Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
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( و قضى ربك ألا تعبدوا إلا إياه وبالوالدين إحسانا إما يبلغن عندك الكبر أحدهما أو كلاهما فلا تقل لهما أف ولا تنهرهما وقل لهما قولاً كريما* واخفض لهما جناح الذل من الرحمة وقل رب ارحمهما كما ربيانى صغيراً*). Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency.

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Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

(و قضى ربك ألا تعبدوا إلا إياه وبالوالدين إحسانا إما يبلغن عندك الكبر أحدهما أو كلاهما فلا تقل لهما أف ولا تنهرهما وقل لهما قولاً كريما* واخفض لهما جناح الذل من الرحمة وقل رب ارحمهما كما ربيانى صغيراً*)



This thesis consists of six parts
This thesis consists of six parts cerebrovascular insufficiency

  • Part I: General introduction

  • Part II: New spectrophotometric method for simultaneous determination of binary mixtures of nicergoline and cinnarizine and stability indicating for vincamine.

  • Part III: Simultaneous determination of nicergoline and cinnarizine.

  • Part IV: Stability indicating methods for the determination of meclophenoxate hydrochloride.

  • Part V: Stability indicating methods for the determination of vinpocetine.

  • Part VI: Stability indicating methods for the determination of Pyritinol dihydrochloride.


Part i general introduction
Part I cerebrovascular insufficiencyGeneral introduction

  • Types of cerebrovascular disease

  • Cerebrovascular insufficiency Etiology and Pathophysiology

  • Complications of cerebrovascular insufficiency

  • Mechanism of action of the selected drugs


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Part II cerebrovascular insufficiency

New spectrophotometric method for simultaneous determination of binary mixtures of nicergoline and cinnarizine and stability indicating for vincamine


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

  • This part includes a general introduction about the chemistry of nicergoline, cinnarizine and vincamine.

  • Review article on the reported methods used for their quantitative determination.

  • This part is subdivided into two sections:

  • Section(A): Determination of vincamine in presence of its acid degradation product by the ratio subtraction method

  • Section(B): Determination of nicergoline and cinnarizine by the ratio subtraction and isosbestic point methods


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Section(A) chemistry of nicergoline, cinnarizine and vincamine.

Determination of vincamine in presence of its acid degradation product by the ratio subtraction method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency


Theory of ratio subtraction method

Theory of ratio subtraction method: chemistry of nicergoline, cinnarizine and vincamine.

The method depends on that, if you have a mixture of two drugs (X) and (Y) with overlapping spectra and the spectrum of (Y) is extended than (X), the determination of (X) can be done by dividing the spectrum of the mixture by a certain concentration of (Y) as a devisor (Y'). The division will give a new curve that represents .If we subtract this constant, then multiply the new curve obtained after subtraction by (Y') (the devisor), therefore we can obtain the original curve of (X). This can be summarized as follows:

The constant can be determined directly from the curve

by the straight line which is parallel to the wavelength axis in the region

where (Y) is extended.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

268nm chemistry of nicergoline, cinnarizine and vincamine.

Figure (2): Absorption spectra of

vincamine 20 µg ml-1 (———)

degradation product 20 µg ml-1 (---------- )

and deg.product 16 µg ml-1 (devisor) (———)

using 0.1N hydrochloric acid as a solvent.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

A (X+Y/Y’) chemistry of nicergoline, cinnarizine and vincamine.

Figure (5): Division spectra of laboratory prepared mixtures of vincamine (X) and its degradation product (Y) using 16 µg ml-1 of degradation product (Y') as a divisor and 0.1 N HCl as a solvent.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

A (X/Y') chemistry of nicergoline, cinnarizine and vincamine.

Figure (6): Division spectra of laboratory prepared mixtures of vincamine (X) and its degradation product (Y) using 16 µg ml-1 of degradation product (Y') as a divisor and 0.1 N HCl as a solvent after subtraction of the constant.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

A (X/Y’*Y’ = X) chemistry of nicergoline, cinnarizine and vincamine.

268nm

268nm

Figure (7): The obtained absorption spectra of vincamine in lab.mixtures 8-32 g.ml-1

Figure (7): The original absorption spectra of vincamine in cal.curve from 8-40 g.ml-1


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Absorbance chemistry of nicergoline, cinnarizine and vincamine.

Figure (4): Linearity of the absorbance of the zero order curve at 268.2 nm to the corresponding concentration of vincamine.



Table ii determination of vincamine in oxybral capsules by the proposed method
Table (II): Determination of vincamine in oxybral capsules by the proposed method.

* Stability indicating spectrophotometric method.

