STANDARD ANALYTICAL PROCEDURE (SAP) OF RAW MATERIAL TWEEN 80 (POLYSORBATE 80) is a not unusual place nonionic surfactant, emulsifier and solubilizer this is utilized in a huge form of industries and applications. TWEEN 80 (POLYSORBATE 80) is generally utilized in meals products, cosmetics, medications, nutrients and vaccines. There may be many motives a organization may also want to decide the precise quantity of polysorbates of their fabric. For instance, groups may also want to decide a limits of their products, validate analytical techniques for product manufacturing or require the records for opposite engineering purposes. Thus, a well-evolved analytical approach is important for correctly quantitating polysorbates.
Due to the chemical shape and chemical houses of TWEEN 80 (POLYSORBATE 80), it can’t be correctly quantitated via way of means of analytical techniques such flowline chromatography or liquid chromatography with ultraviolet absorption. Since polysorbates do now no longer comprise a UV chromophore, this doesn’t offer enough chromatography. However, Charged Aerosol Detection (CAD) can offer a standard detection this is unbiased of the chemical shape for lots chemicals, together with polysorbates. This paper discusses an analytical approach evolved via way of means of Impact Analytical to offer a unmarried height for quantitating polysorbate eighty. The analytical approach that changed into evolved is capable of quantitate the quantity of WEEN 80 (POLYSORBATE 80) in a pharmaceutical system containing TWEEN 80 (POLYSORBATE 80), lively ingredient, microcrystalline cellulose (CMC) and sodium carboxymethyl cellulose.
TABLE OF CONTENTS
6. Quality Records
8. List of recipients
|Sr.#||Date of Amendment||Issue Status||Page No||Page Status||Details of change / review|
|Written by||Checked by||Approved by||Authorized by|
Quality Control Manager
Quality Assurance Manager
General Manager Plant
Instruction: Coping of this document in part or whole is strictly prohibited.
1.1 To ensure the quality of incoming raw material of Tween 80 (Polysorbate 80).
2.1 It is applicable for the analysis of Tween 80 (Polysorbate 80) in the quality control department.
3.1 Quality Control Manager
3.2 Assist. Q.C. Manager
3.3 Q.C Analyst
4.1 SAP: Standard Analytical Procedure
4.2 QC: Quality control
A preceding analytical approach the usage of High Performance Liquid Chromatography (HPLC) with Charged Aerosol Detection (CAD) did now no longer offer enough chromatography to quantitate for TWEEN 80 (POLYSORBATE 80). The TWEEN 80 (POLYSORBATE 80) requirements and samples had been organized in methanol. A Cadenza-C18, three μm, 4.6 x 50 mm column changed into used for the approach with a 0.5 mL/minute flow charge for a complete run time of 23 minutes. A methanol and water gradient changed into used for the analysis. The calibration curve used for quantitation with this approach supplied a linear curve and a R2 fee of 0.9850.
A TWEEN 80 (POLYSORBATE 80) USP preferred changed into used to expand the analytical approach similarly to gain a unmarried height via way of means of the usage of HPLC/CAD. The TWEEN 80 (POLYSORBATE 80) USP preferred changed into used to put together requirements starting from 107 μg/mL to 323 μg/mL in acetonitrile. The calibration curve used to quantitate TWEEN 80 (POLYSORBATE 80) is supplied. The calibration curve suggests linearity and a R2 fee of 0.9988. The device approach used an Inertial ODS-3, 5μm, 4.6 x a 150 mm column managed at a temperature of 40 °C. The cell section changed into isocratic with 50 Acetonitrile and 50% water with 2 ml ammonium acetate. The float charge changed into 1 mL/minute and a complete run time of 6 minutes. An instance chromatogram of a polysorbate eighty USP preferred at a attention of 215 μg/mL is proven in Figure three. The Polysorbate eighty USP preferred supplied a unmarried height to quantitate the quantity of TWEEN 80 (POLYSORBATE 80). A system pattern changed into analyzed with the progressed analytical approach to expose a height.
5.1.1 Analytical Balance
5.1.2 Glass ware
5.1.3 Magnetic Stirrer
5.1.4 pH Meter
5.1.5 Test Tubes
5.1.6 Titration Flask 250 ml
5.1.7 Viscosity meter (Viscometer)
5.1.8 Volumetric Flask 100 ml
5.2 Chemicals / Reagents
- Ethanol Absolute
- Ethyl Acetate
- Ethanol 96%
- Sodium Chloride
- 1 N Sodium Hydroxide
- Hydrochloric Acid 1N & 0.5M
- 0.1M Potassium Hydroxide
- Bromine Water R
- Phenolphthalein Solution R
- 0.1M Potassium Hydroxide VS
- Acetic anhydride solution R1
- 0.05M Potassium Hydroxide Ethanolic
- Phenolphthalein Solution R1
- 0.01M Sodium Thiosulphate
- Saturated Potassium Iodide
5.3 Physical Analysis
5.3.1 Test of Physical Form:
Check the sample and confirm its physical form; it should be clear oily liquid.
5.3.2 Test of Colour:
Confirm the colour of the sample with naked eye; it should be yellowish or brownish yellow.
