G.Sasikala & G.Venkateshwarlu*Dept. Of Chemistry , Osmania University , Hyderabad

Quantitative Determination ofPromethazine Hydrochloride

&

Prasugrel Hydrochloride

By using FC Reagent : A Spectrophotometric Study

Promethazine Hydrochloride

Promethazine hydrochloride[-meth′əzēn]

a phenothiazine antiemetic, antihistamine, and sedative.

Indications It is prescribed in the treatment of motion sickness, nausea, rhinitis, itching, an

d skin rash and as anadjunct to anesthesia and pain control.

Prasugrel Hydrochloride

Prasugrel is a member of the thienopyridine class

of ADP receptor inhibitors, like ticlopidine (trade

name Ticlid) and clopidogrel (trade name Plavix).

These agents reduce the aggregation ("clumping")

of platelets by irreversibly binding to

P2Y12 receptors. Compared to clopidogrel, prasugrel

inhibits adenosine diphosphate–induced platelet

aggregation more rapidly, more consistently, and to a

greater extent than do standard and higher doses of

clopidogrel in healthy volunteers and in patients with

coronary artery disease, including those undergoing

PCI.

Folin–Ciocalteu Reagent

• The Folin–Ciocalteu reagent (FCR) or Folin's

phenol reagent or Folin–Denis reagent, also

called the Gallic Acid Equivalence

method (GAE), is a mixture of

phosphomolybdate and phosphotungstate used

for the colorimetric in vitro assay of phenolic

and polyphenolic antioxidants. It is named

after Otto Folin, Vintilă Ciocâlteu, and Willey

Glover Denis.

Promethazine Hydrochloride

• Because of the physiological significance the drughas been analysed by various methods like:

1. FIA Method- Flow Injection analysis Method

2. Electrophoresis Method

3. Potentiometric Method

4. Oxidative Coupling with sulphonilic acid

5. Titrimetric Method

6. By forming Choloroform extractable complexeswith (BCG) Bromocresol Green

7. UV Visible Spectrophotometric Method

Prasugrel Hydrochloride

• Because of the physiological significance the drug hasbeen analysed by various methods like:

1. Visible Spectrophotometric Method

2. By forming Ion Pair complex with Bromo Phenol Blue( BPB)

3. UV Visible Spectrophotometric Method

4. High Performance Liquid Chromatography Method (HPLC)

5. Liquid Chromatography Method (LC)

6. Reverse Phase Liquid Chromatography Method(RP-LC)

7. Titrimetric Method – Based on the Oxidation of Drugby meta Venadate in acidic medium orCerium(IV)Sulphate.

Analysis

• Thorough Survey of Literature revealed that

Folin–Ciocalteu reagent (FCR) is a sensative ,

Precise , accurate reagent for the analysis of

Phenolic Materials

• As the Above Mentioned drugs possess

Phenolic Nature , the author is prompted to

use this method for the above drugs for their

analysis and validation.

• The reagent does not only measure phenols,

and will react with any reducing substance.

Experiment : Methods & Materials

• A Double Beam systronics

spectrophotometer 117 had

been used along with An

ELICO UV double beam

spectrophotometer SL210

to record the spectra of the

drugs, reagent and the

product formed due to the

interaction of the drug with

the reagent was recorded.

Experiment : Materials

• Drugs were procured from M/s. HarikaDrugs Pvt Ltd , Hyderabad as GiftSamples .

• Folin–Ciocalteu reagent (FCR) wassupplied by Finar Chemicals (AR) grade ,this was used without further Purification.

• NaOH was supplied by SD fine Chem(AR) grade

• Triple Distilled water was used forpreparing Solutions.

Experiment : Materials

Systronic Spectrophotometer Elico Spectrophotometer

Quivites

Construction of Calibration

• A stock solution of 1000μg/ml was

prepared and diluted to a concentration

range of 10 μg/ml to 80 μg/ml , to each

solution 1 ml of the Folin–Ciocalteu reagent

(FCR) , 1ml of NaOH was added and made

up to the mark of 10ml using triple distilled

water.

