cromatografia liquida lc-hplc

8/11/2019 Cromatografia Liquida Lc-hplc

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CROMATOGRAFIA LIQUIDA LC

Cromatografa HPLC


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Comatografa en fase Normal (NPLC).

-cromatografa de particin donde la fase estacionaria es polar

NPLC columna retiene compuestos polar es- fase movil debil es un lquido no-polar: solvente orgnico

- fase movil fuerte es un lquido polar: agua o metanol

- fase etacionaria debe tener una baja miscibilidad con la fase movil para que la fase estacionaria

no se disuelva en la columna.

ejemplos deliquid NPLC stationary phases:

< Dimethylsulfoxide


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Aplicaciones comunes de NPLC:

- purificacin de compuestos organicos e inorganicos provenientes de una mezcla de

reaccion (sntesis)

- separaciones con propsitos generales de compuestos polares/no-polares cuando la

muestra est en un solvente no-polar .

PrepLCMS Analysis (50 mg injection)

4.36

5

2e7

4e7

6e7

8e7

Intensity,cps

Desired

Product

Automated chromatography purification of designed drug combinatorial libraries


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C.) Elucion:

Retencin y elucin de solutos en LC depende de las interacciones de los solutos tanto con la

fase movil como con la fase estacionaria.

- para describir el como los solutos son retenidos en una columna con diferentes solventes,

se usa los terminosfase mvil debil y fase mvilfvuerte.

Fase movil fuerte: solvente que eluye rapidamente los solutos de la columna (i.e., pequeo valor k)

Esto ocurre si la fase movil es muy similar a la fase estacionaria phase en sus interacciones

intermoleculares con los solutos

- Solventes polares sern una fase movil fuerte para una columna que tiene fase

estacionaria polar

Rapid elution in a few minutes for all

compounds in the mixture


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C.) Elution:

Fase movil debil: a solvent that slowly elutes solutes from the column (i.e., high solute

retention or large k)

This occurs if the mobile phase is very different from the stationary phase in itsintermolecular interactions with the solutes

- a non-polar solvent would be a weak mobile phase for a column containing a

polar stationary phase

Note: whether a solvent is a weak or strong mobile phase depends on the stationary phase being

used. Hexane is a weak mobile phase on a polar stationary phase, but a strong mobile phase

on a non-polar stationary phase.

Slow elution (~ 20 minutes) for all

compounds in the mixture


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Common applications of RPLC:

- most popular type of liquid chromatography

separation of a wide variety of non-polar andpolar solutes- popularityweak mobile phase is a polar solvent (e.g., water)

ideal for the separation of solutes in aqueous-based samples, suchas biological compounds

l f ( d)


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Common applications of RPLC (continued):

- purification of biological and organic compounds present in aqueous solutions

- pharmaceutical analysis (drug quantitation and quality control)

- protein & peptide mapping

- analysis of soil and water samples

- clinical analysis of blood and urine samples

RPLC Analysis of Patient blood

serum for presence of drug during

clinical trial


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Common applications of RPLC:

- most popular type of liquid chromatography

separation of a wide variety of non-polar andpolar solutes- popularityweak mobile phase is a polar solvent (e.g., water)

ideal for the separation of solutes in aqueous-based samples, suchas biological compounds

cromatografia liquida lc-hplc

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