Sitagliptin + Metformin HCl Tablet is a combination medication used to treat type 2 diabetes. Testing for this medication typically involves a combination of preclinical and clinical studies to evaluate its safety, efficacy, and pharmacokinetics.
Preclinical studies involve testing the medication in animal models to determine its toxicity, pharmacokinetics, and pharmacodynamics. These studies help researchers identify any potential safety concerns and determine appropriate dosages for humans’ SOP for Metformin Hydrochloride and Sitagliptin Phosphate Tablet Testing by UV and HPLC Method.
Clinical studies involve testing the medication in human subjects to evaluate its safety and efficacy. These studies are typically conducted in several phases, starting with small studies on unhealthy volunteers to determine the medication’s pharmacokinetics and dosing. If the medication is found to be safe and effective, larger studies are conducted in patients with type 2 diabetes to evaluate its clinical effectiveness.
During clinical studies, researchers also monitor adverse events and side effects, as well as drug interactions with other medications. They may also conduct additional tests to evaluate the medication’s effects on various metabolic parameters, such as blood glucose levels, HbA1c levels, and lipid profiles.
Once a medication is approved, post-marketing surveillance may also be conducted to monitor for any potential long-term safety concerns or adverse events.
Overall, testing for Sitagliptin + Metformin HCl Tablet is a rigorous process that involves multiple stages of evaluation to ensure its safety and effectiveness for treating type 2 diabetes.
PURPOSE:
To describes the procedure for analysis at in-process and finished stage of Sitagliptin + Metformin HCl Tablet.
SCOPE:
This SAP gives detailed outline for the finished product analysis of Sitagliptin + Metformin HCl 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 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
- pH meter
- Moisture analyzer
- Mortar and pestle
- Spatula
- Filter Paper
- Magnetic Stirrer & Hot Plate
- Sonicator
- Vacuum Pump
- Glassware
- Methanol HPLC Grade
- Hydrochloric acid
- Distilled water
PROCEDURE:
FINAL MIX
SOP for Metformin Hydrochloride and Sitagliptin Phosphate Tablet Testing by UV and HPLC Method
Description:
A white 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 major peak of the sample preparation corresponds to that 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%) Compression Weight of Powder/Tablet: 1200 mg /tablet
Procedure:
Buffer Solution: Dissolve 2.720 g of KH2PO4 in 1000 mL of water. Adjust with phosphoric acid to a pH of 4.30
Mobile Phase: Methanol and Buffer Solution (45:55)
Diluent: Methanol and Buffer Solution (45:55)
Standard Stock Solution:
0.321 mg/mL of Sitagliptin Phosphate Monohydrate WS ͠= 0.25 mg/mL of Sitagliptin and 5 mg/mL Metformin HCl WS. Take 32.1 mg of Sitagliptin Phosphate Monohydrate and 500mg of Metformin HCl dissolved in Diluent and make up the volume to 100mL with Diluent. Shake, and sonicate for 5 minutes to facilitate dissolution
Standard Solution:
0.0642mg/mL of Sitagliptin Phosphate Monohydrate WS and 1 mg/mL Metformin HCl WS. Take 20mL from standard stock solution and make up the volume to 100mL with Diluent. Pass through a suitable filter.
Sample Stock solution: Nominally, finely powder NLT5 tablets, transfer powder, equivalent to 0.50mg/mL of Sitagliptin and10 mg/mL Metformin HCl prepared as follows; Finally, powdered tablets equivalent to 50mg of Sitagliptin and 1000mg of Metformin HCl dissolved in Diluent and make up the volume to 100mL with Diluent.
Shake by mechanical mean for 10 minutes to homogenize and allow to soak until the sample is fully homogenized and allow any insoluble matter to settle.
Sample Solution: Pass a portion of the sample stock solution through a suitable filter of 0.45um pore size. Transfer the appropriate quantity of supernatant to a suitable volumetric flask to obtained nominally 0.050 mg/mL of Sitagliptin and 1.0 mg/mL Metformin HCl dissolved in Diluent.
Take 10mL from Sample stock solution and make up the volume to 100mL with Diluent.
