Supercritical Fluid Chromatography · Supercritical Fluid Chromatography Theory, Design and...

Supercritical Fluid ChromatographyTheory, Design and Applications

DJ TognarelliChromatography Application Scientist

JASCO Seminar Series

Founding Members

1958

Established at the Optical Research Institute at Tsukuba University, Tokyo

Founding members include: World famous physicist Yoshio Fujioka Nobel Prize winner Shinichiro Tomonaga

(1965 - Physics for QED with Richard Feynman)

1972

JASCO in the USA

HPLC Products

1985

SFC – First dynamic BPR

Dr. Tomonaga

Products

Overview

Section I Introduction to Supercritical Fluids Advantages of SFC

Section II SFC System Preparative Unique Capabilities

Section III Applications

SupercriticalFluid

-56.4

0.51

31.3

7.29

Triple Point

Critical Point

SolidLiquid

Gas

Temperature (oC)

Pres

sure

(MPa

)

Phase Diagram for CO2- Critical temperature (Tc):

the temperature above whicha distinct liquid phase can NOTexist, regardless of pressure

- Critical pressure (Pc): the vapor pressure at the

critical temperature

At T and P above itscritical point (Tc, Pc), a substance exists in

a supercritical fluid state!

Introduction to Supercritical Fluid

Compound Tc (C) Pc (Mpa)

NH3 132 11.28

CO2 31 7.38

N2O 36 7.24

H2O 374 22.06

C3H8 97 4.25

C6H14 234 2.97

CH3OH 239 8.09

C2H5OH 243 6.38

C6H5(CH3) 318 4.11

Critical Temperatures and Pressures

Advantages of SC-CO2

SC-CO2 → Non toxic, non flammable, inexpensive

Low Cp and CtEasy removal of CO2 by decompression

Environmentally friendly (alternative solvent to hazardous organic solvents)Safely used in food and pharmaceutical products.

Gas-liquid 2-phase state

Supercritical State1-phase state

Critical pointRayleigh Scattering

Phase Transition

72.9 atm (bar) / 31.3 C

Diffusivity (cm2/s)

Density (g/cm3)

Viscosity (g/cm · s)

Gas 10-1 10-3 10-4

Supercritical Fluid

10-4 – 10-3Liquid Like

0.2-0.8Liquid Like

10-4Gas Like

Liquid 10-5 - 10-6 1 10-2

Physical Properties

High diffusivity and densitywith low viscosity

Faster flow rates with lower pressure=

1. SFC is a normal phase technique where CO2 replaces hexane

2. Faster separation/analysis time (3 to 5 times faster than HPLC)

SFC Advantages

-1000

100200300400500600700800900

0 1 2 3 4 5 6 7 8 9 10 11 12

Time (min)

mAU

-1000

100200300400500600700800900

0 1 2 3 4 5 6 7 8 9 10 11 12

Time (min)

mAU

-1000

100200300400500600700800900

0 1 2 3 4 5 6 7 8 9 10 11 12

Time (min)

mAU

HPLC

SFC

HT-SFC

Mobile phase: Hexane/Isopropanol (IPA)(60/40)Flow rate: 0.5 mL/minColumn: CHIRALCEL OD (4.6 mm ID x 250 mm L)Column temp.: 40 ºCWavelength: 230 nm

CO2 : 3.0 mL/minModifier: Methanol, 0.1 mL/minColumn: CHIRALCEL OD (4.6 mm ID x 250 mm L)Column temp.: 40 ºCPressure: 20 MPaWavelength: 230 nm

CO2 : 5.0 mL/minModifier: Methanol, 2.5 mL/minColumn: CHIRALPAK OD (4.6 mm ID x 150 mm L)Column temp.: 40 ºCPressure: 20 MPaWavelength: 230 nm

HPLC, SFC, and HT-SFC

System Column ID HPLC Flow Rate SFC Flow Rate

Analytical 3mm, 4.6mm 0.6mL/min, 1mL/min 2mL/min, 5mL/min

Hybrid 4.6mm, 10mm 1mL/min, 5mL/min 5mL/min, 20mL/min

Semi-Preparative 4.6mm, 10mm, 20mm 1mL/min, 5mL/min, 20mL/min 5mL/min, 20mL/min, 90mL/min

