For Restricted Circulation Only
Nixi Laboratories Pvt. Ltd.
VPO: Mouza Ogli, Sadhora Road, Kala Amb (H.P.)
Quality Control Department
Raw Material Standard Testing Procedure
Title : ADRENALINE BITARTRATE IP
STP NO. QC/RM/STP/170 Reference : As per IP
Supersede No. Nil Revision No. : 00
Effective date : 11/03/2022 Review Date: 10/03/2024
1. DESCRIPTION
A white or greyish-white, crystalline powder; odourless. It darkens on exposure to air and light,
decomposition being faster in the presence of moisture and at higher temperatures; (Take a small
quantity of sample in a Petri dish and observe visually for its colour, odour and foreign particles, if
any).
2. SOLUBILITY
Freely soluble in water; slightly soluble in ethanol (95 per cent); practically insoluble in
chloroform and in ether
3. IDENTIFICATION
Procedure: Dissolve 5 g in 50 ml of a 0.5 per cent w/v solution of sodium metabisulphite and
make alkaline by addition of ammonia. Keep the mixture at room temperature for at least 15
minutes and filter. Reserve the filtrate for identification test C. Wash the precipitate with 3
quantities, each of 10 ml of methanol. Evaporate to dryness at 80°. The residue complies with the
following tests.
(A) By Infrared Absorption Spectrophotometer: Compare the spectrum with that obtained with
adrenaline WS or with the reference spectrum of adrenaline.
(B) By Ultraviolet and Visible Absorption Spectrophotometry: When examined in the range
230 nm to 360 nm, a 0.005 per cent w/v solution in 0.01 M hydrochloric acid shows an absorption
maximum only at about 279 nm absorbance at about 279 nm, about 0.4.
(C) By Chemically (Reaction ‘C’ of Tartrate): Heat 0.1 ml of solution containing about 2 mg of
tartaric acid or 0.1 ml of prescribed solution on a water-bath for 5 to 10 minutes with 0.1 ml of a
10 per cent w/v solution of potassium bromide, 0.1 ml of 2 per cent w/v solution of resorcinol and
3 ml of sulphuric acid; a dark blue colour that changes to red when the solution is cooled and
poured into water is produced.
4. APPEARANCE OF SOLUTION
Sample solution: Dissolve 2.5g of the substance under examination in 50ml of carbon dioxide-
free water; (5.0 per cent w/v). The solution is not more opalescent than opalescence standard OS2
and not more intensely coloured than reference solution BYS4.
Preparations: Standard Suspension: Dissolve 1.0 g of hydrazine sulphate in sufficient water to
produce 100.0 ml and set aside for about 6 hours. To 25.0 ml of this solution add 25.0 ml of a 10.0
Prepared By Checked By Approved By
Name Sachin Kumar Parul Mehandiratha Pramod Kumar
Designation Officer QC Executive QC [Link] QC
Signature
Date
Format No.: NL/QC/032/ F01-02 Page 1 of 7
For Restricted Circulation Only
Nixi Laboratories Pvt. Ltd.
VPO: Mouza Ogli, Sadhora Road, Kala Amb (H.P.)
Quality Control Department
Raw Material Standard Testing Procedure
Title : ADRENALINE BITARTRATE IP
STP NO. QC/RM/STP/170 Reference : As per IP
Supersede No. Nil Revision No. : 00
Effective date : 11/03/2022 Review Date: 10/03/2024
per cent w/v solution of hexamine mix well and allow to stand for 24 hours. Keep in a glass
container with a smooth internal surface in which the suspension does not adhere to the glass. Store
in this manner, the suspension is stable for about 2 months. Prepare the standard suspension by
diluting 15 ml of the well mixed suspension to 1000 ml with water. The standard suspension
should be used within 24 hours of preparation.
Opalescence Standards: Prepare opalescence standards by mixing aliquots of the standard
suspension with water.
Clarity or opalescence: Express the degree of opalescence in terms of the opalescence standard. A
liquid is considered clear if its clarity is the same as that of water or of the solvent used for
preparing the solution under examination or if its opalescence is not more than that of opalescence
standard OS1.
Reference Solution Standard Suspension (ml) Water (ml)
OS2 10.0 90.0
Special Reagents: Ferric Chloride Colorimetric Solution (FCS): Dissolve about 46 g of ferric
chloride hexahydrate in enough of a mixture of 25 ml of hydrochloric acid and 975 ml of water to
produce 1000 ml. Pipette 10 ml of this solution into a 250 m1 iodine flask add 15 ml of water, 3 g
of potassium iodide and 5 ml of hydrochloric acid and allow the mixture to stand for 15 minutes.
