Analytical Techniques & Preformulation Studies ANALYTICAL ...shodhganga.inflibnet.ac.in/bitstream/10603/8952/9/10_chapter 3.pdf · _____Analytical Techniques & Preformulation Studies

_____________________________________Analytical Techniques & Preformulation Studies

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ANALYTICAL TECHNIQUES AND

PREFORMULATION STUDIES

3.1 PROCHLORPERAZINE MALEATE

3.1.1 Analytical Methods Established

Gas Chromatography Mass Spectroscopy:

McKay G et al., (1982), compared a gas chromatographic mass spectrometric assay

using selected ion monitoring with a high performance liquid chromatographic assay

using an electrochemical detector for single dose studies of the psychotherapeutic

phenothiazine drug prochlorperazine. Measurements were made after extraction of

prochlorperazine from basified plasma with an isopropanol pentane solvent mixture.

The residual sample was analyzed by high performance liquid chromatography using

an electrochemical detector. This method displayed excellent correlation for plasma

concentration determinations in the range of 0.25-10 ng/ml and will allow for the

study of the pharmacokinetics of prochlorperazine following single low doses of the

drug.1

Liquid Chromatography:

Yan M et al., (2009), developed a sensitive and specific method using a one step

liquid-liquid extraction with dichloromethane followed by liquid chromatographic-

electrospray ionization mass spectrometric was developed and validated to determine

prochlorperazine maleate in human plasma using amitriptyline hydrochloride as an

internal standard. The samples were separated using a Thermo Hypersil-Hypurity C18

reversed-phase column (150 mm x 2.1 mm i.d., 5 µm). A mobile phase containing 10

mM ammonium acetate (pH 3.6)-methanol-acetonitrile (27:68:5, v/v/v) was used

isocratically eluting at a flow rate of 0.22 ml/min. The average extraction recovery of

prochlorperazine and internal standard were 81.8±2.2% and 79.5±3.7%, respectively.

Prochlorperazine maleate and internal standard were measured by electrospray ion

source in positive selective ion monitoring mode. The method demonstrated that good

linearity ranged from 0.20 to 6.40 ng/ml with r2 0.9989. The limit of quantification for

prochlorperazine maleate in the plasma was 0.20 ng/ml. The established method has

successfully applied to a bioequivalence study of two prochlorperazine maleate

formulations in 18 healthy male Chinese volunteers.2


44

Uraisin K et al., (2006), developed a simple and fast flow injection

spectrophotometric method for the determination of bromate in water samples. The

detection system was based on the oxidation of prochlorperazine (PCP) with bromate

in strongly acidic medium. Large amounts of chloride and bromide was found, for the

first time, to act as an activator and to enhance the sensitivity for bromate detection.

The oxidation product of PCP gives pink color, which can be used to monitor the

reaction spectrophotometrically at 525 nm. Under the optimal conditions, the method

was selective; only nitrite, chlorite and hypochlorite can interfere with the

determination of bromate. The elimination of these three ions was discussed. The

calibration graph for bromate determination was linear in the range of 10-130 µg/l

with a detection limit of 2.3 µg/l. The repeatability was satisfactory, with the relative

standard deviation of 1.1% (25 µg/l, n=10). The sample throughput was 44 per hour.

The proposed method was found to be highly reliable for screening drinking waters

containing bromate, which was above or below legislation limit of 10µg/l.3

Marumo A et al., (2005), extracted seven phenothiazine derivatives, perazine,

perphenazine, prochlorperazine, propericiazine, thioproperazine, trifluoperazine, and

flupentixol from human plasma and urine samples using disk solid-phase extraction

(SPE) with an Empore C18 cartridge. Human plasma and urine (1 ml each) containing

the 7 phenothiazine derivatives were mixed with 2 ml of 0.1M NaOH and 7 ml

distilled water and then poured into the disk SPE cartridges. The drugs were eluted

with 1 ml chloroform-acetonitrile (8:2) and determined by liquid chromatography

with ammonium formate/formic acid-acetonitrile gradient elution. The detection was

performed by ultraviolet absorption at 250 nm. The separation of the 7 phenothiazine

derivatives from each other and from impurities was generally satisfactory using a

Symmetry Shield RP8 column (150 x 2.1 mm id, 3.5 µm particle size). The recoveries

of the 7 phenothiazine derivatives spiked into plasma and urine samples were 64.0-

89.9% and 65.1-92.1%, respectively. Regression equations for the 7 phenothiazine

derivatives showed excellent linearity with detection limits of 0.021-0.30 µg/ml for

plasma and 0.017-0.30 µg/ml for urine. The within day and day to day coefficients of

variation for both samples were commonly below 9.0 and 14.9%, respectively.4

Honigberg IL et al., (1975), investigated parameters associated with the separation of

antianxiety-antispasmodic agents using high pressure liquid chromatography. Eight

widely prescribed drugs were studied. The compounds were chromatographed on


45

reversed phase octadecyltrichlorosilane (C18) or diphenyldichlorosilane (phenyl)

columns, using mixtures of absolute methanol and distilled water buffered with

ammonium dihydrogen phosphate, ammonium acid phosphate or ammonium

carbonate. A mixture of phenobarbital-propantheline bromide was selected to

demonstrate the utility of the separation and quantification method. The mixture was

chromatographed on a phenyl column, using absolute methanol-aqueous 1%

ammonium dihydrogen phosphate (60:40) (pH 5.85) at a flow rate of 1.4 ml/min.

Each determination can achieve in approximately 15 min with an accuracy of 1-2%.5

High Performance Thin Layer Chromatography:

Tanaka E et al., (2007), developed a high performance liquid chromatographic

method for the simultaneous analysis of the 12 phenothiazines (chlorpromazine,

fluphenazine, levomepromazine, perazine, perphenazine, prochlorperazine,

profenamine, promethazine, propericiazine, thioproperazine, thioridazine and

trifluoperazine) in human serum using HPLC/UV. The separation was achieved using

a C18 reversed-phase column (250 mm x 4.6 mm i.d., particle size 5 µm, Inersil ODS-

SP). The mobile phase, consisting of acetonitrile-methanol-30 mM NaH2PO4 (pH 5.6)

(300:200:500, v/v/v), was delivered at a flow rate of 0.9 ml/min and UV detection

was carried out at 250 nm. The recoveries of the 12 phenothiazines spiked into serum

samples were 87.6-99.8%. Regression equations for the 12 phenothiazines showed

excellent linearity, with detection limits of 3.2-5.5 ng/ml for serum. The inter-day and

intraday coefficients of variation for serum samples were commonly below 8.8%. The

selectivity, accuracy and precision of this method were satisfactory for clinical and

forensic purposes.6

Mizuno Y et al., (2002), devolved a method to detect phenothiazines compounds by

high-performance liquid chromatography/fast atom bombardment-mass spectrometry

(HPLC/FAB-MS) method. Authentic samples of the compounds were subjected to

our HPLC/FAB-MS system their mass spectra were obtained by positive and negative

modes. Four typical phenothiazines, in the serum samples of two patients, were also

analyzed. All 17 phenothiazines were sufficiently separated on the chromatogram. In

the positive mode, all the base peaks were quasimolecular ions; their main fragment

ions observed were [M-R(1)+CH(2)](+), [R(1)](+), [M-R(1)](+) and [M+H+Gly](+).

In the negative mode, the base peaks were [Cl](-) for chlorpromazine,

prochlorperazine and perphenazine, three compounds containing chloride. For the


46

other compounds, they were [M-R(1)-CH(3)](-), [M-R(1)-CH(2)CH(3)](-) or [M-

R(1)-(CH(3))(2)](-) ions. The present method would useful in forensic toxicological

practice.7

Mou C et al., (1997), developed a high performance liquid chromatographic method

for simultaneous extraction, elution and determination of doxorubicin and

prochlorperazine content in human plasma samples. The procedure consists of

extraction through a conditioned C18 solid phase extraction cartridge, elution from a

Spherisorb C8 reversed-phase column by an isocratic mobile phase (60% acetonitrile,

15% methanol and 25% buffer) followed by detection with electrochemical and

fluorescence detectors. Recovery of doxorubicin and prochlorperazine from pooled

human plasma samples (n=3) containing 100 ng/ml of the two drugs was 77.8 ± 3.5%

and 89.1 ± 6.0%, respectively. The lower limits of quantitation for doxorubicin and

prochlorperazine in plasma samples were 6.25 ng/ml and 10 ng/ml, respectively. A

linear calibration curve was obtained for up to 2 µg/ml of doxorubicin and

prochlorperazine. This combination method may be of particular value in clinical

studies where phenothiazines such as prochlorperazine were used to enhance retention

of doxorubicin in drug resistant tumor cells.8

UV-Visible Spectroscopy:

Kitamura K et al., (1998), measured the absorption spectra of six phenothiazine

derivatives, chlorpromazine, triflupromazine, promazine, promethazine,

trifluoperazine and prochlorperazine, in the solutions containing various amounts of

human erythrocyte ghosts (HEG) showed bathocromic shifts according to the amount

of HEG. Due to the strong background signals caused by HEG, the baseline

compensation was incomplete, even though the sample and the reference solutions

contained the same amount of HEG, hence further spectral information could not be

obtained. The second derivative spectra of these absorption spectra clearly showed the

derivative isosbestic points, indicating that the residual background signal effects

were entirely eliminated. The derivative intensity differences of the phenothiazines

(DeltaD values) before and after the addition of HEG were measured at a specific

wavelength. Using the DeltaD values, the partition coefficients (Kp) of these drugs

were calculated and obtained with RSD of below 10 %. The results indicate that the

derivative method can be applicable to the determination of partition coefficients of

drugs to HEG without any separation procedures.9


47

3.1.2 Drug Analysis

Melting Point: The melting point of the prochlorperazine maleate was determined by

capillary fusion method. A capillary was sealed at one end filled with a small amount

of prochlorperazine maleate and the capillary was kept inverted i.e. sealed end

downwards into the melting point apparatus.13

Reported Melting Point: 229o

Observed Melting Point: 230o

Infrared Spectral Assignment: The FTIR analysis of the sample was carried out for

qualitative compound identification. The pellet of approximately 01 mm diameter of

the prochlorperazine maleate was prepared grinding 3-5 mg of sample with 100-150

mg of potassium bromide in pressure compression machine. The sample pellet was

mounted in FTIR (8400S, Shimadzu) compartment and scanned at wavelength 4000 –

500 cm-1

. On analysis of the FTIR spectra of the reference spectra (Fig. 3.1) given in

Clarke Analysis and pure prochlorperazine maleate (Fig. 3.2), no major differences

were observed in the characteristic absorption peak (751, 1220, 1280, 1569 cm−1

)

pattern.

