Post on 17-May-2018
International Bulletin of Drug Research., 4(7): 66-83, 2014
66
RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR
SIMULTANEOUS ESTIMATION OF DEXTROMETHORPHAN
HYDROBROMIDE AND QUINIDINE SULPHATE
ZAHID A. CHAUDHARY*1
, JIGNASHA DERASARI2
_________________________________________________________________________
ABSTRACT
A simple, accurate, rapid, economical RP-HPLC method have been developed and validated
for estimation of Dextromethorphan Hydrobromide and Quinidine Sulphate in combined
capsuledosage form and can be used in routine analysis. RP-HPLC, method was carried out
by isocratic technique on a reversed-phase: Thermo scientific ODS C18 (250mm X 4.6 mm
i.d., 5 μm particle size) and UV detection at 234 nm with mobile phase containing a mixture
of Phosphate buffer (pH 4.5): Methanol (55: 45 v/v) at a flow rate of 1.0 mL/min. The
average retention times for Dextromethorphan Hydrobromide and Quinidine Sulphate were
3.6 and 5.5 min respectively. The calibration curves were linear in the concentration range of
10-30 μg/mL and 5-15μg/mL for Dextromethorphan Hydrobromide and Quinidine Sulphate
respectively. Correlation co efficient was found to be 0.995 and 0.997 for Dextromethorphan
Hydrobromide and Quinidine Sulphate respectively. Mean assay was found to be 99.5% and
97.00% for Dextromethorphan Hydrobromide and Quinidine Sulphate respectively.
The developed method was validated as per ICH guideline, for its accuracy, precision, LOD
& LOQ and the results were found to be satisfactory, thus the method is specific, rapid and
simple with good sensitivity for estimation of Dextromethorphan Hydrobromide and
Quinidine Sulphate. These analytical methods are also applicable in ordinary laboratories. It
can also be adopted for quality control tests for these drugs in capsule.
KEYWORDS
Dextromethorphan Hydrobromide, Quinidine Sulphate, RP-HPLC method, Validation.
AUTHORS AFFILIATION
*Address for correspondence
Department of Quality Assurance Techniques,Parul Institute of Pharmacy and research,
Vadodara,India-391760: Email: Zahid.chaudhary90@gmail.com.
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INTRODUCTION
Dextromethorphan Hydrobromide (3-methoxy-9-a-methylmorphinan Hydrobromide) is
methyl analog of DEXTRORPHAN that shows high affinity binding to several regions of the
brain, including the medullary cough center. This compound is an NMDA receptor antagonist
(RECEPTORS, N-METHYL-D-ASPARTATE) and acts as a non-competitive channel
blocker. It is one of the widely used antitussives. However Dextromethorphan is a
dextrorotatory analogue of levorphanol. It is σ-1 receptor agonist as well as a noncompetitive
inhibitor of the excitatory NMDA receptor. Blocking σ-1 receptors inhibits glutamate release
and, along with the noncompetitive inhibition of the NMDA glutamate receptor, would lead
to an inhibition of excitatory synaptic transmission, hence it is useful in treatment of
neurological disorders like Pseudobulbar affect (pathological laughing and crying).
Figure 1: structure of Dextromethorphan Hydrobromide.
Quinidine Sulphate (6 - methoxycinchonan-9-01 Sulphate) is a specific inhibitor of CYP2D6-
dependent oxidative metabolism.It increase the Systemic bioavailability of
Dextromethorphan Hydrobromide when given in combination for the treatment of
Pseudobulbar affect.
Figure 2: Structure of Quinidine Sulphate.
Crying associated with Pseudobulbar affect may be incorrectly interpreted as depression;
laughter may be embarrassing. It is therefore critical for families and caregivers to recognize
the pathological nature of Pseudobulbar affect and the reassurance that this is an involuntary
syndrome that is manageable.
Traditionally, antidepressants such as sertraline, fluoxetine, citalopram, nortriptyline
and amitriptyline have been prescribed with some efficacy for the treatment of Pseudobulbar
affect.
Dextromethorphan/Quinidine (trade name: Nuedexta) is the first FDA-approved drug for the
treatment of Pseudobulbar affect. Treatment with Dextromethorphan/Quinidine significantly
decreased laughing and crying episodes in patients with amyotrophic lateral sclerosis (ALS)
or multiple sclerosis (MS) as compared with placebo in a 12-week, randomized, double-blind
study.
