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In this study IR and UV spectroscopic procedures are describe for the quantitative determination of
Zidovudine (AZT) from solid dosage form. For IR spectroscopic method (KBr disc technique) has
been used and Ursodeoxycholic acid (UDCA) was used as an internal standard. The specific absorption
bands at 2105 and 2931 cm1 were chosen for AZT and UDCA respectively. In this method, beer’s law
Published on 05 October 2010 on http://pubs.rsc.org | doi:10.1039/C0AY00341G
was obeyed in the concentration range 0.8–2.0% w/w in KBr disc. The regression equation was found to
be y ¼ 1.088x + 1.194 with correlation coefficient 0.9819, and the assay was found to be 99.8% with
RSD of 2.157%. The UV spectroscopic method was also used in the quantitative determination of AZT
in solid dosage form. 266 nm was chosen as lmax. The regression equation was found to be y ¼ 0.0434x
+ 0.027, and the correlation coefficient 0.9989. The assay was found to be 99.81% with RSD of 0.273%
by the UV spectroscopic method.
1. Introduction
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discs were prepared for each mixture and employed for quanti- respectively. The solutions were scanned in the UV range 200–
tative measurement. Each disk of 125 mg contained 2 mg, 3 mg, 400 nm. The scanning spectrums of the drug are shown in Fig. 3
4 mg, 5 mg and 6 mg AZT respectively so a linear curve could be and Fig. 4 and the wavelength 266 nm was selected for further
plotted. The calibration curve for AZT was prepared using the study.
LogPB–LogPO values of AZT-UDCA in synthetic standard
mixtures. The correlation of coefficient for AZT was found to be 2.5.3 Determination of E (1%, 1 cm). 0.3 ml aliquot of AZT
0.9819 with equation line of y ¼ 1.0883x + 1.1944. from stock solution was transferred to six 10 ml volumetric flaks
and volume was adjusted to the mark with methanol to obtain six
2.4.2.2. Accuracy and precision. Varying concentrations of individual concentrations of 30 mg ml1. The absorbance of each
AZT were prepared with internal standard in KBr disk range solution was measured at 266.0 nm. E (1%, 1 cm) values of drug
from 80%, 100% and 120%. PB and PO points of the absorption were calculated using eqn (1) and results of E (1%, 1 cm) of the
peaks were determined and LogPB–LogPO values of AZT- drug are shown in Table 1:
UDCA calculated by base line technique as depicted in Fig. 2.
The intra-day and inter-day accuracy precision was studied. The E (1%, 1 cm) ¼ Absorbance/Concentration (g/100ml) (1)
percentage RSD of the method was calculated at these three
levels.
2.4.2.3. Recovery study. The recovery study was performed 2.5.4. Method validation
at 80%, 100% and 120%. 4 mg of AZT and concentration 3.2 mg, 2.5.4.1. Linearity. Different concentration of AZT from 10–
4 mg, and 4.8 mg added into the disk containing AZT-UDCA 50 mg mL1 were prepared using the stock solution to prepare the
standard mixture of 4 mg AZT. The recoveries of three levels calibration curve of AZT. The slope and intercept was calculated
were estimated and %RSD was calculated. from the regression line equation.
Absorptivity at 266 nm
standard mixtures. The correlation of coefficient for AZT was
a
Mean 449.83 found 0.9819 with equation line of y ¼ 1.0883x + 1.1944 (Fig. 5).
SDb 0.81 The linearity values of LogPB–LogPO are presented in Table 2.
%RSDc 0.1798
a
Absorptivity values are the mean of six determinations. b
S.D. is 3.1.2. Accuracy and precision. The developed method of AZT
Standard Deviation. c RSD is Relative Standard Deviation. shows high levels of accuracy and precision at 80%, 100% and
120% levels as given in Table 3 and 4. The intra-day and inter-
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Y ¼ TAZT/TUDCA
AZT 2105
2.7576
4.3746
4.9633
3.8574
3.26
80% 124.500 3.200 14.340 0.132
124.300 3.200 14.310 0.131
124.600 3.200 14.370 0.133
Mean 124.467 3.200 14.340 0.132
X ¼ CAZT/CUDCA
SD 0.153 0.000 0.030 0.001
%RSD 0.123 0.000 0.209 0.758
100% 125.900 4.000 15.020 0.166
125.300 4.000 14.990 0.165
125.700 4.000 15.030 0.168
Where, CAZT ¼ AZT conc. in KBr disc, CUDCA¼ UDCA conc. in KBr disc, TAZT¼ LogPB–LogPO values of AZT, TUDCA¼ LogPB–LogPO values of UDCA.