Preparative 10mm, 20mm, 30mm 5mL/min, 20mL/min, 40mL/min 20mL/min, 70mL/min, 150mL/min

HPLC vs SFC Systems

SFC is 3 – 5 times faster



3. Easy removal of mobile phase (typically alcohol)

4. Using up to 95% less solvent – Cost Savings

SFC Advantages

Tocopherol (Vitamin E)

HPLCColumn : Finepak SIL-5 (4.6mm I.D. x 250mmL)Eluent : n-Hexane / IPA / CH3COOH (99 / 1 / 0.1)Flowrate : 1.0 mL/min Temperature : 40 deg.CINJ.VOL. : 10 µLWavelength range (PDA) : 200 - 650 nm

SFCColumn : SFCpak SIL (4.6mm I.D. x 250mmL)CO2 : 3.0 mL/minSolvent (EtOH) : 0.2 mL/min Pressure : 20.0 MPaTemperature : 80 deg.CINJ.VOL. : 10 µLWavelength : 280 nm

0.0

1.5E+05

3.0E+05uV

5.0 10.0 15.0 20.0 [min]

0.0

2.0E+04

4.0E+04

6.0E+04 1: Tocopherol α 2: Tocopherol β

3: Tocopherol γ 4: Tocopherol δ

12

3

4

12

3

4

HPLC (14min) SFC (8min)Amount Price Amount Price

Solvent 26.600L $2356 9.680L $1633Column 2cm 2cmFlow rate 19mL/min 60.5mL/minTime 23 hours 13 hoursSolvent disposal cost is not included

Column : CrestPak C18S (4.6mm I.D. x 150mmL)Eluent : A; MeOH / H2O (75 / 25)

: B; /.EtOAc / CH3CN (50 / 50)Flowrate : 1.5 mL/min Temperature : 30 deg.CINJ.VOL. : 15 µLWavelength : 270 nm

Time(min) A(%) B(%)0.0 90 10

27.0 0 10032.0 0 100

Polymer Additives

Column : SFCpak SIL (4.6mm I.D. x 250mmL)CO2 : 3.0 mL/minSolvent (EtOH) : 0.3 mL/min Pressure : 20.0 MPaTemperature : 60 deg.CINJ.VOL. : 10 µLWavelength : 280 nm

0.0

4.0E+04

8.0E+04

uV

10.0 20.0 30.0 [min]0.0

6.0E+04

1.2E+05

1

2

3

1

23

1: Irganox 1010 2: Irganox 1076 3: Irgafos 168

HPLC (42min) SFC (9min)Amount Price Amount Price

Solvent 267.750L $26,902 21.760L $3,944Column 3cm 3cmFlow rate 42.5mL/min 136mL/minTime 70 hours 15 hoursSolvent disposal cost is not included

30.0

[min]

20.0

10.0

0.0

6.0E+04

1.2E+05

0.0

4.0E+04

8.0E+04

uV





5. Higher selectivity especially in chiral separation with wider array of columns and co-solvents

6. Longer column lifetime

SFC Advantages

Solvents and Columns

Stationary Phase Choices

Silica

2-ethylpyridine

Cyano

Aminopropyl

Diol

Amide

PFP

Phenyl

C18 < C8 Reversed-phase range

Normal phase range

SFC range

Solvent

Solvent Polarity [P’]

Pentane, Hexane, Heptane 0.1

Xylene 2.5

Toluene 2.4

Diethyl ether 2.8

Dichloromethane 3.1

Chloroform 4.1

Acetone 5.1

Dioxane 4.8

THF 4.0

MTBE 2.5

Ethyl acetate 4.4

DMSO 7.2

Acetonitrile 5.8

Isopropanol 3.9

Ethanol 4.3

Methanol 5.1

Water 10.2





5. Higher selectivity especially in chiral separation with wider array of columns and co-solvents

6. Longer column lifetime

7. Similar methodology to HPLC

SFC Advantages

Method Principles

Sample : CaffeineColumn : SCFpak SIL (4.6 mm I.D. x 250 mmL)Pressure : 20.0 MPaTemperature : 60 deg.C

0.0

6.0E+04

1.2E+05uV

0.0

1.0E+05

2.0E+05

4.0 8.0 12.0 16.0 [min]0.0

1.5E+05

3.0E+05

10%

15%

20%

Same methodology used in HPLC applies to SFC Gradient Isocratic Column Solvent (with additives)

Structurally similar compounds

Chiral compounds

Degradants

Metabolites

Increased efficiency makes SFC a superior technique for separating these types of compounds.