Fig. 3.1: FTIR Spectra of prochlorperazine maleate given in Clarke’s Analysis

Fig. 3.2: FTIR Spectra of prochlorperazine maleate determined experimentally


48

Solubility: The solubility of prochlorperazine maleate was determined in different

solvent systems. Small amounts of the prochlorperazine maleate was added to 5 ml of

each solvent in screw capped glass tubes and shaken. The solutions were examined

physically for the absence or presence of prochlorperazine maleate particles.

Qualitative solubility determined by UV- Spectrophotometer at 254 nm.

Table 3.1: Solubility profile of prochlorperazine maleate

Solvent Solubility Solubility (gm/ml)

Distill Water + 0.002±0.01

0.1N Hydro Chloride ++ 0.041±0.016

0.1N Sodium Hydroxide ++ 0.057±0.029

Ethanol +++ 0.231±0.028

Ether ++ 0.049±0.031

Chloroform ++ 0.062±0.023

Buffer (pH 6.8) ++ 0.055±0.011

Acetone - -

Freely soluble +++ Soluble ++ Slightly soluble + Practically insoluble -

Ultraviolet Absorption Maxima:

Preparation of Sorenson’s Buffer (pH 6.8)

24.5 ml of 0.2 M dibasic sodium phosphate and 0.2 M 25.5 ml of monobasic sodium

phosphate was placed in 100 ml volumetric flask, and make up the volume 100 ml by

water. UV spectra absorption in the rage 200 to 400 nm of a 50 g/ml solution in

Sorenson’s buffer (pH 6.8) was measured. The absorption maxima (λmax) of

prochlorperazine maleate (50 µg/ml) in the solution was found to be 254 nm and 305

nm which was concordant with the Clarke Analysis shown in Table 3.2 and Fig. 3.3.

Table 3.2: Determination of wavelength maxima (λmax)

Wavelength (nm) Absorbance

200 0.612

224 0.337

254 0.682

274 0.035

305 0.084

361 0.003

Fig. 3.3: Determination of wavelength maxima (λmax)

0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9

1

200 225 250 275 300 325 350 375 400

Ab

sorb

ance

Wavelength (nm)


49

Preparation of Calibration Curve:

Prochlorperazine maleate (100 mg) was dissolved in small amount of Sorenson’s

buffer (pH 6.8) in a 100 ml of volumetric flask and final volume was made with the

Sorenson’s buffer. 10 ml of this solution was diluted to 100 ml with Sorenson’s buffer

(pH 6.8) in a 100 ml volumetric flask to obtain a stock solution of 100 µg/ml.

Aliquots of 1, 2, 3, 4, 5, 6 and 7 ml were taken from stock solution in 10 ml

volumetric flasks and volume was made up to 10 ml with buffer (pH 6.8). The

absorbance of these solutions was measured at 254 nm. The calibration curve was

plotted between concentration and absorbance.

Table 3.3: Calibration curve of prochlorperazine maleate

Concentration (µg/ml) Absorbance (254 nm)

0 0

10 0.155

20 0.294

30 0.423

40 0.551

50 0.674

60 0.815

70 0.941

Fig. 3.4: Calibration curve of prochlorperazine maleate

3.1.3 Drug-Polymer Interaction Studies

While designing fast dissolving tablets, it was imperative to give consideration to the

compatibility of prochlorperazine maleate and polymer used within the systems. It

was therefore necessary to confirm the interaction between polymer and drug under

y = 0.0133x + 0.0168

R² = 0.9992

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

0 10 20 30 40 50 60 70

Ab

sorb

ance

(2

54

nm

)

Concentration (µg/ml)


50

experimental conditions (40±50 and 75±5% RH) for 4 weeks. The physical changes

like discoloration, liquefaction and clumping of material were observed after regular

interval of a week. The infrared absorption spectra of 4 week aged physical mixture

of polymer and prochlorperazine maleate are run between 4000 - 500 cm-1

. The FTIR

spectra of physical mixture of polymers and prochlorperazine maleate are shown in

Fig. 3.5-3.11. The absorption maxima of the prochlorperazine maleate polymer

mixture were determined to know the any effect on the analysis of formulation

sample. No interaction was seen between prochlorperazine maleate and polymer. The

results are shown in Table 3.4.

Table 3.4: Prochlorperazine maleate polymer(s) interaction studies

Mixture

Week 1

Physical

Changes

Week 2

Physical

Changes

Week 3

Physical

Changes

Week 4

Physical

Changes

FTIR

peaks

(cm−1

)

max

(nm)

PCP - - - - 752, 1281,

1566, 1221 254, 305

PCP+Ac-di-sol - - - - 754, 1281,

1571, 1219 254, 305

PCP+SSG - - - - 750, 1282,

1566, 1220 254

PCP+Crospovidone - - - - 751, 1283,

1569, 1222 254, 305

PCP +Menthol - - - - 746, 1279,

1566, 1220 254

PCP +Camphor - - - - 751, 1278,

1566, 1220 254

PCP +Thymol - - - - 751, 1279,

1566, 1220 254, 304

PCP +NaHCo3

+Citric Acid - - - - 1580, 1220 253

Physical changes: (-) Sign implies – No change

Fig. 3.5: FTIR Spectra of prochlorperazine maleate and Ac-di-sol


51

Fig. 3.6: FTIR Spectra of prochlorperazine maleate and SSG

Fig. 3.7: FTIR Spectra of prochlorperazine maleate and crospovidone

Fig. 3.8: FTIR Spectra of prochlorperazine maleate and camphor


52

Fig. 3.9: FTIR Spectra of prochlorperazine maleate and menthol

Fig. 3.10: FTIR Spectra of prochlorperazine maleate and thymol

Fig. 3.11: FTIR Spectra of prochlorperazine maleate, SBC and CA


53

3.1.4 Results and Discussions

The sample of prochlorperazine maleate was analyzed by various organoleptic,

physicochemical and spectrophotometric methods. The sample of prochlorperazine

maleate possessed similar color, odor, taste and texture as given in officials. The

melting point of the sample was found to be 2290. The FTIR spectra of reference and

sample are shown in Fig. 3.1 and 3.2 respectively. The FTIR spectra verified the

authenticity of the procured sample as the characteristic peaks of the prochlorperazine

maleate was found at 751, 1220, 1280, 1569 cm−1

in concordance to the reference

spectra. The qualitative solubility of prochlorperazine maleate was determined in

various solvents. The maximum solubility was found in ethanol and least in acetone.

The result for solubility of prochlorperazine maleate in various solvents was shown in

Table 3.1. The absorption maxima of prochlorperazine maleate was observed at 254

nm in Sorenson’s buffer (pH 6.8), which was concordant with the value given in

Clarke Analysis of Drug. The UV spectrum of prochlorperazine maleate is shown in

Table 3.2 and Fig. 3.3. The calibration curve of prochlorperazine maleate was

prepared in Sorenson’s buffer (pH 6.8). The plot of different concentrations of

prochlorperazine maleate versus absorbance was found linear in the concentration

range of 0-70 g/ml at 254 nm. The absorbance’s obtained at different concentrations

are shown in Table 3.3. The data of standard curve was linearly regressed. The values

of slope and correlation coefficient were found to be 0.013 and 0.999 respectively.

The intercept on Y-axis was found to be 0.016. The calibration curve is shown in Fig.

3.4.

Prochlorperazine maleate polymer interaction study was carried out for 4 weeks and

samples were evaluated after every week for physical changes, change in absorption

maxima and FTIR studies. Results are shown in Table 3.4 and Fig. 3.5-3.11. There

was not any sign of physical change at the end of study. The FTIR spectra of the

various physical mixtures retained all the peaks of the pure prochlorperazine maleate

and there was no significant shift in the peaks corresponding to the prochlorperazine

maleate was observed on storage. Both the prochlorperazine maleate and polymer(s)

were found to be compatible with each other. Hence, the selected prochlorperazine

maleate and polymer(s) were successfully incorporated in the design fast dissolving

tablets.


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3.2 PROMETHAZINE THEOCLATE

3.2.1 Analytical Methods Established

Gas Chromatography Mass Spectroscopy:

Hasegawa C et al., (2006), extracted and analyzed ten antihistamine drugs, from

human plasma samples using MonoTip C18 tips, inside which C18 bonded monolithic

silica gel was fixed. Human plasma (0.1 ml) containing the ten antihistamines was

mixed with 0.4 ml of distilled water and 25 µl of a 1 M potassium phosphate buffer

(pH 8.0). After centrifugation of the mixture, the supernatant fraction was extracted to

the C18 phase of the tip by 25 repeated aspirating/dispensing cycles using a manual

micropipettor. The analytes retained on the C18 phase were then eluted with methanol

by five repeated aspirating/dispensing cycles. The eluate was injected into a gas

chromatography (GC) injector without evaporation and reconstitution steps and was

detected by a mass spectrometer with selected ion monitoring in the positive-ion

electron impact mode.10

Leelavathi DE et al., (1985), developed highly specific and sensitive method using

automated high performance liquid chromatography with electrochemical detection

(HPLC-ED) and a method using gas chromatograph mass spectrometry (GCMS) for

the quantitative determination of promethazine in plasma. The lowest detectable

concentration by HPLC-ED was 0.1 ng/ml of plasma and by GC-MS 0.5 ng/ml of

plasma. The HPLC-ED method incorporates a valve switching unit to prevent all of

the electroactive impurities from entering the electrode compartment, thus

maintaining the sensitivity of the detector for the analyses of large numbers of

samples. The GC-MS method incorporates the highly specific selected ion monitoring

technique.11

Reddropa CJ et al., (1980), determined unchanged promethazine in biological

material using gas chromatography mass spectrometry by a nitrogen-selective flame

ionization detector.12

Nuclear Magnetic Resonance:

Lutka A, (2002), studied the effect of beta-CD and its substituted derivatives (DM-

beta-CD and HP-beta-CD) on the solubility and photostability of promethazine (PM)

was investigated in solution and in the solid state. Formation of solid inclusion

complexes of PM with CDs was evaluated using FTIR, 13C NMR and DSC methods.

The results obtained indicate that independently of the complexation method in the


55

solid state (kneading or heating), the presence of CD decreases the solubility of PM;

the reason may be that the phenothiazine ring of PM did not enter into the cavity of

beta-CD and its derivatives.13

DeMol NJ and Koenen J, (1985), investigated degradation products of the

promethazine radical cation, generated from promethazine with horseradish

peroxidase/H2O2. Several products were identified which resulted from fission of the

bond between the two ethanamine carbon atoms of the N10 side chain. The main

product was identified as 10-formyl-5-oxophenothiazine. The likely structure of three

minor products was also elucidated. The degradation of the promethazine radical

cation was different from that of radical cations derived from the propanamine side

chain containing phenothiazine drugs.14

High Performance Liquid Chromatography:

Saleha OA et al., (2009), developed simple, rapid and validated method for

separation and determination of promethazine enantiomers. Promethazine was

separated and quantitated on a Vancomycin Chirobiotic V column (250 x 4.6 mm),

using a mixture of methanol, acetic acid and triethylamine (100:0.1:0.1) as a mobile

phase at 20° and at a flow rate of 1 ml/min. The UV detector was set to 254 nm.