Pass through a membrane filter of 0.45-µm pore size.
Chromatographic Conditions:
Detector: UV 260 nm
Column: 4.6-mm x 25-cm; packing L1
Column Temperature: 30±1°C
Flow Rate: 0.9 mL/min
Injection Volume: 20 µL
SYSTEM SUITABILITY:
Samples:
Standard solution and system suitability solution
Tailing Factor: NMT 2.0 for the Sitagliptin and Metformin HCl peak, standard solution.
Theoretical Plates: Not less than 1500
Relative standard deviation: NMT2.0 %standard solution.
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 Metformin HCl & Sitagliptin: 90%-110%of the labeled amount)
Alternate Method By UV
Sitagliptin
Standard Solution:
0.65 mg/mL Sitagliptin phosphate monohydrate WS = 0.5 mg/mL Sitagliptin
Take 65mg of Sitagliptin phosphate monohydrate equivalent to 50 mg of Sitagliptin dissolved in methanol and make up the volume to 100mL with methanol. Shake, and sonicate for 5 minutes to facilitate dissolution
Take 5 mL from standard stock solution and make up the volume to 25mL with methanol.
Sample solution:
0.5 mg/mL Sitagliptin prepared as follows.
Take compression weight of finally grinded granules and dissolved in methanol and make up the volume to 100mL with methanol. 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 5 mL of filtrate to a 25mL volumetric flask make up volume with methanol. Take reading at UV spectrophotometer at 272 nm using methanol as blank.
Calculations:
Sample Absorbance x Standard Concentration x Ave. Weight of Tablet x Potency
Standard Absorbance x Sample Concentration x Label Claim
(Limit: Sitagliptin: 90%-110%of the labeled amount)
Metformin HCl Standard Solution:
1 mg/mL Metformin HCl WS:
Take 100mg of Metformin HCl dissolved in water and make up the volume to 100mL with water. Shake, and sonicate for 5 minutes to facilitate dissolution
Take 1mL from standard solution and make up the volume to 100mL with water
Sample solution:
1 mg/mL Metformin HCl prepared as follows.
Take compression weight of finally grinded granules and equivalent to 100 mg of metformin HCl dissolved in water and make up the volume to 100mL with water. Shake by mechanical mean for10 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 1 mL of filtrate to a 100 mL volumetric make up volume with water. Take reading at UV spectrophotometer at 236 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: Metformin HCl: 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 = (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.
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
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 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 Test:
0.025 M Sodium Chloride:
1.4625g/L of Sodium chloride dissolved in water, make up volume 1000mL with water.
USP Apparatus: USP Apparatus II Peddle
Medium: 900mL (0.025 M NaCl)
Speed: 75 rpm
Time: 30 Minute
Medium Temperature: 37º ± 0.5º
Recommended Sampling Time: Sitagliptin and Metformin HCl: 30 Minute
Diluent: Dissolution medium
Sitagliptin as Phosphate monohydrate & Metformin HCl:
Mobile phase, System suitability solution and System suitability:
Proceed as directed in the assay.
Chromatographic Conditions:
Detector: UV 260 nm
Column: 4.6-mm x 25-cm; packing L1
Column Temperature: 30±1°C
Flow Rate: 0.9 mL/min
Injection Volume: 20 µL
Standard stock solution:
Take 35.7 mg of Sitagliptin Phosphate Monohydrate and 555 mg Metformin HCl in volumetric flask add 5mL dissolution medium and make up the volume 50mLwith dissolution medium. Shake, and sonicate to facilitate dissolution.
Standard solution:
Take 5mL from standard stock solution in dissolution medium and make up the volume to 50mL. Pass through a suitable filter.
Sample solution: Take 10 mL of sample Pass through a suitable filter and use.
Calculations:
Sample AUC x Standard Concentration x Potency
Standard AUC x Sample Concentration
Tolerance:
The amount of drug dissolve in solution for each tablet is not less Than 80% Q of the amount stated on the label for Sitagliptin and Metformin HCl at 30 minutes.
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 Weight variation
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