Isolation and characterization of novel degradation ... · S-1 Isolation and characterization of...
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S-1 Isolation and characterization of novel degradation products in Cilnidipine by LC-QTOF-MS/MS, LCMS n , 2D-NMR and FTIR Ch Krishnam Raju 1,2 , J V Shanmukha Kumar* 1 , Sanath Kumar Goud 2 1 Department of chemistry,K L University, Guntur, Andhra Pradesh-522502, India. 2 Analytical Research and Development, United States Pharmacopeial Convention – India (P) Ltd., Plot No. D6 & D8, IKP, Genome Valley, Shameerpet, Hyderabad, Telangana-500078, India. List of figures: 1. HPLC chromatogram ofCIL degradation products S-6 2. UPLC chromatogram of mixed sample solution of CIL with degradants S-7 3. Final purity of CD1 S-8 4. Final purity of CD2 S-8 5. Final purity of CD3 S-9 6. Chiral purity of CIL S-9 7. High resolution mass spectrum of CIL S-10 8. Mass accuracy of CIL S-11 Electronic Supplementary Material (ESI) for New Journal of Chemistry. This journal is © The Royal Society of Chemistry and the Centre National de la Recherche Scientifique 2017
Transcript of Isolation and characterization of novel degradation ... · S-1 Isolation and characterization of...
S-1
Isolation and characterization of novel degradation products in Cilnidipine by LC-QTOF-MS/MS, LCMSn,
2D-NMR and FTIR
Ch Krishnam Raju1,2 , J V Shanmukha Kumar*1, Sanath Kumar Goud2
1Department of chemistry,K L University, Guntur, Andhra Pradesh-522502, India.
2Analytical Research and Development, United States Pharmacopeial Convention – India (P) Ltd., Plot No. D6 & D8, IKP, Genome
Valley, Shameerpet, Hyderabad, Telangana-500078, India.
List of figures:
1. HPLC chromatogram ofCIL degradation products S-6
2. UPLC chromatogram of mixed sample solution of CIL with degradants S-7
3. Final purity of CD1 S-8
4. Final purity of CD2 S-8
5. Final purity of CD3 S-9
6. Chiral purity of CIL S-9
7. High resolution mass spectrum of CIL S-10
8. Mass accuracy of CIL S-11
Electronic Supplementary Material (ESI) for New Journal of Chemistry.This journal is © The Royal Society of Chemistry and the Centre National de la Recherche Scientifique 2017
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9. High resolution MSMS Spectrum of CIL S-12
10. MSnspectrum of CIL S-13
11. Mass fragmentation pathway of CIL S-14
12. High resolutionmass Spectrum of CD1 S-15
13. Mass accuracy of CD1 S-16
14. High resolutionMSMS Spectrum of CD1 S-17
15. MSn spectrum of CD1 (a) S-18
16. MSn spectrum of CD1 (b) S-19
17. Mass fragmentation pathway of CD1 S-20
18. High resolutionmass Spectrum of CD2 S-21
19. Mass accuracy of CD2 S-22
20. High resolutionMSMS Spectrum of CD2 S-23
21. MSn spectrum of CD2 (a) S-24
22. MSn spectrum of CD2 (b) S-25
23. Mass fragmentation pathway of CD2 S-26
24. High resolution mass Spectrum of CD3 S-27
25. Mass accuracy of CD3 S-28
26. High resolution MSMS Spectrum of CD3 S-29
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27. MSn spectrum of CD3 S-30
28. Mass fragmentation pathway of CD3 S-31
29. High resolution mass Spectrum of KD1 S-32
30. Mass accuracy of KD1 S-33
31. GCMS Spectrum of KD2 S-34
