A
novel validated RP-HPLC-DAD method for the estimation of Eluxadoline in bulk
and pharmaceutical dosage form
Uttam Prasad Panigrahy1*,
A. Sunil Kumar Reddy2,3
1Department of Pharmaceutical
Analysis and Quality Assurance, Malla Reddy College of Pharmacy, Maisammaguda,
Secunderabad-500014, India
2Department of Pharmaceutical
Chemistry, Bharat Institute of Technology-Pharmacy, Ibrahimpatnam,
Hyderabad-501510, India
3APL Research Centre-2,
Aurobindo Pharma Ltd., Sanga Reddy, Medak, Telengana-502329, India
*Corresponding Author E-mail:
uttampanigrahy@gmail.com
ABSTRACT:
The present
study describes about novel approach to develop and validate a rapid, accurate,
precise, simple, efficient and reproducible isocratic Reversed Phase-High
Performance Liquid Chromatographic (RP-HPLC-DAD) method for the estimation of
Eluxadoline in bulk and pharmaceutical dosage form. Eluxadoline were separated
using Hypersil C18 column (250mm×4.6 mm, 5µm particle size), Waters Alliance
e2695 HPLC system with 2998 PDA detector and the mobile phase contained a
mixture of Acetonitrile and Water (70:30, v/v). The flow rate was set to
1ml/min with the responses measured at 260nm. The retention time of Eluxadoline
was found to be 3.481min.Linearity was established for Eluxadoline in the range
of 10-125µg/ml with correlation coefficient (r2=0.999). The percentage
recoveries were between 99.92% to 100.07%. Validation parameters such as
specificity, linearity, precision, accuracy, robustness, limit of detection
(LOD) and limit of quantitation (LOQ) were evaluated for the method according
to the International Conference on Harmonization (ICH) Q2 R1 guidelines. The
developed method was successfully applied for the quantification and hyphenated
instrumental analysis.
KEYWORDS: Eluxadoline,
PDA detector, Hyphenated, ICH.
INTRODUCTION:
Eluxadoline is a novel, orally-active drug for
the treatment of diarrhea and abdominal
pain in individuals with diarrhea-predominant irritable bowel syndrome. Eluxadoline is a μ-and κ-opioid receptor agonist and δ-opioid receptor antagonist that acts locally in the enteric nervous system, possibly decreasing
adverse effects on the central nervous
system.[1] Eluxadoline is chemically known as 5-({[(2S)-2-amino-3-(4-carbamoyl-2,
6-dimethylphenyl) propanoyl] [(1S)-1-(4-phenyl-1 H-imidazol-2-yl)
ethyl] amino} methyl)-2-methoxybenzoic acid
was shown in Figure 1.
Literature review reveals that no analytical methods
have been reported for the determination of Eluxadoline
and few pharmacokinetics and preclinical studies was performed which includes
Eluxadoline benefits patients with irritable bowel syndrome with diarrhea in a
phase 2 study [2], Drug discovery
approaches to irritable bowel syndrome and Eluxadoline approved for irritable bowel syndrome with
diarrhea [3,4], Effect of Uptake Transporters OAT3 and
OATP1B1 and Efflux Transporter MRP2 on the Pharmacokinetics of Eluxadoline [5]
and Molecular characterization of eluxadoline as
a potential ligand targeting mu-delta opioid receptor heteromers
[6]. The present study was aimed to develop a novel, simple, economic and
validated RP-HPLC-DAD method for the estimation of Eluxadoline according to ICH guidelines
[7].
Figure 1: Chemical structure
of Eluxadoline
MATERIALS AND METHODS:
Chemicals and Reagents:
Instrumentation:
The analysis was performed by using a chromatographic system from
Waters Alliance e2695 HPLC system with 2998 PDA detector. The HPLC system was
equipped with Empower 2 software. Semi-micro analytical
balance (India), an Ultrasonic bath sonicator
(Frontline FS 4, Mumbai, India), Digital pH meter (Systronics model 802) and Whatman
filter paper No. 41 (Whatman International Ltd., England) were used in the
study.