** Spectrophotometric method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (III): Application of standard addition for the determination of vincamine by the proposed method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (IV): Statistical comparison for the results obtained by the proposed method and the reported method for the analysis of vincamine in pure powder form

* Stability indicatingspectrophotometric method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Section [B] by the proposed method and the reported method for the analysis of vincamine in pure powder form

Determination of nicergoline and cinnarizine by the ratio subtraction and isosbestic point methods


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

  • Structure of nicergoline: by the proposed method and the reported method for the analysis of vincamine in pure powder form

  • Structure of cinnarizine:


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

235.8 nm by the proposed method and the reported method for the analysis of vincamine in pure powder form

270.2 nm

Figure (9): Absorption spectra of

Nicergoline 20 µg ml-1 (———)

Cinnarizine 20 µg ml-1 (------------- )

and mixture of 10 µg ml-1 of each drug (……….)

using methanol as a solvent.



Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Absorbance 36

Absorbance

Figure (13): Linearity of the absorbance of the zero order curve at 235.8 nm to the corresponding concentration of nicergoline

Figure (12): Linearity of the absorbance of the zero order curve at 270.2 nm to the corresponding concentration of nicergoline.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

A (X+Y/Y’) 36

Figure (15): Division spectra of laboratory prepared mixtures of cinnarizine (X) and nicergoilne (Y) using 6 µg ml-1 of nicergoline (Y') as a divisor and methanol as a solvent. (Scale x 0.1)


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

A (X/Y') 36

Figure (16): Division spectra of cinnarizine (X) and nicergoilne (Y) using 6 µg ml-1 of nicergoline (Y') as a divisor and methanol as a solvent after subtraction of the constant.(scale x 0.1)


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

252nm 36

A (X/Y‘*Y’ = X)

Figure (17): The obtained absorption spectra of cinnarizine in lab.mixtures.

Figure (17): The original absorption spectra of cinnarizine in cal.curve.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Absorbance 36

Figure (14): Linearity of the absorbance of the zero order curve at 252.0 nm to the corresponding concentration of cinnarizine.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (V): Determination of nicergoline and cinnarizine in laboratory prepared mixtures by the proposed methods


Table vi determination nicergoline and cinnarizine in cinibral tablets by the proposed method
Table (VI): Determination nicergoline and cinnarizine in cinibral tablets by the proposed method.

* HPLC method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (VII): Application of standard addition for the determination of nicergoline and cinnarizine by the proposed method.

* Average of four determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (VIII): Statistical comparison for the results obtained by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder form

* HPLC method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Table ix assay parameters and method validation
Table IX: Assay parameters and method validation obtained by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder form

RSD %a, RSD %b the intra-day, inter-day respectively (n=5) relative standard deviation of concentrations (28, 32 µg/ml) for vincamine, (6, 8 µg/ml) for nicergoline and (14, 16 µg/ml) for cinnarizine..


Part iii simultaneous determination of nicergoline and cinnarizine in their binary mixture

Part III obtained by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder form

Simultaneous determination of nicergoline and cinnarizine in their binary mixture


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

  • This part is subdivided into four sections: obtained by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder form

  • Section [A]:Simultaneous determination of nicergoline and cinnarizine by the derivative spectrophotometry

  • Section [B]: Simultaneous determination of nicergoline and cinnarizine by densitometric methods

  • Section [C]: Simultaneous determination of nicergoline and cinnarizine by high-performance liquid chromatography

  • Section [D]: Simultaneous determination of nicergoline and cinnarizine by chemometric methods


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Section [A] obtained by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder form

Simultaneous determination of nicergoline and cinnarizine by the derivative spectrophotometry


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (67): Absorption spectra of obtained by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder form

Nicergoline 22 µg ml-1 (———) and

Cinnarizine 12 µg ml-1 (---------- )

using methanol as a solvent.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

dA/d obtained by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder formλ

307nm

Figure (68): First order spectra of

Nicergoline 22 μg ml-1 (______)

Cinnarizine 12 μg ml-1 (_ _ _ _ _ _)

using methanol as a solvent.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

dA/d obtained by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder formλ

307nm

Figure (69): First – derivative absorption spectra of

6 - 38 µg ml-1 nicergoline.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (70): Linearity of the peak amplitude of the first derivative at 307.6 nm to the corresponding concentration of nicergoline.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

A (cinnarizine/nicergoline) derivative at 307.6 nm to the corresponding concentration of nicergoline.

