Formulation and Evaluation of Fixed Dose Immediate

Release Tablets of
5-Hydroxytryptophan and AET (S,2-aminoethylisothiourea-
Br-HBr).

Presented by- Supervisor

Mona Prof P.K. Sahoo
M.Pharm (Pharmaceutics)
Enrollment No.-03/MPH/DIPSAR/2017

1
Content
Introduction

Drug selection

Objective

Scheme of work

Methods

Results

Conclusion

2
Introduction
o Ionizing radiations are high energy radiations which can remove electrons from outer shells of
atoms and molecules rendering them ionized
o Excellent tool for diagnostic purposes, but these are very harmful to humans in case of
frequent exposure or exposure to high dose of radiations
o Low linear energy transfer radiations (LET) which include X-ray, gamma radiations and beta
particles are the most harmful type of radiations for the human body
oLarge total-body doses of radiation impact most severely on the regenerative cells of the
intestinal epithelium, on the regenerative cells of the bone marrow and on the microvascular
system's endothelial cells

3
 Scavenge free
radicals

Inexpensive Reduce
and readily oxidative
available damage

Facilitate
Long shelf life cellular and
DNA repair

Ideal
radioprotector

Should not
Repopulation of
show any
damaged or
significant toxic
affected organ
reaction

Suitable for oral

administration, Immunomodula
rapid systemic tion
Good
absorption
protection
against both
acute and
chronic
radiation injury.

4
Drug selection
o 5-HTP is immediate precursor of serotonin which can freely cross BBB
o 5-HTP protects cells against radiation by the upsurge of 5-HT molecules in cells, and some
altered physiological state of cells such as metabolic activity or an arrangement of the cell
constituents which is induced by 5HTP penetrating into the cells
oRadioprotective dose of 5-HTP = 33.4mg/kg
oAET is an isothiouranium compound which has been studied extensively for its radioprotective
activity
oRadioprotective dose of AET = 334mg/kg
o Researches have proved that if used in combination of 5 (5-HTP) :1 (AET) their dose to be used
as radioprotectant decrease drastically which is 8.4mg/kg for 5-HTP and 1.7mg/kg for AET

5
Objective
oThere is no marketed formulation of 5-HTP (to be used for radioprotection) and AET either
individually or in combination
o Immediate release tablets are those which disintegrate rapidly and get dissolved to release the
medicaments
oDirect compression is the easiest way to manufacture tablets

Direct Low manufacturing cost

compression
advantages Uses conventional equipment, commonly available excipients, and a limited
number of process steps
Rapid dissolution

6
Scheme of Work
• Preparation of standard curve of 5-HTP and AET in 0.1N HCL
Preformulation • Drug excipient compatibility studies
studies • Flow properties of drug powder

• Preparation of fixed dose immediate release tablets by direct

Formulation of compression method
Tablets • Optimization of tablet formulation

• Tablet dimension
Evaluation of • Hardness test
Tablets • Friability test
• Disintegration test
• In vitro dissolution test

7
Preparation of Tablets
All the excipients and Drug powers were passed through
sieve no.16

Powders were then transferred to air filled polythene

bags and tumbled for 10min

Flow properties of the mixture were evaluated

Mixture was then compressed on rotary punching

machine using 12mm punch to get tablet of 750mg

8
 Composition of Tablets with SSG as disintegrating agent Composition of Tablets with Crospovidone as disintegrating agent

Components Weight (mg/tablet) Percentage per Components Weight (mg/tablet) Percentage per
tablet tablet
5-Hydroxytryptophan 500 66.67%% 5-Hydroxytryptophan 500 66.67%
AET 100 13.33% AET 100 13.33%
Pregelatinized starch 92.1 12.28% Pregelatinized starch 103.1 13.74%
Sodium starch 37.5 5% Crospovidone 26.5 3.54%
glycolate
Talc 14 1.87%
Talc 14 1.87%
Magnesium stearate 1.9 0.25%
Magnesium stearate 1.9 0.25%
Silicon dioxide 4.5 0.6%
Silicon dioxide 4.5 0.6%
Total 750mg 100%
Total 750 mg 100%

9
Drug characterization

UV scan of 5-HTP in 0.1N HCl UV scan of AET in 0.1N HCl

λmax = 274nm λmax = 200nm

10
 DSC scan of 5-HTP and AET
DSC Temp D SC Tem p
mW C mW C
40.00 Start 271.55 0x10
C 400.00 Start 188.770C
x 10
End 288.67 0
x10
C 10.00 End 201.350C
x 10
Peak 281.82 0
x10
C Peak 198.630C
x 10
20.00 200.00
Onset 277.60 0x10
C 300.00 Ons et 190.680C
x 10
Endset 284.77 0x10
C Ends et 200.890C
x 10
0.00 0.00
Heat 1112.53 0x10
mJ H eat 288.620m
x 10
J
200.00
100.00
-20.00

100.00
-10.00
-40.00

0.00 10.00 20.00 30.00 40.00 0.00 10.00 20.00

Time [min] Tim e [m in]

