Bisoprol Fumarate and Hydrochlorothiazide Identification Test using Silica gel G TLC plates

By: Dr. Sanjay Poman, Merck KGaA, Darmstadt, Germany, Life Science Application Centre (GAC), Mumbai, India

Introduction

Bisoprolol Fumarate is used in the treatment of high blood pressure (hypertension), and it reduce blood flow. Thus a preventive treatment before and after heart attacks, decreasing the chances of recurrence. Hydrochlorothiazide is diuretic medication used for treatment of high blood pressure.

Current Indian pharmacopoeia (version 2018) indicates that a TLC silica gel GF254 plate should be used for Bisprolol Fumarate and hydrochlorothiazide tablets Identification Test. TLC silica gel GF254 plate complies with the requirements for a TLC silica gel plate R but with the following modification, it contains calcium sulfate hemihydrate as binder and a fluorescent indicator having a maximum absorbance at 254 nm. This application note show how 60G F254 plates can be used for this test.
 

Bisoprolol Fumarate

(2RS)-1-[4-[[2-(1-Methylethoxy)ethoxy]methyl]phenoxy]-3-[(1-methylethyl)amino]propan-2-ol fumarate

 

Hydrochlorothiazide

6-Chloro-3,4-dihydro-2H-1,2,4-benzothiadiazine-7-sulfonamide 1,1-dioxide

 

Experimental Conditions
TLC Plate Silica Gel 60G F254  20x20 cm  
Injection volume 25 µL
Detection Exposure to UV light at 254 nm and iodine spray
Mobile Phase Dichloromethane:Methanol:Ammonia 43:20:8 (v/v/v) 
Plate development Prewash the plate in methanol and allow to dry in air before use. Use mobile phase described above. Spot standard and sample.Develop the plate for 15 cm. Remove the plate dry in air, spray with 5 g/L potassium ferricyanide in ferric chloride. Examine in day light.
Development   Over 2/3rd of the plate
Diluent Methanol
Drying In current of dry air
Standard Reference Solution A – 0.1% w/v solution of Bisoprolol Fumarate in methanol
Reference Solution B -  0.1% w/v solution of Hydrochlorothiazide in methanol 
Sample Disperse 1 tablet in 5 mL volumetric flask. Dilute with methanol to volume, sonicate for 5 minutes and centrifuge and use clear supernatant liquid

 

TLC Plate after development

TLC Plate after development
 

Chromatogram

Chromatogram
 

Chromatographic Data (Standard Solution)

Chromatographic Data (Standard Solution)   

 

Compound Rf
Area
Bisoprolol Fumarate 0.1 3711

 

Chromatographic Data (Standard Solution)

Chromatographic Data (Standard Solution)

 

Compound Rf
Area
Hydrochlorothiazide 0.78 5871

 

Chromatogram Sample

Chromatogram Sample
 

Compound Rf
Area
Bisoprolol Fumarate 0.1 3488
Hydrochlorothiazide 0.74 5502

 

Conclusion

  1. The two principal spots in the chromatogram obtained with test solution corresponds to those in the chromatogram obtained with reference solution (A) and (B).
  2. Silica gel G plates can be used for the Bisoprolol Fumarate and Hydrochlorothiazide identification test using LiChrosolv grade dichloromethane and methanol solvents, and all other listed products, and comply to set requirements in the Indian Pharmacopoeia.
     

Materials