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Assay of alkaloidal drugs
Chemical nature of alkaloids
• insoluble or slightly soluble in water
• soluble in organic solvents which are
  immiscible with water
• Salts of alkaloids are soluble in water

This character makes it possible to separate the
             alkaloid by partitioning
Preparation of drugs for assay
• The drug should be pulverized to the required
  finesse-required mesh
• Care should be taken to avoid the loss of
  water during the pulverization
Weighing for assay
• The accuracy should be within 10 mg for
  quantities equal to 5g or over
Extraction
• The alkaloid bearing drugs are extracted with
  one of the following methods
  – Maceration
  – Percolation
  – Continuous extraction
Maceration
• Accurately weighed powdered drug is dipped
  in a specified amount of solvent or mixture of
  solvents
• Then made it alkaline with ammonia TS
  solution and mix well and allow to macerate
  for 12 -24 hours with occasional shaking
• Afterwards allow the solvent to settle
• Decant and aliquots of which are used for
  analysis
Percolation
• Place accurately weighed powdered drug is in
  a suitable container, saturate with suitable
  solvent for 5min
• Make it alkaline with Ammonia solution, mix
  thoroughly and put in percolator
• Allow the drug to macerate for 10-12 h
• Percolate it slowly until percolate is alkaloid
  free
Alkaloidal test
• Take 4 ml percolate, evaporate and dissolve
  the residue in 0.5N HCl and add a drop of
  mercuric chloride
• If turbidity is not produced, percolate is free
  from alkaloid
Continuous extraction
• Counter current extraction using soxhlet
  extractor
Purification
• Take the aliquot, and extract with dilute acid
• Then make the acid part alkaline and extract
  with some immiscible solvent
Determination of alkaloids
• Evaporate the alkaloidal solution to dryness
  by steam or with a current of air
• Soften the residue with 1 ml neutral ether or
  alcohol
• Add a volume of standard acid and warm
  gently to make complete solution
• Add the volume of water to make 25 ml
• Titrate the excess of acid by a standard alkali
  using methyl red as an indicator
• If alkaloidal residue is to be weighed, dry it at
  105oC to a constant weight
• If solvent is chloroform
• Remove the traces with few ml neutral ether
  or alcohol followed by evaporation
Special treatment
• Use of adsorbent: for fluidextracts and
  tinctures having alkaloidal drugs, it is
  necessary to evaporate them to dryness
• To facilitate evaporation and reduce loss,
  extracts are added on an adsorbent-previously
  acid or alkali washed-then made neutral by
  washing with water and dried before use
• Emulsions: Avoid shaking of immiscible
  solvent with water
• If emulsion forms, add excess of either of the
  solvents
• It will results in the break down of emulsion
• Emulsion can also be broken down by adding
  anhydrous sodium sulphate
Assignment
• Assay of belladonna leaf as stated in USP

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Assay of alkaloidal drugs

  • 2. Chemical nature of alkaloids • insoluble or slightly soluble in water • soluble in organic solvents which are immiscible with water • Salts of alkaloids are soluble in water This character makes it possible to separate the alkaloid by partitioning
  • 3. Preparation of drugs for assay • The drug should be pulverized to the required finesse-required mesh • Care should be taken to avoid the loss of water during the pulverization Weighing for assay • The accuracy should be within 10 mg for quantities equal to 5g or over
  • 4. Extraction • The alkaloid bearing drugs are extracted with one of the following methods – Maceration – Percolation – Continuous extraction
  • 5. Maceration • Accurately weighed powdered drug is dipped in a specified amount of solvent or mixture of solvents • Then made it alkaline with ammonia TS solution and mix well and allow to macerate for 12 -24 hours with occasional shaking • Afterwards allow the solvent to settle • Decant and aliquots of which are used for analysis
  • 6. Percolation • Place accurately weighed powdered drug is in a suitable container, saturate with suitable solvent for 5min • Make it alkaline with Ammonia solution, mix thoroughly and put in percolator • Allow the drug to macerate for 10-12 h • Percolate it slowly until percolate is alkaloid free
  • 7. Alkaloidal test • Take 4 ml percolate, evaporate and dissolve the residue in 0.5N HCl and add a drop of mercuric chloride • If turbidity is not produced, percolate is free from alkaloid
  • 8. Continuous extraction • Counter current extraction using soxhlet extractor
  • 9. Purification • Take the aliquot, and extract with dilute acid • Then make the acid part alkaline and extract with some immiscible solvent
  • 10. Determination of alkaloids • Evaporate the alkaloidal solution to dryness by steam or with a current of air • Soften the residue with 1 ml neutral ether or alcohol • Add a volume of standard acid and warm gently to make complete solution • Add the volume of water to make 25 ml • Titrate the excess of acid by a standard alkali using methyl red as an indicator
  • 11. • If alkaloidal residue is to be weighed, dry it at 105oC to a constant weight • If solvent is chloroform • Remove the traces with few ml neutral ether or alcohol followed by evaporation
  • 12. Special treatment • Use of adsorbent: for fluidextracts and tinctures having alkaloidal drugs, it is necessary to evaporate them to dryness • To facilitate evaporation and reduce loss, extracts are added on an adsorbent-previously acid or alkali washed-then made neutral by washing with water and dried before use
  • 13. • Emulsions: Avoid shaking of immiscible solvent with water • If emulsion forms, add excess of either of the solvents • It will results in the break down of emulsion • Emulsion can also be broken down by adding anhydrous sodium sulphate
  • 14. Assignment • Assay of belladonna leaf as stated in USP