1.33
3
2
0.048
0.081
0.05
PB PO
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AZT 2105
UDCA 2931 cm1
1.5
Conc./mg
2
3
4
5
6
123.8
123.5
AZT 2105
Analyst-I Analyst-II
LogPB LogPB
Disc Wt Conc./mg PB-PO LogPO Disc Wt Conc./mg PB-PO LogPO
AZT 125.900 4.000 15.756 0.161 AZT 125.900 4.000 15.980 0.175
125.300 4.000 14.834 0.167 125.300 4.000 14.950 0.170
125.700 4.000 15.678 0.169 125.700 4.000 15.890 0.176
Mean 125.633 4.000 15.423 0.166 Mean 125.633 4.000 15.607 0.174
SD 0.306 0.000 0.511 0.004 SD 0.306 0.000 0.570 0.003
%RSD 0.243 0.000 3.315 2.513 %RSD 0.243 0.000 3.655 1.968
35% RH 40% RH
LogPB LogPB
Disc Wt Conc./mg PB-PO LogPO Disc Wt Conc./mg PB-PO LogPO
AZT 128.400 4.000 14.348 0.179 AZT 128.400 4.000 14.798 0.169
122.380 4.000 15.278 0.174 122.380 4.000 15.898 0.177
128.790 4.000 15.350 0.171 128.790 4.000 15.450 0.178
Mean 126.523 4.000 14.992 0.175 Mean 126.523 4.000 15.382 0.175
SD 3.594 0.000 .559 0.004 SD 3.594 0.000 0.553 0.005
%RSD 2.840 0.000 5.295 3.728 %RSD 2.840 0.000 3.596 2.824
Published on 05 October 2010 on http://pubs.rsc.org | doi:10.1039/C0AY00341G
45% RH
AZT 2105
LogPB
Disc Wt Conc./mg PB–PO LogPO
Table 8 Assay results of commercial samples (Retrovir tablet 300 mg of Table 10 Summary of repeatability, precision and ruggedness
AZT) by IR spectroscopy.b
Parameters (UV Spectroscopy)
IR-Spectroscopy AZT %RSD
Parameters AZT
Repeatability 0.416
Label Claim 300 Intra-day 0.225
%Drug Contenta 99.8 Inter-day 0.112
SD 2.152 Ruggedness
%RSD 2.157 Analyst I 0.555
Analyst II 0.337
a b
Value for Drug content (%) is the mean of five estimations. S.D. is
standard deviation and R.S.D. is relative standard deviation.
Table 11 Determination of AZT in bulk
tablets. A linear concentration was obtained as 10–50 mg ml1 in 2 L. B. Laurence, S. L. John and L. P. Keith, Goodman and Gilman’s:
this method. The regression equation was found to be y ¼ The Pharmacological Basis of Therapeutics, McGraw-Hill, Medical
Publishing Division, 11th edn, pp 1283.
0.0434x + 0.027 and the correlation coefficient 0.9984. E (1%, 1 3 V. P. Nagulwar and K. P. Bhusari, A validated UV
cm) was calculated as 449.83 in methanol. The assay was found spectrophotometric method for the simultaneous estimation of
to be 99.81% with %RSD 0.273%. Lamivudine, Nevirapine and zidovudine in combined tablet dosage
Although various methods like HPLC-UV, LC-MS/MS, form, J. Pharm. Res., 2009, 2(4), 666–669.
4 C. Sharada, K. Channabasavaraj and M. T. Tamizh, Development of
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expensive detectors, costly solvents, columns and also more time estimation of Zidovudine in bulk and pharmaceutical dosage forms,
needed to develop and validate the method.14–16 The present IR I.J.P.R.D, 2010, 2(3), 001.
5 K. B. Kenney, S. A. Wring, R. M. Carr, G. N. Wells and J. A. Dunn,
spectroscopy method is a simple, rapid, accurate and cost Simultaneous determination of Zidovudine And Lamivudine in
effective analytical tool for AZT determination in tablet human serum using HPLC with tandem Mass Spectrometry, J.
formulations. Pharma. Bio. Ana., 2005, 22(6), 967–983.
6 S. Anbazhagan, N. Indumathy, P. Shanmugapandiyan and
Published on 05 October 2010 on http://pubs.rsc.org | doi:10.1039/C0AY00341G