Gentamicin Sulfate

Gentamicin Sulfate

ABOUT:

Gentamicin Sulfate is the sulfate of a mixture of aminoglycoside substances having antibacterial activity produced by the growth of Micromonospora purpurea or Micromonospora echinospora.

PHYSICAL APPEARANCE:

Gentamicin Sulfate occurs as a white to light yellowish white powder. It is highly hygroscopic.

CHEMICAL FORMULA:

C21H43N5O7. xH2SO4

CHEMICAL STRUCTURE:

Gentamicin Sulfate
Gentamicin Sulfate

MOLECULAR WEIGHT:

477.60

IUPAC NAME:

(6R)-2-Amino-2,3,4,6-tetradeoxy-6-methylamino-6-methyl-α-D-erythrohexopyranosyl-(1→4)-[3-deoxy-4-C-methyl-3-methylamino-β-L-arabinopyranosyl-(1→6)]-2-deoxy-Dstreptamine sulfate

CONTENTS:

It contains not less than 590 mg (potency) and not more than 775 mg (potency) per mg, calculated on the dried basis. The potency of Gentamicin Sulfate is expressed as mass (potency) of gentamicin C1

SOLUBILITY:

  • Soluble in water.
  • Typically insoluble in ethanol. (96%)

IDENTIFICATION BY PPT. FORMATION:

Dissolve 50 mg of Gentamicin Sulfate in 5 mL of water, and add 0.5 mL of barium chloride TS: a white precipitate is formed.

COLOR OF SOLUTION:

Dissolve 1.0g of Gentamicin Sulfate in 10 mL of water, the solution is clear and colorless to pale yellow.

IDENTIFICATION BY THIN LAYER CHROMATOGRAPHY:

Dissolve 50 mg each of Gentamicin Sulfate and Gentamicin Sulfate Reference Standard in 10 mL of water, and use both of these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under thin layer-Chromatography. Spot 20 µL of the sample solution and standard solution on a plate of silica gel for thin layer-Chromatography. Separately, shake a mixture of chloroform, ammonia solution and methanol in ration of (2:1:1) in a separator, and allow the mixture to stand for more than 1 hour. To 20 mL of the lower layer so obtained add 0.5 mL of methanol, and use this as the developing solvent. Develop the plate with the developing solvent to a distance of about 17cm in a developing container with a cover, having an opening of about 20 mm2, and without putting a filter paper in the container, and air-dry the plate. Allow the plate to stand in iodine vapors: three principal spots obtained from the sample solution are the same with the corresponding spots obtained from the standard solution in color tone and the Rf value, respectively.

pH:

The pH of a solution obtained by dissolving 400 mg of Gentamicin Sulfate in 10 mL of water is between 3.5 and 5.5

LOSS ON DRYING:

Not more than 18.0% when 0.15g of Gentamicin Sulfate at pressure not exceeding 0.67 kPa and temperature 110°C for 3 hours. Handle the sample avoiding absorption of moisture.

RESIDUE ON IGNITION:

Should be not more than 1.0% when 1.0 gram is ignited.

SPECIFIC OPTICAL ROTATION:

+107°   to   +121° When 100mg of Gentamicin Sulfate is dissolved in 10ml of water.

QUANTITATIVE ASSAY (BIO-ASSAY):

Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics according to the following conditions.

Test organism Staphylococcus epidermidis ATCC

Agar media for seed and base layer

  • Glucose 1.0 g
  • Peptone 6.0 g
  • Meat extract 1.5 g
  • Yeast extract 3.0 g
  • Sodium chloride 10.0 g
  • Agar 15.0 g
  • Water 1000 mL

Mix all the ingredients, and sterilize. Adjust the pH of the solution so that it will be 7.8 to 8.0 after sterilization.

Agar medium for transferring test organisms.

Use the medium Agar media for seed and base layer in Medium for other organisms under Agar media for transferring test organisms.

Standard solutions

Weigh accurately an amount of Gentamicin Sulfate Reference Standard, equivalent to about 25 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 25 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 15°C or lower, and use within 30 days. Take exactly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 4 mg (potency) and 1 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively.

Sample solutions

Weigh accurately an amount of Gentamicin Sulfate, equivalent to about 25 mg (potency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 25 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 4 mg (potency) and 1 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.

STORAGE CONDITIONS:

It should be stored in tight and air-free containers and should be keep away from moisture area.

REFERENCES:

Japan Pharmacopia (JP XVII) Official monographs Page 983.

United State Pharmacopia (USP 41) Page 938.

Diclofenac Sodium

Diclofenac Sodium

DESCRIPTION:

Diclofenac Sodium is white or slightly yellowish, slightly hygroscopic, crystalline powder.

