It is important to note that the specifics of the assay method, including the instrumentation, chromatographic conditions (for HPLC), wavelength selection (for UV spectrophotometry), and sample preparation steps, may vary depending on the official compendial method, published literature, or specific requirements of your laboratory. Hplc and UV Testing Method of Montelukast Sodium in Tablet It is recommended to consult relevant references or validated methods for detailed experimental conditions.
PURPOSE:
To describe the procedure for analysis at the in-process and finished stage of Montelukast Sodium Tablet.
SCOPE:
This SAP gives a detailed outline for the finished product analysis of the Montelukast Sodium Tablet and will cover In process testing activities on 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 43, NF 38
MATERIAL AND EQIUPMENT:
- UV Spectrophotometer
- HPLC
- Dissolution apparatus
- Friability apparatus
- Hardness apparatus
- Disintegration Tester
- Vernier Caliper
- Analytical Balance
- Moisture analyzer
- Mortar and pestle
- Spatula
- Filter Paper
- Magnetic Stirrer & Hot Plate
- Sonicator
- Vacuum Pump
- Glassware
- Methanol AR Grade
- Sodium acetate AR Grade
- Sodium hydroxideAR Grade
- Acetonitrile AR Grade
- Distilled water
PROCEDURE:
FINAL MIX
Hplc and UV Testing Method of Montelukast Sodium in Tablet
Description:
A white to off white granular 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 UV SPECTROPHOTOMETER and HPLC)
The spectrum of the sample preparation corresponds to spectrum of the standard preparation as obtained in the assay.
The retention time of the major peak of the sample solution correspond to that of the standard solution 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 3.0% when determined by LOD Apparatus, Use 2.0 g of powder.
Assay:
(Limit: Montelukast Sodium Tablets contain Montelukast Sodium equivalent to NLT 92.5% and NMT 107.5% of the labeled amount of montelukast (C35H36C1NO3S)
Compression Weight of Powder/Tablet:
Procedure:
Diluent: Methanol and water (3:1)
Solution A: 0.2% (v/v) Trifluoroacetic acid in water
Solution B: Methanol and acetonitrile (3:2)
Mobile phase: See Table 1.
Table 1
| Time (Minute) | Solution A (%) | Solution B (%) |
| 0 | 48 | 52 |
| 5 | 45 | 55 |
| 12 | 45 | 55 |
| 22 | 25 | 75 |
| 23 | 25 | 75 |
| 25 | 18 | 52 |
| 30 | 48 | 52 |
Standard solution: 0.52 mg/mL of USP Montelukast Dicyclohexylamine RS in Diluent
System suitability solution: Transfer 10 mL of the Standard solution to a clear 1O-mLvolumetricflask, add 4 uL of hydrogen peroxide, and mix well. Expose the flask for at least 4 h to ambient light or 10 min to a 4 klx cool white light. [Note-Montelukast is partially converted to thecis-isomer under these conditions.]
Sensitivity solution: 0.52 ug/mL of USP Montelukast Dicyclohexylamine RS in Diluent from the Standard solution
Sample solution: Nominally 0.4 mg/mL of montelukast prepared as follows. Transfer a number of Tablets equivalent to 100 mg of montelukast to a suitable volumetric flask, add 70% of the flask volume of Diluent, and sonicate for 30 min. Shake for 30 min on a plat form shaker. Dilute with Diluent to volume and stir for 30 min.
Pass a portion through a suitable filter of 0.45-um pore size, discarding the first mL of filtrate. Use the filtrate.
Chromatographic system
(See Chromatography (621), System Suitability.)
Mode: LC
Detector: UV 255 nm
Columns
Guard: 3.0-mm x 4-mm; packing L11
Analytical: 4.6-mm x 10-cm; 3-um packing L11
Column temperature: 50°
Flow rate: 1.5 mL/min
Injection volume: 15 uL
Run time: 2 times the retention time of montelukast
System suitability
Samples: Standard solution, System suitability solution, and
Sensitivity solution
[Note- The relative retention times for the cis-isomer and montelukast are about 0.92 and 1.0,respectively.]
Suitability requirements
Resolution: NLT 1.5 between the cis-isomer and montelukast, System suitability solution
Relative standard deviation: NMT2% for five injections, Standard solution
Signal-to-noise ratio: NLT 10, Sensitivity solution
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of montelukast (C35H36C1NO3S) in the portion of Tablets taken:
Result = (Ru/Rs) x (Cs/Cu) x (Mr1/Mr2) x 100
Ru= peak response from the Sample solution
Rs=peak response from the Standardsolution
Cs = concentration of USP Montelukast Dicyclohexylamine RS in the Standard solution (mg/mL)
Cs= nominal concentration of montelukast in the Sample solution(mg/mL)
MrI =molecular weight of montelukast, 586.18
Mr2 = molecular weight of montelukast dicyclohexylamine, 767.50
Acceptance criteria: Montelukast as Sodium: 92.5%-107.5% of the labeled amount of montelukast (C35H36CINO3S) is dissolved.
