It is recommended to follow established protocols, official pharmacopoeial methods, and Development of a Simple HPLC Method for the Determination of Azithromycin in Tablets or validated procedures during the development and validation of the HPLC method for the determination of Azithromycin in tablets.
PURPOSE:
To describe the procedure for analysis at the in-process and finished stage of the Azithromycin Tablet.
SCOPE:
This SAP gives a detailed outline for the finished product analysis of Azithromycin tablet and will cover In process testing activities on a physical, chemical & instrumental basis.
RESPONSIBILITY:
QC Analyst is responsible for physical/chemical testing and preparing standard analytical procedures.
It is the responsibility of the QC Manager to assist and ensure the Testing Procedure as per SAP and to make certain that this SAP is followed in its entirety, reviewed regularly, and revised as necessary.
REFERENCE:
USP 44, NF 39
MATERIAL AND EQIUPMENT:
- HPLC
- Dissolution apparatus
- Friability apparatus
- Hardness apparatus
- Disintegration Tester
- Vernier Caliper
- Analytical Balance
- pH meter
- Moisture analyzer
- Mortar and pestle
- Spatula
- Filter Paper
- Magnetic Stirrer & Hot Plate
- Sonicator
- Vacuum Pump
- Glassware Ammonium Acetate
- Triethylamine
- Acetonitrile HPLC Grade
- Methanol HPLC Grade
- Distilled water
- Cupric sulfate
- L-isoleucine
- Ammonium Acetate
PROCEDURE:
FINAL MIX
Development of a Simple HPLC Method for Determination of Azithromycin in Tablet
Description:
White color powder.
Procedure: Take 2.0 g of the test sample in a watch glass and observe visually with black background. Check the appearance of color, nature and any visible foreign particles.
Identification: (By HPLC)
The retention time of the major peak in the chromatogram of the assay sample preparation corresponds to that in the chromatogram of the standard preparation as obtained in the assay.
Loss on Drying: (By Moisture Analyzer)
Procedure:
Turn on instrument 20 minute before the test. Take 2 g powder spread uniformly on Moisture Analyzer plate. Close the lid and press start button.
NMT 5.0% when determined by LOD Apparatus, Use 2.0 g of powder.
Assay: (Limit: 90%-110%)
Procedure:
Buffer: Dissolve 4.6 g of monobasic potassium phosphate anhydrous in 900 mL of water. Adjust with 1 N sodium hydroxide to a pH of 7.5, and dilute with water to 1 L.
Mobile phase: Acetonitrile and Buffer.
Standard solution: 1 mg/mL of USP Azithromycin RS in Mobile phase. Sonicate and shake as needed to dissolve.
Sample solution: Nominally 1 mg/mL of azithromycin in Mobile phase from NLT 20 Tablets, finely powdered. Sonicate and shake as needed to dissolve.
Chromatographic system
Mode: LC
Detector: UV 210 nm.
Column: 4.6mm x 25-cm; 5-µm packing L1
Temperatures: Column: 50°C
Flow rate: 2.0 mL/min
Injection volume: 100 µL
System Suitability:
Samples: Standard solution
Tailing Factor: NMT2.0.
Relative standard deviation: NMT 2.0%.
Calculations:
Average Sample AUC x Standard Concentration x Ave. Weight of Tablet x Potency
Average Standard AUC x Sample Concentration x Label Claim x Factor
(Limit for Azithromycin: 90%-110%of the labeled amount)
Hardness Test & Dimensions:
Perform the hardness test on 10 tablets and take the average. Power on the instrument and regulate zero adjustment and preset the ‘’HARDNESS’’ and “THICKNESS” mode. Place the tablet on the test plate, begin testing and read the hardness thickness and diameter. Clean the shattered tablet and print out test results, perform the test on 10 tablets and take the average.