Figure (71): Zero order of ratio spectra of cinnarizine 6- 22

μg ml-1 using 10 µg ml-1 of nicergoline as a divisor.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

244nm derivative at 307.6 nm to the corresponding concentration of nicergoline.

dA(cinnarizine/nicergoline)/dλ

Figure (72): First order of ratio spectra of cinnarizine 6- 22 μg ml-1

using 10 µg ml-1 of nicergoline as a divisor.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (73): Linearity of the peak amplitude of the first derivative of the

ratio spectra at 244.6 nm to the corresponding concentration

of cinnarizine.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXIII): Simultaneous determination of nicergoline and cinnarizinee in laboratory prepared mixtures by the proposed methods.


Table lxiv determination of nicergoline and cinnarizine in cinibral tablets by the proposed methods
Table (LXIV): Determination of nicergoline and cinnarizine in cinibral tablets by the proposed methods.

* HPLC method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXV): Application of standard addition for the determination of nicergoline and cinnarizine by the proposed method.

* Average of four determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (LXVI): Statistical comparison for the results obtained by the proposed method and the reported method for the analysis of nicergoline andcinnarizine in pure powder form

* HPLC method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Section b simultaneous determination of nicergoline and cinnarizine by densitometric methods

Section [B] obtained by the proposed method and the reported method for the analysis of nicergoline and

Simultaneous determination of nicergoline and cinnarizine by densitometric methods


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (74): TLC chromatogram of nicergoline and cinnarizine obtained by the proposed method and the reported method for the analysis of nicergoline and

A= nicergoline, Rf= 0.505.

B= cinnarizine, Rf = 0.807.

M= mixture of both drugs

Developing system, chloroform : methanol : ethyl acetate(5: 3 :2 v/v/v)


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Reflectance obtained by the proposed method and the reported method for the analysis of nicergoline and

Reflectance

Distance (mm)

Distance (mm)

Figure (76):

Scanning profile of the TLC chromatogram of cinnarizine at 252 nm.

Figure (75):

Scanning profile of the TLC chromatogram of nicergoline at 287 nm.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

spot obtained by the proposed method and the reported method for the analysis of nicergoline and-1

Figure (77): Linearity of the area under the peak to the corresponding

concentration of nicergoline

Figure (78): Linearity of the area under the peak to the corresponding

concentration of cinnarizine


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXVII): Determination of nicergoline and cinnarizine in laboratory prepared mixtures by the proposed method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXVIII): Determination of nicergoline and cinnarizine in cinibral tablets by the proposed method.

* HPLC method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXIX): Application of standard addition for the determination of nicergoline and cinnarizine by the proposed method.

* Average of four determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (LXX): Statistical comparison for the results obtained by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder form

* HPLC method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Section [C] by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder form

Simultaneous determination of nicergoline and cinnarizine by high-performance liquid chromatography


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Detector response by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder form

Time (min)

Figure (79): Liquid chromatographic separation of nicergoline and cinnarizine

using final assay conditions:

Column: RP18

Mobile phase: methanol : acetonitrile : water (4: 4: 2 v/v/v).

Flow rate: 1.5 ml min-1.

Detection: 287 nm for nicergoline and 252 nm for cinnarizine.

Retention time nicergoline: 2.15 min.

Retention time cinnarizine: 4.72 min.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (80): Linearity of the relative peak area to the corresponding

concentration of nicergoline.

Figure (81): Linearity of the relative peak area to the corresponding

concentration of cinnarizine.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXXI): Determination of nicergoline and cinnarizine in laboratory prepared mixtures by the proposed method.



Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXXIII): Determination of nicergoline and cinnarizine in cinibral tablets by the proposed method.

* HPLC method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXXIV): Application of standard addition for the determination of nicergoline and cinnarizine by the proposed method.

* Average of four determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (LXX): Statistical comparison for the results obtained by the proposed method and the reported method for the analysis of nicergoline and cinnarizine in pure powder form

* HPLC method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Table lxxvi assay parameters and method validation for nicergoline and cinnarizine
Table (LXXVI): Assay parameters and method validation for nicergoline and cinnarizine

*RSD%a, RSD%b: the intra-day, inter-day respectively (n=5) relative standard deviation of concentrations (20 and 30 µg/ml nicergoline and 10 and 14 µg/ml cinnarizine) for derivative ratio, (16 and 20 µg/spot nicergoline and 4 and 6 µg/spot cinnarizine) for densitometric method and (20 and 30 µg/ml from both nicergoline and cinnarizine) for HPLC method.


Section d simultaneous determination of nicergoline and cinnarizine by chemometric methods

Section [D] nicergoline and cinnarizine

Simultaneous determination of nicergoline and cinnarizine by chemometric methods


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

  • Chemometrics is the application of mathematical and statistical methods to provide maximum chemical information through analysis of chemical data.