Experiment

• The Experiment was repeated atleast for 6

times to get precise and accurate optical

densities

• Average 95% , 110% points falling within

the range were taken.

Optimization of the factors effecting

Absorbance:

• Promethazine Hydrochloride produced blue

colour with λ max 741 nm immediately after

mixing the solutions . The intensity did not

change even after 1 hr observation.

• Prasugrel Hydrochloride too produced blue

colur at λ max 963 nm after heating the

contents for 50 minutes on a water bath at

60°Celsius , further increase in time and

temperature did not show any effect.

Optimization of the factors effecting

Absorbance:

• Effect of NaOH : The concentration of NaOH

was varied between 0.1 M to 1 M optimum

absorbance was found at 0.5M solution.

• Effect of Concentration of the Reagent: To test

the Reagent Concentration 5ml of the drug

was uniformly taken into 6 standard flasks and

1% to 6% of the reagent was added . It was

observed that the concentration of the reagent

dint effect the OD appreciably

Validation

• The methods developed have been

validated in terms of LOD, LOQ, limits of

Beers law, Robustness, Druggedness,

Accuracy & Precision are presented in the

table.

Table

Name of the Property Promethazine HCL Prasugrel HCL

λmax 741 nm 963 nm

Beer’s law limits (µg/ml) 10-80 10-80

Sandell’s sensitivity (µg/Cm²) 0.0793 0.0854

Std. Dev. of intercepts 0.002723 0.002066

LOD (µg/ml) 0.713 0.582

LOQ (µg/ml) 2.139 1.746

Slope ,b 0.0126 0.0117

Intercept, a 0.06852 0.09028

Correlation coefficient 0.9469 0.9973

Regression equation 0.0685+ 0.0902+

Y=a+bX* ( X= Conc. Of Drug) 0.0126X 0.0117X

Application of the Methods for the

Analysis of Pharmaceuticals

• Promethazine HydroChloride name :

Phenergan Injection 4 voils of 25 mg/2ml

ABBOTT company were purchased .

• Solutions were combined and diluted to the

required concentrations of working

range.Recovery studies have been

performed to determine accuracy and

precision of the method.

Application of the Methods for the

Analysis of Pharmaceuticals

• Prasugrel Hydrochloride : Dry 10mg tablet

of this drug with brand name : Prasusafe

manufactured by MSN laboratories was

ground well into a motor & pestle.

• Powder Equivalent to 25mg of the drug was

dissolved in 100ml of triple distilled water ,

filtered with 41 micron Watt paper and the

solutions was diluted to the concentration

of working range.

Application of the Methods for the

Analysis of Pharmaceuticals

• Precision and accuracy are determined in

terms of %RSD and the reproducibility by

performing 5 replicates.

• The applicability of methods are statistically

analysed by performing t-test and F-test

with a standard method available in

literature.

Table : Parameters Using Pure Sample

Name of the Drug Taken

OD

Found

OD

%

Recov

ery

RSD% t-test F- test

Promethazine

Hydrochloride

0.952

0.875

0.738

0.640

0.961

0.876

0.733

0.643

100.94

100.11

99.32

100.46

0.808 1.425 3.429

Prasugrel

Hydrochloride

1.016

0.896

0.774

0.670

1.021

0.906

0.776

0.675

100.49

101.11

100.25

100.74

0.365 0.6094 2.057

Conclusion

• The Methods Developed are useful for the

analysis of the drugs either in Bulk

Industries or in Pharmaceuticals.

Acknowledgement

• The authors are thankful to the Head ,

Department of Chemistry , Osmania

University , Hyderabad -500007, for

providing facilities.

• Special Thanks to the Chairman & Principal

of Princeton PG College , Ramanthapur ,

Hyderabad for their Support.

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