Linearity and Range

A(n=3) B(n=3)
conc Mean
Range . Mean area SD %RSD conc. area SD %RSD
20% 30 8138.66 26.15 0.321306 24 2353386 26604.07 1.130459
60% 90 19950 51.11 0.25619 72 6583445 98097.5 1.490063
100% 150 32147 256.03 0.796435 120 10539760 36067.33 0.342203
140% 210 46530 274.1 0.589082 168 14898589 31983.51 0.214675
180% 270 56931.33 90.42 0.158823 216 19311761 131401.9 0.680424
220% 330 72644 433.44 0.596663 264 22841124 78489.23 0.343631
260% 390 85624 352.34 0.411497 312 27838639 87265.14 0.313468

Drug A
90000
80000 f(x) = 215.85 x + 666.45
R² = 1
70000
60000
50000
Area

40000
30000
20000
10000
0
0 50 100 150 200 250 300 350 400 450
Concentration

Drug B
30000000

25000000 f(x) = 87606.49 x + 191639.39

R² = 1
20000000
Area

15000000
 Precision:
Mean± SD Acceptance
Conc. % RSD Result Interference
(n=3) criteria
A B A B A B
12 0.1
96 28223.6±163.27 8649885.0±10204.24 0.57 RSD Within
0 1
15 12 10491034.0±28144.1 0.2 should be acceptable
31737.3±146.50 0.46
0 0 3 6 1% limit
18 14 12914986.0±13904.2
45606.0±292.22 0.64 0.1
0 4 7

Mean± SD Acceptance
Conc. % RSD Result Interference
(n=3) criteria
A B A B A B
12
96 28190.3±169.04 8649548.0±10324.63 0.59 0.11 Within
0 RSD should
15 10491009.0±28158.7 acceptabl
120 31725.3±142.50 0.44 0.26 be 1%
0 7
e limit
18 12914901.0±13967.9
144 45584.3±330.83 0.72 0.1
0 8

Accuracy:

Level Test Std. Total

Drug (%) conc. conc. conc. Mean SD % RSD
0.99116
80 150 120 270 57773.33 572.63 7
0.63047
A
100 150 150 300 64482 406.54 1
0.58850
120 150 180 330 71479.33 420.66 6
1878586 15146.0 0.08062
80 120 96 216 9 6 5
B
2085149 104567. 0.50148
100 120 120 240 5 8 8
 LOD AND LOQ:
Drug A
LoD = 3.3*σ/s 10.15
LoQ= 10*σ/s 30.75

Standard deviation of
σ= intercept 663.87
S= Slope of calibration curve 215.86

Drug B
LoD = 3.3*σ/s 7.22
LoQ= 10*σ/s 21.88

σ= Standard deviation of intercept 191639

S= Slope of calibration curve 87606

System suitability:

Time duration A B
Mean area±SD % Assay Mean assay±SD Mean area±SD % Assay
32778.33±29.9
Room temperature Initial 3 99.2 98.86±0.47 10546560±19276.26
After 24 32559.67±38.5
hours 2 98.52 10542803±18340.73
% RSD 0.104808097 0.475419786 0.178368709
Fridge temperature (2- 32778.33±29.9
8˚C) Initial 3 99.2 99.05±0.21 10546560±19276.26
After 24
hours 32681±34.39 98.9 10544100±19200.54
% RSD 0.111767603 0.212014134 0.178030967