Chromatographic conditions:
Eluxadoline was analysed in Hypersil C18 (250mm×4.6 mm, 5mm particle size) column for the chromatographic
separation. The mobile phase was composed of Acetonitrile and Water (70:30,
v/v). Filtered through 0.45µm nylon membrane filter under vacuum filtration and
pumped at ambient temperature, at a flow rate of 1 ml/min with UV detection
wavelength at 260nm. Injection volume was 20μl. The run time was 5min and the retention time of Eluxadoline was found to be 3.481min.
Chromatographic
Parameters:
|
Equipment |
: |
Waters
Alliance e2695 HPLC system with 2998 PDA
detector. |
|
Column |
: |
Hypersil C18
(250mm×4.6 mm,
5mm particle size) |
|
Flow rat |
: |
1ml/min |
|
Wavelength |
: |
260 nm |
|
Injection
volume |
: |
20 ml |
|
Column oven |
: |
Ambient |
|
Run time |
: |
5 Minutes |
Preparation
of mobile phase:
Acetonitrile HPLC grade and Water HPLC grade were mixed in the
proportion of 70:30 v/v and was filtered through 0.45μm nylon membrane
filter and degassed by sonication.
Preparation
of diluent:
Mobile phase was used as diluent.
Preparation
of standard stock solution of Eluxadoline:
Standard stock solution of Eluxadoline were prepared by dissolving 100mg
of Eluxadoline in 100ml of diluent into a 100ml clean dry volumetric flask and
the standard solution was filtered through 0.45 μm nylon membrane filter
and degassed by sonicator to get the concentration of 1000µg/ml of Eluxadoline.
Preparation
of standard solution of Eluxadoline for assay:
From the above standard stock solution of 1000µg/ml of Eluxadoline
further pipette 0.5 ml and transferred into a 10ml volumetric flask and dilute
up to the mark with diluent to get the concentration of 50µg/ml of Eluxadoline.
Selection of wavelength:
Standard stock solution of Eluxadoline were prepared by dissolving
100mg of Eluxadoline in 100ml of diluent into a 100ml clean dry volumetric
flask and the standard solution was filtered through 0.45μm nylon membrane
filter and degassed by sonicator to get the concentration of 1000µg/ml of
Eluxadoline. From the above standard stock solution of 1000µg/ml of Eluxadoline
further pipette 0.5 ml and transferred into a 10ml volumetric flask and dilute
up to the mark with diluent to get the concentration of 50µg/ml of Eluxadoline.
The wavelength of maximum absorption (λmax) of 50µg/ml of Eluxadoline were
scanned using UV-Visible spectrophotometer within the wavelength region of
200–400 nm against mobile phase as blank. The wavelength (λmax) was found
to be 260nm shown in Figure 2.
Figure 2:
UV-Spectrum of Eluxadoline at 260nm
Preparation
of sample solution of Eluxadoline:
Twenty tablets were accurately weighed and powdered and tablet powder
equivalent to 100mg of Eluxadoline were taken into 100ml clean dry volumetric
flask, diluent was added and sonicated to dissolve it completely and volume was
made up to the mark with the same diluent and filtered through 0.45 μm
nylon membrane filter. Further pipette out 0.5ml from the above Eluxadoline
sample stock solution into a 10ml volumetric flask and diluted up to the mark
with diluent to get the concentration of 50µg/ml of Eluxadoline. 20mL from standard and sample solution were injected into
the chromatographic system and the peak areas were measured for Eluxadoline was
shown in Figure 3 and 4 respectively. The % Assay was calculated by comparing
the peak area of standard and sample chromatogram by using the formula given
below and the assay result was shown in Table 1.
