HOT POST

50/recent/ticker-posts

Header Ads Widget

Assay Method for Metformin, Pioglitazone and Glimepiride in bulk samples and in tablet dosage forms

Glimepiride + Metformin + Pioglitazone is a combination of three antidiabetic medicines: Glimepiride , Metformin and Pioglitazone. They work by different mechanisms to provide better control of blood sugar when single or dual therapy is not effective. 
  1. Glimepiride is a sulfonylurea which works by increasing the amount of insulin released by the pancreas in order to lower the blood glucose. 
  2. Metformin is a biguanide. It works by lowering glucose production in the liver, delaying glucose absorption from intestines and increasing the body's sensitivity to insulin. 
  3. Pioglitazone is a thiazolidinedione which further increases insulin sensitivity.

Each Tablet Contains
Metformin Hydrochloride BP . . . . . . . . . . .500 mg 
Pioglitazone BP . . . . . . . . . . . . . . . 15mg
Glimepiride BP . . . . .  . . . . . . . . . . 1mg

Chromatographic Conditions:
Column : 150 mm × 4.6 mm, C18, 5 µm
Flow Rate : 1 mL / minute
Column Temperature : Ambient
Detection Wavelength : 254 nm
Injection Volume : 20 µL
Run time: 7 min.

Phosphate Buffer: 
7.0 grams of potassium dihydrogen ortho phosphate in 1000 mL water [HPLC Grade] pH adjusted with orthophosphoric acid. (pH: 3.0)

Mobile Phase:
Phosphate Buffer: Methanol (25: 75 v/v)

Preparation of Phosphate buffer: 
The buffer solution prepared by dissolving accurately weighed 7.0 grams of potassium dihydrogen ortho phosphate and transferred into a clean and dry 1000 mL volumetric flask, dissolved and diluted with 1000 mL water [HPLC Grade]. The final pH of the buffer is adjusted to 3.0 by using ortho phosphoric acid.

Preparation of mobile phase and diluent: 
The mobile phase was prepared by mixing 250 mL (25%) of the above buffer and 750 mL of methanol [HPLC Grade] (75%) and degassed in an ultrasonic water bath for 10 minutes. Then the resultant solution was filtered through 0.45 μ filter under vacuum filtration. The mobile phase is used as diluent.


Preparation of the metformin, pioglitazone and glimepiride standard and sample Solution:

For Bulk Samples
Preparation of stock solution: 
Weigh accurately 10 mg metformin, pioglitazone and glimepiride working standard and transferred into clean and dry 10mL, 100mL and 100mL volumetric flask respectively. About 7mL, 70mL and 70mL of diluent was added to the flask respectively and sonicated. Make up the volume up to the mark with the same diluent. 
From the above prepared Stock solution pipette out 1.0 mL of metformin, 0.3 mL of pioglitazone and 1.0 mL of glimepiride solution and transferred into a clean and dry 10 mL volumetric flask, add diluent up to the mark to get final concentration.


Preparation of sample solution:
Weigh accurately 10 mg metformin, pioglitazone and glimepiride powder to be examined and transferred into clean and dry 10mL, 100mL and 100mL volumetric flask respectively. About 7mL, 70mL and 70mL of diluent was added to the flask respectively and sonicated. Make up the volume up to the mark with the same diluent. 
From the above prepared Stock solution pipette out 1.0 mL of metformin, 0.3 mL of pioglitazone and 1.0 mL of glimepiride solution and transferred into a clean and dry 10 mL volumetric flask, add diluent up to the mark to get final concentration.

System Suitability: 
The tailing factor for the peaks due to metformin, pioglitazone and glimepiride in standard solution should not be more than 2.0. The Theoretical plates for the metformin, pioglitazone and glimepiride peaks in standard solution should not be less than 2000.


Calculation:


Where,
At = Average Area of Sample solution
As = Average Area of Standard solution
Ws = Weight of Standard
Wt= Weight of Sample
P = Potency of Standard


For Tablet Dosage
Preparation of stock solution: 
Weigh accurately 10 mg metformin, pioglitazone and glimepiride and transferred into clean and dry 10 mL, 100 mL and 100 mL volumetric flask respectively. About 7 mL, 70 mL and 70mL of diluent was added to the flask respectively and sonicated. Make up the volume up to the mark with the same diluent. 
From the above prepared Stock solution pipette out 1.0 mL of metformin, 0.3 mL of pioglitazone and 1.0 mL of glimepiride solution and transferred into a clean and dry 10 mL volumetric flask, add diluent up to the mark to get final concentration.

Preparation of sample solution:
Weigh tablet Powder equivalently 10 mg metformin, pioglitazone and glimepiride and transferred into a 10 mL clean and dry volumetric flask and add 7mL of diluent and sonicate to dissolve it completely and make up the volume with the same solvent. 
From above prepared stock solution pipette out 1.0 mL of solution and transferred into a clean and dry 10 mL volumetric flask, add diluent up to the mark to get final concentration.

System Suitability: 
The tailing factor for the peaks due to metformin, pioglitazone and glimepiride in standard solution should not be more than 2.0. The Theoretical plates for the metformin, pioglitazone and glimepiride peaks in standard solution should not be less than 2000.

Calculation:


Where,
At = Average Area of Sample solution
As = Average Area of Standard solution
Ds = Dilution for Standard Solution
Dt = Dilution for Sample Solution
Ws = Weight of Standard
Wt= Weight of Sample
P = Potency of Standard
ATW = Average Tablet Weight
L = Label Claim

Post a Comment

0 Comments

close