  • In this section, three chemometric techniques were applied for simultaneous determination of nicergoline and cinnarizine

    • Classical Least Squares (CLS) with non zero intercept,

    • principal component regression (PCR)

    • partial least squares (PLS).


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXXVII): The concentration of different mixtures of nicergoline and cinnarizine used in the training set


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (82): RMSEC plot of the cross validation results of the training set as a function of the number of principal components used toconstruct the PCR calibration for nicergoline.

Figure (83): RMSEC plot of the cross validation results of the training set as a functionof the number of principal components used to construct the PLS calibration for nicergoline.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (36): RMSEC plot of the cross validation results of the training set as afunction of the number of principal components used to construct the PLScalibrationfor cinnarizine.

Figure (35): RMSEC plot of the cross validation results of the training set as afunction of the number of principal components used to construct the PCR calibration for cinnarizine.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXXVIII): Results of the analysis of the mixtures of the validation set of nicergoline & cinnarizine by the proposed methods.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (85): Predicted concentration versus actual concentration of cinnarizine in the validation set using CLS method

Figure (84): Predicted concentration versus actual concentration of nicergoline in the validation set using CLS method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (86): Predicted concentration versus actual concentration of nicergoline in the validation set using PCR method

Figure (87): Predicted concentration versus actual concentration of cinnarizine in the validation set using PCR method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (88) : Predicted concentration versus actual concentration of nicergoline in the validation set using PLS method

Figure (89) : Predicted concentration versus actual concentration of cinnarizine in the validation set using

PLS method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (91): Concentration residuals versus actual concentration of cinnarizine in the validation set using CLS method

Figure (90): Concentration residuals versus actual concentration of nicergoline in the validation set using CLS method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (93): Concentration residuals versus actual concentration of cinnarizine in the validation set using PCR method

Figure (92): Concentration residuals versus actual concentration of nicergoline in the validation set using PCR method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (94): Concentration residuals versus actual concentration of nicergolinein the validation set using PLS method

Figure (95): Concentration residuals versus actual concentration of cinnarizine in the validation set using PLS method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXXIX): RMSEP and Q2 values of the validation set analysis of cinnarizine hydrochloride by the proposed methods.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXXX): Quantitative determination of nicergoline and cinnarizine in cinibral tablets by the proposed methods

* (μg.ml –1).


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (LXXXI): Results of the standard addition technique for the simultaneous determination of nicergoline and cinnarizine in cinibral tablets by the proposed methods


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (LXXXII): Statistical comparison for the results obtained by the proposed methods and the reported method for the analysis of nicergoline and cinnarizine in pure powder form.

* HPLC method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Part iv stability indicating methods for the determination of meclophenoxate hydrochloride

Part IV obtained by the proposed methods and the reported method for the analysis of nicergoline and cinnarizine in pure powder form.

Stability indicating methods for the determination of meclophenoxate hydrochloride


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

  • This part includes a general introduction about the chemistry of meclophenoxate hydrochloride.

  • Review article on the reported methods used for its quantitative determination.

  • This part is subdivided into three sections:

  • Section(A): High-performance liquid chromatographic determination of meclophenoxate hydrochloride in presence of its acid degradation product

  • Section(B): Kinetic study on the degradation of meclophenoxate hydrochloride

  • Section (C):Determination of meclophenoxate hydrochloride in presence of its acid degradation product using ion selective electrodes


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Section [A] chemistry of meclophenoxate hydrochloride.

High-performance liquid chromatographic determination of meclophenoxate hydrochloride in presence of its acid degradation product


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

-Structure of meclophenoxate chemistry of meclophenoxate hydrochloride.

-The proposed mechanism for preparing the degradation product:

2 N NaOH

Reflux 25 min.

then neutralize with HCl


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Detector response chemistry of meclophenoxate hydrochloride.

Time (min)

Figure (19): Liquid chromatographic separation of meclophenoxate.HCl and its degradation product using final assay conditions:

Column: RP18

Mobile phase: 0.01 M ammonium carbonate: acetonitrile (7:3 v/v).

Flow rate: 1.0 ml min-1.

Detection: 277 nm.

Retention time meclophenoxate hydrochloride: 5.39 min.

