1. Part III
Stability indicating colorimetric method for the determination of meclophenoxate hydrochloride.

2. -This part includes a general introduction about the chemistry and mode of action of meclophenoxate hydrochloride.
-A review on the reported methods for its quantitative determination.
Stability indicating colorimetric method for the determination of meclophenoxate hydrochloride using ferric hydroxamate complex formation.

3. -Structure of meclophenoxate
-The proposed mechanism for preparing the degradation product:
2 N NaOH
Reflux 25 min.

4. The proposed reaction mechanism:

5. Figure ( 26 ): Absorption spectra of
Meclophenoxate.HCL 100 µg. ml ( )
and colored product 300 µg. ml-1 (———).

6. Figure (30): Effect of volume (ml) of hydroxyl amine hydrochloride
solution on the absorbance of the ferric hydroxamate
complex with meclophenoxate hydrochloride.

7. Figure (31): Effect of volume (ml) of ferric chloride solution on the
absorbance of the ferric hydroxamate complex with
meclophenoxate hydrochloride.

8. Figure (32): Effect of volume (ml)of 1 N sodium hydroxide on the
absorbance of the ferric hydroxamate complex with
meclophenoxate hydrochloride.

9. Figure (28): Absorption spectra of ferric hydroxamate complex of
meclophenoxate μg. ml-1

10. Figure (29): Linearity of the absorbance of ferric hydroxamate
complex of meclophenoxate hydrochloride to the
corresponding concentration of meclophenoxate
hydrochloride.

11. Concentration (µg/ml)
Table (XXI): Determination of meclophenoxatehydrochloride in laboratory prepared mixtures by the proposed procedures.
Concentration (µg/ml)
Percentage %
Ferric hydroxamate
Method
Meclophenoxate.HCl
Degradation product
Recovery %
350 50
87.5%
12.5%
99.85%
300
100
75%
25%
102.50%
250
150
62.5%
37.5%
98.82%
200
50%
99.99%
98.45%
100.40%
Mean
100.00
S.D.
1.431

12. Table (XXII): Determination of meclophenoxatehydrochloride in lucidril tablets by the proposed procedures.
Lucidril tablets claimed to contain 250 mg
Batch number
Ferric hydroxamate method
Compendial method**
% Found
Recovery % ± S.D.*
010132
100.87
99.54 ± 0.632
010156
99.39
99.65 ± 0.951
020512
99.01
± 0.547
* Average of six determinations.
**Spectrophotometric method

13. Ferric hydroxamate method
Table (XXIII): Statistical comparison for the results obtained by the proposed method and the compendial method for the analysis of meclophenoxate hydrochloride in pure powder form.
Ferric hydroxamate method
Compendial method*
Meclophenoxate. HCl
Mean
S.D.
0.755
0.547
Variance
0.570
0.299 N 7 6
F test
1.906 (4.95)
Student’s t test
0.658 (2.201)
The figures in parenthesis are the corresponding tabulated values at P=0.05.
*Spectrophotometric method

14. Ferric hydroxamate method
Table (XXIV): Results of application of standard addition to the determination of meclophenoxate hydrochloride by the proposed method.
Batch number
Standard added
(mg.ml-1)
Ferric hydroxamate method
Meclophenoxate. HCl
Recovery % of added
010132
1.00
1.50
2.00
99.06
100.20
101.06
Mean ± S.D.
± 1.003
010156
98.33
99.07
99.50
Mean ± S.D.
98.96± 0.591
020512
98.50
99.75
99.18
99.14 ± 0.625

15. Table (XXV) : Assay parameters and method validation
Ferric hydroxamate method
Meclophenoxate. HCl.
Range (μg.ml-1)
Slope
0.0022
Intercept
0.0061
Mean
S.D.
0.755
Variance
0.57
Coff. of variation
0.744
Correl. Coef.(r)
0.9996
* RSD%a
*RSD %b
* RSD%a , RSD%b the intraday, interday respectively (n=5) relative standard deviation of concentrations ( µg/ml) for meclophenoxate HCl.

16. Part IV
Different stability indicating methods for the determination of vincamine in presence of its degradation product.

17. -This part includes a general introduction about the chemistry of vincamine, mode of action.
-A review on the reported methods used for vincamine quantitative determination.

18. Section [A]
Determination of vincamine in presence of its acid degradation product by the derivative ratio spectrophotometry.

19. -Structure of vincamine.
-Structure of vincamine.
-The proposed mechanism for degradation of vincamine

20. Figure ( 37 ): Absorption spectra of
  vincamine 20 µg. ml (———) and
its degradation product 20 µg. ml-1 ( )
Using 0.1N hydrochloric acid as a solvent.

21. dA/dλ Figure (38): First order spectra of
Figure (38): First order spectra of
Vincamine 20 μg.ml-1 (______)
Degradation product 20 μg.ml-1 (_ _ _ _ _ _)
Using 0.1N hydrochloric acid as a solvent.

22. A(vincamine/deg.prod.)
Figure (39) :
Zero order of the ratio spectra of vincamine μg.ml-1
using 20 µg.ml-1 of deg. product as a divisor.

