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Group VI
Dri Waskitho
Debby Citra Dewi
Ni Luh Witariani
Tenofovir Disoproxil
IUPAC Name :
({[(2R)-1-(6-amino-9H-purin-9-yl)propan-2-yl]oxy}methyl)phos
phonic acid
Tenofovir
PENDAHULUAN
APAKAH TENOFOVIR DISOPROXIL FUMARATE?
Obat untuk pengobatan infeksi HIV pada orang dewasa dan anak-anak berusia 2 tahun ke atas.
Tenofovir DF selalu digunakan dalam kombinasi dengan obat HIV lainnya.
Tenofovir DF termasuk dalam kelompok (kelompok) obat HIV yang disebut nucleoside reverse
transcriptase inhibitor (NRTI). NRTI memblokir enzim HIV yang disebut reverse transcriptase. (Enzim
adalah protein yang memulai atau meningkatkan kecepatan reaksi kimia.) Dengan menghalangi
reverse transcriptase, NRTI mencegah HIV berkembang biak dan dapat mengurangi jumlah HIV di
dalam tubuh.
BACKGROUND
Obat HIV juga mengurangi risiko penularan HIV. Jika Anda memakai
obat-obatan HIV, termasuk tenofovir DF, jangan mengurangi,
melompati, atau berhenti meminumnya kecuali jika petugas
kesehatan Anda memberi tahu Anda.
(https://aidsinfo.nih.gov/drugs/290/tenofovir-disoproxil-
fumarate/0/patient)
BACKGROUND
Impurities
USP 40-NF 35 General Chapter <1086>
Impurities in Drug Substance and Drug Product
Impurity :
“Any component of a drug substance that is not the chemical entity
defined as the drug substance and in addition, for a drug product, is any
component that is not a formulation ingredient”
BACKGROUND
Reference standards
Name Lot No Purity Loss on Drying
Tenofovir Disoproxil Fumarate USPRS GDM335 0.999 mg/mg -
Tenofovir USPRS R044CD -
Adenine USPRS IDM090 -
Equipment And Materials
Sample Information
Sample Name : Tenofovir Disoproxil Fumarate
Dosage Form : Tablet
Composition : Tenofovir Disoproxil Fumarate 300 mg
Appearance : Light blue, round, biconvex tablets, imprinted with “M”
on one side and “153” on other side
Reg. No. : GKX1340400217A1
Package : Bottle @ 30 tablets
Manufacturer : Mylan Laboratories Ltd.
Importer/Distributor : PT. Kimia Farma Tbk., Jakarta-Indonesia
Batch No. : 3061232
Mfg./Expiry Date : Nov 2016 / Oct 2019
Mobile Phase Preparation
2
1 0.01 M Na2HPO4 Buffer Solution (pH 5.5)
Solution A
Methanol-tertiary butanol-
buffer (11:1:28)
For 1000 mL:
8.9 g Na2HPO4 Methanol = 275 mL
Water Tertiary butanol = 25 mL
0.01 M Na2HPO4
Buffer = 700 mL
5000 mL 5000 mL 5000 mL
a)
About 8.9 g of Na2HPO4
b)
A portion of water is
c)
The mixture is
d)
Additional water is added
3
is weighed on an added to the volumetric swirled until all of up to the mark on the Solution B
analytical balance and flask the solid has volumetric flask and the Methanol-tertiary butanol-
transferred into a 5000 dissolved solution is adjusted to a buffer (27:1:12)
mL volumetric flask completely pH of 5.5 with phosphoric
acid For 1000 mL:
Methanol = 675 mL
Tertiary butanol = 25 mL
Buffer = 300 mL
Figure 1. Preparation of mobile phase
System Suitability Solution
1 2
+
up to 50 mL
3.75 mg a) b) c)
5.0 mL of adenine and The measured volume The measured volume in the
tenofovir stock solution is of stock solution is second flask is then diluted
each measured using transferred into 50 mL with solution A up to the
volumetric pipette volumetric flask volumetric mark
1 2
+
up to 100 mL
5 mg of tenofovir
disoproxil fumarate
a) b) c)
5.0 mL of tenofovir disoproxil The measured volume The measured volume in the
fumarate stock solution is of stock solution is second flask is then diluted
measured using volumetric transferred into 50 mL with solution A up to the
pipette volumetric flask volumetric mark
1 2
a)
NLT 20 tablets equivalent to 6000 mg/mL
tenofovir disoproxil fumarate is add with
about 3500 mL of solution A in 5000 mL
volumetric flask
b)
The mixture is sonicated for 10 minute,
cooled to room temperature, and then
mark 3
diluted with solution A up to the volumetric
a) b) c)