Retention time degradation product: 2.70 min.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (20): Linearity of the relative peak area to the corresponding concentration of meclophenoxate hydrochloride.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (X): Determination of meclophenoxate hydrochloride in laboratory prepared mixtures by the proposed method



Table xii determination of meclophenoxate hydrochloride in lucidril tablets by the proposed method
Table (XII): Determination of meclophenoxate hydrochloride in lucidril tablets by the proposed method

* Stability indicating HPLC method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (XIII): Application of standard addition for the determination of meclophenoxate hydrochloride by the proposed method

* Average of four determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (XIV): Statistical comparison for the results obtained by the proposed method and the reported method for the analysis of meclophenoxate hydrochloride in pure powder form

* Stability indicating HPLC method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Section b kinetic study on degradation of meclophenoxate hydrochloride

Section [B] by the proposed method and the reported method for the analysis of meclophenoxate hydrochloride in pure powder form

Kinetic study on degradation of meclophenoxate hydrochloride


Kinetic study on degradation of meclophenoxate hydrochloride includes
Kinetic study on degradation of meclophenoxate hydrochloride includes:

Study the kinetic order of the reaction

Study the effect of sodium hydroxide concentration on the reaction rate

Study the effect of temperature on the reaction rate

Calculate energy of activation (Ea):

log =


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (22): First order plot of the hydrolysis of meclophenoxate hydrochloride (1000 mg %) with 2 N NaOH at 80 oC


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (23): First order plot of the hydrolysis of meclophenoxate hydrochloride (1000 mg %) with 2 N NaOH at different temperatures.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (24): First order plot of the hydrolysis of meclophenoxate hydrochloride (1000 mg %) with 1.5 N NaOH at different temperatures.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (25): First order plot of the hydrolysis of meclophenoxate hydrochloride (1000 mg %) with 1.0 N NaOH at different temperatures.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (26): Arrhenius Plot for the hydrolysis of meclophenoxate hydrochloride (1000 mg %) with 1.0, 1.5, 2.0 N NaOH

Ea" was found to be 12.331 kilo calories mol-1



Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Section [C] hydrolysis

Determination of meclophenoxate hydrochloride in presence of its acid degradation product using ion selective electrodes


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

  • Ion selective electrodes are electrodes containing membranes having a selective response for a particular ion.

  • Meclophenoxate hydrochloride (Cation) reacted with tetraphenylborate or reineckate (anionic ion exchangers) to form stable 1:1, water insoluble ion association complex.

  • -CD-based sensors form inclusion complexes in the aqueous and in solid state with organic molecules.

  • Three membranes are studied in this part:

    a) meclo-TPB

    b) meclo-RNC

    c) -CD-RNC


The electrode assembly
The electrode assembly having a selective response for a particular ion.

Figure (27): PVC matrix membrane ion selective electrode: [1] Shielded cable. [2] Rubber sheath. [3] Quickfit cone. [4] Quickfit socket. [5] Mercury. [6] Calomel reference electrode. [7] Internal solution. [8] PVC tubing. [9] Sensor membrane.




Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (30): Effect of pH on the response of reineckate electrode CD- reineckate electrode.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (31): Effect of temperature on the response of reineckate electrode CD- reineckate membrane electrode.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (32): Profile of the potential in mV to the –log concentration of meclophenoxate hydrochloride with meclo- TPB, meclo – reineckate and  CD- reineckate.



Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (XIX) Potentiometric selectivity coefficients (K the three investigated electrodesPlotprimary ion, interferent) for the three proposed electrodes.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (XX): Determination of meclophenoxate hydrochloride in laboratory prepared mixtures by the proposed method


Table xxi determination of meclophenoxate hydrochloride in lucidril tablets by the proposed method
Table (XXI): Determination of meclophenoxate hydrochloride in lucidril tablets by the proposed method

* Stability indicating HPLC method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (XXII): Determination of meclophenoxate hydrochloride in spiked human plasma by the proposed electrodes.

* Average of three determinations


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (XXIII): Application of standard addition for the determination of meclophenoxate hydrochloride by the proposed method

* Average of three determinations


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (XXIV): Statistical comparison for the results obtained by the proposed method and the reported method for the analysis of meclophenoxate hydrochloride in pure powder form

* Stability indicating HPLC method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Table xliv assay parameters and method validation for meclophenoxate hydrochloride
Table (XLIV): Assay parameters and method validation for meclophenoxate hydrochloride

* RSD%a, RSD%b the intra-day, inter-day respectively (n=5) relative standard deviation of concentrations (200 and 300 µg ml-1) and (10-3M and 10-4M) for ion selective electrodes method.


Part v stability indicating methods for determination of vinpocetine

Part V meclophenoxate hydrochloride

Stability indicating methods for determination of vinpocetine


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

  • This part includes a general introduction about the chemistry of vinpocetine.

  • Review article on the reported methods used for its quantitative determination.

  • This part is subdivided into four sections:

  • Section [A]: Determination of vinpocetine in presence of its acid degradation product by the derivative ratio spectrophotometry

  • Section [B]: Densitometric determination of vinpocetine in presence of its acid degradation product

  • Section [C]: High-performance liquid chromatographic determination of vinpocetine in presence of its acid degradation product

  • Section [D]: Chemometric determination of vinpocetine in presence of its acid degradation product


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Section [A] chemistry of vinpocetine.

