Amlodipine and Valsartan Tablets Summary Validation Report

Amlodipine and Valsartan Tablets
Summary Validation Report
April 26, 2013
________________
1. Executive Summary
a. Goals: The reference procedure described herein was developed for the MC monograph for Amlodipine and
Valsartan Tablets.
b. Background
i. Amlodipine is calcium channel blocker dihydropyridine class used as an antihypertensive and used in
thetreatment of hypertension and coronary heart desease. Valsartan is an angiotensin II receptor antagonist
and used in the treatment of hypertension.
ii. The active moiety of the medicine is Amlodipine and Valsartan, and all dosing information is based on the
free base.
iii. Quality monographs are not available.
c. The Reference procedure is based upon the USP-MC procedure for Amlodipine and Losartan Tablets.
2. Reference Procedures
a. Assay Procedure: LC-PDA (Both Waters (primary) and Agilent (secondary))
i. Solution A: 25 mM ammonium acetate in water. Adjust with ammonia solution to a pH of 7.5.
ii. Solution B: Acetonitrile
iii. Diluent: Water and methanol (3:7)
iv. System suitability Solution: 1 μg/mL each of USP Amlodipine Besylate RS, USP Valsartan RS,
USP Valsartan Impurity B RS, and USP Valsartan Impurity E RS in Diluent. [NOTE—Integrate the
chromatogram at 237 nm.]
v. Standard Solution: 0.05 mg/mL of USP Amlodipine Besylate RS and 1.6 mg/mL of USP Valsartan
RS in Diluent.
vi. Sample Solution: Weigh and finely powder an adequate number of Tablets, and dissolve a portion
in Diluent. Sonicate and centrifuge to obtain a final concentration of 0.05 mg/mL of amlodipine and 1.6
mg/mL of valsartan.
vii. Chromotographic System (See Chromatography , System Suitability.)
1. Mode: LC
2. Detector: PDA (Scan 200-700 nm) [Note: Calculations are based on the chromatograms collected at
237 nm for amlodipine and 295 nm for valsartan.]
3. Column: 4.6-mm x 15-cm; 5-µm L1 packing, Purosphere Star RP-18
4. Flow rate: 1 mL/min
5. Column Oven Temperature: 40°C
6. Auto sampler Temperature: 10°C
7. Injection volume: 20 µL
viii. Gradient Program:
Time
(min)
Solution A
(%)
Solution B
(%)
0 85 15
5 85 15
15 50 50
27 50 50
30 10 90

35 10 90
36 85 15
40 85 15
b. Organic Impurities Procedure: LC-PDA (Both Waters (primary) and Agilent (secondary)) and MS
i. Solution A: 25 mM ammonium acetate in water. Adjust with ammonia solution to a pH of 7.5.
ii. Solution B: Acetonitrile
iii. Diluent: Water and methanol (3:7)
iv. System suitability Solution: 0.05 mg/mL of USP Amlodipine Besylate RS and 1.6 mg/mL of USP
Valsartan RS in Diluent.
v. Standard Solution: 0.5 μg/mL each of USP Amlodipine Besylate RS, USP Amlodipine Impurity A RS, USP
Amlodipine Impurity B RS, USP Amlodipine Impurity C RS, USP Amlodipine Impurity D RS, USP
Amlodipine Impurity E RS, USP Amlodipine Impurity F RS, USP Valsartan RS, USP Valsartan Impurity B
RS, USP Valsartan Impurity C RS, and USP Valsartan Impurity E RS in Diluent.
vi. Sample Solution: Weigh and finely powder an adequate number of Tablets, and dissolve a portion in
Diluent. Sonicate and centrifuge to obtain a final concentration of 0.5 mg/mL each of amlodipine and
valsartan.
vii. Chromotographic System (See Chromatography , System Suitability.)
1. Mode: LC
2. Detector: PDA (Scan 200-700 nm) [Note: Calculations are based on the chromatograms collected at
237 nm for amlodipine and 295 nm for valsartan.]
3. Column: 4.6-mm x 15-cm; 5-µm L1 packing, Purosphere Star RP-18
4. Flow rate: 1 mL/min
5. Column Oven Temperature: 40°C
6. Auto sampler Temperature: 10°C
7. Injection volume: 20 µL
viii. Chromotographic System (See Chromatography , System Suitability.)
1. Mode: LC
2. Detector: MS Source: ES Scan (+ and -)
3. MS Conditions:
a. Capillary (kv): 3.00
b. Cone (v): 25.0
c. Source temperature: 90°C
d. Desolvation temperature: 500°C
ix. Gradient Program:
Time
(min)
Solution A
(%)
Solution B
(%)
0 85 15
5 85 15
15 50 50
27 50 50
30 10 90
35 10 90
36 85 15
40 85 15
Amlodipine and Valsartan Tablets Summary Validation Report—Page 2 of 24

