UV Spectrophotometric Development and Validation of Zidovudine API and in Marketed Zidovudine Tablet Dosage form

 

Santosh Gada1*, Anand Kumar Y2, Saritha2, C. Mallikarjuna Setty3

1Research Scholar, JNTU Hyderabad and KCT College of Pharmacy, Gulbarga, India.

2Department of Pharmaceutics, V L College of Pharmacy, Raichur. Karnataka, India.

3Department of Pharmaceutics, Oxford College of Pharmacy, Bengaluru. Karnataka, India.

*Corresponding Author E-mail: gadasantosh@yahoo.co.in

 

ABSTRACT:

To develop and validate simple, rapid, linear, accurate, precise and economical UV Spectroscopic method for estimation of zidovudine API and in zidovudine tablet dosage form. The UV spectroscopic method was developed for estimation of zidovudine API and in zidovudine tablet dosage form and also validated as per ICH guidelines. The different solvent blends viz., Saline phosphate buffer pH 7.2(Method-I); Methanol: Water: 0.1N HCl (3:1:1) (Method-II); Methanol: Water: 0.1N NaOH (3:1:1) (Method-III) were selected. It showed absorption maxima 267 nm for all three different methods. The linearity was observed between 1-18 μg/ml in Method-I, 1-20 µg/ml in Method-II and 1-16 µg/ml in Method-III, with correlation coefficient of 0.9998. The results of the analysis were validated by recovery studies. The recovery was found to be 99.9±0.750, 100.2±1.114, 98.8±1.635 with % RSD values of 0.750, 1.111, 1.635 for Method-I, Method-II and Method-III respectively which were within the acceptance limit. The % RSD values of intraday precision for zidovudine in formulations were found to be 0.554, 0.532, 0.437 and % RSD values of inter day precisions for zidovudine were found to be 1.354, 1.659, 1.406 for Method-I; Method-II and Method-III respectively which were within the acceptance limit. A simple, rapid, linear, accurate, precise and economical UV Spectroscopic method has been developed for estimation of zidovudine API and zidovudine tablet dosage form. The method could be considered for the determination of zidovudine in quality control laboratories.

 

KEYWORDS: UV Spectrophotometer, Validation, Accuracy, Linearity, Ruggedness, Precision.

 

 


INTRODUCTION:

Analysis of active pharmaceutical component is an integral part of preformulation and formulation development. It is essential to have a validated, specific method of analysis for the drug.

 

 

UV spectrophotometer technique is one of the earliest and most widely applied detection techniques for drug estimation. UV spectrophotometer method is preferred over other technique for routine analysis as it is less time consuming and also cost effective. Method validation is the process used to confirm that the analytical procedure employed for a specific test is suitable for its intended use. Results from method validation can be used to judge the quality, reliability and consistency of analytical results and it is an integral part of any good analytical practice1.

 

 

 

Antiretroviral drugs like nucleoside reverse transcriptase inhibitors, non nucleoside reverse transcriptase inhibitors and protease inhibitors are essential in the management of HIV infection. Zidovudine is a thymidine analogue2,3. It is phosphorylated in the body to zidovudine triphosphate which is the active form that inhibits HIV replication4. Zidovudine inhibits the key enzyme reverse transcriptase.

 

It has been clearly demonstrated that zidovudine reduces opportunistic infections and prolongs survival in patients with advanced HIV infection5. Zidovudine is chemically 1-(3-azide-2, 3-di deoxy-β-D-ribofuranosyl)-5-methyl pyrimidin- 2,4 (1H, 3H)–dione, (Fig 1) with the molecular formula C10H13N5O4 and molecular weight of 267.25g/mol. Zidovudine is Freely Soluble in water slightly soluble in Acetic acid and methanol, very slightly soluble in dehydrated alcohol6. Since it was first recognized in 1981, the Acquired Immuno Deficiency Syndrome (AIDS) has been a major public health problem. Zidovudine the first anti-HIV compound approved for clinical use is still widely used for antiretroviral therapy in combination with other antiretroviral agents7,8.

 

In the literature, numerous methods to quantify the Zidovudine have been described either alone or in combination with other antiretroviral drugs9-11. A literature survey reveals that HPLC is the most widely used technique for the estimation of zidovudine in human plasma, saliva, cerebrospinal fluid and human blood cells, as well as for studying the drug metabolites in the urine. Most of the methods using HPLC with UV detection are relatively time-consuming or use expensive instrumentation not readily accessible to many groups12-24

 

 

Fig-1: Structure of zidovudine

 

Thus best of our knowledge, there is no specific spectrophotometric method available for determination of Zidovudine in pharmaceutical formulation. Hence in the present work it was aimed to develop and validate accurate, precise, simple and rapid UV spectroscopic methods for the estimation of zidovudine API and zidovudine tablet as per ICH guidelines.