AT WS
DT P
Avg. Wt
Assay % =------
x ------x ------ x -------x---------------- X 100
AS DS WT
100 Label Claim
Where:
AT = Average peak area of sample preparation
AS= Average peak area of standard
preparation
WS = Weight of standard taken in mg
WT=Weight of sample taken in mg
P = Percentage purity of working standard
DS= Dilution factor for standard
preparation
DT=Dilution factor for sample preparation
Figure 3: Standard chromatogram of Eluxadoline
Figure 4: Sample chromatogram of Eluxadoline
Table 1:
Assay of marketed formulation of Eluxadoline
|
Drug |
Viberzi Tablet Label Claim (mg) |
Amount Found (mg) |
% Label Claim ± %
RSD (n=3) |
|
Eluxadoline |
100 |
99.97 |
99.97±0.12 |
Validation
of the proposed method:
The developed method for the simultaneous
estimation of Eluxadoline was validated as per the ICH guidelines for the
parameters like system suitability, specificity, linearity, accuracy,
precision, ruggedness, robustness, limit of detection (LOD) and limit of
quantitation (LOQ) [7].
System
Suitability:
At first the HPLC system was optimized as per the chromatographic
conditions. One blank followed by six replicates of a single calibration
standard solution of 50µg/ml of Eluxadoline was injected to check the system
suitability. To ascertain the system suitability for the proposed method, the
parameters such as retention time, theoretical plates and peak asymmetry were
taken and results were presented in Table 2.
Table 2: System suitability
test parameters for Eluxadoline
|
Parameter (n=6) |
Eluxadoline |
|
Retention
Time (Mins) |
3.481 |
|
Theoretical
plates |
2985 |
|
Tailing
factor |
1.1 |
Specificity:
The effect of excipients and other additives usually present in the
dosage form of Eluxadoline in the determination under optimum conditions was
investigated. The specificity of the RP-HPLC method was established by
injecting the blank and placebo solution into the HPLC system. The
representative chromatogram of blank and placebo was shown in Figure 5 and 6
respectively.
Linearity for Eluxadoline:
Aliquots of 0.1, 0.25, 0.5, 0.75, 1 and 1.25ml of standard working
solution of Eluxadoline was pipetted out from the standard stock solution of
1000µg/ml of Eluxadoline and transferred into a series of 10ml clean dry
volumetric flask and make volume up to the mark with the same diluent to get
the concentration of 10, 25, 50, 75, 100 and 125µg/ml of Eluxadoline. The
calibration standard solutions of Eluxadoline were injected using a 20μl
Hamilton Rheodyne injector and the chromatograms were recorded at 260nm and a
calibration graph was obtained by plotting peak area versus concentration of
Eluxadoline.
Figure 5:
Chromatogram of Blank
Figure 6:
Chromatogram of Placebo
The linearity data is presented
in Figure 7 and Table 3. Acceptance Criteria: Correlation coefficient should be
not less than 0.999
Table 3: Linearity data for Eluxadoline
|
Linearity
of Eluxadoline |
|
|
Concentration (µg/ml) |
Peak Area |
|
10 |
141572 |
|
25 |
348774 |
|
50 |
718146 |
|
75 |
1060463 |
|
100 |
1411231 |
|
125 |
1765875 |
Figure 7:
Linearity graph of Eluxadoline
Accuracy studies for Eluxadoline:
The accuracy of the
method was determined by calculating recovery of Eluxadoline by the method of
standard addition. Known amount of standard solution of Eluxadoline at 50%,
100% and 150% was added to a pre quantified sample solution and injected into the HPLC system.
The mean percentage recovery of Eluxadoline at each level was calculated and
the results were presented in Table 4.
Preparation of pre quantified sample solution for
accuracy studies:
Twenty tablets were accurately weighed
and powdered and tablet powder equivalent to 100mg of Eluxadoline were taken
into 100ml clean dry volumetric flask, diluent was added and sonicated to dissolve
it completely and volume was made up to the mark with the same diluent and
filtered through 0.45 μm nylon membrane filter. Further pipette out 0.5ml
from the above Eluxadoline sample stock solution into a 10ml volumetric flask
and diluted up to the mark with diluent to get the concentration of 50µg/ml of
Eluxadoline.