Acetyl salicylic acid (aspirin) was used as an internal standard. The applied HPLC

method allowed separation and quantification of promethazine enantiomers with good

linearity (r > 0.999) in the studied range. The relative standard deviations were 0.29

and 0.36 for the promethazine enantiomers with accuracy of 100.06 and 100.08. The

limit of detection and limit of quantification of promethazine enantiomers were found

to be 0.04 and 0.07 μg/ml, respectively. The method was validated through the

parameters of linearity, accuracy, precision, and robustness. The HPLC method was

applied for the quantitative determination of promethazine in pharmaceutical

formulations.15

Thumma S, et al., (2008), developed simple and rapid stability indicating HPLC

method for determination of promethazine in hot-melt extruded films and sustained

release tablets. Chromatographic separation was achieved on a 150 mm x 4.6 mm i.d.,

3 µm2 particle size, C8 column with acetonitrile 25 mM phosphate buffer (pH 7.0),

50:50 (v/v) as mobile phase at a flow rate of 1 ml/min. Quantization was achieved

with UV detection at 249 nm based on peak area. The method was validated in terms

of linearity, precision, accuracy, robustness specificity, limits of detection and

quantitation according to ICH guidelines.16


56

Tanaka E et al., (2007), developed high performance liquid chromatographic UV

method for the analysis of the promethazine in human serum. The separation was

achieved using a C18 reversed phase column (250 mm x 4.6 mm i.d., particle size 5

µm, Inersil ODS-SP). The mobile phase, consisting of acetonitrile-methanol-30 mM

NaH2PO4 (pH 5.6) (300:200:500), was delivered at a flow rate of 0.9 ml/min and UV

detection was carried out at 250 nm. The recoveries of the promethazine spiked into

serum samples were 87.6 - 99.8%. Regression equations for the promethazine show

excellent linearity, with detection limits of 3.2 - 5.5 ng/ml for serum.6

Patel RB and Welling PG, (2006), determined plasma levels of promethazine using a

high pressure liquid chromatographic procedure incorporating a fixed wavelength

(254 nm) UV detector, following single 50 mg intravenous, intramuscular and oral

doses to two male dogs. Initial plasma promethazine concentrations following

intravenous doses were 556 and 535 ng/ml in the two dogs. The subsequent decline in

drug levels was satisfactorily described by a triexponential function. Peak

promethazine levels of 76 and 64 ng/ml were obtained at 0.5 h following

intramuscular doses. Peak levels for the oral doses were 10.6 and 11.0 ng/ml

occurring 2 h after dosing. The apparent biological half-life of promethazine, obtained

from only 2-3 data points, varied from 8.5 to 27.7 h. Areas under the promethazine

plasma curves, compared to values obtained from intravenous doses between 0 and 24

h, indicated that systemic availability of intact drug was 55 - 73% following

intramuscular injection and 8.3 - 9.5% following oral administration.17

Liquid Chromatography:

Cruz VM et al., (2009), developed a simple and rapid method for the determination

of promethazine derivatives in human urine samples. The analytes were extracted

from the sample in 50 µl of the ionic liquid 1-butyl-3-methyl-imidazolium

hexafluorophosphate working in an automatic flow system under dynamic conditions.

The chemical affinity between the extractant and the analytes allows a good isolation

of the drugs from the sample matrix achieving at the same time their pre

concentration. The separation and detection of the extracted compounds was

accomplished by liquid chromatography and UV detection. The proposed method was

a valuable alternative for the analysis of these drugs in urine within the concentration

range 0.07-10 µg/ml. Limits of detection were in the range from 21 to 60 ng/ml. The

repeatability of the proposed method expressed as RSD (n=5) varied between 2.2%

and 3.9%.18


57

Ponder GW and Stewart JT, (1995), developed LC method for the concurrent assay

of R(+) and S(-) promethazine from human urine and serum. The method involves the

use of solid-phase extraction for sample clean-up. Chromatographic resolution of the

enantiomers was performed under isoeratic conditions using a mobile phase of hexane

1,2-dichlorethane-absolute ethanol-trifluoroacetic acid (400:150:100:1) at a flow rate

of 1 ml/min on a brush-type column KK-CARNU. The enantiomers were detected by

fluorescence using an excitation wavelength of 250 nm and a 280 nm emission cutoff

filter. Chlorpromazine was used as the internal standard for urine analysis. Standard

addition was used for promethazine analysis from serum. Drug to internal standard

ratios were linear from 0.25 to 10 μg/ml in urine. Serum levels were linear from 2 to

10 ng/ml.19

High Performance Thin Layer Chromatography:

Magdalena WK et al., (2006), described high performance thin-layer

chromatography (HPTLC) method combined with densitometry for determination of

phenothiazine derivatives. Quantitation was performed in reflectance mode by using a

computer controlled densitometer. Established calibration curve (r > 0.999), precision

(RDS values: 0.95 – 2.53%), detection limits as well as recovery values (101.1 –

102.8%) were found to be satisfactory. The presented method was rapid, precise and

sensitive and may be alternative to traditionally used HPLC. The method has

successfully applied in the analysis of pharmaceutical formulations.20

UV-Visible Spectroscopy:

Abdol Am and Khoi E, (2006), developed spectrophotometric method for

determining promethazine complexed with bromcresol green and then extracted with

chloroform. The complex in chloroform showed maximum absorption at 415 nm and

obeyed Beer's law over 1.2-8.5 g/ml. The complex molar absorptivity was 1.93 M.

Complex formation and extraction were complete and quantitative over pH 2.7 - 2.8.

The promethazine -bromcresol green molar ratio was 1:1. Excipients, coloring matter,

flavoring agents and other substances likely to be present in promethazine

preparations did not interfere in the determination. Direct determination in tablet,

syrup, and injection preparations were carried out satisfactorily.21

Liu YM and Yu RQ, (1987), determined promethazine in pure form and in a number

of pharmaceutical in 0.5N sulphuric acid by employing first-derivative at 265 nm and

zero-order at 250 nm spectrophotometric modes. The results obtained by utilizing the

first derivative procedure were 99.98, 101.70, 101.70 and 101.15 for the tablets, oral


58

suspension, drops and suppositories respectively. In a similar way the results obtained

for the zero order technique were 105.38, 101.70, 108.56 and 102.23 in the order.22

3.2.2 Drug Analysis

Melting Point: Melting point of the promethazine theoclate was determined by

capillary fusion method; one sided closed capillary filled with promethazine theoclate

was put into the melting point apparatus. Temperature was noted at which solid

promethazine theoclate converts into liquid.

Reported Melting Point: 232o

Observed Melting Point: 231o

Infrared Spectral Assignment: The FTIR analysis of the sample was carried out for

qualitative compound identification. The pellet of approximately 01 mm diameter of

the promethazine theoclate was prepared grinding 3-5 mg of sample with 100-150 mg

of potassium bromide in pressure compression machine. The sample pellet was

mounted in FTIR compartment and scanned at wavelength 4000 – 500 cm

-1. On

analysis of the FTIR spectra of the reference spectra (Fig. 3.12) given in Clarke

Analysis and pure promethazine theoclate (Fig. 3.13), no major differences were

observed in the characteristic absorption peak (749, 1160, 1220, 1259 cm−1) pattern.

Fig. 3.12: FTIR Spectra of promethazine theoclate given in Clarke’s Analysis


59

Fig. 3.13: FTIR Spectra of promethazine theoclate determined experimentally

Solubility: The solubility of promethazine theoclate was determined in different

solvent systems. Small amount of the promethazine theoclate was added to 5 ml of

each solvent in screw capped glass tubes and shaken. The solutions were examined

physically for the absence or presence of promethazine theoclate particles. Qualitative

solubility determined by UV- Spectrophotometer at 250 nm.

Table 3.5: Solubility profile of promethazine theoclate

Solvent Solubility Solubility (gm/ml)

Distilled Water - -

0.1N Hydro Chloric acid + -

0.1N Sodium hydroxide + -

Ethanol ++ 0.045 ± 0.025

Ether - -

Chloroform +++ 0.112 ± 0.012

Buffer (pH 6.8) + -

Acetone - -

Freely soluble +++ Soluble ++ Slightly soluble + Practically insoluble –

Ultraviolet Absorption Maxima: Ultraviolet absorption in the rage 200 to 400 nm of a

5 g/ml solution in Sorenson’s buffer (pH 6.8) was measured. The absorption

maximum (λmax) of promethazine theoclate (5 µg/ml) in this solution was found to be

250 nm is shown in Table 3.6 and Fig. 3.14.

Table 3.6: Determination of wavelength maxima (λmax)

Wavelength (nm) Absorbance

200 0.472

224 0.302

250 0.557

266 0.198

280 0.264

200 0.472


60

Fig. 3.14: Determination of wavelength maxima (λmax)

Preparation of Calibration Curve: Promethazine theoclate (100 mg) was dissolved in

little amount of Sorenson’s buffer (pH 6.8) in a 100 ml of volumetric flask and final

volume was made with the Sorenson’s buffer. 1.0 ml of this solution was diluted to

100 ml with Sorenson’s buffer (pH 6.8) in a 100 ml volumetric flask to obtain a stock

solution of 10 µg/ml. From this stock solution, aliquots of 1, 2, 3, 4, 5, 6, 7, 8 and 9

ml were taken, transferred to 10 ml volumetric flasks and volume was made upto 10

ml using Sorenson’s buffer (pH 6.8). The absorbance of these solutions was measured

at 250 nm. The calibration curve was plotted between concentration and absorbance.

Table 3.7: Calibration curve of promethazine theoclate

Concentration

(µg/ml)

Absorbance

(250 nm)

0 0.000

1 0.181

2 0.257

3 0.352

4 0.458

5 0.557

6 0.665

7 0.782

8 0.865

9 0.942

0

0.1

0.2

0.3

0.4

0.5

0.6

200 225 250 275 300

Ab

sorb

ance

Wavelength (nm)


61

Fig. 3.15: Calibration curve of promethazine theoclate

3.2.3 Drug Polymer Interaction Studies

To confirm that promethazine theoclate was not interacting with polymer under

experimental conditions (40±50 and 75±5% RH) for 4 weeks. The physical changes

like discoloration, liquefaction and clumping of material were observed after regular

interval of a week. The infrared absorption spectra of 4 week aged physical mixture

of polymer and promethazine theoclate were run between 4000 - 500 cm-1

. The FTIR

spectra of physical mixture of polymers and promethazine theoclate are shown in Fig.

3.16-3.23. The absorption maxima of the promethazine theoclate polymer mixture

were determined to know the any effect on the analysis of formulation sample.