 

MATERIALS AND METHODS:

Materials

Zidovudine API gift sample was obtained from Strides Arco lab, Bengaluru. Zidovudine 100 mg tablets were purchased from local stores. Methanol, Hydrochloric acid and Sodium hydroxide were procured from S.D Fine chemicals Mumbai, double distilled water was used throughout the experiments.

 

METHODS:

Preparation of zidovudine API standard stock solutions (1000µg/ml):

Weighed accurately about 50 mg of zidovudine working standard and transferred to a 50 ml volumetric flask. Add 40 ml of saline phosphate buffer pH 7.2 (Method-I) and shake for 5 minutes to dissolve and dilute to volume with saline phosphate buffer pH 7.2. Similarly prepare standard stock solutions in selected solvent blends viz., Methanol: Water: 0.1N HCl (3:1:1) (Method-II); Methanol: Water: 0.1N NaOH (3:1:1) (Method-III).

 

Determination of absorption maxima of zidovudine API in selected solvent blends:

From the zidovudine API standard stock solution, different aliquots were taken and diluted to 10 ml mark with saline phosphate buffer pH 7.2 (Method-I) to obtain series of concentrations. The solutions were scanned on spectrophotometer in the UV range of 200-380 nm. Similarly find out the absorption maxima in selected solvent blends viz., Methanol: Water: 0.1N HCl (3:1:1) (Method-II); Methanol: Water: 0.1N NaOH (3:1:1) (Method-III).

 

Determination of linearity range of zidovudine API in selected solvent blends:

Standard solutions of zidovudine in the concentration range of 1-30 μg/ml were prepared and absorbance was measured at 267 nm taking saline phosphate buffer pH 7.2 (Method-I) as the blank. Similarly absorbance of zidovudine in the concentration range of 1-30 μg/ml in selected solvent blends viz., Methanol: Water: 0.1N HCl (3:1:1) (Method-II); Methanol: Water: 0.1N NaOH (3:1:1) (Method-III) were measured at respective wavelengths using respective solvent blend as blank.

 

Preparation of calibration curve for zidovudine in selected solvent blends:

Appropriate aliquots from standard zidovudine stock solutions were transferred to series of 10 ml volumetric flasks. The volume was adjusted to the mark with saline phosphate buffer pH 7.2 (Method-I) to obtain concentrations of 2, 4, 6, 8 and 10µg/ml. Similarly a set of same concentrations were prepared in other solvent blends viz., Methanol: Water: 0.1N HCl (3:1:1) (Method-II); Methanol: Water: 0.1N NaOH (3:1:1) (Method-III). Absorbance spectra of each solution against respective solvent blend as blank were measured at respective obtained absorption maxima and concentration Vs absorbance was plotted and interpreted statistically.

 

Preparation of zidovudine sample preparation (for zidovudine tablets):

Weigh accurately 5 tablets and grind in a mortar and transfer equivalent to 50 mg of zidovudine into a 50 ml volumetric flask, add 40 ml of saline phosphate buffer pH 7.2 (Method-I) and shake it for 1h. Dilute to volume with saline phosphate buffer pH 7.2 mix the contents and filter through 0.45 µm membrane filter. Transfer aliquots of the filtrate to 25ml volumetric flask and dilute to volume with the saline phosphate buffer pH 7.2 to get desired concentration. Similarly sample preparations were prepared in other solvent blends viz., Methanol: Water: 0.1N HCl (3:1:1) (Method-II); Methanol: Water: 0.1N NaOH (3:1:1) (Method-III).

 

Accuracy:

The accuracy was evaluated applying the proposed method to the analysis formulations with known amounts of drug. The accuracy was calculated as the percentage of the drug recovered from the formulations studies were carried out by adding known amount of standard drug (40% and 20%) to the sample solution, measure the absorbance and calculate the amount of drug from the calibration curve. The % recovery was calculated in terms of % RSD and it should be less than 2%.