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The present study was designed to develop a simple, precise, and rapid analytical RP-HPLC
procedure, which can be used for the analysis of assay method for simultaneous estimation of
Dextromethorphan Hydrobromide and Quinidine Sulphate as there was only individual
methods reported for both drugs. The combination of these two drugs is not official in any
pharmacopoeia; hence no official method is available for the simultaneous estimation of these
two drugs in their combined dosage forms.
The developed method was validated as per ICH guidelines and its updated international
convention. The linearity of response, precision and robustness of the described method has
been checked.
EXPERIMENTAL
Chemicals and Reagents
Dextromethorphan Hydrobromide and Quinidine Sulphate were obtained from Gitar
laboratory, Ahmedabad. The dosage form (capsule) is manufacture by Avanir
Pharmaceutical, marketed as brand name Nuedexta. All the reagents including potassium
dihydrogen phosphate, ortho phosphoric acid and Triethylene amine were purchased from
Merck and Rankem Ltd. New Delhi, India.
HPLC Instrumentation and Conditions
The analysis was carried out on a HPLC system (Shimadzu-LC 20AT) equipped with UV
detector, pressure controlled by prominence pump and operated by Spinchrom Software. C18
column (250 mm × 4.6 mm i.d., particle size 5 μm) was used for separation. Mobile phase
used for separation was containing Phosphate Buffer: Methanol in the ratio of 55:45
v/vadjusted to pH 4.5 with orthophosphoric acid. The flow rate was kept at 1.0 mL/min,
column temperature was ambient (25°C), eluents were detected by UV detector at 234 nm,
and the injection volume was 20 μL.
Preparation of Phosphate Buffer
Phosphate buffer was used for the estimation of both the drugs in their combined dosage
form. 6.8 gm of potassium dihydrogen phosphate (KH2PO4) was weighed accurately and
transferred in to 1000 mL beaker. 800 mL HPLC grade water was added to dissolve it. The
pH 4.5 was adjusted with diluted ortho phosphoric acid (1%) and made up the volume up to
1000 mL with HPLC grade water to give 0.05 M Phosphate buffer.
Preparation of Mobile Phase
Mobile phase was prepared by mixing 60 volumes of 0.05 M Phosphate buffer and 40
volumes of Methanol was adjusted to pH 4.5 with ortho phosphoric acid. The mobile phase
was ultrasonicated, filtered through 0.45 μm membrane filter, and degassed.
Preparation of mixed standard stock solutions of Dextromethorphan Hydrobromide
and Quinidine Sulphate
20 mg of standard Dextromethorphan Hydrobromide and 10 mg standard Quinidine Sulphate
were accurately weighed and transferred to a 100 mL volumetric flask and dissolved in 25
mL mobile phase. The flask was shaken and volume was made up to the mark with mobile
International Bulletin of Drug Research., 4(7): 66-83, 2014
69
phase to give a solution containing 200 μg/mL Dextromethorphan Hydrobromide and 100
μg/mL Quinidine Sulphate. From this stock solution different aliquots were prepared.
Selection of detection wavelength
In the present study individual drug solutions of 10μg/mL were prepared in different solvent
mixtures of HPLC grade organic and inorganic solvents at different ratio. These drug
solutions were than scanned in the UV region of 200-400 nm and the spectrums were
recorded to get λmax of analytes in Mobile Phase. 234 nm wavelength was selected for
estimation of this combination.
Method Validation
Linearity and Range
The linearity peak area response was determined by analyzing solutions having
concentrations in the range of 10-30 μg/mL and 5-15 μg/mL for Dextromethorphan
Hydrobromide and Quinidine Sulphate respectively from same solution. Peak area of each
solution was measured using developed method. Calibration curve of peak area Vs
concentration was plotted. The correlation coefficient and regression line equations for
Dextromethorphan Hydrobromide and Quinidine Sulphate were determined.
Accuracy
To study the accuracy powdered synthetic mixture analysis of thesame was carried out.
Recovery studies were carried out by addition of standard drugto the sample at 3 different
concentration levels (80, 100 and 120 %) taking into consideration percentagepurity of added
bulk drug samples. It was determined by calculating the recovery of Dextromethorphan
Hydrobromide and Quinidine Sulphateby standard addition method. Accuracy is the
closeness of the test results obtained by the method to the true value.
