March 2, 2023 by KBS.

Trimetazidine Hydrochloride MR 35 mg Tablet assay testing is a process to determine the potency of the active ingredient in the medication. This Trimetazidine Hydrochloride MR 35mg Tablet Testing Procedure SOP ensures that the medication contains the correct amount of the active ingredient, trimetazidine hydrochloride, which is necessary for it to be effective.

Table of Contents

PURPOSE:

To describe the procedure for analysis at in-process and finished stage of Trimetazidine Hydrochloride MR 35 mg Tablet.

SCOPE:

This SAP gives a detailed outline for the finished product analysis of Trimetazidine Hydrochloride MR 35 mg 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 procedure.
It is the responsibility of QC Manager to assist and ensure Testing Procedure as per SAP and to make certain that this SAP is followed in its entirety, reviewed regularly and revised as necessary.

REFERENCE:

INNOVATORS SPECIFICATIONS

MATERIAL AND EQIUPMENT:

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
monobasic sodium phosphate
phosphoric acid
sodium dodecyl sulfate
Acetonitrile
Distilled water

PROCEDURE:

FINAL MIX.

Assay testing typically involves a variety of analytical techniques such as high-performance liquid chromatography (HPLC), gas chromatography (GC), or titration methods to quantitatively determine the amount of active ingredient in the medication.

During the assay testing of Trimetazidine Hydrochloride MR 35 mg Tablet, a sample of the medication is extracted and prepared for analysis. The sample is then analyzed using the chosen analytical technique to determine the amount of trimetazidine hydrochloride present in the tablet. The results are compared with the labeled amount of active ingredient to ensure that it falls within the acceptable range of variation.

Assay testing is an essential quality control measure that ensures the safety and efficacy of medications. It helps to guarantee that patients receive the correct dose of medication, which is crucial for treating their medical conditions effectively.

Description:

white powder.

Trimetazidine Hydrochloride MR 35mg Tablet Testing Procedure SOP

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:

A- The spectrum of the sample preparation corresponds to spectrum of the standard preparation as obtained in the assay.
B- The retention time of the major peak of the Sample solution corresponds 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: 90%-110%)Compression Weight of Powder/Tablet:……mg /tablet

Procedure:

Assay Weigh accurately not less than 20 tablets of Trimetazidine Hydrochloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 3 mg of trimetazidine hydrochloride (C14H22N2O3.2HCl), add about 15 mL of a mixture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1), and treat with ultrasonic waves for10 minutes. Then shake for 10 minutes, add the mixture of0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1) to make exactly20 mL, and centrifuge. To exactly 5 mL of the supernatant liquid add exactly 5 mL of the internal standard solution and 0.1 mol/L hydrochloric acid TS to make 50mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of trimetazidine hydrochloride for assay (separately determine the water <2.48>in the same manner as Trimetazidine Hydrochloride), and dissolve in the mixture of 0.1 mol/L hydrochloric acid TS and ethanol(99.5) (1:1) to make exactly 200 mL. To exactly 5 mL of this solution add exactly 5 mL of the internal standard solution and 0.1mol/L hydrochloric acid TS to make 50mL, and use this solution as the standard solution. Perform the test with10 mL each of the sample solution and standard solution as directed under Liquid Chromatography<2.01>according to the following conditions, and calculate the ratios, QT and QS, of the peak area of trimetazidine to that of the internal standard.
Amount (mg) of trimetazidine hydrochloride
(C14H22N2O3.2HCl) ＝MS × QT/QS × 1/10
MS: Amount (mg) of trimetazidine hydrochloride for assay taken, calculated on the anhydrous basis

Operating conditions

Detector: An ultraviolet absorption photometer (wavelength:230 nm).
Column: A stainless steel column 4.6 mm in inside dia meter and 15 cm in length, packed with octadecylsilane silica gel for liquid chromatography (5 mm in particle diameter).
Column temperature: A constant temperature of about40C.
Mobile phase: A mixture of 0.05 mol/L potassium dihydrogen phosphate TS (pH 3.0) and methanol (17:3).
Flow rate: Adjust so that the retention time of trimetazidine is about 7 minutes.
Limit: Trimetazidine dihydrochloride: 90%-110%of the labeled amount

Alternate Method:

Standard Solution:

Take 35 mg of Trimetazidine dihydrochloride WS dissolved in 10mL of 0.1 normal hydrochloric acid shake, then add 0.1 normal hydrochloric acid and make up the volume to 100mL with 0.1 normal hydrochloric acid. 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 0.1 normal hydrochloric acid.