Dilute with 100 ml of water and titrate the liberated iodine with 0.1 M sodium thiosulphate using
0.5 ml of starch solution, added towards the end of the titration, as indicator. Carry out a blank
titration.
1 ml of 0.1 M sodium thiosulphate is equivalent to 0.02703 g of FeC1 3, 6H2O. Adjust the final
volume of the solution by the addition of enough of the mixture of hydrochloric acid and water so
that each ml contains 0.045 g of FeC1 3, 6H2O. The solutions should be stored protected from light
and standardised before use.
Cobaltous Chloride Colorimetric Solution (CCS): Dissolve about 60 g of Cobaltous chloride in
enough of a mixture of 25 ml of hydrochloric acid and 975 ml of water to produce 1000 ml. Pipette
5 ml of this solution into a 250 ml iodine flask, add 5m1 of hydrogen peroxide solution (10
volume) and 15 ml of sodium hydroxide solution, boil for 10 minutes, cool and add 2 g of
potassium iodide and 60 ml of dilute sulphuric acid. Dissolve the precipitate by gentle shaking, if
necessary, and titrate the liberated iodine with 0.1 M sodium thiosulphate using 0.5 ml of starch
solution, added towards the pink endpoint, as indicator. Carry out a blank titration.
Prepared By Checked By Approved By
Name Sachin Kumar Parul Mehandiratha Pramod Kumar
Designation Officer QC Executive QC [Link] QC
Signature
Date
Format No.: NL/QC/032/ F01-02 Page 2 of 7
For Restricted Circulation Only
Nixi Laboratories Pvt. Ltd.
VPO: Mouza Ogli, Sadhora Road, Kala Amb (H.P.)
Quality Control Department
Raw Material Standard Testing Procedure
Title : ADRENALINE BITARTRATE IP
STP NO. QC/RM/STP/170 Reference : As per IP
Supersede No. Nil Revision No. : 00
Effective date : 11/03/2022 Review Date: 10/03/2024
1ml of 0.1 M sodium thiosulphate is equivalent to 0.02379 g of CoC1 2, 6H2O. Adjust the final
volume of the solution by the addition of enough of the above mixture of hydrochloric acid and
water so that each ml contains 0.0595 g of CoC12, 6H2O.
Cupric Sulphate Colorimetric Solution (CSS): Dissolve about 63 g of cupric sulphate in enough
of a mixture of 25 ml of hydrochloric acid and 975 ml of water to produce 1000 ml. Pipette 10 ml
of this solution into a 250 ml iodine flask, add 40 ml of water, 4 ml of acetic acid, 3 g of potassium
iodide, and 5 ml of hydrochloric acid and titrate the liberated iodine with 0.1 M sodium
thiosulphate using 0.5 ml of starch solution, added towards the pale brown end-point, as indicator.
Carry out a blank titration.
1 ml of 0.1 M sodium thiosulphate is equivalent to 0.02497g of CuSO 4, 5H2O. Adjust the final
volume of the solution by the addition of enough of the above mixture hydrochloric acid and water
so that each ml contains 0.0624 g of CuSO4, 5H2O.
Reference FCS (ml) CCS(ml) CSS(ml) Hydrochloric Acid
Solution (1.0% w/v HCl) (ml)
BYS4 6.0 2.5 1.0 90.5
5. SPECIFIC OPTICAL ROTATION
Test Preparation: Dissolve 2.0 g of residue obtained in identification test in 50 ml 1 M
hydrochloric acid. Determine the specific optical rotation by calibrated instrument; (4.0 per cent
w/v)
Reading + Reading + Reading
Optical Rotation: --------------------------------------------
3
O.R x dilution x 100
Specific Optical Rotation: --------------------------------------------
2 x Sample weight x (100 –LOD)
6. pH
Sample solution: A supernatant liquid obtained by dissolving 0.5g of the substance under
examination in 50ml of carbon dioxide- free water; (1.0 per cent w/v)
Procedure: Take sample solution in a beaker and determine pH by using calibrated pH meter
Prepared By Checked By Approved By
Name Sachin Kumar Parul Mehandiratha Pramod Kumar
Designation Officer QC Executive QC [Link] QC
Signature
Date
Format No.: NL/QC/032/ F01-02 Page 3 of 7
For Restricted Circulation Only
Nixi Laboratories Pvt. Ltd.