5.3.3 Test of solubility:
It should be miscible with water, with absolute ethanol, with ethyl acetate and with methanol.
5.3.4 Acid Value:
Take 5 gm of sample and add 25 ml of a mixture of equal volume of ethanol (96%) and ether that has been neutralized with 0.1M Potassium Hydroxide VS using 0.5 ml of Phenolphthalein Solution R1 as indicator. When the substance has completely dissolved, titrate with 0.1M Potassium Hydroxide VS, shaking constantly until a pink color that persists for at least 15 seconds is produced.
Calculate the acid value from the expression: 5.610 x V/W
Where “V” is the volume, in ml, of Potassium Hydroxide solution required and “W” is the weight, in gm, of substance taken.
5.3.5 Test of Viscosity:
Take 300 ml sample and cool to 25C and then measure its viscosity by using viscometer.
Spindle No. 1
Note the reading on viscometer in mPa s the viscosity should be between
300 – 500 mPa s.
5.3.6 Relative Density:
a) Cool the liquid and water at 20oC.
b) Weigh a dry pycnometer (X gm)
c) Fill the pycnometer with water and weigh (Y gm)
d) Dry and fill the same pycnometer with liquid and weigh (A gm)
Weight of Water = Y – X = A
Weight of Liquid= Z – X = B
d20 = B/A
d20 = Relative Density of Liquid when temperature of both liquid and water is 20oC.
It should be 1.062 to 1.090.
5.3.7 Hydroxyl value
Take about 2.0 g accurately weighed in a 150-ml acetylation flask fitted with an air condenser and add 5.0ml of acetic anhydride solution R1. Heat for 1 hour in a water-bath, maintaining the level of the water about 2.5 cm above the level of the liquid in the flask throughout. Remove the flask and condenser, allow to cool and add 5 ml of water through the top of the condenser; if this causes cloudiness, add sufficient pyridine to produce a clear liquid, recording the volume added. Shake the flask, replace in the water-bath for 10 minutes, remove and allow to cool. Rinse the condenser and the walls of the flask with 5 ml of ethanol (96%), previously neutralised to phenolphthalein solution R1. Titrate with 0.5M ethanolic potassium hydroxide VS using 0.2 ml of phenolphthalein solution R1 as indicator. Repeat the operation without the substance being examined.
Calculate the hydroxyl value from the expression a + 28.05v/w where v is the difference, in ml, between the titrations, a is the acid value and w is the weight, in g, of the substance taken.
It should be 65 to 80
5.3.8 Peroxide value
Place 5.00 g (m g) of the substance being examined in a 250-ml conical flask fitted with a ground-glass stopper. Add 30 ml of a mixture of 2 volumes of chloroform and 3 volumes of glacial acetic acid. Shake to dissolve the substance and add 0.5 ml of saturated potassium iodide solution. Shake for exactly 1 minute then add 30 ml of water. Titrate with 0.01M sodium thiosulphate VS adding the titrant slowly with continuous vigorous shaking until the yellow colour is almost discharged. Add 5 ml of starch solution and continue the titration, shaking vigorously, until the colour is discharged (n1 ml of 0.01M sodium thiosulphate VS). Carry out a blank test under the same conditions (n2 ml of 0.01M sodium thiosulphate VS). The volume of 0. 01M sodium thiosulphate VS used in the blank titration must not exceed 0.1 ml.
it should be not more than 10.
5.3.9 Saponification value
Dissolve 35 to 40 g of potassium hydroxide in 20 ml of water and add sufficient ethanol (96%) to produce 1000 ml. Allow to stand overnight and pour off the clear liquid.
Weigh 2 g of the substance into a 200-ml flask, add 25 ml of the ethanolic solution of potassium hydroxide and boil under a reflux condenser for 1 hour, rotating the contents frequently. While the solution is still hot, titrate the excess of alkali with 0.5M hydrochloric acid VS using 1 ml of phenolphthalein solution R1 as indicator. Repeat the operation without the substance being examined.
Calculate the saponification value from the expression 28.05v/w
where v is the difference, in ml, between the titrations and w is the weight, in g, of substance taken.
It should be 45 to 55.
Use 15.0 ml of 0.5 M alcoholic potassium hydroxide and dilute with 50 ml of water R before carrying out the titration.
5.4 Chemical Analysis:
5.4.1 Identification Test of Tween-80:
a) Dissolve 0.5 g in water R at about 50 °C and dilute to 10 ml with the same solvent. The solution produces copious foam on shaking. Add 0.5 g of sodium chloride R and heat the solution to boiling. The resulting cloudiness disappears during cooling to about 50 °C.
b) Take 2 ml of a 5% w/v of sample in test tube and add 0.5 ml of bromine water; the colour of bromine should be decolorized.
6.0 Quality Record(s)/Form(s):
6.1 The following Quality Records shall be generated and managed in accordance with the Procedure for Control of Company Quality Records.
8.0 List of recipients
|Issued to||Issued on (Date / Signature)||Withdrawn on (Date / Signature)|
|Manager Quality Control|
|Manager Quality Assurance|
|Analyst (Work Station)|