23. First order of the ratio spectra of vincamine 12-48 μg.ml-1
dA(vincamine/deg.product)/dλ
Figure (40):
First order of the ratio spectra of vincamine μg.ml-1
using 20 µg.ml-1 of deg. product as a divisor.

24. Figure (41): Linearity of the peak amplitude of the first derivative of the
ratio spectra at nm to the corresponding concentration
of vincamine.

25. Table (XXVI) : Determination of vincamine in laboratory prepared mixtures by the proposed procedures.
Concentration (µg/ml)
Percentage %
Derivative ratio method
Vincamine
Degradation product
Recovery % 42 6
87.5 %
12.5 %
99.96 36 12
75 %
25 %
99.19 30 18
62.5 %
37.5 %
101.40 24
50 %
100.75
101.30
Mean
100.52
S.D.
0.937

26. Table (XXVII): Determination of vincamine in oxybral capsules by the proposed procedures.
Oxybral capsules claimed to contain 30 mg
Batch number
Derivative ratio method
Compendial method**
% Found
Recovery % ± S.D.* A
99.02
99.32 ± 0.956 A
98.98
98.56 ± 0.857 A
99.52
99.21 ± 0.659
* Average of four determinations.
**Spectrophotometric method

27. Derivative ratio method
Table (XXVIII): Statistical comparison for the results obtained by the proposed method and the compendial method for the analysis of vincamine in pure powder form.
Derivative ratio method
Compendial method*
Vincamine
Mean
99.90
99.58
S.D.
1.041
1.011
Variance
1.084
1.022 N 10 6
F test
1.060 (4.77)
Student’s t test
0.601 (2.145)
The figures in parenthesis are the corresponding tabulated values at P=0.05.
*Spectrophotometric method

28. Derivative ratio method
Table (XXIX): Results of application of standard addition to the determination of vincamine by the proposed method.
Batch number
Standard added
(mg.ml-1)
Derivative ratio method
Vincamine
Recovery % of added A
0.250
0.375
0.500
99.73
101.50
102.80
Mean ± S.D.
± 1.541 A
99.64
100.09
99.19
Mean ± S.D.
99.64 ± 0.450
021554A
98.15
98.98
99.23
98.78 ± 0.565

29. Section (B)
Densitometric determination of vincamine in presence of its acid degradation product

31. Figure ( 44 ):
Scanning profile of the TLC chromatogram of vincamine at 281 nm.

32. Figure (45): Linearity of the area under the peak to the
corresponding concentration of vincamine.

33. Concentration (µg/spot)
Table (XXX): Determination of vincamine in laboratory prepared mixtures by the proposed procedures.
Concentration (µg/spot)
Percentage %
Densitometric
method
Vincamine
Degradation product
Recovery % 9 1
90%
10%
98.18 8 2
80%
20%
99.40 7 3
70%
30%
100.04 6 4
60%
40%
99.13 5
50%
100.01
101.36
99.00
Mean
99.70
S.D.
1.032

34. Oxybral capsules claimed to contain 30 mg
Table (XXXI): Determination of vincamine in oxybral capsules by the proposed procedures.
Oxybral capsules claimed to contain 30 mg
Batch number
Densitometric method
Compendial method**
% Found
Recovery % ± S.D.* A
99.96
99.32 ± 0.956 A
100.31
98.56 ± 0.857 A
99.57
99.21 ± 0.659
* Average of four determinations.
**Spectrophotometric method

35. Table (XXXII): Statistical comparison for the results obtained by the proposed method and the compendial method for the analysis of vincamine in pure powder form.
Densitometric method
Compendial method*
Vincamine
Mean
100.09
99.58
S.D.
0.761
1.011
Variance
0.579
1.022 N 8 6
F test
1.765 (4.362)
Student’s t test
1.08 (2.179)
The figures in parenthesis are the corresponding tabulated values at P=0.05.
*Spectrophotometric method

36. Table (XXXIII): Results of application of standard addition to the determination of vincamine by the proposed method.
Batch number
Standard added
(µg.ml-1)
Densitometric method
Vincamine
Recovery % of added A
1.00
1.50
2.00
97.90
97.40
99.80
Mean ± S.D.
98.36 ± 1.266 A
99.60
99.10
99.59
Mean ± S.D.
99.43 ± 0.285
021554A
100.09
100.68
100.30
± 0.299

37. Section ( C )
Colorimetric determination of vincamine by using p-chloranilic acid reagent ( ion pair complexation ).

38. - The reaction between vincamine and p-chloraanilic acid can be presented as follows:-

39. p-Chloranilic acid ( _ _ _ _ _)
Figure (46): Absorption spectra of
Vincamine 20 µg. ml (…….)
p-Chloranilic acid ( _ _ _ _ _)
Colored product 200 µg. ml (_______).

40. Figure (49): Effect of volume (ml)of p- chloranilic acid solution on the absorbance of the colored product with vincamine.

41. Figure (50): Determination of the stoichiometry of the reaction of
vincamine with p- chloranilic acid by the continuous
variation method.