 

Precision:

The precision was determined by repeatability (intra-day) and intermediate precision (inter day). Repeatability was evaluated assaying 3 determinations at the same concentration (10 µg/ml), during the same day, under the same experimental conditions. Intermediate precision was analyzed comparing the assays in 3 determinations at the same concentration (10µg/ml) during 3 different days. Precision (repeatability and intermediate precision) was expressed as relative standard deviation (RSD).

 

Intraday precision was determined by analyzing zidovudine for three times in the same day (morning, afternoon, evening) at respective absorption maxima using respective solvent blends. Interday precision was determined by analyzing daily once (morning) for three days at respective absorption maxima using respective solvent blends. The % RSD values were calculated and it should be less than 2%. The lowest possible concentration where the drug zidovudine shows response was determined in all the solvent blends. The absorbance at this concentration was measured in triplicate in respective solvent blends at respective absorption maxima. The LOD/LOQ was calculated by using formulae from the data obtained.

 

LOD (µg/ml) =3.3 X σ/S 

LOQ (µg/ml) =10 X σ/S

 

Where

σ-Standard deviation of the response; s–Slope ratio curve

 

Robustness:

Robustness of the proposed methods were determined by the analysis of samples and standard solutions (10 µg/ml) at different wavelengths (±5 nm), at different solution temperatures (refrigerated condition and room temperature). To assess the stability of drug, the stability study was performed maintaining the drug working solution in respective solvent blends for 48 h protected from light, looking for the decrease of absorbance compared with those of freshly prepared solutions. Appropriate concentrations of zidovudine API and tablets were prepared in respective solvent blends. Analysis was carried out at three different wavelengths (actual and ±5 nm). Amount found was calculated at three different wavelengths in terms of % RSD and values should be less than 2%.

 

Ruggedness:

Ruggedness is not addressed in the ICH documents. Ruggedness is a measure of reproducibility of test results under normal, expected operational conditions from analyst to analyst and instrument to instrument. Appropriate concentrations of zidovudine from bulk and formulations were prepared in respective solvent blends. Analysis was carried out by two different analysts and also two instruments. Amount found was calculated at three different wavelengths in terms of % RSD and values should be less than 2%.

 

RESULTS AND DISCUSSION:

Simple, rapid, economic and reproducible UV spectroscopic methods were developed and validated as per ICH guideline and USP2000 for zidovudine API and tablet formulations (Zidovudine 100mg). The different solvent blends viz., Saline phosphate buffer pH 7.2(Method-I); Methanol: Water: 0.1N HCl (3:1:1) (Method-II); Methanol: Water: 0.1N NaOH (3:1:1) (Method-III). The developed methods were further validated for accuracy, precision, LOD, LOQ, specificity, robustness, and ruggedness with statistical data. The λmax with characteristic peak for zidovudine in saline phosphate buffer pH 7.2 (Method-I) at 267nm, in methanol: water: 0.1N HCl (Method-II) at 267nm and in methanol: water: 0.1N NaOH (3:1:1) at 267nm (Method-III) were observed. The linearity ranges for zidovudine were studied for all the selected solvent blends at their respective absorption maxima in the concentration range of 0-30µg/ml. The calibration curve for zidovudine (Fig 2) were prepared in all the selected solvent blends in the concentration range of 2-10 µg/ml and are shown with statistical data in tables 1-3.



Figure 2: calibration curve for zidovudine

 


Table-1: Linearity range data of Zidovudine in Method I, Method II and Method III.

Conc. (mg/ml)

(Method-I)

(Method-II)

(Method-III)

Absorbance

Absorbance

Absorbance

2

0.089

0.076

0.070

4

0.170

0.156

0.138

6

0.248

0.229

0.212

8

0.329

0.312

0.272

10

0.403

0.381

0.354

12

0.488

0.463

0.423

14

0.556

0.532

0.481

16

0.640

0.606

0.561

18

0.727

0.688

0.540

20

0.917

0.763

0.620

22

0.981

0.618

0.600

24

1.001

0.642

0.792

26

1.181

0.988

0.916

28

1.204

1.220

1.064

30

1.296

1.350

1.144

*Average of six determinations

Table-2: Calibration/linearity curve for Zidovudine

Conc

(mg/ml)

Method-I

Method-II

Method-III

 

Absorbance

Mean±SD

Absorbance

Mean±SD

Absorbance

Mean±SD

2

0.082±0.0029

0.078±0.0020

0.070±0.0028

4

0.163±0.0019

0.161±0.0036

0.141±0.0039

6

0.246±0.0037

0.241±0.0064

0.211±0.0039

8

0.326±0.0020

0.322±0.0069

0.282±0.0032

10

0.403±0.0030

0.404±0.0030

0.354±0.0041

 

The beer`s range was found to be 1-18 µg/ml in saline phosphate buffer pH 7.2(Method-I), 1-20 µg/ml in methanol: water: 0.1N HCl (3:1:1) (Method-II) and 1-16 µg/ml in methanol: water: 0.1N NaOH (3:1:1) (Method-III) with correlation coefficient of 0.9998. The linearity range graphs were shown in figures 3.

