Fifth stage Third lab Advanced pharmaceutical analysis

Spectrophotometric Assay for determination of Metformin hydrochloride in pharmaceutical preparations

Objective:-

Determination the amount of Metformin hydrochloride content in a pharmaceutical product.

Metformin hydrochloride (glucophage). chemicals, 1,1 dimethyl biguanide hydrochloride (C4H12ClN5). It

is an oral antidiabetic drug that has been used in the treatment of non-insulin dependent diabetes
which improves control of glycemia primarily by inhibiting hepatic gluconeogenesis and glycogenolysis and
seems to ameliorate hyperglycemia by improving the peripheral sensitivity to insulin. And reducing GIT
glucose absorption and hepatic glucose production.

.
2
MWt= 165.6

Description: A white, odorless, crystalline powder, odorless and has a bitter taste, hygroscopic. It is freely soluble
in water, slightly soluble in alcohol, practically in soluble in acetone. Melting range: 225ºC and stored in an airtight
Container, protected from light.

Chemical Principle

The method is based on the oxidation of metformin by a known excess of sodium hypochlorite for
forming of a yellow-colored chromophore (β-diketone yellow- chromophore) having a maximum
absorbance at 420nm.

Procedure:

1-Reagents:

3% Sodium hypochlorite solution: was prepared by dilution 3 mL of sodium hypochlorite to 100

mL by distilled water.

2- Preparation of standard:

Metformin hydrochloride stock solution (100 ppm)

was prepared by dissolving 0.01 g of metformin hydrochloride in 100 ml distilled water in a

volumetric flask.

1
 Fifth stage Third lab Advanced pharmaceutical analysis

Then transfer (1.2, 2.5, 3.75, 5 mL) of the standard solution to 25ml flasks, and add of 2 ml Sodium
hypochlorite reagent. wait for 5 min; then make up the solution to the mark (25m1) with distilled
water.

3- Blank:

Transfer 2 m1 from 3% Sodium hypochlorite solution to 25m1 volumetric flask and make up to the
mark with D.W.

4- Preparation of sample:

Give amount of metformine HCl to prepare solution of unknown concentration, and then transfer into
25 m1 volumetric flask. And add 2 ml of sodium hypochlorite reagents, wait for 5 min; then make
up the solution to the mark (25m1) with distilled water.

5-Testing the Solutions.

1. Turn on the spectrophotometer.

2. Adjust the wavelength to max= 420 nm and record the absorbance (A).
3. Prepare a standard curve by plotting the (Abs) of standard solution (VS) concentration.
4. Calculate the metformin HCl content of the sample under test from the standard curve.

Required Questions
1. Give the structure of the final yellow-colored chromophore.
2. Based on the experimental results obtained, specify the operation range of Beer’s law?