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: Montelukast as Sodium: 92.5%-107.5%of the labeled amount
ALTERNATE METHOD By UV
Procedure:
Standard Solution:
Take 20.8 mg of Montelukast sodium WS dissolved in 0.1Nmethanolic HCl and make up the volume to 50mL with 0.1Nmethanolic HCl. Shake, and sonicate for 5 minutes to facilitate dissolution
Take 1 mL from standard stock solution and make up the volume to 50 mL with 0.1Nmethanolic HCl.
Sample solution:
Take 5 gram of sample grind them to fine powder. Take compression weight of finally grinded powder equivalent to 10 mg of Montelukast and dissolved in 0.1Nmethanolic HCl and make up the volume to 50 mL with 0.1Nmethanolic HCl. Shake by mechanical mean for 10 minutes to disperse and allow any insoluble matter to settle.
Pass a portion of the sample stock solution through a suitable filter of 0.45um pore size.
Transfer the 1 mL of filtrate to a 25 mL volumetric flask make up volume with 0.1Nmethanolic HCl. Take reading at UV spectrophotometer at 394 nm using 0.1Nmethanolic HCl as blank.
Calculations:
Sample Absorbance x Standard Concentration x Ave. Weight of Tablet x Potency
Standard Absorbance x Sample Concentration x Label Claim x Factor
CORE TABLET
Compression weight/Tablet:
Uniformity of Dosage Units: (By Weight Variation)
Weigh 20 tablets individually and calculate the average weight as:
Average weight = (Weight Of 20 Tablets)/20
Weigh all these tablets individually and mark among these weights maximum (WMax) & minimum (WMin) weight tablets. Calculate the maximum and minimum variation in percent as:
Maximum% age variation =((WMax) – (WAvg.))/WAvg×100
Minimum % age variation ((WMin) – (WAvg.))/WAvg×100
Standard Limit: ± 7.50%
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: Not more than 10.00 Kg
Thickness Standard Limit:
Length/Diameter Standard Limit:
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
FILM COATING
Weight/Tablet:
Uniformity of Dosage Units: (By Weight Variation)
Proceed as directed in Compression stage
Standard Limit: ± 7.50%
Hardness Test & Dimensions: Proceed as directed in Compression stage
Hardness Standard Limit: Not more than 10.00 Kg
Thickness Standard Limit:
Length/Diameter Standard Limit:
Disintegration Test:
Proceed as directed in Compression stage
Disintegration Time: NMT 30 minutes at 370C ± 20C
Identification: (By UV SPECTROPHOTOMETER and HPLC)
The spectrum of the sample preparation corresponds to spectrum of the standard preparation as obtained in the assay.
The retention time of the major peak of the sample solution correspond to that of the standard solution as obtained in the assay.
DISSOLUTION TEST:
USP Apparatus: USP Apparatus II Peddle
Medium: 0.5% (w/v) Sodium dodecyl sulfate in water;900 mL. Do not deaerate.
Speed: 50 RPM
Time: 20 Minute
Medium Temperature: 37º ± 0.5º
Recommended Sampling Time: Montelukast: 20 Minute
Solution A: 0.2% (v/v) Trifluoroacetic acid in water
Solution B: 0.2% (v/v) Trifluoroacetic acid in acetonitrile
Standard-stock solution: 0.35 mg/mL of USP Montelukast Dicyclohexylamine RS in methanol (equivalent to0.27 mg/mL of montelukast)
Standard solution: (L/900) mg/mL of montelukast in Medium from the Standard stock solution, where L is the label claim in mg/Tablet of montelukast
Sample solution: Pass a portion of the solution under testthrough a suitable filter or centrifuge to obtain a clear solution.
Chromatographic system
(See Chromatography (621), System Suitability.)
Mode: LC
Detector: UV 389 nm
Column: 3.0-mm x 10-cm; 5-um packing L11
Column temperature: 50°
Flow rate: 0.9 mL/min
Injection volume: 20 uL
Run time: 1.5 times the retention time of montelukast
System suitability
Sample: Standard solution
Suitability requirements
Tailing factor: NMT 1.5
Relative standard deviation: NMT 2%
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of montelukast (C35H36CINO3S) dissolved:
Result =(Ru/Rs) x Cs x V x (l/L) x 100
Ru=peak response from the Sample solution
Rs= peak response from the Standard solution
Cs =concentration of montelukast in the Standard solution (mg/mL)
V = volume of Medium, 900 mL
L =label claim (mg/Tablet)
Calculations:
Sample Absorbance x Standard Concentration x Potency
Standard Absorbance x Sample Concentration x Factor
Tolerance:
Not less Than 80% (Q) of the labeled amount of Montelukast (C35H36CINO3S) is dissolved,
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.
CONTENT UNIFORMITY TEST: (By HPLC)
Proceed as directed in the Compressed Tablet.
Calculation of Acceptance Value (AV):
Limits:
L1= ≤ 15 (For 10 Unit)
L2= ≤ 25 (For 30 Unit)
General Formula = M-x̄ ±ks
Acceptability Constant = (k)
Standard Deviation (SD) = s
Number of Units = (n)
Reference Value = M
Mean of Individual Content = x̄