VPO: Mouza Ogli, Sadhora Road, Kala Amb (H.P.)
Quality Control Department
Raw Material Standard Testing Procedure
Title : ADRENALINE BITARTRATE IP
STP NO. QC/RM/STP/170 Reference : As per IP
Supersede No. Nil Revision No. : 00
Effective date : 11/03/2022 Review Date: 10/03/2024
7. RELATED SUBSTANCES
By Liquid Chromatography (HPLC):
Note: Prepare the solutions protected from light.
Solvent mixture A: Dissolve 5.0 g of potassium dihydrogen phosphate and 2.6 g of sodium
octanesulphonate in water and dilute to 1000 ml with the same solvent, stir for at least 30 minutes,
adjust to pH 2.8 with orthophosphoric acid.
Solvent mixture B: 13 volumes of acetonitrile and 87 volumes of solvent mixture A.
Test solution: Dissolve 75 mg of the substance under examination in 5 ml of 0.1 M hydrochloric
acid and dilute to 50.0 ml with solvent mixture B.
Reference solution (a): Dilute 1.0 ml of the test solution to 100.0 ml with solvent mixture B.
Reference solution (b): Dissolve 1.5 mg of noradrenaline tartrate WS (adrenaline impurity A WS)
in solvent mixture B, add 1.0 ml of the test solution and dilute to 100.0 ml with solvent mixture B.
Chromatographic system:
Column Dimension: A stainless steel column 10 cm x 4.6 mm, packed with endcapped
octadecylsilane bonded to porous silica (3µm),
Column temperature: 50°
Mobile phase A: A mixture of 5 volumes of acetonitrile and 95 volumes of solvent mixture A,
Mobile phase B: A mixture of 45 volumes of acetonitrile and 55 volumes of solvent mixture A,
Time (in min) Mobile phase A (percent v/v) Mobile phase B(percent v/v)
0 92 8
15 50 50
20 92 8
25 92 8
Flow rate: 2.0 ml per minute,
Spectrophotometer: 210 nm,
Injection volume: 20 µl.
System Suitability: Inject reference solution (b). The test is not a lid unless the resolution between
the peaks due to adrenaline impurity A and adrenaline is not less than 3.0.
Inject reference solution (a) and the test solution. In the chromatogram obtained with the test
solution, the area of any peak at relative retention time of about 3.2 with reference to the principal
peak is not more than 0.3 times the area of the principal peak in the chromatogram obtained with
reference solution (a) (0.3 per cent). The area of any peak at relative retention times of about 0.8
Prepared By Checked By Approved By
Name Sachin Kumar Parul Mehandiratha Pramod Kumar
Designation Officer QC Executive QC [Link] QC
Signature
Date
Format No.: NL/QC/032/ F01-02 Page 4 of 7
For Restricted Circulation Only
Nixi Laboratories Pvt. Ltd.
VPO: Mouza Ogli, Sadhora Road, Kala Amb (H.P.)
Quality Control Department
Raw Material Standard Testing Procedure
Title : ADRENALINE BITARTRATE IP
STP NO. QC/RM/STP/170 Reference : As per IP
Supersede No. Nil Revision No. : 00
Effective date : 11/03/2022 Review Date: 10/03/2024
and 1.3 with reference to the principal peak is not more than 0.2 times the area of the principal
peak in the chromatogram obtained with reference solution (a) (0.2 per cent). The area of any other
secondary peak is not more than 0.1 times the area of the principal peak in the chromatogram
obtained with reference solution (a) (0.1 per cent). The sum of areas of all the secondary peaks is
not more than 0.6 times the area of the principal peak in the chromatogram obtained with reference
solution (a) (0.6 per cent). Ignore any peak with an area less than 0.05 times the area of the
principal peak in the chromatogram obtained with reference solution (a) (0.05 per cent).
8. PHENONES
By Ultraviolet and Visible Absorption Spectrophotometry:
Procedure: Absorbance of a 0.2 per cent w/v solution in 0.1 M hydrochloric acid at the maximum
at about 310 nm, not more than 0.10, calculated on the dried basis.
9. SULPHATED ASH
Instrument: Muffle furnace and Analytical balance.
Apparatus: Silica crucible, pipette and pair of tongs.
Reagent and Solution: Sulphuric acid.
