.4 0.15

Total 21228.933 953.494 100.00 100.0

10/9/2013 2:31 PM Page 2 of 2Chromatogram L:\INTB8259.PRM

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Chromatogram Info:

File Name : L:\INTB8259.PRM File Created : 10/8/2013 3:05:40 PM

Origin : Acquired, Acquisition started 10/8/2013 2:04:46 PM Acquired Date : 10/8/2013 2:44:45 PM

Project : c:\Clarity\Projects\Test.PRJ By : Administrator

Method : Unit11_40_min_run By : Administrator

Description : UV trace of 14C material alone

Created : 11/6/2007 2:08 PM Modified : 10/9/2013 2:30 PM

Column : Detection : 205 nm

Mobile Phase : Temperature :

Flow Rate : Pressure :

Note :

[min.]Time

0 10 20 30 40

[mV]

Volta

ge

0

50

100

150

200L:\ I NTB8259 - Unit 11 UV

10/10/2013 10:35 AM Page 1 of 2Chromatogram L:\INTB8259.PRM

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Result Table (Uncal - L:\INTB8259 - Unit 11 UV)

Reten. Time[min]

Area[mV.s]

Height[mV]

Area[%]

Height[%]

W05[min]

No peak to report

10/10/2013 10:35 AM Page 2 of 2Chromatogram L:\INTB8259.PRM

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Chromatogram Info:

File Name : L:\INTB8258.PRM File Created : 10/8/2013 3:05:40 PM

Origin : Acquired, Acquisition started 10/8/2013 1:12:15 PM Acquired Date : 10/8/2013 1:52:15 PM

Project : c:\Clarity\Projects\Test.PRJ By : Administrator

Method : Unit11_40_min_run By : Administrator

Description : UV trace of standard material

Created : 11/6/2007 2:08 PM Modified : 10/9/2013 11:22 AM

Column : Detection : 205 nm

Mobile Phase : Temperature :

Flow Rate : Pressure :

Note :

[min.]Time

10 20 30 40

[mV]

Volta

ge

0

50

100

150

2.9

1

7.8

2

8.9

3

L:\ I NTB8258 - Unit 11 UV

10/9/2013 2:35 PM Page 1 of 2Chromatogram L:\INTB8258.PRM

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Result Table (Uncal - L:\INTB8258 - Unit 11 UV)

Reten. Time[min]

Area[mV.s]

Height[mV]

Area[%]

Height[%]

W05[min]

1 2.897 8.559 1.570 0.24 0.9 0.07

2 7.777 26.534 0.741 0.75 0.4 0.57

3 8.930 3492.132 179.075 99.01 98.7 0.30

Total 3527.225 181.386 100.00 100.0

10/9/2013 2:35 PM Page 2 of 2Chromatogram L:\INTB8258.PRM

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Chromatogram Info:

File Name : L:\INTB8260.PRM File Created : 10/8/2013 4:05:16 PM

Origin : Acquired, Acquisition started 10/8/2013 3:22:32 PM Acquired Date : 10/8/2013 4:02:31 PM

Project : c:\Clarity\Projects\Test.PRJ By : Administrator

Method : Unit11_40_min_run By : Administrator

Description : Radiochemical trace of 14C material co-injected with standard

Created : 11/6/2007 2:08 PM Modified : 10/9/2013 2:28 PM

Column : Detection : Radiochemical

Mobile Phase : Temperature :

Flow Rate : Pressure :

Note :

[min.]Time

0 10 20 30 40

[mV]

Volta

ge

0

200

400

600

800

9.1

1

10.3

2

10.4

3

10.6

4

10.9

5

11.2

6

11.3

7

11.5

8

11.9

9

L:\ I NTB8260 - Unit 11 Radio

10/9/2013 2:29 PM Page 1 of 2Chromatogram L:\INTB8260.PRM

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Result Table (Uncal - L:\INTB8260 - Unit 11 Radio)

Reten. Time[min]

Area[mV.s]

Height[mV]

Area[%]

Height[%]

W05[min]

1 9.093 21319.564 771.647 99.37 97.4 0.39

2 10.277 5.927 1.775 0.03 0.2 0.04

3 10.443 18.492 3.826 0.09 0.5 0.08

4 10.643 27.650 2.613 0.13 0.3 0.06

5 10.897 30.607 3.291 0.14 0.4 0.12

6 11.227 9.181 1.903 0.04 0.2 0.07

7 11.293 14.720 2.242 0.07 0.3 0.12

8 11.510 13.401 2.398 0.06 0.3 0.06

9 11.877 14.967 2.512 0.07 0.3 0.10

Total 21454.509 792.209 100.00 100.0

10/9/2013 2:29 PM Page 2 of 2Chromatogram L:\INTB8260.PRM

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Chromatogram Info:

File Name : L:\INTB8260.PRM File Created : 10/8/2013 4:05:16 PM

Origin : Acquired, Acquisition started 10/8/2013 3:22:32 PM Acquired Date : 10/8/2013 4:02:31 PM

