Analyzing Mecobalamin 500mcg tablets at both the in-process and finished stages is essential to ensure the quality, safety, and efficacy of the product. The analysis involves various tests and quality control measures. Below is a general Procedure for Analysis at the In-process and Finished Stage of the Mecobalamin 500mcg Tablet
Have the data and results reviewed by authorized personnel, including quality control and quality assurance teams, before approving the product for release or further processing?
Always follow approved procedures and protocols specific to your laboratory and adhere to Good Manufacturing Practices (GMP) to ensure the accuracy and reliability of the analysis. This procedure is a general guideline and may need to be adapted based on the specific requirements and regulations applicable to your location or company.
PURPOSE:
To describe the procedure for analysis at the in-process and finished stage of the Mecobalamin 500mcg Tablet.
SCOPE:
This SAP gives a detailed outline for the finished product analysis of the Mecobalamin 500mcg 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:
JAPANESE PHARMACOPOEIA
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
- Distilled water
PROCEDURE:
FINAL MIX
Description:
Maroon 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 UV SPECTROPHOTOMETER)
Method of preparation:
Prepare as directed under Tablets, with Mecobalamin.
The spectrum of the sample preparation corresponds to spectrum of the standard preparation as obtained in the assay.
Conduct this procedure without exposure to light, using light-resistant vessels. To a quantity of powdered Mecobalamin Tablets, equivalent to 1 mg of Mecobalamin, add 10 mL of hydrochloric acid-potassium chloride buffer solution (pH 2.0), treat with ultrasonic waves, and add hydrochloric acid-potassium chloride buffer solution (pH 2.0) to make 20 mL. Centrifuge this solution, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.8 mm.
Determine the absorption spectrum of the filtrate as directed under Ultraviolet visible Spectrophotometry: It exhibits maxima between 262 nm and 266 nm, between 303 nm and 307 nm, and between 461 nm and 465 nm.
Conduct this procedure without exposure to light, using light-resistant vessels. To a quantity of powdered Mecobalamin Tablets, equivalent to 1 mg of Mecobalamin, add 10 mL of phosphate buffer solution (pH 7.0), treat with ultrasonic waves, and add phosphate buffer solution (pH 7.0) to make 20 mL. Centrifuge this solution, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.8 mm.
Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry it exhibits maxima between 264 nm and 268 nm, between 339 nm and 343 nm, and between 520 nm and 524 nm.
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: 92%-108%)Compression Weight of Powder/Tablet: …mg /tablet
Procedure:
Conduct this procedure without exposure to light, using light-resistant vessels.
Disintegrate 20 tablets of Mecobalamin Tablets with V/5 mL of water. Add methanol to make exactly V mL so that each mL contains about 50 µg of mecobalamin (C63H91CoN13O14P).
After shaking for 5 minutes, allow to stand for not less than 10 minutes.
Filter thus obtained supernatant liquid through a membrane filter with a pore size not exceeding 0.45 µm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution.
Separately, weigh accurately about 25 mg of Mecobalamin RS (separately determine the water <2.48>in the same manner as Mecobalamin), and dissolve in water to make exactly 100 mL.
To exactly 10 mL of this solution add methanol to make exactly 50 mL, and use this solution as the standard solution.
Perform the test with exactly 10 µL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>according to the following conditions, and determine the peak areas, AT and AS, of mecobalamin in each solution.
Amount (mg) of mecobalamin (C63H91CoN13O14P) in 1 tablet ＝MS × AT/AS × V/10000
MS: Amount (mg) of Mecobalamin RS taken, calculated on the anhydrous basis
Operating conditions:
Detector: An ultraviolet absorption photometer (wavelength: 266 nm).
Column: A stainless-steel column 4.6 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about 40oC.
Mobile phase: To 200 mL of acetonitrile add 800 mL of 0.02 mol/L phosphate buffer solution (pH 3.5), then add 3.76 g of sodium 1-hexane sulfonate to dissolve.