CHEMICAL FORMULA:

C14H10Cl2NNaO2

CHEMICAL STRUCTURE:

Diclofenac Sodium
Diclofenac Sodium

MOLECULAR WEIGHT:

318.1 Dalton

USE AS:

Cyclo-oxygenase inhibitor; analgesic; anti-inflammatory.

IUPAC NAME:

Sodium [2-[(2,6-dichlorophenyl)amino]phenyl]acetate.

CONTENTS:

It contains not less than 99.0% and not more than 101.0% of C14H10Cl2NNaO2.

SOLUBILITY:

  • Sparingly soluble in water.
  • Freely soluble in Methanol.
  • Soluble in ethanol.
  • Slightly Soluble in acetone.

MELTING POINT:

Determine the melting point of sample by using M.P instrument. It decomposes at 280°C.

IDENTIFICATION BY FTIR SPECTROPHOTOMETER:

Measure the FTIR spectra of sample and compare with standard.

IDENTIFICATION BY SODIUM REACTION:

Dissolve 60mg in 0.5mL of methanol and add 0.5 mL of water R. The solution gives reaction of sodium.

IDENTIFICATION BY COLOR OF SOLUTION:

Dissolve 1.25 gm in 25 mL methanol. And check its color & measure absorbance at 440 nm its absorbance should not be greater than 0.05

pH:

Dissolve 1.0 gm in Distilled water & dilute to 100ml and determine the pH. It should be in between 7.5 to 9.0.

LOSS ON DRYING:

Take 1.0gm of sample & dry in oven at 105-110oC for 3.0 hours. It should not be greater than 0.5%.

QUANTITATIVE ASSAY: (By Non-Aqueous TITRATION)

Dissolve 450 mg of Diclofenac Sodium in 25mL Glacial acetic acid. Add 2 drops of crystal violet indicator. Titrate with 0.1N Perchloric acid till change in color of indicator. If necessary, carry out blank titration.

Each ml of 0.1N HClO4 is equivalent to 31.81 mg of C14H10Cl2NNaO2.

CALCULATIONS:

Calculate the Percentage of Diclofenac Sodium by using following Expression.

%age assay = ((R x 31.81) /  (W-(100-X)) x 100

Where

R = Reading at Burette (Vol. of 0.1N HClO4)

W = Weight of sample taken

X = Water contents

STORAGE CONDITION:

It should be packed in airtight containers. Containers should not be transparent to light.

REFERENCE:

British Pharmacopia 2016. Volume-I, Page 740

Pregabalin

Pregabalin

White Crystalline powder, Slightly hygroscopic. On dried basis, It contains 98.0% to 102.0% of Pregabalin.

Chemical Formula:

C8H17NO2

Chemical Structure

Pregabalin
Pregabalin

SOLUBILITY 

  • Soluble in Water
  • Slightly Soluble in Methanol
  • Very slightly soluble in ethanol
  • Practically insoluble in acetone, ethyl acetate, heptane and in methylene chloride

IDENTIFICATION BY FTIR SPECTROPHOTOMETER

Measure the FTIR spectra of sample and compare with standard.

IDENTIFICATION BY HPLC

The retention time of peak of Pregabalin in sample solution is similar to the retention time of peak of pregabalin in standard solution

APPEARANCE OF SOLUTION

Dissolve 0.5 grams in CO2 Free water and dilute 20ml with same solvent (solution S)

The solution is clear and not more intensely colored than GY5

WATER CONTENTS

NMT 0.5%, determined on 0.200 gram

QUANTITATIVE ASSAY BY HPLC

  • Mobile Phase:

Mix 75 volumes of methanol and 25 volumes of a 6.805 grams/liter solution of potassium dihydrogen phosphate previously adjusted to pH 6.8.

  • Standard solution:

Dissolve 100 mg of pregabalin CRS in the mobile phase and dilute to 100ml with same solvent

  • Sample Solution:

Dissolve 100 mg of substance to examind in the mobile phase and dilute to 100ml with same solvent

 

CHROMATOGRAPHIC SYSTEM

  • Mode    :   LC 100
  • Detector   :   UV 210 nm
  • Column   :   A stainless steel column 0.25 m ­x 4.6mm x 5 µm C18 packing; Octadecylsilanized silica gel (5µm particle diameter).
  • Column temperature :  Ambient
  • Flow rate   :   35 ml/min
  • CHROMATOGRAPHIC PROCEDURE

  • Inject 10µl of mobile phase (blank) and record the chromatograph
  • Inject 10µl of standard solution two times and record the chromatograph
  • Inject 10µl of sample solution and record the chromatograph
  • Inject 10µl of standard solution and record the chromatograph
  • Inject 10µl of mobile phase (blank) and record the chromatograph