All compounds typically used on Normal Phase HPLC

Polar Compounds

Those that are difficult to retain in HPLC

Orthogonality

When other separation techniques don’t work.

When To Try SFC?

Overview




SFC1

23

MS

1. Cooled CO2 Pump2. High Pressure Flow Cell3. Back Pressure Regulator

SFC System

HPLCMS

SFC Hardware Requirement 1

Pump

Liquid CO2 is required for SFC Typically from a cylinder for analytical SFC Bulk CO2 for Prep SFC

To maintain the liquid CO2, the pump head must be cooled Peltier cooled for analytical pumps Circulated chiller for Prep pumps

Otherwise it is a HPLC pump

1. CO2 Pump

Pump heads (-10 °C)

Peltier cooling

Recirculating Chiller Head (-10 °C)

Insulating Cooling Block

Automatic

Shut-Off Valves

CO2 Physical Properties


Detector Flow Cell

HPLC pressure is between the pumps and column Flow cell is low pressure

SFC pressure is between the pumps and back pressure regulator Flow cell must be high pressure

Detector is identical, just a different flow cell

2. High Pressure Flow Cell

HPLC Detector + High Pressure Flow Cell

25.0

15.4

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 Retention Time [min]

-1000

0

1000

2000

Inte

nsity

[µV]

Performance and Sensitivity

# Peak Name tR Area Height S/N NTP Resolution Symmetry Factor1 Peak-001 1.662 2688 858 25.0 7250 7.341 1.3672 Peak-002 2.472 2751 530 15.4 4767 N/A 1.076

10uL Injection

0.05% flurbiprofen test

SFC Detectors

Circular DichroismVariable Wavelength UV Photodiode Array Fluorescence

Flame Ionization(FID)

Evaporative Light Scattering(ELSD)

Mass Spectrometry


Back Pressure Regulator

SFC requires a back pressure regulator This maintains the critical pressure for CO2 to stay in a supercritical fluid state The performance of this is critical to the separation efficiency and reproducibility

If this restrictor is open or bypassed, then you would have HPLC

• Pressure control without changing flow rate of SCF.

• Pressure programming vs. time

• Self cleaning of precipitated substances

• Low dead-volume

• Patented

Valve seatValve needle

Needle-drive solenoid

Needle seal Return spring

Heater

FLOW

Gap adjustment screw

Electrical-Feedback Regulator

3. Back-Pressure Regulator

Back-Pressure Regulator Performance

Caffeine ConditionsFlowrate 3mLs/minCO2/MeOH 85/15Back pressure 150 barColumn Silica 4.6x150, 5um5uL Injection

0.169% Retention Time RSD

Preparative SFC Fraction Collection

(Prep Column ID)2

(Analytical Column ID)2Prep Flow Rate

Analytical Flow Rate=Scale-Up

CO2

Sample（and solvent）

Sample solutiondissolved in CO2+Modifier

Open-Bed Fraction Collection

Recovery 95%+

Unique Capabilities

Back Pressure Regulation

Parallel SFC

Stacked Injections

Back Pressure Regulation

0.0

1.5E+04

3.0E+04uV

0.0

5.0E+04

1.0E+05

1.5E+05

5.0 10.0 15.0 [min]0.0

1.0E+05

2.0E+05

10.0 MPa

15.0 MPa

20.0 MPa

Sample : CaffeineColumn : SCFpak SIL (4.6mm I.D. x 250mmL)CO2 : 3.0 mL/minSolvent (EtOH) : 0.5 mL/min Temperature : 60 deg.C

Back Pressure Regulation is unique to SFC and can provide additional separation optimization as the pressure changes the density.

Parallel SFC

4X or 5X Throughput

-1000

100200300400500600700800900

0 1 2 3 4 5

mAU

-1000

100200300400500600700800900

0 1 2 3 4 5

mAU

1 2 3 4 5

1 2 3 4 5

Stacked Injections

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17

-100

1 2 3 4 5

1 2 3 4 5

Stacked injections eliminate the time between injection and start of the first peak for all injections except the first significantly reducing the total purification time.