Flow rate: (Almost 0.95mL/minute) Adjust so that the retention time of mecobalamin is about 12 minutes.
System suitability:
System performance:
When the procedure is run with 10 µL of the standard solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of mecobalamin are not less than 3000 and 0.8 to 1.1, respectively.
System repeatability:
When the test is repeated 6 times with 10 µL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of mecobalamin is not more than 1.0%.
Limit: Mecobalamin: 92%-108%of the labeled amount
ALTERNATE METHOD BY UV Procedure for Analysis at the In-process and Finished Stage of the Mecobalamin 500mcg Tablet
Standard Solution:
Take 25 mg of mecobalamin WS dissolved in water and make up the volume to 50mL with water. Shake, and sonicate for 5 minutes to facilitate dissolution
Take 2 mL from standard stock solution and make up the volume to 50 mL with water.
Sample solution:
Take compression weight of finally grinded granules equivalent to 5mg (Compression weight x 10) of mecobalamin and dissolved in water and make up the volume to 250mL with water. 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.
Take reading at UV spectrophotometer at 268 nm using water as blank.
Calculations:
Sample Absorbance x Standard Concentration x Ave. Weight of Tablet x Potency
Standard Absorbance x Sample Concentration x Label Claim
Limit: Mecobalamin: 92%-108%of the labeled amount
CORE TABLET
Description:
White 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
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: ……
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: …..
Acceptance criteria of disintegration for tablets:
If 1 or 2 tablets fail to disintegrate completely repeat the test on 12 additional tablets, not less than 16 of the totals of 18 tablets tested disintegrate completely.
DISSOLUTION <6.10>
When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Mecobalamin Tablets is not less than 80%.
USP Apparatus: USP Apparatus II Paddle
Medium: 900mL water
Speed: 50 rpm
Time: 45 Minutes
Medium Temperature: 37º ± 0.5º
Conduct this procedure without exposure to light, using light-resistant vessels. Start the test with 1 tablet of Mecobalamin Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 µm.
Sample Preparation:
Discard first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, and add water to make exactly V- mL so that each mL contains about 0.28 ug of mecobalamin (C63H91CoN13O14P). Use this solution as the sample solution.
Standard Preparation:
Separately, weigh accurately about 28 mg of Mecobalamin RS (separately determine the water <2.48> in the same manner as Mecobalamin), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL.
Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution.
Perform the test with exactly 100 µL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>according to the following conditions, and determine the peak areas, AT (Area of test solution) and AS (Area of Standard solution), of mecobalamin in each solution.
Dissolution rate (z) with respect to the labeled amount of mecobalamin (C63H91CoN13O14P)＝MS × AT/AS× V-/V × 1/C × 9/10
MS: Amount (mg) of Mecobalamin RS taken, calculated on the anhydrous basis
C: Labeled amount (mg) of mecobalamin (C63H91CoN13O14P) in 1 tablet
Operating conditions:
Detector: An ultraviolet absorption photometer (wavelength: 264 nm).
Column: A stainless-steel column of 4.6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about 40OC.
Mobile phase: Adjust to pH 3.0 of a solution of 6.0 g of L-tartaric acid in 1000 mL of water with a solution of 14.3 g of disodium hydrogen phosphate dodecahydrate in 1000 mL of water. To 630 mL of this solution add 370 mL of methanol.
Flow rate: Adjust so that the retention time of mecobalamin is about 8 minutes.
System suitability:
System performance:
When the procedure is run with 100 µL of the standard solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of mecobalamin are not less than 3000 and not more than 1.5, respectively.
System repeatability:
When the test is repeated 6 times with 100 µL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of mecobalamin is not more than 2.0%.
Tolerance:
The amount of drug dissolve in solution for each tablet is not less Than 80% of the amount stated on the label for Mecobalamin in 45 minutes.
Containers and storage Containers: Tight containers. Storage—Light-resistant.
BLISTER STAGE:
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.