Preparation of sample solution for % recovery
Marketed formulation equivalent to 20 mg Dextromethorphan Hydrobromide and 10 mg
Quinidine Sulphate was accurately weighed and transferred to volumetric flask of 100mL
capacity and aliquate them to make final concentration 10 μg/mL Dextromethorphan
Hydrobromide and 5 μg/mL Quinidine Sulphate. The resulting solution was filtered through
Whatman filter paper. Absorbance of sample solutions was measured at selected wavelength
of Dextromethorphan Hydrobromide and Quinidine Sulphate and concentration is calculated
which is known as pre-analyzed sample. In pre-analyzed sample 80, 100 and 120 % of
Dextromethorphan Hydrobromide and Quinidine Sulphate was spiked. Absorbance of spiked
samples was measured and total amount of drug was calculated and from which % recovery
was calculated.
Precision
Repeatability
6 replicates of standard mixture solution having Dextromethorphan Hydrobromide (20
μg/mL) and Quinidine Sulphate (10 μg/mL) were prepared and chromatograms were
recorded and RSD was calculated.
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Intraday precision
Standard solutions containing 10, 20 and 30 μg/mL Dextromethorphan Hydrobromide and 5,
10 and 15 μg/mL Quinidine Sulphate were analyzed 3 times on the same day as per the
procedure. Chromatogram of each sample was taken. SD and RSD were calculated.
Interday precision
Standard solutions containing 10, 20 and 30 μg/mLDextromethorphan Hydrobromide and 5,
10 and 15 μg/mLQuinidine Sulphate were analyzed on three different daysas per the
procedure. Chromatogram of each sample was taken. SD and RSD were calculated.
Limit of Detection
Calibration curve was repeated for 6 times and the standard deviation (SD) of the intercepts
was calculated than LOD was calculated as follow from the formula:
LOD= (3.3*SD)/Slope
Where, SD = the standard deviation of Y- intercept of 6 calibration curves.
Slope = the mean slope of the 6 calibration curves.
Limit of Quantitation
Calibration curve was repeated for 6 times and the standard deviation (SD) of the intercepts
was calculatedthan LOQ was calculated as follow from the formula:
LOQ= (10*SD)/Slope
Where, SD = the standard deviation of Y- intercept of 6 calibration curves.
Slope = the mean slope of the 6 calibration curves.
Robustness Study
To evaluate robustness of the method few parameters were deliberately varied. The
parameters included were variation of flow rate, change in pH of mobile phase and change of
mobile phase ratio. The change was made at 3 levels and replicate for 3 times. The system
suitability parameters were calculated for Dextromethorphan Hydrobromide and Quinidine
Sulphate.
System suitability
20 μg/mLDextromethorphan Hydrobromide and 10 μg/mLQuinidine Sulphate in mixture
was injected and the chromatograms was recorded. The resolution, number of theoretical
plates, and peak asymmetry were calculated to determine whether the result complies with
the recommended limit.
Analysis of Pharmaceutical Preparations: %Assay of Dextromethorphan Hydrobromide
and Quinidine Sulphate in their Capsule Dosage Form (Formulation: NUEDEXTA capsule)
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Sample preparation
For estimation of Dextromethorphan Hydrobromide and Quinidine Sulphate, 20 capsules was
taken and removed their shells, Powder was accurately weighed. Powder equivalent to 20 mg
Dextromethorphan Hydrobromide and 10 mg Quinidine Sulphate was accurately weighed
and transferred to volumetric flask of 100 mL capacity. 100 mL of methanol was transferred
to this volumetric flask and sonicated for 15 min. The flask was shaken and volume was
made up to the mark with methanol. The above solution was filtered through whatman filter
paper (0.45μ). From this solution 1 mL was transferred to volumetric flask of 10 mL
capacity. Volume was made up to the mark to give a solution containing 20 μg/mL
Dextromethorphan Hydrobromide and 10 μg/mL Quinidine Sulphate (solution A). This
solution was used for the estimation of Dextromethorphan Hydrobromide and Quinidine
Sulphate.
RESULT AND DISCUSSION
Optimization of Chromatographic Conditions
To develop suitable RP-HPLC method for simultaneous estimation of Dextromethorphan
Hydrobromide and Quinidine Sulphate, different chromatographic conditions were applied
and optimized chromatographic conditions were developed.(see figure 3)
Figure 3: Optimized chromatogram of Dextromethorphan Hydrobromide and
Quinidine Sulphate.
Optimized chromatographic conditions are as follows (see table 1)
Table 1: Optimized chromatographicconditions.
S.