Determination of vinpocetine in presence of its acid degradation product by the derivative ratio spectrophotometry


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (34): Absorption spectra of chemistry of vinpocetine.

vinpocetine 12 µg ml-1 (———) and

degradation product 10 µg ml-1 (---------- )

using 0.1N hydrochloric acid as a solvent.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

dA/d chemistry of vinpocetine.λ

Figure (35): First order spectra of

vinpocetine 12 μg ml-1 (______)

degradation product 10 μg ml-1 (_ _ _ _ _ _)

using 0.1N hydrochloric acid as a solvent.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

A (vinpocetine/deg.prod.) chemistry of vinpocetine.

Figure (36): Zero order of ratio spectra of vinpocetine 4-32 μg ml-1 using 10

µg.ml-1 of degradation product as a divisor.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

311nm chemistry of vinpocetine.

dA(vinpocetine/deg.product)/dλ

Figure (37): First order of ratio spectra of vinpocetine 4-32 μg ml-1

using 10 µg ml-1 of degradation product as a divisor.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (38): Linearity of the peak amplitude of the first derivative of the

ratio spectra at 311 nm to the corresponding concentration

of vinpocetine.


Table xxv determination of vinpocetine in laboratory prepared mixtures by the proposed method
Table (XXV): Determination of vinpocetine in laboratory prepared mixtures by the proposed method.


Table xxvi determination of vinpocetine in vinporal tablets by the proposed method
Table (XXVI): Determination of vinpocetine in vinporal tablets by the proposed method.

* Spectrophotometric method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (XXVII): Application of standard addition for the determination of vinpocetine in its pharmaceutical preparation by the proposed method.

* Average of four determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (XXVIII): Statistical comparison for the results obtained by the proposed method and the reported method for the analysis of vinpocetine in pure powder form

* Spectrophotometric method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Section b densitometric determination of vinpocetine in presence of its acid degradation product

Section [B] obtained by the proposed method and the reported method for the analysis of vinpocetine in pure powder form

Densitometric determination of vinpocetine in presence of its acid degradation product


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (40): TLC chromatogram of vinpocetine and its obtained by the proposed method and the reported method for the analysis of vinpocetine in pure powder form

degradation product.

A= Vinpocetine Rf = 0.80.

B= Degradation product Rf = 0.54

M= mixture of vinpocetine and its deg. product

Developing system, methanol: chloroform: ethyl acetate

(2:1:1 v/v/v).


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Reflectance obtained by the proposed method and the reported method for the analysis of vinpocetine in pure powder form

Distance (mm)

Figure (41):

Scanning profile of the TLC chromatogram of vinpocetine at 268 nm.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (42): Linearity of the area under the peak to the corresponding

concentration of vinpocetine


Table xxix determination of vinpocetine in laboratory prepared mixtures by the proposed method
Table (XXIX): Determination of vinpocetine in laboratory prepared mixtures by the proposed method.


Table xxx determination of vinpocetne in vinporal tablets by the proposed method
Table (XXX): Determination of vinpocetne in vinporal tablets by the proposed method

* Spectrophotometric method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (XXXI): Application of standard addition for the determination of vinpocetine in its pharmaceutical preparation by the proposed method.

* Average of four determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (XXXII): Statistical comparison for the results obtained by the proposed method and the reported method for the analysis of vinpocetine in pure powder form.

* Spectrophotometric method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Section [C] obtained by the proposed method and the reported method for the analysis of vinpocetine in pure powder form.

High-performance liquid chromatographic determination of vinpocetine in presence of its acid degradation product


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Detector response obtained by the proposed method and the reported method for the analysis of vinpocetine in pure powder form.

Time (min)

Figure (43): Liquid chromatographic separation of vinpocetine and its degradation product using final assay conditions:

Column: RP18

Mobile phase: methanol: 0.01 M ammonium bicarbonate (60:40 v/v).

Flow rate: 1.6 ml min-1.

Detection: 268 nm.

Retention time vinpocetine (II): 6.12 min.

Retention timedegradation product (I): 2.24 min.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (44): Linearity of the relative peak area to the corresponding

concentration of vincamine.


