1900

, Mikhail S. Tswett
( )
( )
)
( )
, ,
( )

( )
,
Graph Chroma
tsweet ,
 (3 ) (2 1 )

FIGURE A

( ) 1
[B ]
,
( TLC ) ,

FIGURE B
, ,
) ( ) C :2
(
)
. TLC

FIGURE C
 TLC

( ) 3
,
SPE
Tswett ,
,

FIGURE D-1 Column Chromatography – Solid-Phase Extraction SPE]

 50

Tsweet

solid-phase extraction [SPE]

cartridge
, 10
)
,

HPLC

HPLC

Casaba Horváth HPLC

1970

35) psi 500

HPLC
, 1970
( 400 ) 6000
1970 HPLC

HPLC

(PPT )
HPLC
nutraceuticals

FIGURE D-2

UPLC

2004

7
, 1000) 15000
UPLC
1
6800) 100000
 HPLC

E
( ) ,
( ) , ,
,
,
,

,
preparative
chromatography

FIGURE E
 HPLC

,
, UV
,
evaporative-light-scattering detector (ELSD)
MS UV / ELSD ,

LC/MS

FIGURE F
 HPLC

, ,
,
,

Figure G: Understanding How a Chromatographic Column Works – Bands

 ,

Chromatogram

,
, baseline

FIGURE H

, (Cell ) H
,
 , baseline

baseline

baseline
,H
,
,( )
( )
( )

)
 H
)
)

Figure I-1: Identification

I-1
( ) 85 acrylamide

( I-2 B ) 85
)
 baseline

Figure I-2: Identification and Quantitation

, B A
85 ,
, A

B A
 10 A ( ) standards
( 1) B
8

isocratic elution ,

Figure J-1: Isocratic LC System

gradient elution
,
,
Figure J-2: High-Pressure-Gradient System

, J-2

(A)
( B)
Figure J-2

J-3
 Figure J-3: Low-Pressure-Gradient System

K preparative chromatography
)
 Figure K: HPLC System for Purification: Preparative Chromatography

( ) ,

HPLC scale
A

Table A: Chromatography Scale

)

[Vc]

[ i.d] [L]
(L )

Figure L: HPLC Column Dimensions

100 mm 1 mm ,500 mm 20 mm
 ,
, backpressure

15 - 5 40–10 mm X

Table B: Chromatography Scale vs. Column Diameter and Particle Size

)
backpressure
dead volume )
(
outlet inlet
( ) PEEK ( polyetheretherketone )

Figure M-1: Column Hardware Examples

M-2
,

Figure M-2: A Look inside a Column

( Resolution )

, resolution -RS
(resolution)
, )

,
 Efficiency –

,
(N) plate number
(backpressure )

Figure N: Column Length and Mechanical Separating Power [Same Particle Size

[Figure O: Particle Size and Mechanical Separating Power [Same Column Length
 Selectivity -

)
( )
,

HPLC

- -
 ,

Figure P: Chromatographic Polarity Spectrum by Analyte Functional Group

)
(
Figure Q: Proper Combination of Mobile and Stationary Phases Effects Separation Based on Polarity

(Q )
)

R-1, R-2, R-3

Figure R-1: Mobile Phase Chromatographic Polarity Spectrum

R-1
eluotropic series
 ( )

Figure R-2: Stationary Phase Particle Chromatographic Polarity Spectrum

)
R-2 ,(

n-octadecylsilyl- C18, ODS n-octylsilyl- C8 cyanopropylsilyl- CN

C18 ,

Figure R-3: Compound/Analyte Chromatographic Polarity Spectrum

P R-3
,

, ,

( ) ,

) Tswett

Figure S-1: Normal-Phase Chromatography

 S-1
,

, ( )
% 100

,
S-2

Figure S-2: Reversed-Phase Chromatography

,
)
% 75
,
 (ODS ) C18
, C
( )

Table C: Phase Characteristics for Separations Based on Polarity

HILIC

, HILIC
% 100

aprotic ) (% 20 )
) (solvent
(
,

, HILIC

,
 HIC

HIC

HIC
Silica C18 C4
,

IEC ( )

( )

Cation exchange
anion exchange ,
,
 Figure T: Ion-Exchange Chromatography

(pH)

)
, ,
 Table D: Ion-Exchange Guidelines

)
)
(R-1 ) )
)
( ) , D
,7
 3 pH

, pH pKa
( )
pKa pH

SPE HPLC

GPC SEC

Flodin Porath 1950

( )
,gel filtration

gel-permeation GPC

[size-exclusion chromatography [SEC]

GPC 1963
 )

GPC

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