 


 

Fig: 3 Linearity range curve of zidovudine Method-I, Method-II and Method-III.

 

Table-3: Statistical data of calibration curve for zidovudine

Method-I

Method-II

Method-III

λmax(nm)

267

267

267

Beer’s law limits (μg / ml)

1-18

1-20

1-16

Molar Absorptivity (mol-1cm-1)

10.9 x 103

10.7x103

9.3 x 103

Sandell’s sensitivity

0.024

0.024

0.028

Best-fit values

 

 

 

Slope

0.04043±0.0002274

0.04042±0.0001184

0.03529±0.0001650

Y-intercept when X=0.0

0.001190 ± 0.001377

-0.001017 ± 0.0006650

-4.762e-005±0.0004994

X-intercept when Y=0.0

-0.02945

0.02516

0.001350

1/slope

24.73

24.74

28.34

95% Confidence Intervals

 

 

 

Slope

0.03980 to 0.04106

0.04012 to 0.04073

0.03483 to 0.03574

Y-intercept when X=0.0

-0.002632 to 0.005013

-0.002727 to 0.0006927

-0.001434 to 0.001339

X-intercept when Y=0.0

-0.1256 to 0.06427

-0.01724 to 0.06706

-0.03835 to 0.04021

Goodness of Fit

 

 

 

R square

0.9998

0.9999

0.9999

P value

< 0.0001

< 0.0001

< 0.0001

 


All the solvent blends shows correlation coefficient of 0.9998-0.9999 in the concentration range of 2-10 µg/ml; Molar absorptivity was found to be 10.9 x 103, 10.7 x 103 and 9.3 x 103. (Fig 4), Sandell’s sensitivity was found to be 0.024, 0.024 and 0.028 ; Best fit value slope was found to be 0.04043±0.0002274, 0.04042±0.0001184 and 0.03529±0.0001650; 95% confidence interval slope was found to be 0.03980-0.04106, 0.04012-0.04073 and 0.03483-0.03574 for saline phosphate buffer pH 7.2 (Method-I), methanol: water: 0.1N HCl (3:1:1) (Method-II) and methanol: water: 0.1N NaOH (3:1:1) (Method-III) respectively. In all the solvent blends the P value is<0.0001 indicate proposed methods were statistically significant.

 

 

Fig-4: Absorption Maxima of Zidovudine in Method-I, Method-II and Method-III

 

The accuracy was found to be 99.5 %-100.7% in all the proposed methods for the estimation of Zidovudine API. The % recovery of zidovudine in formulations were found to be 99.9±0.750, 100.2±1.114, 98.8±1.635 with % RSD values of 0.750, 1.111, 1.635 for saline phosphate buffer pH 7.2 (Method-I), methanol: water: 0.1N HCl (3:1:1) (Method-II) and methanol: water: 0.1N NaOH (3:1:1) (Method-III) respectively which were within the acceptance limit. The results suggest that proposed methods were accurate in estimation. The data were shown in table 4 and 5.

 

Table-4: Data showing accuracy of Zidovudine API in all solvent blends

METHOD I- saline phosphate buffer pH 7.2

Sample No

Concentration of Zidovudine (mg/ml)

% Recovery

Theoretical

Experimental

1

2

3

4

5

2

4

6

8

10

1.99

4

5.95

8

10.07

99.5

100

99.1

100

100.7

METHOD II- Methanol: Water: 0.1NHCl (3:1:1)

Sample No

Concentration of Zidovudine (mg/ml)

% Recovery

Theoretical

Experimental

1

2

3

4

5

6

1

2

4

6

8

10

1.01

1.98

4.04

6.04

8

10.02

101.0

99.0

101.0

100.6

100

100.2

METHOD III- Methanol: Water: 0.1N NaOH (3:1:1)

Sample No

Concentration of Zidovudine (mg/ml)

% Recovery

Theoretical

Experimental

1

2

3

4

5

1

2

3

4

5

0.99

2.02

2.97

3.98

4.96

99.0

101.0

99.0

99.5

99.2

 

Table-5: Data showing recovery studies of Zidovudine tablet in all solvent blend.