No. PARAMETER CONDITIONS
1. Mobile phase Phosphate buffer (pH 4.5) : Methanol (55 : 45)
2. Pump mode Isocratic
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3. Stationary phase C-18 (250mm x 4.6 mm, i.d. 5μm particle size)
4. Flow rate (mL/min) 1.0
5. Run time (min) 10.0
6. Volume of Injection (μl) 20.0
7. Detection wavelength (nm) 234
8.
Retention
time
(min)
Dextromethorphan
Hydrobromide 3.620
Quinidine Sulphate 5.540
Validation
Linearity
Linearity was performed on synthetic mixture of Dextromethorphan Hydrobromide and
Quinidine Sulphate as per marketed formulation composition.(see figure 4)
Figure 4: Chromatograms of Dextromethorphan Hydrobromide andQuinidine
Sulphate for Linearity.
Calibration curve for the Dextromethorphan Hydrobromide (10 - 30μg/mL)
Chromatogram for following concentrations 10, 15, 20, 25 and 30 μg/mL for
Dextromethorphan Hydrobromide at 234 nm at flow rate of 1 mL/min. Peak area were
obtained as tabulated in Table 2 and the graph of calibration curve was obtained as shown in
Figure 5.
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73
Table 2: Calibration curve for Dextromethorphan Hydrobromide.
Concentrations
(µg/mL)
Peak area of Dextro
(Mean ± S.D.)
10 642.776 ±6.11
15 952.502 ±8.85
20 1297.749 ±15.04
25 1697.989 ± 17.42
30 1944.364 ±22.17
Fig.5 Graph of Calibration curve for Dextromethorphan Hydrobromide
Discussion:Linearity range for Dextromethorphan Hydrobromide was found to be 10-30
µg/mL in Mobile Phase. Regression Equation for Dextromethorphan Hydrobromide at 234
nm: y = 66.973x - 32.38.R2 value: 0.995.
Calibration curve for the Quinidine Sulphate (5-15 μg/mL)
Chromatogram for concentrations 5, 7.5, 10, 12.5 and 15 μg/mL for Quinidine Sulphate at
234 nm at flow rate of 1 mL/min were used for making calibration curve. Peak area were
obtained as tabulated in Table 3 and the graph of calibration curve was obtained as shown in
Figure 6.
Table 3: Calibration curve for Quinidine Sulphate.
Concentrations
(µg/mL)
Peak area of QUINI
(Mean ± S.D.)
5 734.767 ± 9.46
7.5 1089.515 ± 9.82
10 1487.685 ± 8.25
12.5 1704.18 ± 12.58
15 2231.935 ± 14.20
y = 66.97x - 32.38R² = 0.995
0
1000
2000
3000
0 5 10 15 20 25 30 35
a
r
e
a
Conc.(ppm)
dextro
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Figure 6: Graph of Calibration curve for Quinidine.
Discussion
Linearity range for Quinidine Sulphate was found to be 30-150 µg/mL in mobile phase.
Regression Equation for Quinidine Sulphate at 234 nm.y = 144.3x + 6.016.R2 value: 0.997.
Accuracy (% Recovery study)
Dextromethorphan Hydrobromide
Sample was taken equivalent to 10 μg/mL Dextromethorphan Hydrobromide
Table 4: %Recovery data for Dextromethorphan Hydrobromide.
Level of
recovery
Amt of
std
spiked
(μg/mL)
Total amt
of sample
(μg/mL)
Amt of
std found
(μg/mL)
Peak
area of
sample
Amount of
std
recovered
(μg/mL)
% recovery
mean ± SD
(n=3)
80%
8 18 17.88 1142.032 7.88 99.77
±
1.12
8 18 18.06 1153.618 8.06
8 18 18 1149.621 8.00
100%
10 20 19.88 1272.571 9.88 99.48
±
0.65
10 20 20.01 1281.058 10.01
10 20 19.94 1276.524 9.94
120%
12 22 21.99 1410.27 11.99 99.47
±
0.48
12 22 21.87 1402.635 11.87
12 22 21.93 1406.68 11.93
(n=3 Determination)
y = 144.3x + 6.016R² = 0.997
0
1000
2000
3000
0 2 4 6 8 10 12 14 16
a
r
e
a
Conc. (ppm)
quini
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75
Discussion
Result obtained reveals that % recovery of Dextromethorphan Hydrobromide was within
acceptance criteria given in ICH i.e. 98-102%.