CALCULATIONS

Calculate the Percentage of Pregabalin sample by using following formula 

%age Assay = (At/As)× (Cs/Ct) × P

Where

At = Av. Area of the peaks in sample chromatograms

As = Av. Area of the peaks in standard chromatograms

Cs = Concentration (mg/ml) of standard

Ct = Concentration (mg/ml) of sample

P   =  Potency of reference standard

 

Tranexamic Acid

Tranexamic Acid

Physical Appearance :

Tranexamic Acid occurs as white, crystals or crystalline powder. It belongs to antifibrinolytic family.

Tranexamic Acid, when dried, contains not less than 98.0% and not more than 101.0% of tranexamic acid (C8H15NO2)

Structural formula:

 

Tranexamic Acid
Tranexamic Acid

Chemical Formula

(C8H15NO2).

Solubility:

It is freely soluble in water, and practically insoluble in ethanol (99.5).

Identification : BY FTIR

Determine the infrared absorption spectrum of Tranexamic Acid as directed in the potassium bromide disk method under Infrared Spectrophotometry, and compare the spectrum with the Reference Spectrum or the spectrum of Tranexamic Acid RS: both spectra exhibit similar intensities of absorption at the same wave numbers.

pH:

The pH of a solution prepared by dissolving 1.0 g of Tranexamic Acid in 20 mL of water is between 7.0 and 8.0.

Color of solution:

Dissolve 1.0 g of Tranexamic Acid in 10 mL of water: the solution is clear and colorless.

LOD: (LOSS ON DRYING)

Not more than 0.5% (1 g, for 2 hours).

Residue on ignition :

Not more than 0.1%

ASSAY : ( HPLC-Method)

Mobile Phase:

Dissolve 11.0 g of anhydrous sodium dihydrogen phosphate in 500 mL of water, and add 5 mL of triethylamine and 1.4 g of sodium lauryl sulfate. Adjust the pH to 2.5 with phosphoric acid or diluted phosphoric acid (1 in 10), add water to make 600 mL, and add 400 mL of methanol.

Standard Preparation:

Weigh accurately about 50 mg of Tranexamic Acid RS, previously dried, dissolve in water to make exactly 25 mL, and use this solution as the standard solution.

Sample Preparation:

Weigh accurately about 50 mg of Tranexamic Acid, previously dried, dissolve in water to make exactly 25 mL, and use this solution as the sample solution.

Chromatographic conditions:

Sample Volume: 20µL
Detector: UV ʎmax=220nm
Column: A stainless steel column 6.0 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter).
Column temperature: 25°C
Flow rate: Adjust so that the retention time of tranexamic acid is about 20 minutes.

Calculations:

Contents of Tranexamic acid can be calculated by given formula

%age Assay =   Hs/Ha × Ca/Cs × 100

Hs = Peak height ratio from the Sample solution

Ha = peak height ratio from the Standard solution

Ca = concentration of Alclometasone Dipropionate in the Standard solution (mg/ml)

Cs = concentration of the Sample solution (mg/ml)

Assay: (BY TITRATION) (non-aqueous )

Weigh accurately 10mg of tranexamic acid in a round 250ml conical flask and add 50ml of glacial acetic acid. Dissolve in magnetic stirrer. Now simply titrate it 0.1M perchloric acid. Each ml of 0.1M perchloric acid is equivalent to 15.72 mg of tranexamic acid.

Calculation:

Contents of Tranexamic acid can be calculated by given formula

%age of tranexamic acid = (volume of 0.1M perchloric acid × 15.72) ÷ weight of sample taken.

ALTERNATIVE METHOD:(BY UV/vis Spectrophotometer)

STANDARD PREPARATION:

Weigh accurately about 50mg of Tranexamic acid of working standard in to 50ml volumetric flask and dilute it with distilled water. Take 1ml of this solution and dilute upto 100ml with the same solvent.

SAMPLE PREPARATION:

Weigh accurately about 50mg of Tranexamic acid of sample in to 50ml volumetric flask and dilute it with distilled water. Take 1ml of this solution and dilute upto 100ml with the same solvent.

Take Reading at 210nm

Contents of Tranexamic acid can be calculated by given formula

%age of tranexamic acid= At/As × Cs/Ct × 100

At =  Absorbance Of  Tranexamic acid in Sample Solution.

As =  Absorbance Of  Tranexamic acid in Standard Solution.

Cs =  Concentration of standard solution (mg/ml)

Ct =  Concentration of sample solution (mg/ml)