32. GCMS Library match spectrum for KD2 S-35
33. High resolution mass Spectrum of KD3 S-36
34. Mass accuracy of KD3 S-37
35. High resolution mass Spectrum of KD4 S-38
36. Mass accuracy of KD4 S-39
37. 1H NMR Spectrum of CD1 S-40
38. 13C NMR Spectrum of CD1 S-41
39. DEPT90 NMR Spectrum of CD1 S-42
40. DEPT135 NMR Spectrum of CD1 S-43
41. COSY NMR Spectrum of CD1 S-44
42. TOCSY NMR Spectrum of CD1 S-45
43. HSQC NMR Spectrum of CD1 S-46
44. HMBC NMR Spectrum of CD1 S-47
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45. NOESY NMR Spectrum of CD1 S-48
46. ROESY NMR Spectrum of CD1 S-49
47. ROESY NMR Zoomed Spectrum of CD1 S-50
48. 1H NMR Spectrum of CD2 S-51
49. 13C NMR Spectrum of CD2 S-52
50. DEPT90 NMR Spectrum of CD2 S-53
51. DEPT135 NMR Spectrum of CD2 S-54
52. COSY NMR Spectrum of CD2 S-55
53. TOCSY NMR Spectrum of CD2 S-56
54. HSQC NMR Spectrum of CD2 S-57
55. HMBC NMR Spectrum of CD2 S-58
56. NOESY NMR Spectrum of CD2 S-59
57. ROESY NMR Spectrum of CD2 S-60
58. ROESY NMR Zoomed Spectrum of CD2 S-61
59. 1H NMR Spectrum of CIL S-62
60. 13C NMR Spectrum of CIL S-63
61. DEPT90 NMR Spectrum of CIL S-64
62. DEPT135 NMR Spectrum of CIL S-65
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63. COSY NMR Spectrum of CIL S-66
64. TOCSY NMR Spectrum of CIL S-67
65. HSQC NMR Spectrum of CIL S-68
66. HMBC NMR Spectrum of CIL S-69
67. NOESY NMR Spectrum of CIL S-70
68. 1H NMR Spectrum of CD3 S-71
69. 13C NMR Spectrum of CD3 S-72
70. DEPT90 NMR Spectrum of CD3 S-73
71. DEPT135 NMR Spectrum of CD3 S-74
72. COSY NMR Spectrum of CD3 S-75
73. TOCSY NMR Spectrum of CD3 S-76
74. HSQC NMR Spectrum of CD3 S-77
75. HMBC NMR Spectrum of CD3 S-78
76. IR Spectrum of CD1 S-79
77. IR Spectrum of CD2 S-80
78. IR Spectrum of CD3 S-81
79. Prep HPLC chromatogram for the isolation of CD3 S-82
80. Prep HPLC chromatogram for the isolation of CD1 and CD2. S-83
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Figure 1: HPLC chromatograms Base degradation (a), Acid degradation (b), Photolytic degradation (c), Oxidative degradation (d),Heat degradation (e),Humid degradation (f)
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Figure2:UPLC chromatogram of mixed sample solution of CIL with degradants.
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Figure3:Final purity of CD1
Figure4:Final purity of CD2
Pea
k1 -
15.9
91
Pea
k2 -
17.4
30C
D-I
- 18.
221
Pea
k4 -
19.0
00P
eak5
- 19
.952
AU
-0.010
0.000
0.010
0.020
0.030
0.040
0.050
0.060
0.070
0.080
0.090
0.100
0.110
0.120
Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00 36.00 38.00 40.00 42.00 44.00
Pea
k1 -
13.3
70
Pea
k2 -
16.1
52
Pea
k3 -
18.1
90C
D-II
- 18
.979
Pea
k5 -
20.9
06P
eak6
- 21
.031
Pea
k7 -
22.0
67
AU
0.00
0.02
0.04
0.06
0.08
0.10
0.12
Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00 36.00 38.00 40.00 42.00 44.00
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Figure5:Final purity of CD3
Figure6: Chiral purity of CIL
CD
3 - 2
9.10
6P
eak2
- 30
.299
AU
0.00
0.02
0.04
0.06
0.08
0.10
0.12
0.14
0.16
0.18
0.20
Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00 36.00 38.00 40.00 42.00 44.00
CIL
-Isom
er-1
- 5.
404
CIL-
Isom
er-2
- 6.