Table 3.8: Promethazine theoclate polymer(s) interaction studies

Mixture

Week 1

Physical

Changes

Week 2

Physical

Changes

Week 3

Physical

Changes

Week 4

Physical

Changes

FTIR peaks

(cm−1

) max

(nm)

PMT - - - - 749, 1160,

1220, 1259 250

PMT+Ac-di-sol - - - - 751, 1161,

1220, 1259 250

PMT+SSG - - - - 752, 1160,

1218, 1260 250

PMT+Crospovidone - - - - 750, 1160,

1220, 1259 250

PMT +Menthol - - - - 751, 1158,

1219, 1263 250

PMT +Camphor - - - - 752, 1161,

1220 250

PMT +Thymol - - - - 752, 1160,

1270 251

PMT +NaHCo3

+Citric Acid - - - -

750, 1162,

1222, 1260 249

PMT +β-CD - - - - 757, 1158,

1220 250

Physical changes: (-) Sign implies – No change

y = 0.1008x + 0.0495

R² = 0.9953

0

0.2

0.4

0.6

0.8

1

0 1 2 3 4 5 6 7 8 9

Ab

sorb

ance

(2

50

nm

)

Concentration (µg/ml)


62

Fig. 3.16: FTIR Spectra of promethazine theoclate and Ac-di-sol

Fig. 3.17: FTIR Spectra of promethazine theoclate and SSG

Fig. 3.18: FTIR Spectra of promethazine theoclate and crospovidone


63

Fig. 3.19: FTIR Spectra of promethazine theoclate and camphor

Fig. 3.20: FTIR Spectra of promethazine theoclate and menthol

Fig. 3.21: FTIR Spectra of promethazine theoclate and thymol


64

Fig. 3.22: FTIR Spectra of promethazine theoclate, SBC and CA

Fig. 3.23: FTIR Spectra of promethazine theoclate and β-cyclodextrin

3.2.4 Results and Discussions

The sample of promethazine theoclate was analyzed by various organoleptic,

physicochemical and spectrophotometric methods. The sample of promethazine

theoclate possessed similar color, odor, taste and texture as given in officials. The

melting point of the sample was analyzed by capillary fusion method and found to be

2310. The FTIR spectrum of promethazine theoclate sample was concordant with

reference spectra as given in Clarke Analysis of Drug. The FTIR spectra of reference

and sample are shown in Fig. 3.12 and 3.13 respectively. The FTIR spectra verified

the authenticity of the procured sample as the characteristic peaks of the promethazine

theoclate was found at 749, 1160, 1220, 1259 cm-1

in concordance to the reference

spectra. The qualitative solubility of promethazine theoclate was determined in


65

various solvents. The maximum solubility was found in chloroform and least in ether

and acetone. The result for solubility of promethazine theoclate in various solvents is

shown in Table 3.5. The absorption maxima of promethazine theoclate was observed

at 250 nm in Sorenson’s buffer (pH 6.8), which was concordant with the value given

in Clarke Analysis of Drug. The UV spectrum of promethazine theoclate is shown in

Table 3.6 and Fig. 3.14. The calibration curve of promethazine theoclate was prepared

in Sorenson’s buffer (pH 6.8). The plot of different concentrations of promethazine

theoclate versus absorbance was found linear in the concentration range of 0 - 9 g/ml

at 250 nm. The absorbances obtained at different concentrations are shown in Table

3.7. The data of standard curve was linearly regressed. The values of slope and

correlation coefficient were found to be 0.100 and 0.995 respectively. The intercept

on Y-axis was found to be 0.049. The calibration curve is shown in Fig. 3.15.

Promethazine theoclate polymer interaction study was carried out for 4 weeks and

samples were evaluated after every week for physical changes, change in absorption

maxima and by FTIR studies. Results are shown in Table 3.8 and Fig. 3.16-3.23.

There was not any sign of physical change at the end of study. The FTIR spectra of

the various physical mixtures retained all the peaks of the pure promethazine theoclate

and there was no significant shift in the peaks corresponding to the promethazine

theoclate was observed on storage. Both the promethazine theoclate and polymers

were found to be compatible with each other. As the promethazine theoclate and

polymer(s) were compatible and thus were found to be suitable for dosage form

design.

3.3 PREPARATION OF DRUG FREE TABLETS

Drug free fast dissolving tablets were prepared by direct compression method because

of their several advantages.23-25

• Easiest way to manufacture tablets.

• High doses can be accommodated.

• Use of conventional equipment.

• Use of commonly available excipients.

• Limited number of processing steps.

The tablets were prepared by using single punch tablet machine (Cadmach,

Ahemdabad) to produce flat faced tablets weighing 100 mg each with a diameter of 5


66

mm. A minimum of 50 tablets were prepared for each batch. Before compression

tablet blends were evaluated for mass-volume relationship (bulk density, tapped

density, Hausner’s ratio, compressibility index) and flow properties (angle of repose).

The formulations were developed by using different techniques.

3.3.1 Technology Followed – Superdisintegrant Addition

The superdisintegrants (Ac-di-sol, sodium starch glycolate and crospovidone) in

varying concentration (1-5% w/w) are used to develop the tablets. All the ingredients

are shown in Table 3.9 were passed through sieve no. 60 and were co-grounded in a

glass pestle motor.25-27

Table 3.9: Formulation of drug free tablets with superdisintegrants

Ingredients

(mg) F1 F2 F3 F4 F5 F6 F7 F8 F9 F10 F11 F12 F13 F14 F15

Ac-di-sol 1 2 3 4 5 - - - - - - - - - -

Sodium starch

glycollate - - - - - 1 2 3 4 5 - - - - -

Crospovidone - - - - - - - - - - 1 2 3 4 5

Avicel PH102 55 54 53 52 51 55 54 53 52 51 55 54 53 52 51

Lactopress 25 25 25 25 25 25 25 25 25 25 25 25 25 25 25

Mannitol 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15

Talc 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2

Magnesium

stearate 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2

3.3.2 Technology Followed - Sublimation

Another technology employed for developing fast dissolving tablets were

incorporating subliming agents (camphor, thymol and menthol) in varying

concentration (5-20% w/w). Ingredients shown in Table 3.10 were co-grounded in

glass pestle glass mortar. The mixed blends of excipients were compressed using a

single punch machine to produce flat faced tablets weighing 100 mg. Tablets were

subjected for drying for 6 h under vacuum (30 kpa) at 50o for sublimation to make

tablets porous.28-30


67

Table 3.10: Formulation of drug free tablets with sublimating agents

Ingredients

(mg) F16 F17 F18 F19 F20 F21 F22 F23 F24 F25 F26 F27

Camphor 5 10 15 20 - - - - - - - -

Thymol - - - - 5 10 15 20 - - - -

Menthol - - - - - - - - 5 10 15 20

Avicel PH102 51 46 41 36 51 46 41 36 51 46 41 36

Lactopress 25 25 25 25 25 25 25 25 25 25 25 25

Mannitol 15 15 15 15 15 15 15 15 15 15 15 15

Talc 2 2 2 2 2 2 2 2 2 2 2 2

Magnesium

stearate 2 2 2 2 2 2 2 2 2 2 2 2

3.3.3 Technology Followed - Effervescent

Fast dissolving tablets were prepared by using citric acid and sodium-bi-carbonate in

combination in (1:2 ratio) with other excipients shown in Table 3.11 was co-grounded

in glass pestle glass mortar. These tablets contain (1-5% w/w) effervescent agent.31-33

Table 3.11: Formulation of drug free tablet with effervescent technology

Ingredient

(mg) F28 F29 F30 F31 F32

Citric Acid 0.33 0.66 1.00 1.32 1.65

NaHCO3 0.67 1.34 2.00 2.68 3.35

Avicel PH 102 55 54 53 52 51

Lactopress 25 25 25 25 25

Mannitol 15 15 15 15 15

Talc 2 2 2 2 2

Magnesium stearate 2 2 2 2 2

3.4 PRE-COMPRESSION CHARACTERIZATION

The quality of tablet, once formulated by rule, was generally dictated by the quality of

physicochemical properties of blends. There were many formulations and process

variables involved in mixing steps and all these can affect the characteristics of blend

produced. The characterization parameters for evaluating the flow property of mixed


68

blends includes bulk density, tapped density, Hausner’s ratio, compressibility index

and angle of repose.

3.3.1 Bulk Density

Apparent bulk density (ρb) was determined by pouring the blend into a graduated

cylinder. The bulk volume (Vb) and weight of powder (M) was determined.

34-37 The

bulk density was calculated using the formula

b

bV

M

3.3.2 Tapped Density

The measuring cylinder containing a known mass of blend was tapped 100 times

using density apparatus. The constant minimum volume (Vt) occupied in the cylinder

after tappings and the weight (M) of the blend was measured.34-37

The tapped density

(ρt) was calculated using the formula

t

tV

M

3.3.3 Compressibility Index

The simplest way for measurement of flow of the powder was its compressibility, an

indication of the ease with which a material can be induced to flow. 34-37

It is

expressed as compressibility index (I) which can be calculated as follows

100

t

btI

where, ρt = Tapped density; ρb = Bulk density

Table 3.12: Compressibility index as an indication of powder flow properties

Compressibility Index (%) Type of flow

>12 Excellent

12-16 Good

18-21 Fair to passable

23-35 Poor

33-38 Very poor

>40 Extremely poor


69

3.3.4 Hausner’s Ratio

Hausner’s ratio (HR) is an indirect index of ease of powder flow. It was calculated by

the following formula

b

tHR

where, ρt is tapped density and ρb is bulk density.

Lower Hausner’s ratio (<1.25) indicates better flow properties than higher ones.34

3.3.5 Angle of Repose

Angle of Repose was determined using funnel method. The blend was poured through

a funnel that can be raised vertically until a specified cone height (h) was obtained.

Radius of the heap (r) was measured and angle of repose (θ) was calculated using the

formula38-40

r

htan ; Therefore;

r

h1tan

where, θ is angle of repose; h is height of cone; r is radius of cone.

Table 3.13: Angle of repose as an indication of powder flow properties

Angle of repose(o) Type of flow

<25 Excellent

25-30 Good

30-40 Passable

>40 Very poor


70

Table 3.14: Characterization of drug free tablets blend

±SD, n=6.