5-HTP AET

11
FTIR

5-HTP AET

12
 Calibration curve of 5-HTP in 0.1N HCl
Calibration curve of 5-HTP at 274nm
1
0.9
Concentration (µg/ml) Absorbance y = 0.0277x + 0.0062
0.8 R² = 0.9999
2 0.060 0.7

absorbance
4 0.113 0.6
0.5
8 0.233
0.4
16 0.450 0.3
0.2
32 0.890
0.1
0
0 5 10 15 20 25 30 35
concentration

13
 Calibration curve of AET in 0.1N HCl
Calibration curve of AET at 200 nm
1.4
Concentration (µg/ml) Absorbance

1.2
y = 0.0277x + 0.2738
2 0.239 R² = 0.9246
1
4 0.346

Absorbance
0.8
8 0.565
0.6
16 0.849
0.4
32 1.078
0.2

0
0 5 10 15 20 25 30 35
concentration

14
 Drug excipient compatibility study
DSC Temp DSC Temp
mW C mW C
50.00 400.00 400.00
Start 202.29 0x10
C Start 280.43 0x10
C
4.00
End 213.80 0x10
C End 293.78 0x10
C
Peak 208.60 0
x10
C
300.00 Peak 287.13 0x10
C300.00
2.00
Onset 204.49 0x10
C Onset 282.59 0C
x10
Endset 212.19 0
x10
C Endset 292.24 0x10
C
0.00 0.00
200.00 Heat 56.56 0mJ
x10 Heat 200.00
115.43 0mJ
x10

-2.00
100.00 100.00

-4.00
-50.00
0.00 10.00 20.00 30.00 40.00 50.00 0.00 10.00 20.00 30.00 40.00
Time [min] Time [min]

Sodium starch glycolate SSG+ 5-HTP+AET

15
 DSC Temp DSC Temp
mW C mW C
400.00 400.00
40.00
0
205.86x10
C
10.00
300.00 0
278.56x10
C 300.00

20.00

200.00 200.00
0.00
0.00
100.00 100.00

-10.00
-20.00
0.00 10.00 20.00 30.00 40.00 0.00 10.00 20.00 30.00 40.00
Time [min] Time [min]

Crospovidone K-30 Crospovidone K-30 +5-HTP+ AET

DSC Temp
DSC Temp
mW C
mW C
3.00 400.00
400.00 Start 88.91 0x10
C Start 269.63 0x10
C
10.00
End 108.36 0x10
C End 283.28 0x10
C
2.00 Peak 98.53 0x10
C Peak 275.87 0x10
C
Onset 90.57 0x10
C Onset C 300.00
271.45 0x10
300.00
1.00 Endset 105.81 0x10
C Endset 280.88 0x10
C
Heat 70.14 0x10
mJ Heat 55.64 0x10
mJ

0.00 200.00
0.00 200.00
Start 182.36 0x10
C
End 193.95 0
x10
C
-1.00 Peak 189.04 0x10
C
100.00 Onset 183.67 0x10
C 100.00
-2.00 Endset 193.25 0x10
C
Heat 30.75 0x10
mJ
-10.00
0.00 10.00 20.00 30.00 40.00 0.00 10.00 20.00 30.00 40.00
Time [min] Time [min]

Pregelatinized starch Pregelatinized starch + 5-HTP + AET

16
 Thermal Analysis Result
DSC Temp DSC Temp
mW C mW C
3.00 400.00
Start 88.91 0x10
C Start 269.63 0x10
C
Start 88.05 0
x10
C
2.00 End 108.36 0x10
C End 283.28 0x10
C
0
2.00 98.53 0x10
End 98.08 x10
C Peak C Peak 275.87 0x10
C
200.00 90.57 0x10
Peak 93.20 0C
x10
Onset C Onset C 300.00
271.45 0x10
1.00 Endset 105.81 0x10
C Endset 280.88 0x10
C
0.00 Onset 89.71 0C
x10 Heat 70.14 0
x10
mJ Heat 55.64 0x10
mJ
Endset 96.45 0
x10
C
0.00 200.00
Heat 46.30 0mJ
x10 Start 182.36 0x10
C

-2.00 100.00 End 193.95 0x10

C
-1.00 Peak 189.04 0x10
C
Onset 183.67 0x10
C 100.00
-2.00 Endset 193.25 0x10
C
Heat 30.75 0x10
mJ
-4.00
0.00 10.00 20.00 0.00 10.00 20.00 30.00 40.00
Time [min] Time [min]

Magnesium stearate Magnesium stearate + 5-HTP + AET

17
Flow Properties of Powder
API
Properties Sodium starch glycolate Crospovidone K-30
(5-HTP+AET)

Bulk density 140.21g/mL 108.69g/mL 100g/mL

Tapped density 249.5g/mL 144.23g/mL 133.92g/mL

Compressibility index 43.8% 24.64% 25.32%

Hausner ratio 1.78 1.32 1.34

Angle of repose 71.21 38.21 40.29
Flow Very very poor Passable Passable-poor

18
19
Tablet Evaluation
1. Tablet Dimension
Sodium starch glycolate Crospovidone K-30
Parameter
(Batch A) (Batch B)
Thickness 4.64 ± 0.02mm 4.69 ±0.12mm