Method-I

Amount present in formulation (mg/ml)

Amount added

Amount recovered (mg/ml)

Mean % Recovery ± SD

RSD

mg

%

10

4

2

40

20

13.99

11.99

99.9±0.7506

99.9±0.5805

0.750

0.580

Method-II

Amount present in formulation (mg/ml)

Amount added

Amount recovered (mg/ml)

Mean % Recovery±SD

RSD

mg

%

10

4

2

40

20

14.01

12.01

100.2±1.114

100.1 ± 0.568

1.111

0.567

Method-III

Amount present in formulation (mg/ml)

Amount added

Amount recovered (mg/ml)

Mean % Recovery±SD

RSD

mg

%

10

4

2

40

20

13.8

11.8

98.7±4.614

98.8±4.895

1.635

1.918

 

Based on the standard deviation of the response and the slope the limit of detection values for zidovudine were found to be 0.036µg/ml, 0.026 µg/ml, 0.067 µg/ml and limit of quantitation values for zidovudine were found to be 0.113 µg/ml, 0.080 µg/ml, 0.202 µg/ml for saline phosphate buffer pH 7.2(Method-I), methanol: water: 0.1N HCl (3:1:1) (Method-II) and methanol: water: 0.1N NaOH (3:1:1) (Method-III) respectively. The data were shown in table 6.

 

The % RSD values of intraday precision for zidovudine in formulations were found to be 0.554, 0.532, 0.437 and % RSD values of inter day precisions for zidovudine were found to be 1.354, 1.659, 1.406 for saline phosphate buffer pH 7.2(Method-I); methanol: water: 0.1N HCl (3:1:1) (Method-II) and methanol: water: 0.1N NaOH (3:1:1) (Method-III) respectively which were within the acceptance limit. The results suggest the proposed methods were precise and reproducible for the estimation. The data was shown in table 7.


Table-6: Data showing LOD/LOQ of zidovudine in all solvent blends


Method-I

 

Mean±SD

SEM

Limit of Detection

0.18±0.063

0.036

Limit of Quantitation

0.56±0.196

0.113

Method-II

 

Mean±SD

SEM

Limit of Detection

0.38±0.04

0.026

Limit of Quantitation

1.16±0.144

0.08

Method-III

 

Mean±SD

SEM

Limit of Detection

0.41±0.116

0.067

Limit of Quantitation

1.28±0.35

0.202

 

 

 


 


Table-7: Data showing precision intraday and inter day trials with RSD values for Zidovudine in all solvent blends

Method-I

Trial

Label Claim (mg/tab)

% Label Claim Mean±SD

SEM

RSD

Day-1

100

100.2±0.556

99.6±1.332

99.4±1.206

Intra day

trials

0.321

0.768

0.696

0.554

1.337

1.213

Day-2

100

100.3±0.96

99.6±0.938

99.4±1.054

 

0.554

0.541

0.608

0.957

0.941

1.060

Day-3

100

99.9±1.353

99.8±1.513

99.9±1.234

0.781

0.873

0.712

1.354

1.516

1.235

Method-II

Day-1

100

99.8±0.550

99.7±1.266

99.4±0.529

Intra day

trials

0.318

0.731

0.305

0.551

1.269

0.532

Day-2

100

100.36±1.66

100.1±1.305

99.9±0.750

0.961

0.753

0.433

1.659

1.303

0.751

Day-3

100

100.2±0.90

100.4±0.929

100.1±1.415

0.523

0.536

0.817

0.905

0.925

1.413

Method-III

Day-1

100

100.3±0.950

99.9±1.082

99.6±0.436

Intra day

trials

0.548

0.624

0.252

0.949

1.083

0.437

Day-2

100

99.9±1.405

99.4±0.500

99.7±0.709

0.811

0.288

0.409

1.406

0.503

0.711

Day-3

100

100.8±1.249

100.8±0.850

98.7±0.644

0.721

0.491

0.373

1.239

0.843

0.652

 


Change in the λmax of ±5nm to the actual λmax in robust analysis the % recovery of zidovudine was found to be significantly different which clearly indicates change in the λmax of ±5nm affected the method so proposed methods were not robust. Similarly change in the storage conditions robust analysis the % recovery of zidovudine is found to be significantly different which clearly indicates the storage condition is also affecting the method so proposed methods were not robust. The robust data were given in table 8 and 9. The % recovery of Zidovudine in ruggedness analysis by different analyst and change of instrument viz., Analyst-1; Analyst-2 and Instrument-1; Instrument-2 shows the proposed methods were significantly rugged. The ruggedness data were shown in table 10 and 11.