4.0 mL of sample stock solution The measured volume The measured volume in the
is measured using volumetric of stock solution is second flask is then diluted
pipette transferred into 25 mL with solution A up to the
sample solution
Sample solution: 500 µg/mL
Chromatographic Conditions
Instrument: HPLC Shimadzu Prominence LC 20AD
Chromatographic system:
Table 1 Table 2
Detector 260 nm Time (min) % Solution A % Solution B
Column XBridge C18; 4,6 x 25 cm; 5 0.00 – 2.00 100 0
µm 2.01 – 30.0 0 100
Lot No. 0173352721 (Code
067) 30.01 – 45.00 100 0
Column 35 °C 45.01 – 65 100 0
temperature
Table 3
Sample 4 °C (freshly taken out from
temperature refrigerator prior to Parameters Requirements
injection) Tailing factor NMT 2,0
Flow rate 1.0 mL/min Resolution NLT 1.5 (tenofovir-
Injection volume 10 µL adenine)
Gradient system See Table 2 RSD NMT 10.0%
System Suitability
Table 3
Parameters Requirements Samples
Tailing factor NMT 2,0 Standard solution
Resolution NLT 1.5 (tenofovir-adenine) System suitability solution
RSD NMT 10.0% Standard solution
Bracketing
1.0
mV
0.5
0.0
0.0 5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.0 50.0 55.0 60.0 min
Adenine/2.880
Tenofovir/2.609
Detector A:260nm
5.0
2.5
0.0
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 min
TDF/19.827
Fumaric Acid/2.378
0.0 5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.0 50.0 55.0 60.0 min
Quality Assurance
Working Std Control Std
Results Sample Analysis
mV
RT2.382/2.382
RT4.090/4.090
TDF/19.787
Detector A:260nm
RT35.165/35.165
RT28.598/28.598
RT25.312/25.312
1.5
1.0
0.5
0.0
0.0 5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.0 50.0 55.0 60.0 min
Sample 3
Acceptance
Tailing Theoretical C Spl C Std 0,05% % each
No Name RT Area Resolution RRT F Keterangan criteria
factor Plate (ug/mL) (ug/mL) main peak impurity
NMT (%)
1 RT2.381 2.381 445200 1.339 -- 3.695.295 0.120 480 0,56462 3046,42 fumaric acida -
tenofovir isoproxil
2 RT4.091 4.091 70828 1.232 9.096 5.567.396 0.207 480 0,56462 1,5 3046,42 0,770002 3,0
monoester
3 TDF 19.798 6238063 1.222 54.341 48.713.952 1.000 480 0,56462 3046,42 TDF
4 RT25.317 25.317 1751 1.181 16.947 119.837.778 1.279 480 0,56462 3046,42 disregard
5 RT28.608 28.608 2566 1.173 10.762 128.614.602 1.445 480 0,56462 3046,42 disregard
tenofovir
6 RT35.172 35.172 17542 0.766 15.181 66.482.002 1.776 480 0,56462 1 3046,42 0,28606 0,75
disoproxil dimer
Total impurities 1,056063 4,0
Results
Sample Analysis
% Acceptance
RRT
No. Peak Name F RT Area RT Impurit Crit. Conclusion
refer.
y NMT (%)
1 Fumaric acid 0.12 - 2.381 438585 0.120 - - -
2 Tenofovir isoproxil
0.14 1.500 4.090 63809 0.207 0.6937 3.0 Meets req.
monoester
3 Tenofovir disoproxil 1.0 - 19.792 6145937 0.999 - - -
4 Disregard peak - - 25.312 1712 1.278 - - -
5 Disregard peak - - 28.601 2464 1.444 - - -
6 Tenofovir disoproxil
1.74 1.000 35.167 17122 1.776 0.2792 0.75 Meets req.
dimer
Total impurities -- -- -- -- -- 0.9729 4.0 Meets req.
Discussion
Results
Sample Analysis
% Acceptance
RRT
No. Peak Name F RT Area RT Impurit Crit. Conclusion
refer.
Thank You
y NMT (%)
1 Fumaric acid 0.12 - 2.381 438585 0.120 - - -
2 Tenofovir isoproxil
0.14 1.500 4.090 63809 0.207 0.6937 3.0 Meets req.
monoester
3 Tenofovir disoproxil 1.0 - 19.792 6145937 0.999 - - -
4 Disregard peak - - 25.312 1712 1.278 - - -
5 Disregard peak - - 28.601 2464 1.444 - - -
6 Tenofovir disoproxil
1.74 1.000 35.167 17122 1.776 0.2792 0.75 Meets req.
dimer
Total impurities -- -- -- -- -- 0.9729 4.0 Meets req.
Conclusion
Tablet Tenofovir Disoproxil Fumarate : meets
requirement for impurities
Pay attention to :
(GLP Implementation)
1. Develop workplan
2. Protect yourself with the personal protection stuff (APD)
3. Make sure all the instrument are calibrated and in a good condition
4. Make sure all the reagent is correct and not expired
5. Documenting every process and result in a good documentation
practise