3. Validation Results
a. Assay for Amlodipine
i. Precision and Accuracy
1. 7 independent solutions compared to Amlodipine Besylate Standard.
2. Acceptable Criteria: NLT 0.95 (Probability)
3. Calculated probability: 1.00 (Passes)
a. Mean = 99.61
b. Standard Deviation = 0.60
c. % RSD = 0.60
d. Lower = 98.0, Upper =102.0%
Solution
name
ii. Intermediate Precision
Table 1 – Assay Precision and Accuracy
Table 2 – Assay Ruggedness – Data Summary
Mean (%)
Standard
deviation
% RSD Probability
Day 1 99.61 0.60 0.60 1.00
Day 2 99.57 0.28 0.28 1.00
Day 3 99.81 0.09 0.09 1.00
Combined 99.66 0.38 0.38 1.00
4. Day 2 & 3 – Assay Intermediate Precision
Solution
name
Weight of the
sample (mg)
Response
Inj. 1
Response
Inj. 2
Average
response
% Assay
1 333.14 2669145 2646719 2657932 100.44
2 335.77 2680838 2672638 2676738 100.35
3 337.98 2678239 2682904 2680572 99.84
4 337.53 2664576 2660351 2662464 99.30
5 337.85 2662422 2661900 2662161 99.19
6 337.73 2656688 2664418 2660553 99.17
7 339.21 2662477 2672589 2667533 98.99
Weight of the
sample (mg)
Table 3 - Ruggedness (Day 2)
Response
Inj. 1
Response
Inj. 2
Average
response
% Assay
1 336.06 2630662 2628300 2629481 99.19
2 336.29 2639002 2638647 2638825 99.48
3 336.68 2646054 2655910 2650982 99.82
4 336.19 2646038 2650932 2648485 99.87
5 336.54 2656544 2646323 2651434 99.88
6 336.02 2640465 2629441 2634953 99.41
7 336.62 2625329 2651445 2638387 99.36
Amlodipine and Valsartan Tablets Summary Validation Report—Page 3 of 24

Solution
name
b. Assay for Valsartan
i. Precision and Accuracy
1. 7 independent solutions compared to Valsartan Stanadard.
2. Acceptable Criteria: NLT 0.95 (Probability)
3. Calculated probability: 1.00 (Passes)
a. Mean = 100.30
b. Standard Deviation = 0.46
c. % RSD = 0.46
d. Lower = 98.0, Upper =102.0%
ii. Intermediate Precision
Weight of the
sample (mg)
Table 6 – Assay Ruggedness – Data Summary
Mean (%)
Table 4 - Ruggedness (Day 3)
Response
Inj. 1
Standard
deviation
Response
Inj. 2
Average
response
% Assay
1 333.79 2487941 2490249 2489095 99.90
2 333.46 2484347 2480008 2482178 99.72
3 333.02 2478888 2477407 2478148 99.69
4 333.21 2485229 2476278 2480754 99.74
5 333.85 2490061 2484981 2487521 99.82
6 333.12 2480534 2488049 2484292 99.91
7 335.68 2505644 2499580 2502612 99.88
Solution
name
Weight of the
sample (mg)
Table 5 – Assay Precision and Accuracy
Response
Inj. 1
Response
Inj. 2
Average
response
% Assay
1 333.14 1932337 1952261 1942299 99.35
2 335.77 1981923 1980237 1981080 100.54
3 337.98 1990522 1996244 1993383 100.50
4 337.53 1986266 1988679 1987473 100.34
5 337.85 1982695 1985095 1983895 100.06
6 337.73 1996266 1991842 1994054 100.61
7 339.21 2009534 1998522 2004028 100.67
% RSD Probability
Day 1 100.30 0.46 0.46 1.00
Day 2 100.18 0.18 0.18 1.00
Day 3 100.16 0.32 0.32 1.00
Combined 100.21 0.33 0.33 1.00
Amlodipine and Valsartan Tablets Summary Validation Report—Page 4 of 24

