Spectrophotometric Research PPT

1. G.Sasikala & G.Venkateshwarlu*
Dept. Of Chemistry , Osmania University , Hyderabad
Quantitative Determination of
Promethazine Hydrochloride
&
Prasugrel Hydrochloride
By using FC Reagent : A Spectrophotometric Study

2. 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.

3. 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.

4. 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.

5. Promethazine Hydrochloride
• Because of the physiological significance the drug
has 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 complexes
with (BCG) Bromocresol Green
7. UV Visible Spectrophotometric Method

6. Prasugrel Hydrochloride
• Because of the physiological significance the drug has
been 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 Drug
by meta Venadate in acidic medium or
Cerium(IV)Sulphate.

7. 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.

8. 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.

9. Experiment : Materials
• Drugs were procured from M/s. Harika
Drugs Pvt Ltd , Hyderabad as Gift
Samples .
• Folin–Ciocalteu reagent (FCR) was
supplied 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 for
preparing Solutions.

10. Experiment : Materials
Systronic Spectrophotometer Elico Spectrophotometer
Quivites

11. 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.

12. 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.

13. 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.

14. 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

15. 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.

16. 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

17. 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.

18. 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.

19. 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.

20. 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

21. Conclusion
• The Methods Developed are useful for the
analysis of the drugs either in Bulk
Industries or in Pharmaceuticals.

22. 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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25. Thank You