cromatografia liquida lc-hplc
TRANSCRIPT
-
8/11/2019 Cromatografia Liquida Lc-hplc
1/34
CROMATOGRAFIA LIQUIDA LC
Cromatografa HPLC
-
8/11/2019 Cromatografia Liquida Lc-hplc
2/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
3/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
4/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
5/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
6/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
7/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
8/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
9/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
10/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
11/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
12/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
13/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
14/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
15/34
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
-
8/11/2019 Cromatografia Liquida Lc-hplc
16/34
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
-
8/11/2019 Cromatografia Liquida Lc-hplc
17/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
18/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
19/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
20/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
21/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
22/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
23/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
24/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
25/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
26/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
27/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
28/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
29/34
-
8/11/2019 Cromatografia Liquida Lc-hplc
30/34
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
-
8/11/2019 Cromatografia Liquida Lc-hplc
31/34
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
-
8/11/2019 Cromatografia Liquida Lc-hplc
32/34
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)
-
8/11/2019 Cromatografia Liquida Lc-hplc
33/34
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
-
8/11/2019 Cromatografia Liquida Lc-hplc
34/34
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