Preparation
of standard solution of Eluxadoline for accuracy studies:
Standard stock solution of Eluxadoline were prepared by dissolving
100mg of Eluxadoline in100ml of diluent into a 100ml clean dry volumetric flask
and the standard solution was filtered through 0.45μm nylon membrane
filter and degassed by sonicator to get the concentration of 1000µg/ml of
Eluxadoline.
Preparation
of 50% standard solution:
From the standard stock solution of
1000µg/ml of Eluxadoline further pipette 0.25ml and transferred into a 10ml
volumetric flask and dilute up to the mark with diluent to get the
concentration of 25µg/ml of Eluxadoline.
Preparation
of 100% standard solution:
From the standard stock solution of
1000µg/ml of Eluxadoline further pipette 0.5ml and transferred into a 10ml
volumetric flask and dilute up to the mark with diluent to get the
concentration of 50µg/ml of Eluxadoline.
Preparation of 150% standard solution:
From the standard stock solution of
1000µg/ml of Eluxadoline further pipette 0.75ml and transferred into a 10ml
volumetric flask and dilute up to the mark with diluent to get the
concentration of 75µg/ml of Eluxadoline.
Acceptance
Criteria: The % Recovery for each
level should be between 98.0 to 102.0%.
Table 4: Recovery study data of Eluxadoline
|
Sample name |
Amount added (µg/ml) |
Amount found (µg/ml) |
% Recovery |
Statistical
Analysis |
|
S1:50% |
25 |
24.99 |
99.96 |
Mean-99.92 S.D-0.16 %RSD-0.16 |
|
S2:50% |
25 |
24.94 |
99.75 |
|
|
S3:50% |
25 |
25.01 |
100.06 |
|
|
S4:100% |
50 |
49.98 |
99.96 |
Mean-100.06 S.D-0.09 %RSD=0.09 |
|
S5:100% |
50 |
50.04 |
100.09 |
|
|
S6:100% |
50 |
50.07 |
100.14 |
|
|
S |
75 |
75.03 |
100.04 |
Mean-100.07 S.D-0.05 %RSD-0.05 |
|
S8:150% |
75 |
75.10 |
100.13 |
|
|
S9 :150% |
75 |
75.04 |
100.05 |
Precision studies for Eluxadoline:
Method precision (Repeatability):
Twenty tablets were accurately weighed
and powdered and tablet powder equivalent to 100mg of Eluxadoline were taken
into 100ml clean dry volumetric flask, diluent was added and sonicated to
dissolve it completely and volume was made up to the mark with the same diluent
and filtered through 0.45 μm nylon membrane filter. Further pipette out
0.5ml from the above Eluxadoline sample stock solution into a 10ml volumetric
flask and diluted up to the mark with diluent to get the concentration of
50µg/ml of Eluxadoline. A homogenous sample of a single batch analysed six
times and was checked whether the method is giving consistent results. The %RSD
for the area of six replicate injections was calculated as mentioned in Table
5. Acceptance Criteria: The % RSD
for the peak area of six sample injections should not be more than 2%.
