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

 

Santosh Gada^1*, 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.

 

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.

 

 

 

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 analogue^2,3. 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 agents^7,8.

 

In the literature, numerous methods to quantify the Zidovudine have been described either alone or in combination with other antiretroviral drugs^9-11. 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 groups^12-24

 

 

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

 

 

 

 

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