370
AU
-0.10
0.00
0.10
0.20
0.30
0.40
0.50
0.60
0.70
0.80
0.90
1.00
1.10
1.20
1.30
1.40
1.50
Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00
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Figure7:High resolution mass spectrum of CIL
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Figure 8:Mass accuracy of CIL
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Figure9: High resolution MSMS Spectrum of CIL
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Figure 10: MSn spectrum of CIL
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H2N
O OO
OO
NO2C27H29N2O7
+, 493.1969Nitrogen rule: Even, RDBE: 14.5
CIL
C27H27N2O6+, 475.1864
Nitrogen rule: Even, RDBE: 15.5
N
O OO
O
NO2
-H2O
C9H9+, 117.0699
Nitrogen rule: Even, RDBE: 5.5
-C18H20N2O7
N
HOHO O
OO
NO2
C18H19N2O7+, 375.1187
Nitrogen rule: Even, RDBE: 10.5
-C9H10
N
HO OH2
OO
NO2
C15H13N2O6+, 317.0768
Nitrogen rule: Even, RDBE: 10.5
-C3 H
6 O
N
HO
O
NO2
C14H11N2O4+, 271.0713
Nitrogen rule: Even, RDBE: 10.5
-CH2 O
2
-C3H8O2
-C9H8
N
H2O
O
C14H12NO2+, 226.0863
Nitrogen rule: Odd, RDBE: 9.5
N
O
OO
NO
C24H21N2O5+, 417.1445
Nitrogen rule: Even, RDBE: 15.5
OH2
N
HO
OO
NOC15H13N2O5
+, 301.0819Nitrogen rule: Even, RDBE: 10.5
OH2
-CHN
O 4
HN
O
C14H12NO+, 210.0913Nitrogen rule: Odd, RDBE: 9.5
-C10H
8 NO4
N
HO
O
NO
C14H11N2O3+, 255.0764
Nitrogen rule: Even, RDBE: 10.5
-CH2 O
2
Figure 11: Mass fragmentation pathway of CIL
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Figure 12:High resolution mass spectrum of CD1
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Figure13: Mass accuracy of CD1
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Figure14:High resolution MSMS Spectrum of CD1
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Figure15:MSn Spectrum of CD1 (a)
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Figure16:MSn Spectrum of CD1 (b)
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C27H31N2O10+, 543.1973
Nitrogen rule: Even, RDBE: 13.5CD1
O O
O
O
O
O
O
NO2
NH2HO
C26H31N2O8+, 499.2075
Nitrogen rule: Even, RDBE: 12.5
O O
O
O
O
NO2
NH2HO
C26H29N2O6+, 465.2020
Nitrogen rule: Even, RDBE: 13.5
HO
O
O
NO2
HON
NH2
NO2
O
O
OO
OO
H
C18H21N2O8+, 393.1292
Nitrogen rule: Even, RDBE: 9.5
NH
NO2
OOH
OO
H
C17H21N2O6+, 349.1394
Nitrogen rule: Even, RDBE: 8.5
C23H21N2O4+, 389.1496
Nitrogen rule: Even, RDBE: 13.5
HO
O
NO2
N
C14H16NO+, 214.1226Nitrogen rule: Odd, RDBE: 7.5
H2O
N
N
C12H14N+, 172.1121Nitrogen rule: Odd, RDBE: 6.5
C12H11+, 155.0855
Nitrogen rule: Even, RDBE: 7.5
C26H30N2O7•+, 482.2048
Nitrogen rule: Even, RDBE: 13.0
O O
O
O
O
NO2
NH2
NH2
OO
H
C7H12NO2+, 142.0863
Nitrogen rule: Odd, RDBE: 1.5
HN
NO2
O
C15H17N2O3+, 273.1234
Nitrogen rule: Even, RDBE: 8.5
HN
NO2
C14H17N2O2+, 245.1285
Nitrogen rule: Even, RDBE: 7.5
C9H9+, 117.0699
Nitrogen rule: Even, RDBE: 5.5
-C18H22N2O10
-CO2
-H2O2
-OH.