Quinidine Sulphate
Sample was taken equivalent to 5 μg/mL Quinidine Sulphate.
Table 5: % Recovery data for Quinidine Sulphate.
Level of
recovery
Amt of
std
spiked
(μg/mL)
Total
amt of
sample
(μg/mL)
Amt of std
found
(μg/mL)
Peak
area of
sample
Amount
of std
recovered
(μg/mL)
% recovery
mean ± SD
(n=3)
80%
4 9 8.95 1299.854 3.95 99.88
±
1.02
4 9 8.99 1305.491 3.99
4 9 9.01 1308.477 4.01
100%
5 10 9.96 1449.909 4.96 99.67
±
0.64
5 10 10 1456.264 5.00
5 10 9.99 1454.383 4.99
120%
6 11 10.98 1602.617 5.98 99.68
±
0.49
6 11 10.96 1599.223 5.96
6 11 10.99 1603.829 5.99
(n=3 Determination)
Discussion
Result obtained reveals that % recovery of Quinidine Sulphate was within acceptance criteria
given in ICH i.e. 98-102%.
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Precision
Repeatability
Table 6: Repeatability data for Dextromethorphan Hydrobromide and Quinidine Sulphate.
Standard Drug
Target
Concentration
(μg/mL)
Peak
Area of
Sample
Found
Concentration
(μg/mL)
Mean SD %
RSD
Dextromethorphan
Hydrobromide
20 1292.539 19.78
19.79 0.13 0.67
20 1276.196 19.53
20 1297.748 19.86
20 1300.353 19.9
20 1296.428 19.84
20 1296.417 19.84
Quinidine Sulphate
10 1481.705 10.22
10.21 0.08 0.81
10 1484.711 10.24
10 1456.459 10.05
10 1490.674 10.28
10 1486.162 10.25
10 1486.195 10.25
Discussion
The % RSD for Repeatability of both the drugs was found to be less than 2. So, it was
concluded that proposed method for estimation of Dextromethorphan Hydrobromide and
Quinidine Sulphate is précised in nature.
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Intraday precision
Table 7: Intraday precision data for Dextromethorphan Hydrobromide and Quinidine Sulphate.
Standard Drug
Target
Concentration
(μg/mL)
Peak
Area of
Sample
Found
Concentration
(μg/mL)
Mean SD %
RSD
Dextro-
methorphan
Hydrobromide
10 629.763 9.89
9.89 10.03 0.91 10 639.554 10.03
10 641.508 10.06
20 1278.208 19.56
19.71 0.136 0.69 20 1291.241 19.76
20 1295.134 19.82
30 1907.13 28.96
29.26 0.266 0.91 30 1934.612 29.37
30 1940.479 29.46
Quinidine
Sulphate
5 729.626 5.01
5.00 0.03 0.70 5 724.399 4.97
5 733.323 5.04
10 1477.263 10.20
10.15 0.07 0.71 10 1459.152 10.19
10 1484.684 10.07
15 2216.275 15.32
15.29 0.11 0.77 15 2193.198 15.16
15 2227.505 15.39
Discussion
The % RSD for Intraday precision of both the drugs was found to be less than 2. So, it was
concluded that proposed method for estimation of Dextromethorphan Hydrobromide and
Quinidine Sulphate is précised in nature.
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Interday precision
Table 8: Interday precision data for Dextromethorphan Hydrobromide and Quinidine Sulphate.
Standard Drug
Target
Concentration
(μg/mL)
Peak
Area of
Sample
Found
Concentration
(μg/mL)
Mean SD %
RSD
Dextromethorphan
Hydrobromide
10 629.226 9.88
9.98 0.08 0.87 10 640.195 10.04
10 638.942 10.02
20 1273.085 19.49
19.66 0.15 0.76 20 1292.536 19.78
20 1287.394 19.70
30 1914.636 29.07
29.27 0.18 0.60 30 1936.552 29.40
30 1932.718 29.34
Quinidine Sulphate
5 730.354 5.02
5.0 0.03 0.53 5 724.021 4.97
5 730.391 5.01
10 1478.739 10.20
10.16 0.08 0.83 10 1457.289 10.06
10 1478.746 10.21
15 2218.498 15.33
15.25 0.12 0.81 15 2185.983 15.11
15 2218.587 15.32
Discussion
The SD and % RSD for Interday precision of both the drugs was found to be less than 1 and
less than 2 respectively. So, it was concluded that proposed method for estimation of
Dextromethorphan Hydrobromide and Quinidine Sulphate is précised in nature.