Table xxxiii determination of vinpocetine in laboratory prepared mixtures by the proposed method
Table (XXXIII): Determination of vinpocetine in laboratory prepared mixtures by the proposedmethod



Table xxxv determination of vinpocetine in vinporal tablets by the proposed method
Table (XXXV): Determination of vinpocetine in vinporal tablets by the proposed method

* Spectrophotometric method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (XXXVI): Application of standard addition for the determination of vinpocetine in vinporal tablets by the proposed method

* Average of four determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (XXXVII): Statistical comparison for the results obtained by the proposed method and the reported method for the analysis of vinpocetine in pure powder form

* Spectrophotometric method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Table xxxviii assay parameters and method validation for vinpocetine
Table (XXXVIII): Assay parameters and method validation for vinpocetine

*RSD%a, RSD%b: the intra-day, inter-day respectively (n=5) relative standard deviation of concentrations (16 and 20 µg/ml) for derivative ratio, (9 and 11µg/spot) for densitometric method and (8 and 12 µg/ml) for HPLC method.


Section d chemometric determination of vinpocetine in presence of its acid degradation product

Section [D] vinpocetine

Chemometric determination of vinpocetine in presence of its acid degradation product


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (XXXIX): The concentration of different mixtures of vinpocetine and its degradation product used in the training set.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (45): RMSEC plot of the cross validation results of the training set as a function of the number of principal components used to construct the PCR calibration for vinpocetine.

Figure (46): RMSEC plot of the cross validation results of the training set as a function of the number of principal components used to construct the PLS calibration for vinpocetine.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (XXXX): Results of the analysis of the mixtures of the validation set of vinpocetine and its degradation product by the proposed methods


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (47): Predicted concentration versus actual concentration of vinpocetine in the validation set using CLS method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (48): Predicted concentration versus actual concentration of vinpocetine in the validation set using PCR method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (49): Predicted concentration versus actual concentration of vinpocetine in the validation set using PLS method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (50): Concentration residuals versus actual concentration of vinpocetine in the validation set using CLS method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (51): Concentration residuals versus actual concentration of vinpocetine in the validation set using PCR method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (52): Concentration residuals versus actual concentration of vinpocetine in the validation set using PLS method


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (XXXXI): RMSEP and Q2 values of the validation set analysis of vinpocetine by the proposed methods.


Table xxxxii quantitative determination of vinpocetine in vinporal tablets by the proposed methods
Table (XXXXII): Quantitative determination of vinpocetine in vinporal tablets by the proposed methods.

* Spectrophotometric method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (XXXXIII): Results of the standard addition technique for the determination of vinpocetine in vinporal tablets by the proposed methods


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (XXXXIV): Statistical comparison for the results obtained by the proposed methods and the reported method for the analysis of vinpocetine in pure powder form.

* Spectrophotometric method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Part vi stability indicating methods for the determination of pyritinol dihydrochloride

Part VI obtained by the proposed methods and the reported method for the analysis of vinpocetine in pure powder form.

Stability indicating methods for the determination of Pyritinol dihydrochloride


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

  • This part includes a general introduction about the chemistry of pyritinol dihydrochloride.

  • Review article on the reported methods used for its quantitative determination.

  • This part is subdivided into three sections:

  • Section [A]:Determination of pyritinol dihydrochloride in presence of its degradation product by the derivative ratio spectrophotometry

  • Section [B]: High-performance liquid chromatographic determination of pyritinol dihydrochloride in presence of its degradation product

  • Section [C]: Determination of pyritinol dihydrochloride in presence of its degradation product using ion selective electrodes


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

CH chemistry of pyritinol dihydrochloride.

CH

CH

N

N

N

3

3

3

H

O

30% v/v

SO3H

2

2

S -S

2

HO

HO

OH

1 hr at room temp

HO

OH

HO

mol.wt 233

  • Structure of pyritinol.2HCl

  • The proposed scheme for its degradation


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Section [A] chemistry of pyritinol dihydrochloride.

Determination of pyritinol dihydrochloride in presence of its degradation product by the derivative ratio spectrophotometry


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (54): Absorption spectra of chemistry of pyritinol dihydrochloride.

pyritinol dihydrochloride 20 µg ml-1 (———) and

degradation product 20 µg ml-1 (---------- )

using distelled water as a solvent.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

dA/d chemistry of pyritinol dihydrochloride.λ

Figure (55): First order spectra of

pyritinol dihydrochloride 20 µg ml-1 (———) and

degradation product 20 µg ml-1 (---------- )

using distelled water as a solvent.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

A (pyritinol.2HCl/deg.prod.) chemistry of pyritinol dihydrochloride.