42. p-chloranilic acid) 75 – 250 μg. ml-1
Figure (47): Absorption spectra of colored product ( vincamine with
p-chloranilic acid) 75 – 250 μg. ml-1

43. Figure (48): Linearity of the absorbance of the colored product of
vincamine with p-chloranilic acid to the corresponding
concentration of vincamine.

44. Oxybral capsules claimed to contain 30 mg
Table (XXXIV): Determination of vincamine in oxybral capsule by the proposed procedures.
Oxybral capsules claimed to contain 30 mg
Batch number
Colorimetric method
Compendial method**
% Found
Recovery % ± S.D.*
012261A
97.00
99.32 ± 0.956
021554A
98.49
98.56 ± 0.857
011345A
100.45
99.21 ± 0.659
* Average of six determinations.
**Spectrophotometric method

45. Table (XXXV): Statistical comparison for the results obtained by the proposed method and the compendial method for the analysis of vincamine in pure powder form.
Colorimetric method.
Compendial method*
vincamine
Mean
100.13
99.58
S.D.
1.096
1.011
Variance
1.201
1.022 N 8 6
F test
(4.88)
Student’s t test
(2.179)
The figures in parenthesis are the corresponding tabulated values at P=0.05.
*Spectrophotometric method

46. Table (XXXVI): Results of application of standard addition to the determination of vincamine by the proposed method.
Batch number
Standard added
(mg.ml -1)
Colorimetric method
Vincamine
Recovery % of added
012261A
0.50
0.75
1.00
94.60
97.20
97.80
Mean ± S.D.
± 1.700
021554A
96.51
95.44
98.02
Mean ± S.D.
96.65 ± 1.296
011345A
99.52
98.88
99.25
99.22 ± 0.321

47. Section (D)
 High performance liquid chromatographic determination of vincamine in presence of its acid degradation product.

48. Rt degradation product: 3.67 min
Final assay conditions of Liquid chromatographic separation of vincamine and its degradation product:
Column: RP18
Mobile phase: acetonitrile: 0.01 M ammonium carbonate (70:30 v/v).
Flow rate: 1.6 ml. min-1.
Detection:U.V.at 280 nm.
Rt vincamine: 6.61 min.
Rt degradation product: 3.67 min

49. Figure (53): Linearity of the area under the peak to the corresponding
concentration of vincamine.

50. Concentration (µg.ml-1)
Table (XXXVII): Determination of vincamine in laboratory prepared mixtures by the proposed procedures.
Concentration (µg.ml-1)
Percentage %
HPLCmethod
Vincamine
Degradation product
Recovery % 18 2
90%
10%
99.13 16 4
80%
20%
98.45 14 6
70%
30%
100.89 12 8
60%
40%
98.85 10
50%
101.33
100.24
99.76
98.63
99.75
Mean
99.67
S.D.
1.007

51. Oxybral capsules claimed to contain 30 mg
Table (XXXVIII): Determination of vincamine in oxybral capsules by the proposed procedures.
Oxybral capsules claimed to contain 30 mg
Batch number
HPLC method
Compendial method**
% Found
Recovery % ± S.D.* A
98.38
99.32 ± 0.956 A
99.36
98.56 ± 0.857 A
100.95
99.21 ± 0.659
* Average of four determinations.
**Spectrophotometric method

52. Table (XXXIX): Statistical comparison for the results obtained by the proposed method and the compendial method for the analysis of vincamine in pure powder form.
HPLC method
Compendial method*
Vincamine
Mean
100.16
99.58
S.D.
1.026
1.011
Variance
1.054
1.022 N 10 6
F test
1.031 (4.77)
Student’s t test
(2.145)
The figures in parenthesis are the corresponding tabulated values at P=0.05.
*Spectrophotometric method

53. Table (XXXX): Results of application of standard addition to the determination of vincamine by the proposed method.
Batch number
Standard added
(mg.ml-1)
HPLC method
Vincamine
Recovery % of added A
0.10
0.15
0.20
99.90
98.40
101.36
Mean ± S.D.
99.89 ± 1.480 A
98.54
101.56
99.79
Mean ± S.D.
99.96 ± 1.517
021554A
97.96
102.32
100.87
100.38± 2.220

54. Table (XXXXI ) : Assay parameters and method validation
Derivative ratio spectrophotometric method
Densitometric method
Colorimetric method
HPLC method
Vincamine
Range (µg/ml)
12-48
3-17(µg/spot)
75 –250
2-20
Slope
-0.04
0.324
0.0032
0.3643
Intercept
-0.021
0.0958
Mean
99.90
100.09
100.13
100.16
S.D.
1.0413
0.761
1.096
1.026
Variance
1.084
0.579
1.201
1.054
Coff. of variation
1.042
0.760
1.094
1.024
Correl. Coef.(r)
0.9992
0.9998
0.9991
0.9994
* RSD%a
0.183 – 0.233
*RSD %b
0.357 – 0.316
* RSD%a , RSD%b the intraday, interday respectively (n=5) relative standard deviation of concentrations (20-32 µg/ml) for derivative ratio, (9-11µg/spot) for densitometric method,
( 150 – 175 µg/ml) for colourimetric method and (10-14 µg/ml) for HPLC method .

55. THANK YOU