Principle of HPLC (Liquid Chromatography) : Pharmaceutical Guidelines

Editable Pharmaceutical Documents in MS-Word Format

View

8

Principle of HPLC (Liquid Chromatography)

Learn how High Performance Liquid Chromatography works (HPLC principle) and how chromatographic column separates the compounds from a mixture.
High-performance liquid chromatography is an effective type of column chromatography which is widely used in pharmaceuticals. It is very useful to determine the assay and related substances in drug substances. In general, HPLC is used to separate the components of a mixed drug substance.

In HPLC chromatography, column plays a significant role in the separation of different compounds because it contains stationary phase. The stationary phase is a bad of polar or non-polar particles according to the type of column. Polar and non-polar columns are used according to the nature of the sample to be analyzed.


HPLC flow chart

Mechanical pumps are used to pump the mobile phase into the system and injector introduces the sample into the mobile phase which enters in the column at a constant flow rate. Mobile phase acts as a carrier in the whole process.

When a mixture of compound enters in a column, it separates the compounds on the basis of their polarity. If stationary phase is non-polar then it attracts the non-polar compounds and a polar compound elutes first then a non-polar compound and if the stationary phase is polar then non-polar compound elutes first.


HPLC system

Detectors are used to determine the separated compounds by ultraviolet absorption. The light absorption depends upon the concentration of the compound in the mobile phase. These response signals are recorded by the computer software in the form of peaks and purity of the compound is calculated by the peak area of different compounds. Computer system and software should be validated.






Ankur Choudhary is India's first professional pharmaceutical blogger, author and founder of Pharmaceutical Guidelines, a widely-read pharmaceutical blog since 2008. Sign-up for the free email updates for your daily dose of pharmaceutical tips.
.moc.enilediugamrahp@ofni :liamENeed Help: Ask Question


Click Here

6 comments: Post Yours! Read Comment Policy ▼

  1. Anonymous15 July

    Thanks very informative.... :)

    ReplyDelete
  2. Anonymous16 July

    informative

    ReplyDelete
  3. Anonymous16 July

    informative

    ReplyDelete
  4. Thank's a lot Choudhary; Tell me hoe to expand capability of HPLC for amino acids analysis

    ReplyDelete
  5. Thankyou sir ... It helped me with my assignment

    ReplyDelete


Popular Categories

QA SOPs QC SOPs Micro SOPs HVAC Production SOPs Stores SOPs Checklists Maintenance SOPs HPLC Sterile GLP Validation Protocols Water System GDP Regulatory Maintenance Calibration Warning Letters

Follow Pharmaguideline


DOCUMENTS

PHARMACEUTICAL DOCUMENTS




Editable Pharmaceutical Documents in MS-Word Format. Ready to use SOPs, Protocols, Master Plans, Manuals and more...

View


adsbypg

Discussion - Latest Topics

Show All ❭❭Pharmaguideline Forum



GET APP FOR NEWS UPDATES

Pharmaceutical Updates




✔ Worldwide Regulatory Updates
✔ Pharmaceutical News Updates
✔ Interview Questions and Answers
✔ All Guidelines in One Place

Install


Recent Posts