Formulation

Codes

Parameters

Bulk

Density

(gm/cc)

Tapped Density

(gm/cc)

Hausner’s

Ratio

Compressibility

Index (%)

Angle of

Repose (o)

F1 0.396±0.012 0.424±0.013 1.071±0.012 6.604±1.330 23.34±1.363

F2 0.403±0.015 0.429±0.012 1.065±0.024 5.621±1.233 25.19±1.221

F3 0.398±0.023 0.417±0.021 1.048±0.013 4.556±1.422 27.35±1.007

F4 0.386±0.004 0.409±0.002 1.059±0.015 5.623±1.221 24.44±1.126

F5 0.398±0.013 0.427±0.005 1.073±0.010 6.792±1.012 25.99±1.096

F6 0.371±0.025 0.395±0.006 1.065±0.003 6.076±1.231 23.56±1.132

F7 0.408±0.034 0.436±0.014 1.069±0.006 6.422±1.086 26.59±1.165

F8 0.383±0.013 0.405±0.017 1.057±0.016 5.432±1.097 26.32±1.136

F9 0.389±0.017 0.421±0.023 1.082±0.027 7.601±1.242 25.22±1.432

F10 0.396±0.006 0.434±0.023 1.095±0.010 8.756±1.134 23.59±1.243

F11 0.405±0.023 0.429±0.012 1.059±0.015 5.594±1.123 25.62±0.968

F12 0.402±0.005 0.429±0.007 1.067±0.023 6.294±1.324 23.54±0.847

F13 0.381±0.013 0.401±0.016 1.052±0.004 4.987±1.354 24.65±1.166

F14 0.378±0.008 0.396±0.004 1.047±0.007 4.545±1.087 22.67±1.124

F15 0.408±0.021 0.436±0.012 1.068±0.016 6.422±1.035 25.22±1.068

F16 0.418±0.013 0.449±0.008 1.074±0.006 6.904±1.046 26.62±1.035

F17 0.399±0.046 0.438±0.012 1.097±0.034 8.904±1.143 28.61±1.241

F18 0.401±0.035 0.443±0.010 1.105±0.023 9.481±1.135 25.32±1.146

F19 0.395±0.023 0.439±0.022 1.111±0.013 10.022±1.146 27.69±1.253

F20 0.403±0.012 0.432±0.034 1.071±0.017 6.713±1.234 27.54±0.846

F21 0.399±0.031 0.435±0.032 1.090±0.024 8.276±1.124 28.87±0.955

F22 0.407±0.014 0.441±0.023 1.084±0.032 7.709±1.146 29.21±0.866

F23 0.371±0.043 0.415±0.042 1.119±0.043 10.602±1.134 28.34±1.244

F24 0.389±0.023 0.423±0.034 1.087±0.022 8.038±1.152 27.52±1.136

F25 0.391±0.005 0.429±0.013 1.065±0.020 8.858±1.098 26.45±0.957

F26 0.401±0.024 0.439±0.022 1.095±0.019 8.656±1.153 27.61±0.697

F27 0.379±0.021 0.419±0.041 1.106±0.023 10.554±1.136 29.64±0.957

F28 0.584±0.023 0.666±0.039 1.140±0.024 12.281±1.906 23.29±0.897

F29 0.610±0.027 0.695±0.035 1.140±0.015 12.292±1.202 23.86±0.801

F30 0.625±0.030 0.721±0.028 1.153±0.026 13.307±2.018 25.58±0.856

F31 0.658±0.024 0.749±0.031 1.139±0.024 12.203±1.925 27.69±1.041

F32 0.635±0.014 0.742±0.011 1.168±0.010 14.427±0.775 29.62±0.925


71

3.5 POST-COMPRESSION CHARACTERIZATION

After compression of powder blends, the prepared tablets were evaluated for

organoleptic characteristics like color, odor, taste, diameter, thickness and physical

characteristics like hardness, friability, disintegration time, wetting time, dispersion

time. The results are shown in Table 3.16.

3.5.1 General Appearance

The general appearance of a tablet, its visual identification and over all ‘elegance’ is

essential for consumer acceptance. This includes tablet’s size, shape, color, presence

or absence of an odor, taste, surface texture, physical flaws etc.41

3.5.2 Tablet Thickness

Ten tablets were taken and their thickness was recorded using micrometer (Mityato,

Japan).

3.5.3 Weight Variation

The weight variation test would be satisfactory method of determining the drug

content uniformity. As per USP42

, twenty tablets were taken and weighted

individually, calculating the average weight, and comparing the individual tablet

weights to the average. The average weight of one tablet was calculated.

Table 3.15: Weight variation limits for tablets as per USP

Average Weight of Tablets (mg) Maximum % Difference Allowed

130 or less 10

130-324 7.5

More than 324 5

3.5.4 Hardness

Hardness of tablet is defined as the force applied across the diameter of the tablet in

order to break the tablet. The resistance of the tablet to chipping, abrasion or breakage

under condition of storage transformation and handling before usage depends on its

hardness. Hardness of the tablet of each formulation was determined using Pfizer

Hardness Tester.41, 43


72

3.5.5 Friability

Friability of the tablets was determined using Roche friabilator. This device subjects

the tablets to the combined effect of abrasions and shock in a plastic chamber

revolving at 25 rpm and dropping the tablets at a height of 6 inch in each revolution.

Preweighed sample of tablets was placed in the friabilator and were subjected to 100

revolutions. Tablets were dedusted using a soft muslin cloth and reweighed. The

friability (F %) was determined by the formula

100.1%

W

WoF

Where, W0 is initial weight of the tablets before the test and W is the weight of the

tablets after test.41, 44

3.5.6 Wetting Time

Wetting time of the tablets was measured using a piece of tissue paper (12 cm x10.75

cm) folded twice, placed in a small petridish (ID = 6.5 cm) containing 6 ml of

Sorenson’s buffer (pH 6.8). A tablet was put on the paper, and the time for the

complete wetting was measured.35, 45-47

Fig. 3.24: In vitro wetting property

3.5.7 In Vitro Dispersion Time

In vitro dispersion time was measured by dropping a tablet in a glass cylinder

containing 6 ml of Sorenson’s buffer (pH 6.8). Six tablets from each formulation were

randomly selected and in vitro dispersion time was performed.46, 48, 49

Fig. 3.25: In vitro dispersion property


73

3.5.8 Disintegration Test

Disintegration of fast disintegrating tablets is achieved in the mouth owing to the

action of saliva, however amount of saliva in the mouth is limited and no tablet

disintegration test was found in USP and IP to simulate in vivo conditions.50-53

A

modified disintegrating apparatus method was used to determine disintegration time

of the tablets. A cylindrical vessel was used in which 10-mesh screen was placed in

such way that only 2 ml of disintegrating or dissolution medium would be placed

below the sieve (Fig. 3.26). To determine disintegration time, 6 ml of Sorenson’s

buffer (pH 6.8), was placed inside the vessel in such way that 4 ml of the media was

below the sieve and 2 ml above the sieve. Tablet was placed on the sieve and the

whole assembly was then placed on a shaker. The time at which all the particles pass

through the sieve was taken as a disintegration time of the tablet. Six tablets were

chosen randomly from the composite samples and the average value was

determined.43

Fig. 3.26: Modified disintegration test apparatus


74

Table 3.16: Post-compression characterization

Formulation

Codes

Parameters

Thickness

(mm)

Weight

(mg)

Hardness

(kg/cm2)

Friability

(%)

Wetting

Time

(s)

Dispersion

Time

(s)

Disintegration

Time

(s)

F1 2.436±0.012 97.1±3.512 3.2±0.128 0.421±0.069 103±2.25 105±1.07 110±1.69

F2 2.421±0.015 95.4±3.746 3.1±0.133 0.484±0.046 84±2.47 88±3.59 91±1.37

F3 2.414±0.011 98.2±4.341 3.2±0.142 0.644±0.073 61±1.48 66±3.19 72±2.48

F4 2.425±0.011 96.1±3.134 3.2±0.123 0.765±0.063 40±3.43 47±3.58 50±1.63

F5 2.437±0.009 98.6±3.561 3.1±0.134 0.873±0.057 28±2.42 39±2.10 41±3.26

F6 2.412±0.011 97.3±2.891 3.1±0.122 0.412±0.025 112±1.48 125±1.96 128±1.83

F7 2.445±0.008 96.6±3.140 3.1±0.097 0.465±0.023 87±1.69 94±2.59 96±2.41

F8 2.425±0.017 98.1±2.971 3.2±0.124 0.526±0.054 66±2.65 72±2.18 81±2.06

F9 2.431±0.014 102.1±4.128 3.1±0.132 0.766±0.013 45±1.58 51±3.51 66±3.14

F10 2.408±0.012 99.4±3.671 3.0±0.116 0.923±0.025 40±3.58 45±3.72 58±2.95

F11 2.421±0.018 98.1±2.982 3.0±0.134 0.584±0.032 98±1.07 100±2.50 99±1.09

F12 2.396±0.013 97.5±3.656 3.0±0.121 0.509±0.053 82±1.86 86±1.06 84±2.38

F13 2.426±0.014 101.5±4.413 3.1±0.143 0.456±0.014 56±2.60 58±1.18 61±1.48

F14 2.401±0.019 99.4±3.140 3.2±0.068 0.412±0.017 31±2.78 34±2.42 36±3.48

F15 2.417±0.016 101.7±2.414 3.1±0.089 0.404±0.024 22±1.12 25±2.47 27±2.30

F16 2.385±0.014 94.4±0.128 3.0±0.132 0.573±0.032 82±2.59 88±3.17 94±1.69

F17 2.409±0.017 90.1±1.124 3.0±0.141 0.606±0.037 59±1.48 61±2.75 63±2.08

F18 2.414±0.009 86.7±2.317 2.9±0.137 0.984±0.026 34±1.08 36±3.72 42±2.16

F19 2.426±0.017 80.4±3.146 3.0±0.131 1.119±0.021 18±3.44 20±1.49 32±3.27

F20 2.412±0.008 95.7±0.149 3.0±0.213 0.576±0.024 91±2.26 99±2.06 102±1.30

F21 2.396±0.012 92.2±2.426 2.9±0.146 0.613±0.054 64±2.59 68±2.59 75±1.95

F22 2.379±0.015 88.3±0.107 2.9±0.135 0.997±0.042 46±1.92 49±1.07 51±2.16

F23 2.371±0.012 84.8±1.216 2.8±0.145 1.246±0.027 28±2.48 30±1.49 35±2.59

F24 2.424±0.009 95.2±0.141 3.1±0.124 0.668±0.015 69±1.55 73±3.48 85±1.37

F25 2.417±0.016 90.3±0.019 3.0±0.186 0.789±0.019 55±2.70 59±2.38 63±1.19

F26 2.394±0.014 84.4±1.126 3.0±0.136 0.969±0.013 29±3.64 32±1.68 39±3.41

F27 2.375±0.011 80.3±0.219 3.0±0.142 1.396±0.026 15±1.69 18±3.84 30±2.48

F28 2.344±0.034 97.9±1.176 3.1±0.252 0.67±0.143 75±3.51 78±4.50 78±3.05

F29 2.363±0.035 99.6±3.765 3.0±0.276 0.78±0.129 64±2.08 65±3.05 69±3.05

F30 2.343±0.016 98.4±3.551 2.8±0.226 0.96±0.159 38±2.51 41±3.51 43±2.30

F31 2.366±0.041 98.5±3.654 2.8±0.234 1.19±0.134 29±2.08 32±2.08 35±1.52

F32 2.521±0.339 100.4±2.246 2.7±0.257 1.27±0.172 20±1.52 22±2.51 29±1.00

±SD, n=6.