-C 12H 13NO5
-C2H2O -C2H5NO
-C3H8O2
-C3 H
10O3
-C9H10O-C10H10O2
-C9H10O2
-CO2-C11H9NO6
-C2H4O3-C3H4O4
O
NO2
N
C14H11N2O4+, 271.0713
Nitrogen rule: Even, RDBE: 10.5
HO
NO2
N
C13H11N2O3+, 243.0764
Nitrogen rule: Even, RDBE: 9.5
HO
O
O
NO2
N
C23H21N2O5+, 405.1445
Nitrogen rule: Even, RDBE: 14.5
HO
Figure17: Mass fragmentation pathway of CD1
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Figure18:High resolution mass Spectrum of CD2
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Figure19:Mass accuracy of CD2
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Figure20:High resolution MSMS Spectrum of CD2
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Figure21: MSn spectrum of CD2 (a)
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Figure 22: MSn spectrum of CD2 (b)
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C27H31N2O10+, 543.1973
Nitrogen rule: Even, RDBE: 13.5CD2
O O
O
O
O
O
O
NO2
NH2 OH
C9H9+, 117.0699
Nitrogen rule: Even, RDBE: 5.5
C26H31N2O8+, 499.2075
Nitrogen rule: Even, RDBE: 12.5
O O
O
O
O
NO2
NH2 OH
-CO2
C26H29N2O6+, 465.2020
Nitrogen rule: Even, RDBE: 13.5
HO
O
O
NO2
N
OH
-H 2O2
C23H21N2O5+, 405.1445
Nitrogen rule: Even, RDBE: 14.5
O
O
NO2
N
OH
-C3H10O3
O O
O
O
NH2
OH
C18H22NO5+, 332.1492
Nitrogen rule: Odd, RDBE: 8.5
-C9H9NO5
O
C9H9O+, 133.0648Nitrogen rule: Even, RDBE: 5.5
-C9H13NO4
C8H9+, 105.0699
Nitrogen rule: Even, RDBE: 4.5
-CO
-C18H22N2O10
C27H29N2O9+, 525.1868
Nitrogen rule: Even, RDBE: 14.5
O O
O
O
O
O
O
NO2
NH2
-H2O
C26H29N2O7+, 481.1969
Nitrogen rule: Even, RDBE: 13.5
O O
O
O
O
NO2
NH2
-CO2
C18H19N2O7+, 375.1187
Nitrogen rule: Even, RDBE: 10.5
H2O
O
O
O
O
NO2
N
-C9H10O2
O
NO2
N
C14H11N2O4+, 271.0713
Nitrogen rule: Even, RDBE: 10.5
HO
-C4H8O3
C26H27N2O8+, 495.1762
Nitrogen rule: Even, RDBE: 14.5
O O
O
O
O
O
NO2
NH2
-CH2O
C15H12NO6+, 302.0659
Nitrogen rule: Odd, RDBE: 10.5
O O
O
O OH
O
NH2
-C11H15NO2
C13H10NO4+, 244.0604
Nitrogen rule: Odd, RDBE: 9.5
O O
OH
O
NH2
-C2H2O2
NH2
O
OO
C10H18NO3+, 200.1281
Nitrogen rule: Odd, RDBE: 2.5
-C17H11NO6
NH2
OO
C7H12NO2+, 142.0863
Nitrogen rule: Odd, RDBE: 2.5
-C3H6O
C18H23N2O10+, 427.1347
Nitrogen rule: Even, RDBE: 8.5
O O
HO
O
O
O
O
NO2
NH2 OH -C9H8
Figure 23: Mass fragmentation pathway of CD2
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Figure24:High resolution mass Spectrum of CD3
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Figure 25: Mass accuracy of CD3
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Figure26:High resolution MSMS Spectrum of CD3
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Figure27: MSn spectrum of CD3
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-C3H6O
Figure28: Mass fragmentation pathway of CD3
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Figure29:High resolution mass Spectrum of KD1
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Figure30: Mass accuracy of KD1
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Figure31: GCMS spectrum of KD2
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Figure32: GCMS Library match spectrum for KD2
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Figure 33:High resolution mass Spectrum of KD3
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Figure 34: Mass accuracy of KD3
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Figure 35:High resolution mass Spectrum of KD4
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Figure36: Mass accuracy of KD4
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Figure37: 1H NMR Spectrum of CD1 (500 MHz, DMSO-d6, 298K)
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Figure38:13C NMR Spectrum of CD1 (126 MHz, DMSO-d6, 298K)
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Figure39:DEPT90 NMR Spectrum of CD1 (126 MHz, DMSO-d6, 298K)
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Figure 40: DEPT135 NMR Spectrum of CD1 (126 MHz, DMSO-d6, 298K)
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Figure 41: COSY NMR Spectrum of CD1 (500 MHz, DMSO-d6, 298K)
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Figure 42: TOCSY NMR Spectrum of CD1 (500 MHz, DMSO-d6, 298K)
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Figure 43: HSQC NMR Spectrum of CD1 (500 MHz, DMSO-d6, 298K)