4. Day 2 & 3 – Assay Intermediate Precision
Solution
name
Weight of the
sample (mg)
Table 7 - Ruggedness (Day 2)
Response
Inj. 1
Response
Inj. 2
Average
response
% Assay
1 336.06 2025016 2023427 2024222 99.92
2 336.29 2031480 2032359 2031920 100.23
3 336.68 2036285 2036833 2036559 100.35
4 336.19 2032073 2034001 2033037 100.32
5 336.54 2035417 2035330 2035374 100.33
6 336.02 2022883 2025593 2024238 99.93
7 336.62 2020325 2044689 2032507 100.16
Solution
name
Weight of the
sample (mg)
Table 8 - Ruggedness (Day 3)
Response
Inj. 1
Response
Inj. 2
Average
response
% Assay
1 333.79 2177673 2177637 2177655 100.40
2 333.46 2158408 2159626 2159017 99.64
3 333.02 2170296 2176332 2173314 100.43
4 333.21 2172728 2164360 2168544 100.15
5 333.85 2167570 2176234 2171902 100.11
6 333.12 2161150 2163138 2162144 99.88
7 335.68 2190613 2194466 2192540 100.51
Amlodipine and Valsartan Tablets Summary Validation Report—Page 5 of 24

i. Range for Amlodipine
1. Acceptable criteria: NLT 0.95 (Probability over 80-120%)
a. 80% - 1.00 (Pass)
b. 90% - 1.00 (Pass)
c. 100% - 1.00 (Pass)
d. 110% - 1.00 (Pass)
e. 120% - 1.00 (Pass)
Linearity
level (%)
80
90
100
110
120
Table 9 – Assay Range
Sol
’n
Weight of
sample (mg)
Injection
1
Injection
2
Average
Mean
Recovery (%)
%
RSD
% Assay
1 268.92 2104628 2101136 2102882
79.31
2 268.76 2102577 2111828 2107203 79.52
3
4
268.62
268.52
2091771
2098034
2107942
2097455
2099857
2097745
99.81 0.15
79.28
79.23
5 268.80 2099089 2096933 2098011 79.16
6 268.43 2101945 2098277 2100111 79.35
1 303.92 2368293 2383833 2376063
89.20
2 303.82 2387938 2388586 2388262 89.69
3
4
303.72
303.89
2382644
2395121
2386087
2393757
2384366
2394439
100.20 0.26
89.57
89.90
5 303.47 2383933 2380024 2381979 89.56
6 303.82 2387762 2378719 2383241 89.50
1 336.06 2630662 2628300 2629481
99.19
2 336.29 2639002 2638647 2638825 99.48
3
4
336.68
336.19
2646054
2646038
2655910
2650932
2650982
2648485
100.28 0.29
99.82
99.87
5 336.54 2656544 2646323 2651434 99.88
6 336.02 2640465 2629441 2634953 99.41
1 371.66 2905688 2890593 2898141
108.74
2 371.89 2896318 2908730 2902524 108.84
3
4
371.94
371.83
2878371
2908391
2904419
2901177
2891395
2904784
99.57 0.26
108.41
108.94
5 371.53 2879288 2909196 2894242 108.63
6 371.93 2917172 2909864 2913518 109.24
1 405.89 3190435 3185595 3188015
119.49
2 406.52 3183239 3194483 3188861 119.33
3
4
406.17
406.12
3189514
3191267
3175997
3194996
3182756
3193132
100.15 0.11
119.21
119.61
5 406.34 3185777 3190301 3188039 119.36
6 405.55 3183499 3179744 3181622 119.35
Amlodipine and Valsartan Tablets Summary Validation Report—Page 6 of 24

