214 / 227
1$785$/ 5(0(',(6
3DJH RI
4.7) Range of quantification:
4.7.1) Definition:
The range of the method is the interval between the lower and upper
concentration level that have been demonstrated to be linear.
4.7.2) Assessment:
It was calculated from the graph. A graph is plotted using the concentration
on the X-axis and response factor on Y-axis. Response factor is the ratio between the
area and the concentration in mcg/ml.
4.7.3) Acceptance Criteria:
Response factors of concentrations fall within the horizontal zone
±
7.5%
.
4.7.4) Experiment:
a)
Procedure:
20
µ
l of each individual standard solution (refer section 4.2.4) were injected in
replicates and the chromatograms were recorded. Mean area was determined. Response
factors were calculated for all the Withanoside IV, Withanoside V, Withaferin A, 12-Deoxy
withastramonolide, Withanolide A and Withanolide B by dividing the mean area with the
concentration.
The mean response factor for Withanoside IV between the concentration 20.64 to
330.3 mcg/ml is 7838.43
Table-68:
Different Concentrations
Withanoside IV
(mcg/ml)
Dilution -6
(20.64)
Dilution -5
(41.29)
Dilution -4
(82.58)
Dilution -3
(165.15)
Dilution -2
(330.30)
Dilution -1
(660.60)
S. solution
(1321.20)
Mean
166024
329761
651132
1276177
2493821
4816148
9587357
Response
factor
Area /
concentration
8042.32
7986.95
7885.34
7727.38
7550.17
7290.57
7256.55
Mean
(Dilution 2 to 6)
7838.43
± 5.0% of 7838 is 7446 to 8229