 

 

Table-8: Data showing robustness of zidovudine at different wavelength in different solvents

METHOD

Conc (mg/ml)

Wave length

Amount found

Mean % ± SD

SEM

RSD

Method-I

10

272

277

267

9.98

8.27

8.41

99.8±0.862

82.7±1.615

84.1±1.021

0.452

0.912

0.711

0.915

1.417

1.213

Method-II

10

282

287

277

9.97

8.32

8.5

99.7±0.612

83.2±1.007

85±1.059

0.517

0.912

0.612

0.712

1.520

1.311

Method-III

10

272

277

267

9.99

8.41

8.52

99.9±1.241

84.1±1.112

85.±0.978

0.721

0.802

0.662

1.256

1.112

1.126

 

Table-9: Data showing robustness of zidovudine at refrigerated conditions and room temperature in all solvent blends.

 

Trial

Label Claim (mg/tab)

REFREGERATED CONDITIONS

Amount Found (mg/tab)

% Label Claim  Mean±SD

SEM

RSD

METHOD -I

Day-1

100

99.1

98.7

98.4

Intra day

trials

99.1±0.404

98.7±0.500

98.4±0.251

Intra day

trials

0.233

0.288

0.145

0.407

0.506

0.255

Day-2

100

99.2

98.9

98.8

99.2±0.435

98.9±0.529

98.8±0.793

0.458

0.305

0.251

0.534

0.802

0.438

Day-3

100

98.8

98.5

98.4

98.8±0.360

98.5±0.404

98.4±0.251

0.208

0.233

0.145

0.364

0.410

0.255

METHOD -II

Day-1

100

100.2

100.2

99.6

Intra day

trials

100.2±0.200

100.2±0.200

99.6±0.251

Intra day

trials

0.115

0.115

0.145

0.199

0.199

0.252

Day-2

100

99.8

99.8

99.6

99.8±0.529

99.8±0.366

99.6±0.404

0.305

0.208

0.233

0.530

0.366

0.405

Day-3

100

99.1

98.8

98.5

99.1±0.602

98.8±0.655

98.5±0.500

0.348

0.378

0.288

0.607

0.662

0.507

METHOD -III

Day-1

100

100.5

100.2

100.3

Intra day

trials

100.5±0.300

100.2±0.251

100.3±0.168

Intra day

trials

0.173

0.145

0.166

0.298

0.250

0.254

Day-2

100

100.6

99.8

49.5999.6

100.06±0.40

99.8±0.709

99.6±0.550

0.233

0.409

0.318

0.403

0.552

0.710

Day-3

100

99.4

99.3

99.0

99.4±0.300

99.3±0.346

99±0.458

0.173

0.200

0.264

0.348

0.301

0.462

 

Table-9. Continued……

 

Trial

Label Claim (mg/tab)

ROOM TEMPERATURE

Amount Found (mg/tab)

% Label Claim  Mean±SD

SEM

RSD

METHOD-I

Day-1

100

100.2

99.6

99.4

Intra day

trials

100.2±0.556

99.6±1.332

99.4±1.206

Intra day

trials

0.321

0.768

0.696

0.554

1.337

1.213

Day-2

100

100.3

99.6

99.4

100.3±0.96

99.6±0.938

99.4±1.054

0.554

0.541

0.608

0.957

0.941

1.060

Day-3

100

99.9

99.8

99.9

99.9±1.353

99.8±1.513

99.9±1.234

0.781

0.873

0.712

1.354

1.516

1.235

METHOD-II

Day-1

100

99.8

99.7

99.4

Intra day

trials

99.8±0.550

99.7±1.266

99.4±0.529

Intra day

trials

0.318

0.731 0.305

0.551

1.269

0.532

Day-2

100

100.36

100.1

99.9

100.36±1.66

100.1±1.305

99.9±0.750

0.961

0.753 0.433

1.659

1.303

0.751

Day-3

100

100.2

100.4

100.1

100.2±0.90

100.4±0.929

100.1±1.415

0.523

0.536  

 0.817

0.905

0.925

1.413

METHOD-III

Day-1

100

100.3

99.9

99.6

Intra day

trials

100.3±0.950

99.9±1.082

99.6±0.436

Intra day

trials

0.548

0.624

0.252

0.949

1.083

0.437

Day-2

100

99.9

99.4

99.7

99.9±1.405

99.4±0.500

99.7±0.709

0.811

0.288 0.409

1.406

0.503

0.711

Day-3

100

100.8

100.8

98.7

100.8±1.249

100.8±0.850

98.7±0.644

0.721

0.491 0.373

1.239

0.843

0.652

 