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Limit of Detection (LOD) and Limit of Quantitation (LOQ)
Table 9: LOD and LOQ data for Dextromethorphan Hydrobromide and Quinidine Sulphate.
Parameters Dextromethorphan
Hydrobromide
Quinidine Sulphate
Mean Slope (n=6) 66.94 147.91
SD of Y intercept (n=6) 15.15 6.59
LOD (μg/mL) 0.75 0.15
LOQ (μg/mL) 2.26 0.44
Discussion
The proposed method can detect Dextromethorphan Hydrobromide and Quinidine Sulphate at
very low level (0.75 μg/mL Dextromethorphan and 0.15 μg/mL) and can quantify small
amount of drugs (2.26 μg/mL Dextromethorphan Hydrobromide and 0.44 μg/mL Quinidine
Sulphate) with precisely So, it was concluded that the proposed method is very sensitive in
nature.
Robustness Study
Table 10: Robustness data for Dextromethorphan Hydrobromide.
S.No. Flow rate(mL/min) Mobile phase pH
Level 1.2 0.8 57:43 53:47 4.7 4.3
1 1242.249 1326.394 1234.982 1322.269 1217.077 1313.674
2 1267.863 1345.825 1265.322 1328.814 1241.89 1332.821
3 1275.653 1350.97 1274.379 1336.645 1247.116 1340.666
Mean 1261.922 1341.063 1258.228 1329.243 1235.361 1329.054
SD 17.47 12.96 20.63 7.19 16.04 13.88
%RSD 1.38 0.96 1.64 0.54 1.29 1.04
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Table 11: Robustness data for Quinidine Sulphate.
S.No. Flow rate(mL/min) Mobile phase pH
Level 1.2 0.8 57:43 53:47 4.7 4.3
1 1444.551 1538.72 1441.651 1522.284 1417.614 1520.76
2 1434.06 1512.734 1419.96 1496.789 1397.463 1523.056
3 1462.349 1550.579 1460.889 1534.102 1429.675 1538.72
Mean 1446.987 1534.11 1440.833 1517.725 1414.917 1527.512
SD 14.30 19.35 20.47 19.06 16.27 9.77
%RSD 0.98 1.26 1.42 1.25 1.15 0.63
Discussion
The % RSD for Robustness of both the drugs was found to be less than 2 respectively. So, It
was concluded that proposed method for estimation of Dextromethorphan Hydrobromide and
Quinidine Sulphate is précised in nature as per ICH guideline.
System suitability
Table 12: System suitability Parameters.
Name Rt (min) Area Tf Resolution Theoretical
Plate#
Dextromethorphan
Hydrobromide 3.620 1297.749 1.391
8.804 6799
Quinidine Sulphate 5.540 1487.685 1.400 7232
Analysis of Pharmaceutical Preparations
%Assay of Dextromethorphan Hydrobromide (20 μg/mL) and Quinidine Sulphate (10
μg/mL) in their Capsule Dosage Form
Figure 7: Chromatograph of Sample (NEUDEXTA Capsule) Equivalent to 20 μg/mL
of Dextromethorphan Hydrobromide and 10 μg/mL of Quinidine Sulphate.
International Bulletin of Drug Research., 4(7): 66-83, 2014
81
Drug
Concentration
in marketed
formulation
(mg)
Conc.
taken
for
%Assay
Peak
area of
Sample
solution
Concentration
found from
Mixture
%
Assay
±
S.D.
Dextromethorphan
Hydrobromide 20
20
μg/mL 1245.956 19.9μg/mL
99.5
±0.69
Quinidine Sulphate 10 10
μg/mL 1403.354 9.7 μg/mL
97.00
±1.09
Discussion
% Assay of Dextromethorphan Hydrobromide and Quinidine Sulphate was found in an
acceptance limit so this method could be used for analysis of this combination.
CONCLUSION
The proposed RP-HPLC method was used for the simultaneous estimation of
Dextromethorphan Hydrobromide and Quinidine Sulphate was found to be sensitive,
accurate, precise, simple, and rapid. Hence the present RP-HPLC method may be used for
routine analysis of the raw materials, in combinational dosage formulations containing
Dextromethorphan Hydrobromide and Quinidine Sulphate.
ACKNOWLEDGEMENTS
The authors are thankful to all teaching and non-teaching staff ofParul institute of pharmacy
& research,Vadodara, for providing necessary facilities.
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