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Figure 44: HMBC NMR Spectrum of CD1 (500 MHz, DMSO-d6, 298K)
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Figure45: NOESY NMR Spectrum of CD1 (500 MHz, DMSO-d6, 298K)
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Figure46:ROESYNMR Spectrum of CD1 (500 MHz, DMSO-d6, 298K)
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Figure47: ROESY NMR Zoomed Spectrum of CD1 (500 MHz, DMSO-d6, 298K)
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Figure48: 1H NMR Spectrum of CD2 (500 MHz, DMSO-d6, 298K)
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Figure49:13C NMR Spectrum of CD2 (126 MHz, DMSO-d6, 298K)
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Figure 50:DEPT90 NMR Spectrum of CD2 (126 MHz, DMSO-d6, 298K)
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Figure 51:DEPT135 NMR Spectrum of CD2 (126 MHz, DMSO-d6, 298K)
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Figure 52: COSY NMR Spectrum of CD2 (500 MHz, DMSO-d6, 298K)
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Figure 53: TOCSY NMR Spectrum of CD2 (500 MHz, DMSO-d6, 298K)
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Figure 54: HSQC NMR Spectrum of CD2 (500 MHz, DMSO-d6, 298K)
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Figure55: HMBC NMR Spectrum of CD2 (500 MHz, DMSO-d6, 298K)
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Figure56: NOESY NMR Spectrum of CD2 (500 MHz, DMSO-d6, 298K)
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Figure57:ROESYNMR Spectrum of CD2 (500 MHz, DMSO-d6, 298K)
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Figure58: ROESY NMR Zoomed Spectrum of CD2 (500 MHz, DMSO-d6, 298K)
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Figure59: 1H NMR Spectrum of CIL (500 MHz, DMSO-d6, 298K)
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Figure60:13C NMR Spectrum of CIL (126 MHz, DMSO-d6, 298K)
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Figure61:DEPT90 NMR Spectrum of CIL (126 MHz, DMSO-d6, 298K)
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Figure62:DEPT135 NMR Spectrum of CIL (126 MHz, DMSO-d6, 298K)
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Figure63: COSY NMR Spectrum of CIL (500 MHz, DMSO-d6, 298K)
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Figure64: TOCSY NMR Spectrum of CIL(500 MHz, DMSO-d6, 298K)
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Figure65: HSQC NMR Spectrum of CIL(500 MHz, DMSO-d6, 298K)
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Figure66: HMBC NMR Spectrum of CIL(500 MHz, DMSO-d6, 298 K)
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Figure67: NOESY NMR Spectrum of CIL(500 MHz, DMSO-d6, 298K)
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Figure68: 1H NMR Spectrum of CD3 (500 MHz, DMSO-d6, 298K)
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Figure69:13C NMR Spectrum of CD3 (126 MHz, DMSO-d6, 298K)
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Figure70:DEPT90 NMR Spectrum of CD3 (126 MHz, DMSO-d6, 298K)
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Figure71:DEPT135 NMR Spectrum of CD3 (126 MHz, DMSO-d6, 298K)
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Figure72: COSY NMR Spectrum of CD3 (500 MHz, DMSO-d6, 298K)
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Figure73: TOCSY NMR Spectrum of CD3(500 MHz, DMSO-d6, 298K)
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Figure74: HSQC NMR Spectrum of CD3(500 MHz, DMSO-d6, 298K)
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Figure75: HMBC NMR Spectrum of CD3(500 MHz, DMSO-d6, 298 K)
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Figure76:IRSpectrumofCD1
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Figure77:IRSpectrumofCD2
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Figure78:IR Spectrum of CIL
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Figure 79: Prep HPLC chromatogram for the isolation of CD3.
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Figure 80: Prep HPLC chromatogram for the isolation of CD1 and CD2.
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