Cilindipine Final Edited
Cilindipine Final Edited
Cilindipine Final Edited
G.Deepthi, M.Pharm,
Assistant Professor
Department of Pharmaceutical analysis
protection.
• It is reported to be useful in elderly patients and in those with diabetes and
albuminuria.
• Cilnidipine has been increasingly used in patients with chronic kidney disease.
• So far, several analytical methods using high-performance liquid chromatography (HPLC) combined
with ultraviolet/visible (UV) and have been reported for the quantitative determination of Cilnidipine.
• However, these methods have some limitations and need for improvements
• So in this study we aim to develop a reliable validated analytical method for estimation of Cilnidipine using UHPLC.
LITERATURE REVIEW:
• Kadam, A. & Hamrapurkar, P. & Patil, S. & Manoharan, M. & Suryagandha, A..
(2015). Development and validation of stability indicating RP-HPLC method for
the estimation of cilnidipine in bulk and pharmaceutical dosage form.
International Journal of Pharmaceutical Sciences Review and Research. 30. 177-
181.
• Parihar, Y. & Kotkar, T. & Mahajan, M.P. & Sawant, S.D.. (2014). Development
and validation of RP-HPLC method for simultaneous estimation of telmisartan
and cilnidipine in bulk and tablet dosage form. Pharmanest. 5. 2321-2325.
Dosage
Brand Name Dose Manufacturer
Form
• Distilled water,
• Acetonitrile,
Ortho-phosphoric acid.
•Ultrasonicator-BVK enterprises
•Electronics Balance-Denver
Software:
CHEM STATION_open lab
Chromatographic condition
Chromatographic condition:
Preparation of buffer:
Validation:
The validation of the optimized method was carried out using the validation parameters
1. System suitability
2. Specificity
3. Linearity
4. Precision
5. Accuracy
6. Robustness
7. Assay
RESULTS AND DISCUSSION: METHOD DEVELOPMENT :
Method development was done by changing various, mobile phase ratios, buffers etc.
Trials:
Chromatograph Trail-1 Trail-2 Trail-3 Trail-4
ic conditions (optimized)
Mobile phase Buffer :Acetonitrile Buffer :Acetonitrile Buffer :Acetonitrile Buffer :Acetonitrile
(40:60) (50:50) (40:60) (30:70)
Flow rate 1.0ml/min 0.6ml/min 0.6ml/min 0.6ml/min
Column Supelcosil LC18; 3.6 cm Supelcosil LC18; 3.6 cm Supelcosil LC18; 3.6 cm Supelcosil LC18; 3.6 cm
X 4.6 mm; 3.0 microns X 4.6 mm; 3.0 microns X 4.6 mm; 3.0 microns X 4.6 mm; 3.0 microns
Results Peak was eluted but In this Trail peak In this Trail peak Retention time and
peak shape was not was eluted but more was eluted but more peak shape is good
good and more void tailing factor and retention time and
volume. so, Retention time. So Less plate count so,
further trial is further trail was further trail was
carried out carried out. Carried out.
Trial chromatogram 1 Trial chromatogram 2
Chromatographic conditions:
The validation of the optimized method was carried out using the
validation parameters
1. System suitability
2. Specificity
3. Linearity
4. Precision
5. Accuracy
6. Robustness
7. Assay
1.System suitability:
• All the system suitability parameters were within the range
and satisfactory as per ICH guidelines
Cilnidipine
Sample ID
RT AREA
Chromatogram of blank.
Chromatogram of placebo
Standard Chromatogram
Cilnidipine
Discussion: Retention time of Sample ID
Cilnidipine was eluted at RT AREA
1000.000
900.000 916.826
f(x) = 6.09 x + 1.82
Discussion: Five linear 800.000 R² = 1
concentrations of Cilnidipine 700.000
761.148
SD: 0.63
% RSD : 0.63
Discussion: From a single volumetric flask of working standard solution six injections
were given and the obtained areas were mentioned above. Average area, standard
deviation and % RSD was calculated for Cinidipine. % RSD obtained as 0.63% for
Cilnidipine.As the limit of Precision was less than “2” the system precision was
passed in this method
4.2.Intermediate precision :
Calculated Assay
Conc. level Sample ID Sample wt. (mg) Sample Area
(in percentage )
SD. : 0.73
% RSD : 0.73
Discussion: Multiple sampling from a sample stock solution was done and six
working sample solutions of same concentrations were prepared, each injection
from each working sample solution was given and obtained areas were mentioned
in the above table. Average area, standard deviation and % RSD was calculated for
Cilnidipineand obtained as 0.73% . As the limit of Precision was less than “2” the
system precision was passed in this method
5.Accuracy:
Sample
Conc. level Sample wt. (mg) Sample Area Calculated Assay (in percentage )
ID
Sample
51.18 319.786 101.10
-01
Sample
LOW LEVEL (50%) 52.34 321.460 99.40
-02
Sample
51.66 322.667 101.10
-03
Sample
101.79 627.188 99.70
-01
Sample
MIDDLE LEVEL (100%) 101.34 628.474 100.40
-02
Sample
100.69 630.618 101.40
-03
Sample
150.13 940.413 101.40
-01
Sample
HIGH LEVEL (150%) 152.66 941.413 99.80
-02
Sample
150.99 947.844 101.60
-03
Average : 100.66
SD. : 0.84
% RSD : 0.83
%RSD of
S.No Condition
Cilnidipine
Average : 100.42
SD. : 0.52
% RSD : 0.52
ROBUSTNESS - FLOW RATE INCREASE
Calcula
ted
Sample wt. Assay
Conc. level Sample ID Sample Area
(mg) (in
percent
age )
Sample -01 100.76 534.910 100.00
Sample -02 100.67 534.196 99.90
Sample -03 102.09 534.891 98.70
MIDDLE LEVEL (100%)
Sample -04 100.46 538.136 100.90
Sample -05 102.04 537.966 99.30
Sample -06 101.47 540.887 100.40
Average : 99.87
SD. : 0.78
% RSD : 0.78
ROBUSTNESS - WAVE LENGTH DECREASE
Sample Sample wt. Sample Calculated Assay (in
Conc. level
ID (mg) Area percentage )
Sample
100.76 625.782 101.00
-01
Sample
100.67 628.438 101.50
-02
Sample
102.09 627.665 100.00
MIDDLE LEVEL -03
(100%) Sample
100.46 623.890 101.00
-04
Sample
102.04 633.627 101.00
-05
Sample
101.47 629.161 100.90
-06
Average : 100.90
SD. : 0.49
% RSD : 0.49
ROBUSTNESS WAVE LENGTH INCREASE
Sample wt. Sample Calculated Assay
Conc. level Sample ID
(mg) Area (in percentage )
SD. : 0.81
% RSD : 0.81
CALCULATION :
Content in mg
602.076 50.28 5 100 99.36
X X X X X 100.00
597.604 50 50 101.06 100
.= 9.96 mg
.= 99..96 %
Assay Chromatogram