Overview




Applications

Pharmaceuticals Natural products Foods Pesticides and herbicides Surfactants Lipids Cosmetics Polymers Petroleum Explosives Propellants Gasoline, diesel and bio-diesel fuels

Pharmaceuticals

Achiral and Chiral Molecules

LC/MS vs SFC/MS Pinkston et. al. screened 2153 compounds. Eluted and Detected. 87% by SFC/MS 90% by LC/MS Ammonium Acetate aid in retention of very

polar and ionic analytes

SFC method time of 4 minutes

LC method time of 6 minutes

*LC screen required nearly 3 additional days.

0.0 2.0 4.0 6.0 8.0 10.0 Retention Time [min]

-5000

0

5000

Inte

nsity

[µV]

Fraction1Fraction2 Fraction3

Pharmaceutical – Chiral Purification

0.0 2.0 4.0 6.0 8.0 10.0 12.0 Retention Time [min]

0

100000

200000

300000

Inte

nsity

[µV]

Fraction1 Fraction2


-10000

0

10000

20000

Intensit

y [µV]

UV

CD

∆CD/ ∆UV

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 Retention Time [min]

0

50000

100000

Intensi

ty [µV

]


0

50000

100000

Intensi

ty [µV

]

Enantiopurity 99.8 ± 0.5% Enantiopurity 99.6 ± 0.5%

Fraction 1 Fraction 3


0

50000

100000

150000

Inte

nsity

[µV

]

Fraction 1 Fraction 2

Purity 98 ± 0.5% Purity 93 ± 0.5%

Pharmaceutical – Chiral

Enantiomer 1Isomer B

Enantiomer 2 isomer B

16391

14342

Time (min)15 20 25 30 35 40

mAU

Time (min)15 20 25 30 4035

0

45

Isomer A (25 min)

Impurity (37 min)

Enantiomer 2 of isomer B(45.4 min)

Enantiomer 1 of isomer B(41.1 min)

10

20

15

5

100 200 300 400 500 600 700 800 m/z

%

%

%

0

100

%

100

100

100

0

0

0

393452

785

393

785

393

785

389

778448

N

F

X

N

F

* X

Isomer BMz = 392

Isomer AMz = 392

MS

UV

CD

Pharmaceutical – In Vitro Metabolism

VerapamilVerapamil is an L-type calcium channel blocker ofthe phenylalkylamine class. It has been used as avasodilator during cryopreservation of blood vessels,and is a class 4 antiarrhythmic, more effective thandigoxin in controlling ventricular rate. Verapamil hasalso been used in the treatment of hypertension,angina pectoris, cardiac arrhythmia, and mostrecently, cluster headaches.

C27H38N2O4 MW = 454.2832

Sample PrepA sample of in vitro verapamil 25uM and 10uM wasincubated for 30 minutes at 37°C with 2mg/mL protein and aNADPH regenerating system. The reaction was quenchedwith an equal volume of acetonitrile and the sample wascentrifuged. The sample supernatant was transferred to a200uL sample vial.

Metabolism

TIC and Enhanced Product Ion Spectra TIC and Enhanced Product Ion Spectra

Verapamil Norverapamil 1st and 2nd pair (active metabolite)

SFC-MSMS

Pharmaceutical – Potency

WarfarinWarfarin is a coumarin anti-coagulant and is marketed as the racemate. The entantiomers are differentially metabolized by human cytochromes P450(CYP). R-warfarin is metabolized primarily by CYP1A2 to 6- to 8-hydroxywarfarin and by CYP3A4 to 10-hydroxywarfarin. S-warfarin is metabolized primarily by CYP2C9 to 7-hydroxywarfarin. The S(-)-form is the more potent isomer.

O O

OH

CH3

O

C19H16O4 MW = 308.1049

Continued

The blood sample (approximately 4 mL) was collected from a patient that had taken 1.75 mg Q.D. Taro Pharmaceutical’s warfarin sodium USP. The whole blood was mixed with 10 mL of acetonitrile-isopropanol mix (1:1), vortexed for 20 minutes, and centrifuged for 20 min at 5,000 rpm at ambient temperature. The supernatant was filtered with 0.2-mm syringe filter and transferred to a 1.8-mL auto-sampler vial for SFC/MS/MS analysis.