75

3.6 DEVELOPMENT OF COMBINATIONAL DRUG FREE TABLETS

The fast dissolving tablets were prepared by the combination of two disintegrants to

check their influence on the pre and post compression characteristics of the tablets.

These tablets were prepared as methods described earlier. Only the least concentration

of the disintegrants was used in tablets to evaluate their combined effect. The blends

and tablets were characterized as described earlier. The formulation of the tablet is

tabulated in Table 3.17.

Table 3.17: Combined formulation of drug free tablet

Ingredients

(mg) F33 F34 F35 F36 F37 F38

Ac-di-sol 1 - - - - -

SSG - 1 - - - -

Camphor - - 2.5 - - -

Menthol - - - 2.5 - -

Thymol - - - - 5 -

Effervescent - - - - - 1

Crospovidone 1 1 1 1 1 1

Avicel PH102 54 54 52.5 52.5 50 54

Lactopress 25 25 25 25 25 25

Mannitol 15 15 15 15 15 15

Talc 2 2 2 2 2 2

Magnesium

stearate 2 2 2 2 2 2

Table 3.18: Pre-compression tablet characterization

Characterization F33 F34 F35 F36 F37 F38

Bulk Density

(g/cc)

0.587

±0.013

0.599

±0.014

0.429

±0.012

0.360

±0.005

0.426

±0.007

0.629

±0.010

Tapped Density

(g/cc)

0.759

±0.039

0.857

±0.032

0.511

±0.005

0.413

±0.003

0.471

±0.009

0.682

±0.015

Hausner’s

Ratio

1.392

±0.055

1.431

±0.051

1.192

±0.025

1.148

±0.019

1.106

±0.014

1.084

±0.008

Compressibility

Index (%)

22.481

±3.295

30.066

±2.537

16.064

±1.782

12.910

±1.489

16.231

±0.326

7.759

±0.658

Angle of

Repose (o)

36.533

±0.501

38.557

±0.505

25.820

±0.459

25.020

±0.761

25.940

±0.516

24.533

±0.616

±SD, n=6.


76

Table 3.19: Post-compression characterization of drug free tablets

Characterization F33 F34 F35 F36 F37 F38

Weight

(mg)

100.20

±0.966

99.86

±0.266

94.083

±0.878

94.940

±1.195

93.380

±0.960

100.030

±0.121

Hardness

(kg/cm2)

3.0

±0.058

2.9

±0.116

2.9

±0.141

3.0

±0.042

3.0

±0.011

3.1

±0.014

Friability (%) 1.225

±0.059

1.375

±0.029

0.525

±0.032

0.659

±0.095

0.608

±0.032

0.626

±0.041

Disintegration

Time (s)

98

±3.25

92

±2.14

78

±1.19

68

±3.84

81

±2.13

70

±1.58

±SD, n=6.

From this study, it was clears that the combined effect of disintegrants with

crospovidone shows the better results on the properties of the tablets.54, 55

The

friability of the tablets was decreased by the incorporation of the crospovidone. The

disintegration time of the prepared tablets was also decreased by the crospovidone.

In the batches, F33 and F34 fair to passable flow of blends were observed. The

Hausner’s ratio was found greater than 1.25 and compressibility index was found

more than 16%. The poor flow of the blends were also evidenced by the angle of

repose, the values were higher than 30o. Hence it was clears, if a physical mixture of

superdisintegrant was used in high speed tabletting; the problem of segregation of the

disintegrants may be encountered. The attempt was made to overcome these problems

by the coprocessing of superdisintegrants (Ac-di-sol with crospovidone and sodium

starch glycolate with crospovidone).

3.7 DEVELOPMENT OF FDT BY COPROCESSED SUPERDISINTEGRANTS

Coprocessing is defined as combining two or more established excipients by an

appropriate process. Coprocessing of excipients by could lead to formation of

excipients with superior properties compared with simple physical mixture of their

components or with individual components.55

3.7.1 Preparation of Coprocessed Disintegrant Blends

The coprocessed superdisintegrant was prepared as follows. Blends of Ac-di-sol/SSG

and crospovidone in different ratios (1:1, 1:2, 1:3, 2:1, 2:3, 3:1, and 3:2) total weight

of 10 g was added to 50 ml of isopropyl alcohol. The content of beaker was stirred on

a magnetic stirrer at 50 rpm. The temperature was maintained between 65-700 and

stirring was continued till most of isopropyl alcohol evaporated. The wet coherent

mass was sieved through sieve number 100. The wet powder was dried in a tray drier


77

at 600 for 20 min. The dried powder sifted on 120 mesh sieve and stored in airtight

container until further used. For the preliminary study and evaluation only

coprocessed superdisintegrant was prepared in 1:1 ratio. Rest of ratio was prepared

for the factorial design batch/optimization.

3.7.2 Evaluation of Coprocessed Disintegrant Blends

Particle size analysis

The microscopic technique was used to test the particle size distribution of

superdisintegrants and their blends. The particle size of the disintegrants was

evaluating to prepare the slides of powder and observes under the microscope. To test

the swelling of superdisintegrant in water and Sorenson’s buffer (pH 6.8, saliva pH),

disintegrant powder were first dispersed in a small volume of liquid and the

ultrasonicated for 10 min. The suspension transferred with a pipette to a small volume

on the glass slide. The ratio of particle diameter in the dispersing medium to the dry

powders was used as an intrinsic swelling capacity of super disintegrant in the test

medium.

Fig. 3.27: Particle size analysis

0

50

100

150

200

250

Ac-

di-

sol

So

diu

m S

tarc

h G

lyco

late

Cro

spo

vid

on

e

PM

Ac-

di-

sol+

Cro

spo

vid

on

e (1

:1)

PM

SS

G+

Cro

spo

vid

on

e (1

:1)

Co

pro

cess

ed A

c-d

i-

sol+

Cro

spo

vid

on

e(1

:1)

Co

pro

cess

ed S

SG

+C

rosp

ov

ido

ne(

1:1

)

Vo

lum

e m

edia

n d

iam

eter

(μ

m)

Dry powder

Water

pH 6.8


78

Mass- volume relationship and flow properties

For the mass-volume relationship bulk density (ρb), tapped density (ρt), Hausner’s

ratio (RH = ρt / ρb) and compressibility index (Ic =100 (ρt – ρb) / ρb) was determined

with the bulk/tapped densitometer. The angle of repose was determined using funnel

method. The blend was poured through a glass funnel that can be raised vertically

until a specified cone height (h) was obtained. Radius of the conical pile (r) was

measured and angle of repose (θ) was calculated using the formula tan θ = h/r.34-40

The results are shown in Table 3.20.

Table 3.20: Evaluation of superdisintegrant

Batch Ratio

Bulk

Density

(g/cc)

Tapped

Density

(g/cc)

Hausner’s

Ratio

Compressibility

Index

(%)

Angle of

Repose

(0)

Ac-di-sol - 0.742

±0.019

0.911

±0.034

1.235

±0.011

21.059

±0.119

38.18

±0.106

SSG - 0.759

±0.005

0.945

±0.004

1.250

±0.004

20.029

±0.234

36.18

±0.174

Crospovidone - 1.244

±0.020

1.858

±0.015

1.494

±0.034

33.039

±1.519

44.02

±1.010

Physical Mixture

(Ac-di-sol

+Crospovidone)

1:1 0.785

±0.004

1.131

±0.009

1.312

±0.016

25.946

±1.153

39.96

±1.623

Physical Mixture

(SSG+

Crospovidone)

1:1 0.891

±0.008

1.157

±0.040

1.299

±0.039

22.946

±2.268

37.83

±1.714

Coprocessed

(Ac-di-sol

+Crospovidone)

1:1 0.512

±0.080

0.601

±0.017

1.173

±0.023

14.801

±0.218

24.16

±0.529

Coprocessed

(SSG+

Crospovidone)

1:1 0.624

±0.002

0.700

±0.004

1.122

±0.004

10.856

±0.332

22.42

±0.626

±SD, n=6.


79

Scanning electron micrographs

Finally to investigate the morphology of SSG, crospovidone and prepared

coprocessed superdisintegrant, scanning electron micrographs were taken using

(JOEL, JSM-35, CF) scanning electron microscope; where the samples were

previously sputter coated with gold.56

Fig. 3.28: Scanning electron micrographs

A. Crospovidone; B. Ac-di-sol; C. Sodium starch glycolate; D. Coprocessed Ac-di-Sol + Crospovidone;

E. Coprocessed Sodium starch glycolate + Crospovidone

3.7.3 Preparation of FDT with Coprocessed Superdisintegrants

The fast dissolving tablets were prepared with coprocessed superdisintegrants (Ac-di-

sol with crospovidone and sodium starch glycolate with crospovidone) and evaluated

for pre and post-compression properties. The evaluated parameters were compared

with the tablets prepared by physical mixture of superdisintegrants. The formulation

and evaluations are tabulated in Table 3.21.


80

Table 3.21: Development of tablets with coprocessed superdisintegrants

FORMULATION

Ingredients (mg) F39 F40

Ac-di-sol 1 -

Sodium starch glycolate - 1

Crospovidone 1 1

Avicel PH102 54 54

Lactopress 25 25

Mannitol 15 15

Talc 2 2

Magnesium stearate 2 2

PRE-COMPRESSION CHARACTERIZATION

Bulk Density (gm/cc) 0.407

±0.012

0.438

±0.021

Tapped Density (gm/cc) 0.453

±0.011

0.510

±0.009

Hausner’s Ratio 1.112

±0.007

0.587

±0.013

Compressibility Index (%) 10.097

±0.552

1.167

±0.041

Angle of Repose (o) 23.217

±0.901

22.470

±1.265

POST-COMPRESSION CHARACTERIZATION

Weight (mg) 100.01

±0.388

99.76

±0.188

Hardness (kg/cm2) 3.5

±0.100

3.4

±0.091

Friability (%) 0.629

±0.018

0.623

±0.015

Disintegration Time (s) 72

±1.18

76

±1.26

±SD, n=6.

Fig. 3.29: Comparison of tablets prepared by physical mixture and coprocessed

superdisintegrants

0

20

40

60

80

100

120

T33 T39 T35 T40

Dis

inte

grat

ion

tim

e (

s)

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

T33 T39 T35 T40

Fria

bili

ty (

%)


81

3.8 RESULTS AND DISCUSSIONS

During preliminary studies, thirty two blind formulations were prepared by employing

different concentrations of superdisintegrants (1-5% w/w), sublimating agents (5-20%

w/w) and effervescent agents (1-5% w/w). The pre-compression characterizations of

mixed blends were done for determination of mass volume relationship and flow

properties. The evaluated parameters were bulk density, tapped density, Hausner’s

ratio, compressibility index and angle of repose.

For drug free tablets prepared by using various disintegrating agents, the bulk density

of blend varied between 0.371-0.658 gm/cc. The tapped density was found in the

range of 0.395-0.749 gm/cc. The results indicated good packaging capacity of tablets.