 

 

 

 

Table-10: Data showing ruggedness of zidovudine by different Analysts in all solvent blends

METHOD

Conc (mg/ml)

Analyst

Amount found

Recovery ± SD

SEM

RSD

Method-I

10

Analyst 1

Analyst 2

10.01

9.96

100.03±0.23

99.9±0.87

0.095

0.365

0.229

0.878

Method-II

10

Analyst 1

Analyst 2

9.96

9.92

99.6±0.367

99.4±0.21

0.149

0.086

0.368

0.223

Method-III

10

Analyst 1

Analyst 2

10.01

9.99

100.01±0.60

99.9±0.433

0.248

0.177

0.599

0.433

 

Table-11: Data showing ruggedness of zidovudine using different Instruments in all solvent blends

METHOD

Conc (mg/mL)

Instrument

Amount found

Recovery ± SD

SEM

RSD

Method-I

10

Instrument 1

Instrument 2

9.83

9.93

98.3±0.862

99.3±0.755

0.497

0.435

0.876

0.760

Method-II

10

Instrument 1

Instrument 2

9.94

9.85

99.4±0.702

98.5±0.793

0.405

0.458

0.746

0.805

Method-III

10

Instrument 1

Instrument 2

9.79

9.82

97.9±0.305

98.2±0.503

0.176

0.290

0.311

0.512

 


CONCLUSION:

The proposed UV spectroscopic methods were found to be simple, rapid, accurate, precise and economic. From the above data it was observed that all validation parameters meet the predetermined acceptance criteria and validated in terms of linearity, accuracy, precision, reproducibility, robustness, and ruggedness as per the ICH guidelines. Thus it has been concluded that the proposed methods were validated for the analysis of zidovudine API and its tablet formulations.

 

ACKNOWLEDGEMENT:

The authors are thankful to Strides-Arco Lab-Bangalore-Karnataka for providing gift sample of zidovudine. The authors are also grateful to the Principal, staff and Management of V L College of Pharmacy Raichur for providing necessary facilities to carry out the research work.

 

REFERENCES:

1.     Santosh G, Anandkumar Y, Saritha and Mallikarjuna CS. Development and validation of new UV spectroscopic methods for the estimation of lamivudine in active pharmaceutical ingredient and in its tablet formulation. International Journal of Chemical and Pharmaceutical Analysis 2017; 5(1): 2395-2466.

2.     Goodman S, Gilman A. The Pharmacological Basis of Therapeutics, New York, NY: Macmillan Publishing Company; 1985; 1357.

3.     Goodman S, Gilman A. The Pharmacological Basis of Therapeutics. New York, NY In: Joel GH, editor. 10th ed. Mc-Graw Hill: Medical publishers Div; 2006:1349.

4.     US Pharmacopoeia. Asian Edition 30. United States Pharmacopoeial Convention, Inc: 2007: 2447.

5.     Langtry HD and Campoli-Richards DM. Zidovudine. A review of its pharmacodynamic and pharmacokinetic properties and therapeutic efficacy. Drugs 1989; 37(4): 408-450.

6.     Indian Pharmacopoeia. Vol. 2. Government of India: Controller of Publications; Government of India Ministry of Health & Family Welfare 2014:3003-3004.

7.     Yarchoan R, Mitsuya H, Myers CE and Broder S: Clinical pharmacology of 3'-azido-2,3-dideoxythymidine (zidovudine) and related dideoxynucleosides. New England Journal of Medicine; 1989; 321: 726-738.

8.     Chien YW, Wearley LL. Aids and Chemotherapy. Drugs of Today 1989; 25: 19-25

9.     Basavaiah K and Anil Kumar UR. Spectrophotometric Determination of Zidovudine in Pharmaceuticals Based on Charge-Transfer Complexation Involving N-Bromosuccinimide, Metol and Sulphanilic Acid as Reagents. E-Journal of Chemistry 2007; 4(2): 173-179.