Project : c:\Clarity\Projects\Test.PRJ By : Administrator

Method : Unit11_40_min_run By : Administrator

Description : UV trace of 14C material co-injected with standard

Created : 11/6/2007 2:08 PM Modified : 10/9/2013 2:28 PM

Column : Detection : 205 nm

Mobile Phase : Temperature :

Flow Rate : Pressure :

Note :

[min.]Time

0 10 20 30 40

[mV]

Volta

ge

0

50

100

150

200

250

5.1

1

8.8

2

13.0

3

L:\ I NTB8260 - Unit 11 UV

10/9/2013 2:28 PM Page 1 of 2Chromatogram L:\INTB8260.PRM

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Result Table (Uncal - L:\INTB8260 - Unit 11 UV)

Reten. Time[min]

Area[mV.s]

Height[mV]

Area[%]

Height[%]

W05[min]

1 5.127 27.673 3.021 0.91 1.8 0.14

2 8.793 2884.510 164.495 94.91 97.6 0.27

3 12.973 127.030 0.967 4.18 0.6 2.20

Total 3039.213 168.483 100.00 100.0

10/9/2013 2:28 PM Page 2 of 2Chromatogram L:\INTB8260.PRM

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Chromatogram Info:

File Name : L:\INTB8256.PRM File Created : 10/8/2013 3:05:39 PM

Origin : Acquired, Acquisition started 10/8/2013 10:39:17 AM Acquired Date : 10/8/2013 11:19:16 AM

Project : c:\Clarity\Projects\Test.PRJ By : Administrator

Method : Unit11_40_min_run By : Administrator

Description : UV trace of blank injection

Created : 11/6/2007 2:08 PM Modified : 10/9/2013 2:36 PM

Column : Detection : 205 nm

Mobile Phase : Temperature :

Flow Rate : Pressure :

Note :

[min.]Time

0 10 20 30 40

[mV]

Volta

ge

0

50

100

150

200L:\ I NTB8256 - Unit 11 UV

10/9/2013 2:36 PM Page 1 of 2Chromatogram L:\INTB8256.PRM

MC-365

Citric acid, [1,5-14C]-

Lot 742-137-112-A-20131009-SBA

Result Table (Uncal - L:\INTB8256 - Unit 11 UV)

Reten. Time[min]

Area[mV.s]

Height[mV]

Area[%]

Height[%]

W05[min]

No peak to report

10/9/2013 2:36 PM Page 2 of 2Chromatogram L:\INTB8256.PRM

C:\Xcalibur\data\MC365-20131009-SBA 10/4/2013 11:42:16 AM MC365-20131009-SBA

RT: 0.00 - 1.98

0.0 0.5 1.0 1.5

Time (min)

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

Relative Abundance

0.410.30 1.96

1.821.620.181.49

0.73 1.05

NL:3.08E6

TIC MS MC365-20131009-SBA

MC365-20131009-SBA#1-89 RT: 0.01-1.98 AV:T: - c NSI SIM ms [161.00-221.00]m/z= 188.98-202.13

m/z Intensity Relativem/z Intensity Relative

189.59 3049.0 0.22 190.63 700.1 0.05 191.58 27027.4 1.96 192.67 1978.1 0.14 193.58 282875.7 20.51 194.62 10503.3 0.76 195.52 1379467.5 100.00 196.57 53078.2 3.85 197.44 49719.4 3.60 198.41 29432.0 2.13 199.50 5212.5 0.38 200.53 1383.5 0.10 201.45 3360.8 0.24

MC365-20131009-SBA #1-5 RT: 0.01-0.09 AV: 5 NL: 1.62E6T: - c NSI SIM ms [161.00-221.00]

170 180 190 200 210 220

m/z

0

10

20

30

40

50

60

70

80

90

100

Relative Abundance

195.52

193.61

217.42

181.50167.50 197.44171.52 215.44191.60 207.41165.67 182.49 218.57

98

76

54

32

1pp

m

2.7022.7372.8772.917

2.162.00

PC 1.00

GB 0

LB 0.00 Hz

SSB 0

WDW no

SF 400.1300030 MHz

SI 32768

SFO1 400.1324710 MHz

PL1W 10.75174427 W

PL1 −1.00 dB

P1 14.00 usec

NUC1 1H

======== CHANNEL f1 ========

TD0 1

D1 1.00000000 sec

TE 293.8 K

DE 6.50 usec

DW 60.400 usec

RG 574.7

AQ 3.9584243 sec

FIDRES 0.126314 Hz

SWH 8278.146 Hz

DS 2

NS 128

SOLVENT D2O

TD 65536

PULPROG zg30

PROBHD 5 mm PABBO BB−

INSTRUM spect

Time 13.45

Date_ 20131009

PROCNO 1

EXPNO 1

NAME MC365−20131009−SBA

Batch 20131009−SBA

MC365 1H NMR in D2O

Data sheet for MC365Table of contents for MC365 Citric Acid, [1,5-14C]- 742-137-112-A-20131009-SBAINTB8259radINTB8259uvINTB8258uvstdINTB8260radCoINTB8260uvCoINTB8256uvblkMC365-20131009-SBA-MS-1