SFC MS TIC and EPI. TIC of Racemic Warafin 10ng/ul, 10uL injection (top) and enhanced product ion spectra of each enantiomer (bottom 2) . Conditions: AD-H 4.6 x 250nm. Gradient 10% to 50% Methanol with 0.3% TFA over 18minutes at 3mL/min at 30°C.

warfarin MRM result.rdb (warfarin 1): "Linear" Regression ("1 / (x * x)" weighting): y = 33.2 x + 245 (r = 0.9894)

1000.0 2000.0 3000.0 4000.0 5000.0 6000.0 7000.0 8000.0 9000.0 1.0e4Concentration, ng/mL

0.0

2.0e4

4.0e4

6.0e4

8.0e4

1.0e5

1.2e5

1.4e5

1.6e5

1.8e5

2.0e5

2.2e5

2.4e5

2.6e5

2.8e5

3.0e5

3.2e5

3.4e53.5e5

Area, c

ounts

MRM Calibration Curve using Peak 1

SFC MS MRM. 2 MRMs of Taro warfarin sodium USP 1 mg pill dissolved in10 mL methanol, 0.45mm-filtered, 10-uL injection (top). 2 MRMs of the patient’s blood that had taken 1.75mg of O.D. Taro Pharaceutical’swarfarin sodium USP pill (bottom)

1.63mg warfarin in blood

Polymers – SFC & SFEQualitative and Quantitative

Chimassorb 81

Tinuvin 327

Irgafos 168 Phosphate

Irgafos 168

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0

Retention Time [min]

0

500000

1000000

1500000

Inte

nsity

[µV]

SFCColumn : Luna C18 (4.6mm x 150mmL)CO2 : 2.7 mL/minSolvent (MeOH): 0.3 mL/min Pressure : 250 barTemperature : 75 deg.CINJ.VOL. : 20 µLWavelength : 220 nm

Irgafos 168 Tinuvin 326

Chimassorb 81

0.00 0.10 0.20 0.30 0.40 0.50 0.60 0.70 0.80 0.90 1.00

Concentration[mg/mL]

0

10000000

20000000

Inte

nsity

Chimassorb 81 - CH9

Calibration for Chimassorb 81

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0


0

500000

1000000

1500000

Inten

sity [

µV]

Contour Plot

Standard 2

Standard 3

Chimassorb 81

Standard 2

Standard 3

Tinuvin 327

Standard 2Standard 3

Irgafos 168

Extraction Results

Chimassorb 81

Tinuvin 326

Unknown

Irganox 1010

Irgafos 168

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0


0

500000

1000000

Inte

nsity

[µV

]

0.11% Irgafos 1680.45% Tinuvin 3260.30% Chimassorb 81

Fraction collected from extraction

SFE extraction. 500mgs of polymer film was cut into squares and packed in a 10mL extraction vessel.ConditionsPressure: 250 barStatic Extraction Time: 5 minutesDynamic Extraction Time: 5 minutes

Petrochemicals – Olefin Analysis

8.0 8.5 9.0 9.5 10.0 Retention Time [min]

10000

20000

Inte

nsity

1.0% Olefins-3 - CH13.5% Olefins-3 - CH16.0% Olefins-3 - CH18.5% Olefins-3 - CH112% Olefins-3 - CH117% Olefins-3 - CH125% Olefins-3 - CH1

0.0 5.0 10.0 15.0 20.0 25.0 Concentration [%]

0

200000

400000

600000

800000

Inte

nsity

Olefins - CH1

ASTM D6550

Column : Silica 4.6 x 250mm, 5um: Silver 4.6 x 50mm, 5um

CO2 : 3.0 mL/min Pressure : 100 barTemperature : 35 CINJ.VOL. : 0.5 µLDetection : Flame Ionization (FID) 1


20000

40000

60000

Commercial gasoline contained 1.0% olefins

Petrochemicals – Diesel

ASTM D5186Column : Silica 4.6 x 250mm, 5umCO2 : 3.0 mL/min Pressure : 100 barTemperature : 35 CINJ.VOL. : 0.5 µLDetection : Flame Ionization (FID)

1

2

0.0 1.0 2.0 3.0 4.0 5.0 6.0 Retention Time [min]

20000

40000

60000

80000

Commercial gasoline contained 59.3% non-aromatics and 40.7% aromatics.