10.   Basavaiah K and Anil Kumar UR. Simple Spectrophotometric Methods for the Determination of Zidovudine in Pharmaceuticals Using Chloramine-T, Methylene Blue and Rhodamine-B as Reagents. E-Journal of Chemistry 2006; 4(12): 173-181.

11.   Bengi U, Sibel AO. Determination of Lamivudine and Zidovudine in binary mixtures using first derivative spectrophotometric, first derivative of the ratio-spectra and high-performance liquid chromatography–UV methods. Analytica Chimica Acta 2002; 466: 175-185.

12.   Bala SC, Bhogela SS, Shaik M, Vadlamudi CS, Chappa M and Maddirala NS. Simple and Sensitive Spectrophotometric Methods for the Analysis of Mesalamine in Bulk and Tablet Dosage Forms. Quimica Nova 2011; 34(6): 1068-1073.

13.   Vaishali PN and Bhusari KP. A Validated UV Spectrophotometric Method for the Simultaneous Estimation of Lamivudine, Nevirapine and Zidovudine in Combined Tablet Dosage Form. Journal of Pharmacy Research 2009; 2(4): 666-669.

14.   Kothamasu PK, Chindanuru S, Tejaswini J, Vutapalli SKR, Devarakonda S, Reddy CR and Kothuru R. Comparison of Analytical Spectrophotometric Methods for the Determination of Tetrabenazine in Tablets. Pharmaceutical Methods 2018; 9(1): 33-38.

15.   Gunji R, Nadendla RR and Ponnuru VS. Simultaneous UV-Spectrophotometric determination and validation of Diclofenac Sodium and Rabeprazole Sodium using hydrotropic agents in its tablet Dosage Form. International Journal of Drug Development and Research 2012: 4(1): 316.324.

16.   Nanda RK, Gaikwad J and Prakash A. Simultaneous Spectrophotometric Estimation of Tamsulosin in Pharmaceutical Dosage Form. Asian Journal of Research Chemistry 2009: 2(1): 63-65.

17.   Jain P, Patel M and Surana S. Development and validation of UV-Spectrophotometric method for determination of Cefuroxime Axetil in bulk and in Formulation. International Journal of Drug Development & Research 2011; 3 (4): 318-322.

18.   Shelke S, Dongre S, Rathi A,Dhamecha D, Maria S and Mohd HGD. Development and Validation of UV Spectrophotometric Method of Cefuroxime Axetil in Bulk and Pharmaceutical Formulation. Asian Journal of Research Chemistry 2009; 2(2): 222-224.

19.   Pandya HB and Patel PB. Development and validation of analytical methods for the simultaneous estimation of metoprolol succinate, chlorthalidone and cilnidipine in tablet dosage form. World Journal of Pharmacy and Pharmaceutical Sciences 2018; 7(5): 945-96.

20.   Jain S, Maheshwari RK, Nema RK and Singhvi I. Simultaneous estimation of ofloxacin and tinidazole in solid dosage form by UV spectrophotometry using mixed solvency concept. European Journal of Pharmaceutical and Medical Research 2017; 4(12): 413-418.

21.   Reddy JS, Ahmed SM, Chakravarthy IE and Prabhavathi K. Spectrophotometric Determination of Zidovudine in Pharmaceutical Dosage Forms. E-Journal of Chemistry 2012; 9(1): 89-92.

22.   Rajath U, Sikha PK, Charan AS, Kalyani MPY and RajaSekhar KK. Spectrophotometric method for the estimation of zidovudine in bulk, tablet and capsule dosage forms. Journal of Pharmaceutical Research 2012; 5(7):3834-3836.

23.   Nevin Erk. Derivative-differential UV spectrophotometry and compensation technique for the simultaneous determination of zidovudine and lamivudine in human Serum. Pharmazie 2004; 59 (2): 106-111.

24.   Kaur M, Bhardwaj P, Kaur B, Sharma A, Kaur C and Kumar R. Development and Validation of a Novel Stability Indicating UV Spectrophotometric Method for Estimation of Febuxostat in Bulk and Pharmaceutical Formulation (Tablets). Pharmaceutical Methods 2018; 9(1): 24-29.

 

 

 

 

Received on 18.08.2018       Accepted on 12.10.2018     

© Asian Pharma Press All Right Reserved

Asian J. Pharm. Ana. 2018; 8(4): 195-202.

DOI: 10.5958/2231-5675.2018.00036.4