Diameter 11.40± 0.02mm 11.41±0.02mm

2. Hardness Testing
Sodium starch glycolate Crospovidone
(Batch A) (Batch B)
Observed value 5 kg 5.5kg

20
• Weight variation
Sodium starch glycolate Crospovidone

(Batch A) (Batch B)
Observed values ± 5.7mg ±3.2 mg

(average wt = 754mg) (average wt = 752.82mg)

Acceptance limit =±5% Pass Pass

• Friability Test

Sodium starch glycolate Crospovidone

(Batch A) (Batch B)
Observed values 0.60 0.49
Acceptance limit = 1% Pass Pass

21
• Disintegration Test

Sodium starch glycolate Crospovidone

Data for 6 tablets
tablet disintegrated within 7 minutes tablet disintegrated within 11 minutes
Acceptable limit= tablets should
pass Pass
disintegrate within 15 minutes

22
In-vitro dissolution test
oBatch A Batch A
100
Time Cumulative % drug release
90

5 31.67 ± 1.5 80

70

cumulative% drug release

10 54.34 ±2.0
60

20 70 ±1 50

40
30 80.5 ±3.4
30

45 88.34 ±3.2 20

10
60 95 ±1
0
0 10 20 30 40 50 60 70
Time(minute)

23
In-vitro dissolution test
Batch B
oBatch B
100

Time Cumulative % drug release 90

80
5 41.34 ± 1.5
70

cumulative % drug release

10 55 ±1 60

20 65.33 ±3.5 50

40
30 72.67 ±1.5
30

45 82.7 ±1.5 20

10
60 86.67±1.5
0
0 10 20 30 40 50 60 70
Time(minute)

24
 Optimization of Tablet Formulation
oFormulation of Batches for optimization

Batch C Batch D
Components Weight (mg/tablet) Percentage Components Weight (mg/tablet) Percentage
5-HTP 500mg 66.67 5-HTP 500mg 66.67
AET 100mg 13.34 AET 100mg 13.34
Pregelatinized starch 69.6 mg 9.28 Pregelatinized starch 80.6 mg 10.74
Sodium starch glycolate 60 mg 8 Sodium starch glycolate 49 mg 6.5
Magnesium stearate 1.9 mg 0.25 Magnesium stearate 1.9 mg 0.25
Silicon dioxide 4.5 mg 0.6 Silicon dioxide 4.5 mg 0.6
Talc 14 mg 1.8 Talc 14 mg 1.8
Total 750mg 100 Total 750mg 100

25
Disintegration test of optimization Batches

Batch C Batch D

Data for 6 tablets tablet disintegrated within5 minutes tablet disintegrated within3 minutes

Acceptable limit= tablets should

Pass Pass
disintegrate within 15 minutes

26
Dissolution testing of Optimization Batches
Batch C
oBatch C 120

Time (minute) Cumulative % drug release 100

5 44.34 ± 1.8

cumulative % drug release

80
10 66.4 ±0.5
20 75.7 ±0.7 60

30 85.34 ±1.3 40

45 91.34 ±1.6
20
60 96 ±1

0
0 10 20 30 40 50 60 70
time (min)

27
Dissolution testing of Optimization Batches
oBatch D Batch D
120

Time (min) Cumulative % drug release 100

5 52.34± 1.5

cumulative% drug release

80

10 70 ±1
60
20 80.7 ±0.5
30 91.67 ±0.02 40

45 97.45 ±0.5
20

60 98.02 ±0.15
0
0 10 20 30 40 50 60 70
Time (min)

28
Conclusion
oThese investigations demonstrated that it is possible to prepare immediate release tablets
containing fixed dose of 5- HTP and AET by direct compression method, using all the excipients
mentioned
oTablets containing sodium starch glycolate showed promising results to be selected for
preparing immediate release tablets.
oAlthough all the optimization batches passed the dissolution testing the tablets containing 8%
of sodium starch glycolate gave the desired release profile of an immediate release formulation

29
Future Prospects
These tablets further need to be tested for their in vivo release
pattern to understand their effectiveness in the living system. These
tablets could be a very beneficial tool for mankind against the
harmful ionizing radiations once their safety and effectiveness are
proved in humans.

30
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32
Thank you

33
Acknowledgment
I am greatly indebted to my supervisor Dr. P.K Sahoo for their guidance, support and time
throughout this research work
I would also like pay my sincere thanks to Prof. (Dr.) D. P. Pathak, Director DIPSAR
A sincere vote of thanks to my senior, Mr. Aswini, Ms Bhumika and Ms Monika for their
invaluable insights and encouragement during the completion of the project
Also, a vote of thanks to my friends and lab mates Aayush, Dev, Himanshu and Aysha whose
help made my work easy and enjoyable
I would also like to acknowledge the laboratory staff and other teaching staff for helping me
directly or indirectly in completion of my work.

34