Hardness Standard Limit: NMT 10 Kg
Friability Test:
For tablets with a unit mass equal to or less than 650 mg, take a sample of whole tablets corresponding to 6.5 g. For tablets with a unit mass of more than 650 mg, take a sample of 10 whole tablets. The tablets should be carefully dedusted prior to testing. Accurately weigh the tablet sample, and place the tablets in the drum. Rotate the drum 100 times, and remove the tablets. Remove any loose dust from the tablets as before, and accurately weigh. Generally, the test is run once. If obviously cracked, cleaved, or broken tablets are present in the tablet sample after tumbling, the sample fails the test. If the results are doubtful or if the weight loss is greater than the targeted value, the test should be repeated twice and the mean of the three tests determined. A maximum mean weight loss from the three samples of not more than 1.0% is considered acceptable for most products.
% FRIABILITY= W1-W2 x 100/W1
Disintegration Test:
Place the beaker filled with water into the beaker stand inside the bath and. Switch on the instrument, set the temperature at 37ºC± 2º C and wait till the temperature of the beaker reaches to the 37ºC± 2º C. Engage the basket on basket hook put the tablets in 6 tubes individually and place the discs then start the oscillations and timer. Note the disintegration time of all the six tablets.
Note the Disintegration time of 1st tablet to the last tablet.
Disintegration Time: NMT 15 minutes at 370C ± 20C
Acceptance criteria of disintegration for tablets:
If 1 or 2 tablet fail to disintegrate completely repeat the test on 12 additional tablets, not less than 16 of the total of 18 tablets tested disintegrate completely.
Leak Test:
Take 06 blisters of tablet 03 filled and 03 empty. Dip these blisters in the bowl of leak test apparatus containing colored water. Create vacuum 250 mmHg and hold for one minute. Release vacuum and check the penetration of water inside the blisters. All blisters should be free of water.
DISSOLUTION (711 )
Medium: pH 6.0 phosphate buffer; 900 mL
Apparatus 2: 75 rpm
Time: 30 min
Solution A: 4.4 mg/mL of dibasic potassium phosphate and0.5 mg/mL of sodium l-octanesulfonate; adjusted withphosphoric acid to a pH of 8.20 ± 0.05
Mobile phase: Acetonitrile, methanol, and Solution A (9:3:8)
Diluent: 17.5 mg/mL of dibasic potassium phosphate. Adjust with phosphoric acid to a pH of 8.00 ± 0.05. Prepare a mixture of this solution and acetonitrile (80:20).
Standard stock solution: Dissolve USP Azithromycin RS in Medium to obtain a solution with a known concentration of about (L/l 000) mg/mL, where L is the label claim in mg/Tablet.
Standard solution: Dilute the Standard stock solution with Diluent to obtain a solution with a known concentration of about (L/2000) mg/mL, where L is the label claim in mg/Tablet.
Sample solution: Pass a portion of the solution under test through a suitable filter of 0.45um pore size. Dilute a portion of the filtrate with Diluent to obtain a solution with a theoretical concentration of about (L/2000) mg/mL, where L is the label claim in mg/Tablet, assuming complete dissolution.
Chromatographic system
(See Chromatography (621), System Suitability.)
Mode: LC
Detector: UV 210 nm
Column: 4.6-mm x 15-cm; 5um packing L1
Column temperature: 50°
Flow rate: 1.5 mL/min
Injection volume: 50 uL
System suitability
Sample: Standard solution
Suitability requirements
Tailing factor: NMT 2.0
Relative standard deviation: NMT2.0%
Analysis
Samples: Standard solution and Sample solution Calculate the percentage of the labeled amount of azithromycin (C38H72N2O12) dissolved:
Result = (Au/As) x (Cs/L) x (M r1/M r2) x Vx 100
Au =AUC of the Sample solution
As = AUC of the Standardsolution
Cs = concentration of the Standardsolution (mg/mL)
L = Label claim (mg/Tablet)
Mr1 = molecular weight of azithromycin,
Mr2 =molecular weight of azithromycindehydrate,
V = volume of Medium, 900 mL
Tolerances: NLT 80% (Q) ofthe labeled amount of azithromycin (C38H72N2O12) is dissolved.
CONTENT UNIFORMITY TEST: (By HPLC)
Proceed as directed in the Assay of Tablet.
Calculation of Acceptance Value (AV):
Limits:
L1= ≤ 15 (For 10 Unit)
L2= ≤ 25 (For 30 Unit)
General Formula = M-x̄ ±ks