Figure (56): Zero order of ratio spectra of pyritinol dihydrochloride 10-26 μg ml-1 using 20 µg.ml-1 of degradation product as a divisor.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

dA(pyritinol.2HCl/deg.product)/d chemistry of pyritinol dihydrochloride.λ

357nm

Figure (57): First order of ratio spectra of pyritinol dihydrochloride 10-26 μg ml-1 using 20 µg ml-1 of degradation product as a divisor.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (58): Linearity of the peak amplitude of the first derivative of the ratio spectra at 357.4 nm to the corresponding concentration of pyritinol dihydrochloride.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (XXXXV): Determination of pyritinol dihydrochloride in laboratory prepared mixtures by the proposed method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (XXXXVI): Determination of pyritinol dihydrochloride in Encephabol tablets by the proposed method.

* Stability indicating spectrophotometric method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (XXXXVII): Application of standard addition for the determination of pyritinol dihydrochloride by the proposed method.

* Average of four determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (XXXXVIII): Statistical comparison for the results obtained by the proposed methods and the reported method for the analysis of pyritinol dihydrochloride in pure powder form.

* Stability indicating spectrophotometric method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Section [B] obtained by the proposed methods and the reported method for the analysis of pyritinol dihydrochloride in pure powder form.

High-performance liquid chromatographic determination of pyritinol dihydrochloride in presence of its oxidative degradation product


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Detector response obtained by the proposed methods and the reported method for the analysis of pyritinol dihydrochloride in pure powder form.

Time (min)

Figure (60): Liquid chromatographic separation of pyritinol dihydrochloride and its degradation product using final assay conditions:

Column: RP 8

Mobile phase:acetonitrile: solution pH 4(58:42 v/v).

Flow rate: 1.0 ml min-1.

Detection: 296 nm.

Retention time pyritinol dihydrochloride: 1.62 min.

Retention time degradation product: 2.97 min.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (61): Linearity of the relative peak area to the corresponding concentration of pyritinol dihydrochloride.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (IL): Determination of pyritinol dihydrochloride in laboratory prepared mixtures by the proposed method



Table li determination of pyritinol dihydrochloride in encephabol tablets by the proposed method
Table (LI): Determination of pyritinol dihydrochloride in Encephabol tablets by the proposed method.

* Stability indicating spectrophotometric method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LII): Application of standard addition for the determination of pyritinol dihydrochloride by the proposed method.

* Average of four determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (LIII): Statistical comparison for the results obtained by the proposed methods and the reported method for the analysis of pyritinol dihydrochloride in pure powder form.

* Stability indicating spectrophotometric method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Section [C] obtained by the proposed methods and the reported method for the analysis of pyritinol dihydrochloride in pure powder form.

Determination of pyritinol dihydrochloride in presence of its degradation product using ion selective electrodes


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

  • Pyritinol dihydrochloride (Cation) reacted with tetraphenylborate or reineckate (anionic ion exchangers) to form stable 1:2, water insoluble ion association complex.

  • -CD-based sensors form ion seiving membranes.

  • Three membranes are studied in this part:

    a) Pyrit-TPB

    b) Pyrit-RNC

    c) -CD-RNC




Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (64): Effect of pH on the response of reineckate electrode CD- reineckate electrode


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (65): Effect of temperature on the response of reineckate electrode CD- reineckate membrane electrode


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Figure (66): Profile of the potential in mV to the –log concentration of pyritinol dihydrochloride with pyrit- TPB, pyrit – reineckate and  CD- renikate.



Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LVII) Potentiometric selectivity coefficients (K three investigated electrodesPlotprimary ion, interferent) for the three proposed electrodes.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LVIII): Determination of pyritinol dihydrochloride in laboratory prepared mixtures by the proposed method


Table lix determination of pyritinol diydrochloride in encephabol tablets by the proposed method
Table (LIX): Determination of Pyritinol diydrochloride in encephabol tablets by the proposed method

* Stability indicating spectrophotometric method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LX): Determination of pyritinol dihydrochloride in spiked human plasma by the proposed electrodes.

* Average of three determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency
Table (LXI): Application of standard addition for the determination of pyritinol dihydrochloride by the proposed method

* Average of three determinations.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

Table (LXII): Statistical comparison for the results obtained by the proposed method and the reported method for the analysis of Pyritinol dihydrochloride in pure powder form

* Stability indicating spectrophotometric method.

The figures in parenthesis are the corresponding tabulated values at P=0.05


Table lxxxii assay parameters and method validation for pyritinol dihydrochloride
Table (LXXXII): Assay parameters and method validation for pyritinol dihydrochloride

* RSD%a, RSD%b the intra-day, inter-day respectively (n=5) relative standard deviation of concentrations (18 and 20 µg/ml) for derivative ratio method, (40 and 50 µg ml-1) for HPLC method and (10-4M and 10-5M) for ion selective electrodes method.


Contribution for the analysis of certain drugs which treat cerebrovascular insufficiency

THANK YOU pyritinol dihydrochloride