Table 5: Method precision
data for Eluxadoline
|
Eluxadoline |
||||
|
S. No. |
Concentration
(μg/ml) |
Retention
time (min) |
Peak Area |
% Assay |
|
1 |
50 |
3.481 |
720811 |
99.97 |
|
2 |
50 |
3.473 |
721121 |
100.01 |
|
3 |
50 |
3.473 |
719528 |
99.79 |
|
4 |
50 |
3.473 |
719982 |
99.85 |
|
5 |
50 |
3.481 |
719291 |
99.76 |
|
6 |
50 |
3.481 |
721187 |
100.02 |
|
Average |
3.477 |
720320 |
99.90 |
|
|
SD |
0.004382 |
828.8208 |
0.11495 |
|
|
%RSD |
0.13 |
0.12 |
0.12 |
|
System precision:
The system precision was carried out to ensure that the analytical
system is working properly. The standard preparation concentration of 50µg/ml
of Eluxadoline was injected six times into the HPLC and the %RSD for the area
of six replicate injections was calculated as mentioned in Table 6. Acceptance
Criteria: The % RSD for the peak
area of six standard injections should not be more than 2%.
TABLE 6: System precision
data for Eluxadoline
|
Eluxadoline |
|||
|
S.No. |
Concentration
(μg/ml) |
Retention time (min) |
Peak Area |
|
1 |
50 |
3.481 |
721976 |
|
2 |
50 |
3.498 |
720442 |
|
3 |
50 |
3.498 |
720177 |
|
4 |
50 |
3.481 |
720371 |
|
5 |
50 |
3.473 |
721997 |
|
6 |
50 |
3.473 |
722911 |
|
Average |
3.484 |
721312 |
|
|
SD |
0.011419 |
1131.152 |
|
|
%RSD |
0.33 |
0.16 |
|
TABLE 7: Ruggedness data for
Eluxadoline
|
Ruggedness Data for Eluxadoline |
||||||||
|
Laboratory-1
(% Assay)-HPLC-1 |
Laboratory-2
(% Assay)-HPLC-2 |
|||||||
|
|
Analyst-1 |
Analyst-2 |
Analyst-1 |
Analyst-2 |
||||
|
Conc. (μg/ml) |
Day-1 |
Day-2 |
Day-1 |
Day-2 |
Day-1 |
Day-2 |
Day-1 |
Day-2 |
|
50 |
99.87 |
100.04 |
100.12 |
100.03 |
100.06 |
100.08 |
100.09 |
100.07 |
|
50 |
99.90 |
100.02 |
99.98 |
100.01 |
100.09 |
100.01 |
99.89 |
100.03 |
|
50 |
99.73 |
99.74 |
99.76 |
99.62 |
99.67 |
100.07 |
99.87 |
99.35 |
|
50 |
99.74 |
99.83 |
99.83 |
99.78 |
99.84 |
99.85 |
100.08 |
100.02 |
|
50 |
99.75 |
99.77 |
99.76 |
99.74 |
99.79 |
99.72 |
100.04 |
99.65 |
|
50 |
99.99 |
100.11 |
100.12 |
100.09 |
100.07 |
100.06 |
100.15 |
99.91 |
|
Average |
99.83 |
99.92 |
99.93 |
99.88 |
99.92 |
99.97 |
100.02 |
99.84 |
|
SD |
0.1079 |
0.15728 |
0.16869 |
0.1906 |
0.1754 |
0.148044 |
0.113313 |
0.283402 |
|
%RSD |
0.11 |
0.16 |
0.17 |
0.19 |
0.18 |
0.15 |
0.11 |
0.28 |
|
Intermediate precision within-laboratories
variations (n=24) |
||||||||
|
Laboratory-1
(% Assay)-HPLC-1 |
Laboratory-2
(% Assay)-HPLC-2 |
|||||||
|
Average |
99.89 |
Average |
99.94 |
|||||
|
SD |
0.045461 |
SD |
0.076757 |
|||||
|
%RSD |
0.05 |
%RSD |
0.08 |
|||||
|
Reproducibility between laboratories (n=48) (% Assay) |
||||||||
|
Average |
99.92 |
|||||||
|
SD |
0.035355 |
|||||||
|
%RSD |
0.04 |
|||||||
Table 8: Summary of validation parameter for Eluxadoline
|
Parameters |
RP-HPLC method |
|
|
Eluxadoline |
||