Sample solution:

Take weight of finally grinded granules equivalent to 35mg of Trimetazidine dihydrochloride dissolved in 10mL of 0.1 normal hydrochloric acid shake, then add 0.1 normal hydrochloric acid and make up the volume to 200mL with 0.1 normal hydrochloric acid. 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 2mL of filtrate to a 25mL volumetric flask make up volume with 0.1 normal hydrochloric acid. Take reading at UV spectrophotometer at 233 nm using 0.1 normal hydrochloric acid as blank.

Analytical wavelength: 233 nm

Calculations:
Sample Absorbance x Standard Concentration x Ave. Weight of Tablet x Potency
Standard Absorbance x Sample Concentration x Label Claim

Limit: Trimetazidine dihydrochloride: 90%-110%of the labeled amount

Uniformity of Dosage Units: (By Weight Variation)
Weigh 20 tablets individually and calculate the average weight as:
Average weight = (WeightOf 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.

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/W1 x 100

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 TEST:

Preparation of 0.05M Phosphate Buffer at 6.8:

Dissolve 136.1g of Potassium dihydrogen phosphate and 17.92g of sodium hydroxide pellets in an appropriate container. Add 2L of water and stir to dissolve. Transfer to a 20-L flask and dilute to volume with water. Mix. Record the pH of the dissolution medium. Adjust the pH to 6.8 ± 0.05 with 85% m/m orthophosphoric acid or with a 1N sodium hydroxide solution. Degas the dissolution medium prior to use.

USP Apparatus: USP Apparatus II Peddle
Medium: 0.05M Phosphate Buffer at 6.8
Speed: 50 rpm± 2rpm
Time: 8 hours
Volume: 500mL± 15mL
Medium Temperature: 37º ± 0.5º
Recommended Sampling Time: 0,1, 2, 8 hours
Number of vessels per tests: 6 (for Level 1)
Number of Units per vessel: 1
Medium Replacement: None
Diluent: Dissolution medium

Standard solution:

Take 70.0 mg of Trimetazidine dihydrochloride WS in 1000mL volumetric flask. Dissolve and dilute to volume with dissolution medium. Mix for 30min using a magnetic stirrer.

Sample solution:

Withdraw the initial sampling aliquot, then drop one tablet to each vessel. After each sampling time, remove a 10mL aliquot of medium from each vessel. Filter the sample solution through a 10µm polypropylene filter.

Instrumental Conditions
(See Ultraviolet-Visible Spectroscopy (857)
Mode: UV
Absorbance wavelength: 267 nm
Reference wavelength: 300nm

Calculations:
C(i)= (Au/As) x Cs x V x (1/L) x 100
Au= Absorbance of sample
As = Absorbance of WS
Cs= concentration in mg/ml of the working standard solution.
V = volume in ml of medium (900mL),
L = label claim (mg/Tablet).
Calculate the percentage of the labeled amount (Q,) of Trimetazidine dihydrochloride dissolved at each time point (i) in the Buffer stage medium:

Result 1 = C1 x V x (l/L) x 100
Result 2 = {[C2 x (V- Vs)] ± (C1 x Vs)} x (l/L) x 100
Result 3= ({C3 x [V – (2 x Vs)]}± [(C2± C1) x Vs]) x (l/L) x 100

Ri= concentration of Trimetazidine dihydrochloride in the Buffer stage medium in the portion of sample withdrawn at time point i (mg/mL)
V= volume of the Buffer stage medium, 1000 mL
L=Label claim (mg/Tablet)
Vs= volume of the Sample solution withdrawn from the Buffer stage medium (mL)

Tolerances
Buffer stage: See Table 1.

Table 1

Time Point (I)	Time in hour	Amount Dissolved
1	1	15%-35%
2	2	40%- 60%
3	4	70%- 95 %
4	8	NLT 80%