ii. Range for Valsartan
1. Acceptable criteria: NLT 0.95 (Probabiliy over 80-120%)
a. 80% - 1.00 (Pass)
b. 90% - 1.00 (Pass)
c. 100% - 1.00 (Pass)
d. 110% - 1.00 (Pass)
e. 120% - 1.00 (Pass)
Linearity
level (%)
80
90
100
110
120
Table 10 – Assay Range
Sol
’n
Weight of
sample (mg)
Injection
1
Injection
2
Average
Mean
Recovery (%)
%
RSD
% Assay
1 268.92 1626471 1625647 1626059
80.25
2 268.76 1620700 1628693 1624697 80.23
3
4
268.62
268.52
1617139
1621880
1629400
1624392
1623270
1623136
100.05 0.20
80.20
80.22
5 268.80 1634127 1625381 1629754 80.46
6 268.43 1630972 1628856 1629914 80.58
1 303.92 1841279 1837841 1841279
90.45
2 303.82 1839232 1831578 1839232 90.38
3
4
303.72
303.89
1832108
1836983
1838044
1830269
1832108
1836983
99.98 0.19
90.06
90.25
5 303.47 1830269 1830134 1830202 90.04
6 303.82 1837553 1832863 1835208 90.18
1 336.06 2025016 2023427 2024222
99.92
2 336.29 2031480 2032359 2031920 100.23
3
4
336.68
336.19
2036285
2032073
2036833
2034001
2036559
2033037
99.96 0.20
100.35
100.32
5 336.54 2035417 2035330 2035374 100.33
6 336.02 2022883 2025593 2024238 99.93
1 371.66 2251643 2243584 2247614
110.35
2 371.89 2248712 2247775 2248244 110.32
3
4
371.94
371.83
2236576
2245996
2247627
2246284
2242102
2246140
99.99 0.14
110.00
110.23
5 371.53 2230712 2248413 2239563 110.00
6 371.93 2242781 2250176 2246479 110.22
1 405.89 2453253 2452696 2452975
120.30
2 406.52 2457999 2461192 2459596 120.44
3
4
406.17
406.12
2457709
2454305
2452096
2442307
2454903
2448306
100.03 0.16
120.32
120.01
5 406.34 2445800 2451737 2448769 119.97
6 405.55 2456427 2438442 2447435 120.13
Amlodipine and Valsartan Tablets Summary Validation Report—Page 7 of 24

iii. Specificity
a. Amlodipine and Valsartan Tablets – Diluent
b. Amlodipine Besylate and Valsartan – Total Ion Count
Amlodipine and Valsartan Tablets Summary Validation Report—Page 8 of 24

c. Amlodipine Besylate and Valsartan – MS Spectra of Amlodipine (At RT 15.17)
d. Amlodipine Besylate and Valsartan – MS Spectra of Valsartan (At RT 10.41)
Amlodipine and Valsartan Tablets Summary Validation Report—Page 9 of 24

e. Amlodipine and Valsartan Tablets – Mixture of Amlodipine Besylate, Amlodipine Impurity A,
Amlodipine Impurity B, Amlodipine Impurity C, Amlodipine Impurity D, Amlodipine Impurity E,
Amlodipine Impurity F, Valsartan, Valsartan Impurity B, Valsartan Impurity C and Valsartan Impurity
E
f. Amlodipine – UV Spectra of Impurity A
Amlodipine and Valsartan Tablets Summary Validation Report—Page 10 of 24

g. Amlodipine – UV Spectra of Impurity F
h. Amlodipine – UV Spectra of Amlodipine
i. Amlodipine – UV Spectra of Impurity E
j. Amlodipine – UV Spectra of Impurity B
Amlodipine and Valsartan Tablets Summary Validation Report—Page 11 of 24

k. Amlodipine – UV Spectra of Impurity C
l. Amlodipine – UV Spectra of Impurity D
m. Valsartan – UV Spectra of Impurity B
n. Valsartan – UV Spectra of Valsartan
Amlodipine and Valsartan Tablets Summary Validation Report—Page 12 of 24

o. Valsartan – UV Spectra of Impurity E
p. Valsartan – UV Spectra of Impurity C
q. Amlodipine and Valsartan Tablets Solution – 0.5 mg/mL Amlodipine
Amlodipine and Valsartan Tablets Summary Validation Report—Page 13 of 24