|
Linearity range
(µg/ml) |
10-125 |
|
|
Slope |
14128 |
|
|
Intercept |
973.6 |
|
|
Correlation
coefficient |
0.999 |
|
|
LOD (µg/ml) |
0.08 |
|
|
LOQ (µg/ml) |
0.25 |
|
|
Method Precision (%
RSD, n=6) |
0.12 |
|
|
System precision (%
RSD, n=6) |
0.16 |
|
|
Ruggedness (% RSD,
n=24) |
Lab-1 |
Lab-2 |
|
0.05 |
0.08 |
|
|
Reproducibility (% RSD, n=48) |
0.04 |
|
|
% Accuracy |
99.92-100.07 |
|
|
Robustness (% RSD,
n=6) |
Less Flow rate |
More Flow rate |
|
|
0.09 |
0.03 |
|
Less Organic phase |
More Organic phase |
|
|
0.02 |
0.15 |
|
Table 9: Summary of
Robustness (Change in Flow Rate) for Eluxadoline
|
Drug |
Change in Flow rate (ml/min) |
Retention Time (Mins) |
Robustness (0.9 ml/min to 1.1 ml/min) |
||||
|
Average peak area (n=6) |
SD |
% RSD |
USP Plate Count |
Asymmetry |
|||
|
Eluxadoline |
0.9 |
3.634 |
721812 |
633.013 |
0.09 |
2923 |
1.1 |
|
1.0 |
3.481 |
721336 |
1148.885 |
0.16 |
2985 |
1.1 |
|
|
1.1 |
3.110 |
721699 |
233.7991 |
0.03 |
2841 |
1.1 |
|
Table 10: Summary of
Robustness (Change in Mobile Phase) for Eluxadoline
|
Drug |
Change in Mobile Phase |
Retention Time (Min) |
Change in mobile phase (Acetonitrile and water) (63:37
v/v to 77:23 v/v) |
||||
|
Average peak area (n=6) |
SD |
% RSD |
USP Plate Count |
Asymmetry |
|||
|
Eluxadoline |
10% less Organic (63:37 v/v) |
3.147 |
721839 |
145.3113 |
0.02 |
2832 |
1.1 |
|
Actual (70:30 v/v) |
3.481 |
721336 |
1148.885 |
0.16 |
2985 |
1.1 |
|
|
10% more Organic (77:23 v/v) |
3.510 |
724352 |
1072.527 |
0.15 |
2758 |
1.1 |
|
Table 11: Summary of solution
stability-effect of mobile phase (Acetonitrile
and Water (70:30, v/v) for Eluxadoline
for 48 hours at room temperature.
|
|
Solution stability for Eluxadoline |
|||||
|
S.No. |
Concentration (μg/ml) |
Retention time (min) |
Peak Area |
%Assay |
USP Plate Count |
Asymmetry |
|
1 |
50 |
3.481 |
721432 |
100.06 |
2955 |
1.1 |
|
2 |
50 |
3.473 |
721284 |
100.04 |
2971 |
1.11 |
|
3 |
50 |
3.473 |
721120 |
100.01 |
2879 |
1.1 |
|
4 |
50 |
3.473 |
719432 |
99.78 |
2961 |
1.11 |
|
5 |
50 |
3.481 |
721701 |
100.09 |
2946 |
1.1 |
|
6 |
50 |
3.481 |
718057 |
99.59 |
2931 |
1.1 |
|
Average |
3.477 |
720504 |
99.93 |
2941 |
1.1 |
|
|
SD |
0.004382 |
1443.526 |
0.200203 |
33.05601 |
0.005164 |
|
|
%RSD |
0.13 |
0.2 |
0.2 |
1.12 |
0.468 |
|
Intermediate precision/ruggedness:
The intermediate precision (also known as Ruggedness) of the method was
evaluated by performing precision on different laboratory by different analyst
and different days. The sample preparation concentration of 50µg/ml of
Eluxadoline was injected six times into the HPLC and the %RSD for the area of
six replicate injections was calculated as mentioned in Table 7. Acceptance
Criteria: The % RSD for the peak
area of six standard injections should not be more than 2%.