Procedure: Ignite a silica crucible to redness for 10 minutes. Allow to cool in desiccators. Weigh
& note the weight of the crucible w 1 & weigh accurately about 1.0 gm of sample in crucible
[Link] gently first until the substance is thoroughly charred residue being moistened with 2 ml of
nitric acid and 5 drops of sulphuric acid. Heat gently until the white fumes no longer evolved and
ignites at 800°C ± 25°C until the black particles disappear. Allow to cool the crucible in
desiccators and weigh the crucible W3.
Calculation: Sulphated Ash % = (W3 – W1) / (W2 – W1) ×100
Where, W1 = Weight of empty crucible in gm
W2 = Weight of crucible with sample in gm
W3 = Weight of crucible with residue after ignition in gm
10. LOSS ON DRYING
Instrument: Vacuum oven and Analytical balance.
Apparatus: LOD bottle, desiccators and pair of tongs.
Procedure: Keep LOD bottle and stopper in the oven and maintain temp. 105°C for ½ hr. cool the
bottle to room temperature and weigh accurately with stopper. Take the sample around 1.0 gm in
Prepared By Checked By Approved By
Name Sachin Kumar Parul Mehandiratha Pramod Kumar
Designation Officer QC Executive QC [Link] QC
Signature
Date
Format No.: NL/QC/032/ F01-02 Page 5 of 7
For Restricted Circulation Only
Nixi Laboratories Pvt. Ltd.
VPO: Mouza Ogli, Sadhora Road, Kala Amb (H.P.)
Quality Control Department
Raw Material Standard Testing Procedure
Title : ADRENALINE BITARTRATE IP
STP NO. QC/RM/STP/170 Reference : As per IP
Supersede No. Nil Revision No. : 00
Effective date : 11/03/2022 Review Date: 10/03/2024
the Bottle and weigh accurately with stopper. Drying over phosphorus pentoxide at a pressure of
1.5 to 2.5 kPa for 18 hours. Take out the Bottle put the stopper and Allow to cool to room
temperature. Take the weight of bottle again.
Calculation: LOD % - (B–C) / (B–A) x100
Where, A = Weight of empty bottle with stopper.
B = Weight of the bottle and the sample with stopper before drying.
C = Weight of the bottle and the sample with stopper after drying.
11. ASSAY
Procedure: Dissolve 0.3 g in 50 ml of anhydrous glacial acetic acid warming slightly, if necessary,
to effect solution. Titrate with 0.1 M perchloric acid, using crystal violet solution as indicator.
Carry out a blank titration.
1 ml of 0.1 M perchloric acid is equivalent to 0.03333 g of C9H13NO3,C4H6O6
Assay (OSB):
Volume consumed x factor x Actual Normality x 100
: ---------------------------------------------------------------------
Standard Normality x Spl. Wt.
Assay (ODB):
%Assay (OSB) X 100
: --------------------------------
(100-LOD)
12. ABBREVIATIONS
IP: Indian Pharmacopeia STP: Standard Testing Procedure
STS: Standard Test Specification QC: Quality Control
RM: Raw Material No.: Number
NA: Not Applicable CCF: Change Control Format
OSB: On such Basis ODB: On Dried Basis
13. REFERENCE
As per Indian Pharmacopoeia 2018
14. DISTRIBUTION
14.1 Master Copy : Quality Assurance Documentation Cell
Prepared By Checked By Approved By
Name Sachin Kumar Parul Mehandiratha Pramod Kumar
Designation Officer QC Executive QC [Link] QC
Signature
Date
Format No.: NL/QC/032/ F01-02 Page 6 of 7
For Restricted Circulation Only
Nixi Laboratories Pvt. Ltd.
VPO: Mouza Ogli, Sadhora Road, Kala Amb (H.P.)
Quality Control Department
Raw Material Standard Testing Procedure
Title : ADRENALINE BITARTRATE IP
STP NO. QC/RM/STP/170 Reference : As per IP
Supersede No. Nil Revision No. : 00
Effective date : 11/03/2022 Review Date: 10/03/2024
14.2 Controlled Copy-1 : Quality Control
Change History:
Effective Date Version no. Superseded no. CCF no. Reason for Review
00 Nil NA New documents
Prepared By Checked By Approved By
Name Sachin Kumar Parul Mehandiratha Pramod Kumar
Designation Officer QC Executive QC [Link] QC
Signature
Date
Format No.: NL/QC/032/ F01-02 Page 7 of 7