Peaks: 1. n-hexadecane, 2. toluene, 3. tetralin, 4. naphthalene.

Environmental – Pesticides

CO2 : 5.0 mL/minCo-Solvent : MeOH w/ Amm. Ac. gradient to 50% over 7minPressure : 100 barTemperature : 40 CWavelength : 220nm

CarbarylMW 201.22

CarbofuranMW 221.25

DiuronMW 233.09

ChlorpyriphosMW 350.59

MalathionMW 330.36

CoumaphosMW 362.77

1ppm

10ppm

100ppm

Chlopyriphos

Malathion

Coumaphos

Carbofuran

Carbaryl

Diuron

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5


2000

3000

4000

5000

Inten

sity

6 MIX 0.5PPM - CH1

6 MIX 0.1PPM - CH1

6 MIX 0.05PPM - CH1

6 MIX 0.02PPM - CH1

6 MIX 2PPB - CH1

Carbaryl

MS - SIM

0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0


0

2000

4000

6000

8000

229 - CH1241 - CH1257 - CH1281 - CH1284 - CH1288 - CH1297 - CH1302 - CH1303 - CH1304 - CH1313 - CH1317 - CH1328 - CH1330 - CH1331 - CH1335 - CH1345 - CH1372 - CH1394 - CH1407 - CH1503 - CH1

26 pesticides

US and EU Require

Environmental – PAHs

Detection limit(S/N=3)

UV 7040 fg

FL 26 fg

-100

0

100

Inte

nsity

[µV]

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0


-10000

0

10000

Inte

nsity

[µV]

AnthraceneApprox. 300 times

more sensitive

0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0 16.0 18.0 20.0Retention Time [min]

0

10000

20000

30000

Inte

nsity

[µV]

1

23+4

5

67 8 9

1011 12 13 14 15 16

2500 pg each


0

10000

20000

30000

Inte

nsity

[µV]

1

2

3+4

5

6

7

8

9

10

11

12

13

14

15

16

25 pg each

POLYAROMATIC HYDROCARBONS ANTHRACENE

Cannabis – Potency and Purification

CO2 +EtOH : 6.0 mL/min (Gradient)Pressure : 100 barTemperature : 25 CINJ.VOL. : 10 µLDetection : 220nm

Terpenes_C6 C18 x2 + IB N-5 40C 80bar - CH1


0

1. CBDV2. CBN3. Delta-8-THC4. CBC enantiomer 15. CBD6. Delta-9-THC7. CBC enantiomer 28. THCV9. CBG10. CBDA11. CBDVA12. THCA13. CBGA

MS < 3min

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Slide Number 1Founding Members�Products�Overview�Overview�Introduction to Supercritical Fluid�Critical Temperatures and Pressures�Advantages of SC-CO2�Phase Transition�Physical Properties�SFC Advantages �HPLC, SFC, and HT-SFC �HPLC vs SFC Systems�SFC Advantages �Tocopherol (Vitamin E)�Polymer Additives�SFC Advantages �Solvents and Columns�SFC Advantages �Method Principles�When To Try SFC?�Overview�SFC System�SFC Hardware Requirement 11. CO2 Pump�CO2 Physical Properties�SFC Hardware Requirement 22. High Pressure Flow Cell��Performance and Sensitivity��SFC Detectors�SFC Hardware Requirement 33. Back-Pressure Regulator�Back-Pressure Regulator PerformancePreparative SFC Fraction Collection��Unique Capabilities�Back Pressure Regulation�Parallel SFC�Stacked Injections�Overview��Applications�Pharmaceuticals�Pharmaceutical – Chiral Purification�Pharmaceutical – Chiral�Pharmaceutical – In Vitro Metabolism�Metabolism��Pharmaceutical – Potency �Continued�Polymers – SFC & SFE�Qualitative and Quantitative�Extraction Results�Petrochemicals – Olefin Analysis�Petrochemicals – Diesel�Environmental – Pesticides�Environmental – PAHs�Cannabis – Potency and Purification�JASCO Educational Resources�Thanks for attending!��Questions?

Supercritical Fluid Chromatography · Supercritical Fluid Chromatography Theory, Design and...

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