By using these two density data, Hausner’s ratio and compressibility index was

calculated. If the bed particle was more compressible then the powder will be less

flowable and vice versa. The value of compressibility index was found between

4.545-14.427%. The powder blends of all formulation had Hausner’s ratio less than

1.25 indicating good flow characteristics.34

The compressibility–flowability

correlation data indicating a good flowability of the powder blend. The flowability of

the powder was also evidenced by the angle of repose. The angle of repose below 30o

range indicates good to excellent flow properties of powder. Lower the friction

occurring within the mass, better the flow rate.39

The angle of repose was found to be

in range 23.29-29.62o. The results showed good flow property of the formulated

mixed blends due to the addition of talc as lubricant and magnesium stearate as

glidant in the 2% w/w and 2% w/w of the tablet weight respectively. The results for

pre-compression characterization of blend are shown in Table 3.14.

After compression of powder blends, the tablets were evaluated for their post-

compression properties like organoleptic, physical and quality control parameters

(diameter, thickness, hardness, friability, disintegration time, wetting time and

dispersion time). All the formulations are white in color, odorless, flat in shape with

smooth surface. The prepared tablets were elegant and lot-to-lot tablet uniformity and

also free from any surface texture problems.

The thickness of the tablets varied between 2.343-2.521 mm. The average weight of

the prepared tablets with superdisintegrants and effervescent agents were found

between 95.4-102.1 mg. The average weight of the tablets prepared by vacuum drying


82

technique was found 80.3-95.7 mg due to the elimination of the sublimating agents

from the tablets. So it was predicted that all the tablets exhibited uniform weight with

low standard deviation values within the acceptable variation as per USP.42

The friability of the formulations was less than 1.0%, showed the durability of the

tablets; resistance to loss of weight indicates the tablet’s ability to withstand abrasion

in handling, packaging and shipment.39

The friability of all the formulations was

found to be less than 1.0 % except those containing higher concentrations of

subliming agents (F19, F21, F27, F31 and F32). It was clear from the study that as the

concentration of sublimating agents was increasing the percent friability was also

increasing. The hardness of the prepared tablet varied from 2.7-3.2 kg/cm2, which has

satisfactory strength to withstand with the applied mechanical shocks.

A disintegrant was found in all the formulations to facilitate a breakup or

disintegration of the tablet when it contacts with water or saliva in mouth. The

disintegration process of the tablet was fully dependable on nature and concentration

of superdisintegrant used. Disintegrants drawing the water into the tablet causes

wicking, swelling and burst apart. The tablets with crospovidone disintegrate faster

than the tablets with the citric acid and sodium-bi-carbonate, sodium starch glycolate

and Ac-di-sol and camphor, menthol and thymol. The tablets prepared with

superdisintegrants disintegrate in 27-128 s. The tablets prepared with effervescent

technology elaborates the carbon-di-oxide gas when the tablet comes in contact with

little amount of saliva or water due to reaction between citric acid and sodium-bi-

carbonate which results in breakup of tablets. The tablets prepared with effervescent

agents disintegrate in 29-78 s. The porous structure of the tablets prepared with

sublimating agents was responsible for the for fast water uptake, which facilitates the

disintegration of tablets. The tablets prepared with sublimating agents disintegrate in

29-102 s.

The in vitro wetting time was also studied to know the time required for complete

wetting of tablets when placed on tong. The in vitro wetting time of all the

formulations were varied between 20-125 s. The swelling properties of the tablets

were depending upon their concentration and type of superdisintegrants. The result

shows that swelling time was reduced with increase in the concentration of the

superdisintegrant. The results are tabulated in Table 3.16.


83

The tablets with crospovidone showed the best results on compare to others and hence

it was selected as one factor of the optimization of the fast dissolving tablets.54

Thus,

it was decided to carryout optimization studies with other disintegrants in

combination with the superdisintegrant crospovidone.

The tablets prepared by combination of two disintegrants gave better results in terms

of disintegration time and friability than using single disintegrant. The tablet may

disintegrate by two properties. In the tablets, F35, F36 and F37 the porous structure

developed by sublimating agents (camphor, menthol and thymol) was responsible for

water uptake, hence it facilitates wicking action of crospovidone in bringing faster

disintegration.54

By the incorporation of crospovidone the friability of the tablets was

also decreased. In the tablets, batch F38 the capillary action of crospovidone raises the

medium, for the completion of reaction between sodium-bi-carbonate and citric acid.

The faster uptake of medium in the tablets faster the tablet disintegrates. In tablet

batch F33 and F34 pre-compression and post-compression evaluations not found in

limits, due to the great difference between shape, size and compressibility properties

of two superdisintegrants (Ac-di-sol/sodium starch glycolate and crospovidone). The

poor flow property and compressibility properties of the physical mixture was

observed (Table 3.18). Due to the less compressibility the formed tablets were friable

in nature. Thus, it was decided to prepare coprocessed disintegrants (Ac-di-sol with

crospovidone and sodium starch glycolate with crospovidone). Coprocessing of

excipients could lead to formation of excipients with superior properties compared

with the simple physical mixture of their components or individual components.55

In preliminary investigation, water, ethyl alcohol, dichloromethane and isopropyl

alcohol were used for coprocessing of the superdisintegrant. Water was ruled out for

further experiment because gel formation occurs due to the presence of starch in Ac-

di-sol and sodium starch glycolate. Dichloromenthane was omitted because of

floating of crospovidone and sedimentation of Ac-di-sol and sodium starch glycolate.

Ac-di-sol and sodium starch glycolate was sparingly soluble in ethyl alcohol.

Isopropyl alcohol was selected considering the absence of gel formation and phase

separation.

The diameters of superdisintegrants in different media was determined are given in

Fig. 3.27. A significant reduction in swelling capacity was also observed in physical


84

mixture as well as coprocessed superdisintegrants in Sorenson’s buffer (pH 6.8). The

sudden decrease in swelling capacity of chemically modified starch may attribute to

the converting of the carboxymethyl sodium moieties to its free acid form in acidic

medium. Since the acid medium form has less hydration capacity than its salt form,

the liquid holding capacity of the disintegrant particle reduces after deionization in the

slightly acidic medium.26

Therefore, the total degree of substitution and the ratio of

basic to acidic substituent’s were potential factors determining the extent of influence

of medium pH on the swelling properties of disintegrants and blends particles. Unlike

the other superdisintegrant, there was no apparent change in the swelling capability of

the nonionic polymer crospovidone in both media. The results illustrated that the

physical mixing and coprocessing give better swelling than used alone.

The bulk density, tapped density, compressibility index, Hausner’s ratio and angle of

repose studied all batches shown in Table 3.20. According to literature the powder

compressibility index between 5 to 16% was suitable for punching tablets and those

Hausner’s ratio below 1.25 exhibited good flowability.39

Only coprocessed

superdisintegrant batches were fallen in the limit/range. On the evaluation of

superdisintegrant angle of repose of the physical mixture and coprocessed

disintegrants (1:1) was found to be 37.83-39.360 and 22.42-24.16

0 respectively.

According to literature, good flow (angle of repose between 200 and 35

0) was shown

by coprocessed superdisintegrants. It was concluded that the particle size distribution

and shape of the excipients would be kept the same to avoid the tabletting problem.

The morphology and surface properties of Ac-di-sol, sodium starch glycolate,

crospovidone and coprocessed superdisintegrant were visualized using scanning

electron microscopy (SEM) shown in Fig. 3.28. All powder batches were presented in

magnificence of X150. From these micrographs we observed clear difference between

the structure and size of superdisintegrants.

The tablets were prepared by the coprocessed superdisintegrants showed better results

than the tablets prepared by using physical mixture or by using individual components

(Table 3.21 and Fig. 3.29). The results clears that the disintegration time and percent

friability have a great difference in physical mixture and coprocessed

superdisintegrants. When, Ac-di-sol and sodium starch glycolate was used, higher

water uptake swelling and deformation of disintegrants take place, which gives

internal pressure to tablet to disintegrate. It was obvious that in the presence of


85

crospovidone, wicking was facilitated.57

The use of a physical mixture

superdisintegrant resulted in increased friability probably due to low compressibility

of excipients. By the coprocessing technology the friability of tablets was also

decreased.

By evaluating these tablets, the levels for the optimization of the independent factors

were to be set. The three levels (-1, low; 0, medium; +1, high) of different

disintegrating agents were selected.

Table 3.22: Selected independent factors with their levels

Category Group Independent Factor Level (mg)

-1 0 1

Coprocessing

Technology

G-1 X1 Ac-di-sol 1 2 3

X2 Crospovidone 1 2 3

G-2 X1 Sodium Starch Glycolte 1 2 3


Pore forming

Technology

G-3 X1 Camphor 2.5 5 7.5


G-4 X1 Menthol 2.5 5 7.5


G-5 X1 Thymol 5 10 15


Effervescent

Technology

G-6 X1 Effervescent Agent 1 2 3



86

REFERENCES

1. McKay G, Hall K, Cooper JK, Hawes EM, Midha KK. Gas chromatographic

mass spectrometric procedure for the quantitation of prochlorperazine in plasma

and its comparison with a new high performance liquid chromatographic assay

with electrochemical detection. J Chromatogr 1982; 232(2): 275-282.

2. Yan M, Zhu YG, Li HD, Chen BM, Ma N, Lei YQ, Liu YP. Quantification of

prochlorperazine maleate in human plasma by liquid chromatography-mass

spectrometry: application to a bioequivalence study. J Chromatogr B Analyt

Technol Biomed Life Sci 2009; 877(27): 3243-3247.

3. Uraisin K, Takayanagi T, Nacapricha D, Motomizu S. Novel oxidation reaction

of prochlorperazine with bromate in the presence of synergistic activators and its

application to trace determination by flow injection/spectrophotometric method.

Anal Chim Acta 2006; 580(1): 68-74.

4. Marumo A, Kumazawa T, Lee XP, Fujimaki K, Kuriki A, Hasegawa C, Sato K,

Seno H, Suzuki O. Analysis of phenothiazines in human body fluids using disk

solid-phase extraction and liquid chromatography. J AOAC Int 2005;

88(6):1655-1660.

5. Honigberg IL, Stewart JT, Smith AP, Plunkett RD, Justice EL. Liquid

chromatography in pharmaceutical analysis IV: determination of antispasmodic

mixtures. J Pharm Sci 1975; 64(8): 1389-1393.

6. Tanaka E, Nakamura T, Terada M, Shinozuka T, Hashimoto C, Kurihara K,

Honda K. Simple and simultaneous determination for 12 phenothiazines in

human serum by reversed phase high performance liquid chromatography. J

Chromatogr B Analyt Technol Biomed Life Sci 2007; 854(1-2): 116-120.

