RP-HPLC METHOD DEVELOPMENT AND...

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: [email protected].


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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.

http://en.wikipedia.org/wiki/Antidepressant

http://en.wikipedia.org/wiki/Sertraline

http://en.wikipedia.org/wiki/Fluoxetine

http://en.wikipedia.org/wiki/Citalopram

http://en.wikipedia.org/wiki/Nortriptyline

http://en.wikipedia.org/wiki/Amitriptyline

http://en.wikipedia.org/wiki/Dextromethorphan/quinidine


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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


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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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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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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.


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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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