Limit of Detection (LOD)
and Limit of Quantification (LOQ):
Limit of Detection
(LOD) and Limit of Quantification (LOQ) were calculated as 3.3×SD/S and 10×SD/S
respectively as per ICH guidelines, Where
SD is the standard deviation of the response (Y-intercept) and S is the slope
of the calibration curve. The LOD is the
smallest concentration of the analyte that gives a measurable response (signal
to noise ratio of 3). The LOD of Eluxadoline was calculated and shown in Table
8. The LOQ is the smallest concentration of the analyte which gives response
that can be accurately quantified (signal to noise ratio of 10). The LOQ of
Eluxadoline was calculated and shown in Table 8.
Robustness:
As part of the
Robustness, deliberate change in the flow rate and mobile phase proportion of
±10% was made to evaluate the impact on the method. The
results reveal that the method is robust. The results are summarized in Table 9
and 10.
Stability of solution:
The %RSD of the assay of Eluxadoline from the solution stability and
mobile phase stability experiments was within 2%. The results of the solution
and mobile phase stability experiments confirm that the sample solutions and
mobile phase used during the assays were stable upto 48hours at room
temperature was calculated and shown in Table 11.
RESULTS AND DISCUSSION:
To optimize the
RP-HPLC parameters, several mobile phase compositions were tried. A
satisfactory separation and good peak symmetry for Eluxadoline were obtained
with a mobile phase containing a mixture of Acetonitrile and water (70:30, v/v)
was delivered at a flow rate of 1ml/min to get better reproducibility and
repeatability. Quantification was achieved with PDA detection at 260nm based on
peak area. The retention time of Eluxadoline
was found to be 3.481min. Linearity was established for Eluxadoline
in the range of 10-125µg/ml with correlation coefficient 0.999
and mean accuracies were found to be
is 99.92% to 100.07% for
Eluxadoline, which indicates accuracy of the proposed method. The % RSD values
of accuracy for Eluxadoline were found to be < 2 %. The % RSD value of
method precision was 0.12% for
Eluxadoline and % RSD value of system precision was 0.16% for Eluxadoline. The % RSD value of reproducibility is 0.04%
for Eluxadoline reveal that the proposed method is precise. LOD value for
Eluxadoline was found to be 0.08µg/ml
and LOQ value for Eluxadoline were found to be 0.25µg/ml. The % RSD values of robustness studies were found to be
< 2% reveal that the method is robust enough. These data show that the
proposed method is specific and sensitive
for the determination of Eluxadoline.
CONCLUSION:
RP-HPLC method for the estimation of
Eluxadoline in their bulk and pharmaceutical dosage form was established and
validated as per the ICH guidelines. Linearity
was achieved for Eluxadoline in
the range of 10-125µg/ml with
correlation coefficient 0.999. The percentage recovery of drug was
achieved in the range of 98-102% which was within the acceptance criteria. The
percentage RSD was NMT 2 % which proved the precision of the developed method.
The developed method is simple, sensitive, rapid, linear, precise, rugged,
accurate, specific, and robust. Hence it can be used for the routine analysis
of Eluxadoline in their bulk and pharmaceutical dosage form.
ACKNOWLEDGEMENT:
The authors are thankful
to Malla Reddy College of Pharmacy for providing the chemicals and instruments
and Manus Aktteva Biopharma LLP, Ahmedabad, India and Eisai Pharmaceuticals
India Pvt. Ltd, Mumbai, India for providing the samples for research.
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Received on 02.11.2015 Modified on 18.11.2015
Accepted on 21.11.2015 © RJPT All right reserved
Research J. Pharm. and Tech. 8(11): Nov., 2015; Page
1469-1476
DOI: 10.5958/0974-360X.2015.00263.2