. Amlodipine and Valsartan Tablets Solution – 0.5 mg/mL Amlodipine – PDA Spectra
s. Amlodipine and Valsartan Tablets Solution – 0.5 mg/mL Valsartan
Amlodipine and Valsartan Tablets Summary Validation Report—Page 14 of 24

t. Amlodipine and Valsartan Tablets Solution – 0.5 mg/mL Valsartan – PDA Spectra
u. Amlodipine and Valsartan Tablets – Mixture of Amlodipine Besylate, Amlodipine Impurity A,
Amlodipine Impurity B, Amlodipine Impurity C, Amlodipine Impurity D, Amlodipine Impurity E,
Amlodipine Impurity F, Valsartan, Valsartan Impurity B, Valsartan Impurity C and Valsartan Impurity
E – Total Ion Count
Amlodipine and Valsartan Tablets Summary Validation Report—Page 15 of 24

v. Amlodipine – MS Spectra of Impurity A (At RT 13.88)
w. Amlodipine – MS Spectra of Impurity F (At RT 14.18)
Amlodipine and Valsartan Tablets Summary Validation Report—Page 16 of 24

x. Amlodipine – MS Spectra of Amlodipine (At RT 15.20)
y. Amlodipine – MS Spectra of Impurity E (At RT 16.32)
Amlodipine and Valsartan Tablets Summary Validation Report—Page 17 of 24

z. Amlodipine – MS Spectra of Impurity B (At RT 19.66)
aa. Amlodipine – MS Spectra of Impurity C (At RT 20.75)
Amlodipine and Valsartan Tablets Summary Validation Report—Page 18 of 24

. Amlodipine – MS Spectra of Impurity D (At RT 31.34)
cc. Valsartan – MS Spectra of Impurity B (At RT 9.67)
Amlodipine and Valsartan Tablets Summary Validation Report—Page 19 of 24

dd. Valsartan – MS Spectra of Valsartan (At RT 10.74)
ee. Valsartan – MS Spectra of Impurity E (At RT 15.58)
Amlodipine and Valsartan Tablets Summary Validation Report—Page 20 of 24

ff. Valsartan – MS Spectra of Impurity C (At RT 18.70)
gg. Amlodipine and Valsartan Tablets Solution – 0.5 mg/mL Amlodipine – Total Ion Count
Amlodipine and Valsartan Tablets Summary Validation Report—Page 21 of 24

hh. Amlodipine and Valsartan Tablets Solution – 0.5 mg/mL Amlodipine – MS Spectra of Amlodipine (At
RT 15.03)
ii. Amlodipine and Valsartan Tablets Solution – 0.5 mg/mL Valsartan – Total Ion Spectra
Amlodipine and Valsartan Tablets Summary Validation Report—Page 22 of 24

jj. Amlodipine and Valsartan Tablets Solution – 0.5 mg/mL Valsartan – MS Spectra (At RT 8.96)
kk. Amlodipine and Valsartan Tablets Solution – 0.5 mg/mL Valsartan – MS Spectra of Impurity B (At
RT 9.53)
Amlodipine and Valsartan Tablets Summary Validation Report—Page 23 of 24

ll. Amlodipine and Valsartan Tablets Solution – 0.5 mg/mL Valsartan – MS Spectra of Valsartan (At RT
10.57)
mm. Amlodipine and Valsartan Tablets Solution – 0.5 mg/mL Valsartan – MS Spectra (At RT 11.29)
Amlodipine and Valsartan Tablets Summary Validation Report—Page 24 of 24