7. Mizuno Y, Sato K, Sano T, Kurihara R, Kojima T, Yamakawa Y, Ishii A,

Katsumata Y. Identification and characterization of 17 phenothiazine

compounds by capillary high-performance liquid chromatography/fast atom

bombardment mass spectrometry. Leg Med 2002; 4(4): 207-216.

8. Mou C, Ganju N, Sridhar KS, Krishan A. Simultaneous quantitation of plasma

doxorubicin and prochlorperazine content by high performance liquid

chromatography. J Chromatogr B Biomed Sci Appl 1997; 703(1-2): 217-224.

9. Kitamura K, Goto T, Kitade T. Second derivative spectrophotometric

determination of partition coefficients of phenothiazine derivatives between


87

human erythrocyte ghost membranes and water. Talanta 1998; 46(6): 1433-

1438.

10. Hasegawa C, Kumazawa T, Lee XP, Fujishiro M, Kuriki A, Marumo A, Seno

H, Sato K. Simultaneous determination of ten antihistamine drugs in human

plasma using pipette tip solid phase extraction and gas chromatography/mass

spectrometry. Rapid Commun Mass Spectrom 2006; 20(4): 537-543.

11. Leelavathi DE, Dressler DE, Soffer EF, Yachetti SD, Knowles JA.

Determination of promethazine in human plasma by automated high

performance liquid chromatography with electrochemical detection and by gas

chromatography mass spectrometry. J Chromatogr 1985; 339(1):105-115.

12. Reddropa CJ, Riessa W, Slatera TF. Determination of unchanged promethazine

by gas chromatography mass spectrometry. J Chromatogar A 1980; 192(2):

375-386.

13. Lutka A. Investigation of interaction of promethazine with cyclodextrins. Acta

Pol Pharm 2002; 59(1): 45-51.

14. DeMol NJ, Koenen J. Degradation products of the promethazine radical cation.

Pharm World Sci 1985; 7(3): 121-124.

15. Saleha OA, El-Azzounya AA, Aboul-Eneina HY, Badawyb AM. A validated

HPLC method for separation and determination of promethazine enantiomers in

pharmaceutical formulations. Drug Dev Ind Pharm 2009; 35(1): 19-25.

16. Thumma S, Zhang SQ, Repka MA. Development and validation of a HPLC

method for the analysis of promethazine hydrochloride in hot-melt extruded

dosage forms. Pharmazie 2008; 63(8): 562-567.

17. Patel RB, Welling PG. High pressure liquid chromatographic determination of

promethazine plasma levels in the dog after oral, intramuscular, and intravenous

dosage. J Pharm Sci 2006; 71(5): 529-532.

18. Cruz VM, Lucena R, Cardenas S, Valcarcel M. Determination of phenothiazine

derivatives in human urine by using ionic liquid based dynamic liquid-phase

microextraction coupled with liquid chromatography. J Chromatogr B Analyt

Technol Biomed Life Sci 2009; 877(1-2): 37-42.

19. Ponder GW, Stewart JT. A liquid chromatographic method for the

determination of promethazine enantiomers in human urine and serum using

solid-phase extraction and fluorescence detection. J Pharm Biomed Ana 1995;

13(9): 1161-1166.


88

20. Magdalena WK, Skalskaa A, Matysikb A. Determination of phenothiazine

derivatives by high performance thin layer chromatography combined with

densitometry. J Pharm Biomed Ana 2006; 41(1): 286-289.

21. Abdol AM, Khoi E. Spectrophotometric promethazine hydrochloride

determination using bromcresol green. J Pharm Sci 2006; 72(6): 704-705.

22. Liu YM, Yu RQ, (1987), UV spectrophotometric simultaneous determination of

promethazine hydrochloride. Yao Xue Xue Bao 1987; 2(12): 913-917.

23. Mishra DN, Vijaya KSG. Rapidly disintegrating oral tablets of meloxicam.

Indian Drugs 2006; 43(2): 117-121.

24. Bi YX, Sunda Y, Yonezawa Y, Danjo K. Evaluation of rapidly disintegrating

tablets prepared by a direct compression method. Drug Dev Ind Pharm 1999; 25

(5): 571-581.

25. Sreenivas SA, Gadad AP, Dandagi PM, Mastiholimath VS, Patil MB.

Formulation and evaluation of ondeanesetron hydrochloride directly compressed

mouth disintegrating tablets. Indian Drugs 2006; 43(1): 35-38.

26. Zhao N, Augsburger LL. Functionally compression of 3 classes of

superdisintegrants in promoting aspirin tablet disintegration and dissolution.

AAPS Pharm Sci Tech 2005; 6(4): 634-640.

27. Kuchekar BS, Mahajan HS, Bandhan AC. Mouth dissolve tablets of salbutamol

sulphate: a novel drug delivery system. Indian Drugs 2004; 41(10): 592-598.

28. Aly AM, Semreen M, Qato MK. Superdisintegrants for solid dispersion to

produce rapidly disintegrating tenoxicam tablets via camphor sublimation.

Pharm Tech 2005; 20: 68-78.

29. Koizumi K, Watanabe Y, Morita K, Utoguchi N, Matsumoto M. New method of

preparing high-porosity rapidly saliva soluble compressed using mannitol with

camphor: a subliming material. Int J Pharm 1997; 152: 127-131.

30. Mane AR, Kusum DV, Asha AN. A novel technique for the preparation of

mouth dissolving tablets of domperidone. Indian Drugs 2003; 40(9): 544-546.

31. Sallam E, Ibrahim H, Abu DR, Shubair M, Khalil E. Evaluation of fast

disintegrants in terfenadine tablets containing a gas evolving disintegrant. Drug

Dev Ind Pharm 1998; 24(6): 501-507.


89

32. Nayak SM, Gopalkumar P. Design and optimization of fast dissolving tablets for

promethanine. Indian Drugs 2004; 41(9): 554-556.

33. Kaushik D, Dureja H, Saini TR, Formulation and evaluation of olanzapine

mouth dissolving tablets by effervescent formulation approach. Indian Drugs

2004; 41(7): 410-412.

34. Khan S, Kataria P, Nakhat P, Yeole P. Taste masking of ondansetron

hydrochloride by polymer carrier system and formulation of rapid disintegrating

tablets. AAPS Pharm Sci Tech 2007; 8(2): E1-E7.

35. Sunada H, Bi Y. Preparation, evaluation and optimization of rapidly

disintegrating tablets. Powder Technology 2002; 122: 188-198.

36. Marshall K, Compression and consolidation of powdered solid. In: Lachman L,

Liberman HA and Kaning JL, Eds., The Theory and Practice of Industrial

Pharmacy, 3rd

Edn., Varghese publishing house, Mumbai, 1987; 66-99.

37. Gorman EA, Rhodes CT, Rudnic EM. An evaluation of croscarmellose as a

tablet disintegrant in direct compression systems. Drug Dev Ind Pharm 1982;

8(4): 397–410.

38. Chaudhari PD, Chaudhari SP, Kolhe SR, Dave KV, More DM. Formulation and

evaluation of fast dissolving tablets of famotidine. Indian Drugs 2005; 42(10):

641-649.

39. Lindberg N, Palsoson M, Pihl A, Freeman R, Zetzener H, Enstad G. Flowability

measurements of pharmaceutical powder mixtures with poor flow using five

different techniques. Drug Dev Ind Pharm 2004; 30(7): 785-791.

40. Cooper J, Gunn C. Powder flow and compaction. In: Carter SJ, Eds., Tutorial

Pharmacy, 3rd

Edn., CBS publishers and distributors, New Delhi, 1986; 211-

233.

41. Banker SB, Anderson NR, Tablets. In: Lachman L, Liberman HA and Kaning

JL, Eds., The Theory and Practice of Industrial Pharmacy, 3rd

Edn., Varghese

publishing house, Mumbai, 1987; 293-345.

42. United States Pharmacopoeia 30/ NF25. The Official Compendia of Standards.

Electronic edition. Rockville, MD: United States Pharmacopoeial Convention

Inc. 2007.


90

43. Late SG, Yi-Ying Y, Banga AK. Effect of disintegration promoting agent,

lubricants and moisture treatment on optimized fast disintegrating tablets. Int. J

Pharm 2009; 365: 4-11.

44. Pandey PV, Amarnath R. Formulation and evaluation of chlorquine phosphate

tablets using some disintegrants. The Indian Pharmacist 2007; 6(66): 75-79.

45. Bi Y, Sunada H, Yonezawa Y, Danjo K, Otsuka A, Lida K. Preparation and

evaluation of a compressed tablet rapidly disintegrating in oral cavity. Chem

Pharm Bull 1996; 44: 2121-2127.

46. Simone S, Peter CS. Fast dispersible ibuprofen tablets. Eur J Pharm Sci 2002;

15(3): 295-305.

47. Fukani J, Yonemochi E, Yoshihashi Y, Terada K. Evalution of rapidly

disintegrating tablets containing glycine and carboxy methyl cellulose. Int J

Pharm 2006; 310: 101-109.

48. Kuno Y, Kojima M, Ando S, Nakagami H. Evaluation of rapidly disintegrating

tablets manufactured by phase transition of sugar alcohols. J. Control Release

2005; 105: 16-22.

49. Sharma S, Gupta GD. Formulation and characterization of fast dissolving tablets

containing promethazine theoclate solid dispersion with PEG 4000. Pharma

Review 2008; 4: 164-169.

50. Wantabe Y, Yamamoti Y, Fuji M, Kondoh M, Sibata Y. A novel method for

predicting disintegration time in mouth for rapid disintegrating tablet by

compaction analysis. Chem Pharm Bull 2004; 52: 1394-1395.

51. Narazaki R, Harada T, Takami N, Kato T, Ohwaki T, A new method for

disintegration studies of rapid disintegrating tablet. Chem Pharm Bull 2004; 52:

704-707.

52. Schneider. Disintegration testing device. 1988. United States Patent, Patent

number: 4,754,657.

53. El-Arini SK, Clas SD. Evaluation of disintegration testing of different fast

dissolving tablets using the texture analyzer. Pharm Develop Tech 2002; 7: 361-

371.


91

54. Gohel M, Patel M, Amin A, Agarwal R, Dave R, Bariya N. Formulation design

and optimization of mouth dissolving tablets of nimesulide using vacuum drying

technique. AAPS Pharm Sci Tech 2004; 5(3): Article 36.

55. Gohel MC, Parikh RK, Brahmbhatt BK, Shah AR. Preparation and assessment

of novel coprocessed superdisintegratnt consisting of crospovidone and sodium

starch glycolate. AAPS Pharm Sci Tech 2007; 8(1): Article 9.

56. Jeong SH, Park K. Development of sustained release fast disintegrating tablets

using various polymer-coated ion-exchange resin complexes. Int J Pharm 2008;

353: 195–204.

57. Caramella C, Colmbo P, Conte U. Water uptake and disintegration force

measurements: towards a general understanding of disintegration mechanism.

Drug Dev Ind Pharm 1986; 12: 1749-1766.

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