SofTA Corporation ELSD Application

Monitoring of a Synthesized Compound Column: C18, 4.6 x 150mm Mobile Phase: A: Water, B: Methanol Gradient: 0 min: 40% B, 7: 100%B, 12: 100%B Flowrate: 1mL/min Peaks: 1: Glycine Methylester (source), 2: Butoxy phenylalanineglycine methylester, 3 & 4: By Product of synthesis SofTA ELSD (blue line) Spray Chamber: 45°C Drift Tube: 60°C Filter: 0 UV (red line): 254nm

Monitoring the synthesis of this compound is difficult with UV detection because the source material did not have a chromaphore. ELS detection more accurately monitors the synthesis since it can “ see” both the product and the source without the baseline drift from the gradient seen with UV. The ELSD also gives a more accurate representation of concentration of the by products.

SofTA Corporation ELSD Application Underivatized Amino Acids Column: HyperCarb, 4.6 x 100mm Mobile Phase: A: 15mM Nonafluoropentanoic acid B: 70% ACN/30% 15mM Nonafluoropentanoic acid Gradient: (min:%B) 0: 1%, 9: 3%, 10: 11%, 14: 16%, 20: 35%, 26: 65%, 27: 85, 30:85% Flowrate:0.85mL/min Sample: 10nM Each Amino Acid Peaks: 1:Gly, 2: Ser 3: Ala, 4: Thr, 5: Asn, 6: Pro, 7: Asp 8: Gaba, 9: Gln, 10: E, 11: V, 12: Lys, 13: Leu, 14: Cys, 15: Met, 16: Ile, 17: His, 18: Arg, 19: Phe, 20: Tyr, 21: Impurity, 22: Trp SofTA ELSD Spray Chamber: 30°C Drift Tube: 55°C Filter: 6

SofTA Corporation ELSD Application

Gradient Ion Chromatography Column: BioBasic SCX, 2.1 x 250mm, 35°C Mobile Phase: A: Water B: 250mM Ammonium Formate Gradient: (min:%B) 0: 12%, 2:12%, 5:80%, 10:80% Flowrate: 0.35mL/min Peaks: 1: Cl, 2: Na, 3: K, 4: Mg, 5: Ca SofTA ELSD Spray Chamber: 10°C Drift Tube: 55°C Filter: 6

SofTA Corporation ELSD Application Ceramide Mixture Column: Betasil Diol, 4.6 x 150mm Mobile Phase: A: Chloroform B: 5% Water in Methanol Gradient: (min:%B) 0: 7%, 1:7%, 10:50% 13:50%, 14: 7% Flowrate: 1.2mL/min Sample: Ceramide Mixture (approx. 500ng each) Peaks: 1: Glc, 2: Gal, 3: Lac, 4: Gb3, 5: Gb4, 6: GalNacGb4 SofTA ELSD Spray Chamber: 45°C Drift Tube: 55°C Filter: 3

SofTA Corporation ELSD Application

Triglycerides Column: BetaBasic C18, 5µ, 4.6 x 150mL, 25°C Mobile Phase: A: Acetonitrile B: Dicloromethane Gradient: (min:%B) 0:30%, 16:40% Flowrate: 1ml/min Peaks: 1: LnLnLn 2: LLL, 3: OOO SofTA ELSD Spray Chamber: 60°C Drift Tube: 60°C Filter: 3

SofTA Corporation ELSD Application Macadamian Nut Oil Column: BetaBasic C18, 5µ, 4.6 x 150mL, 25°C Mobile Phase: A: Acetonitrile B: Dicloromethane Gradient: (min:%B) 0:30%, 20:50% Flowrate: 1ml/min Sample: 3µl of 0.5% SofTA ELSD Spray Chamber: 60°C Drift Tube: 60°C

SofTA Corporation ELSD Application Japanese Sake Column: Hypercarb, 4.6 x 100mm, 10°C Mobile Phase: A: 20mM NFPA, B: 70% Acetronitrile with 20mM NFP Gradient: (min:%B) 0:0%B, 20:40% B, 30:75%B Flowrate: 0.8mL/min Sample: Japanese Sake SofTA ELSD Spray Chamber: 15°C Drift Tube: 50°C Filter: 6

SofTA Corporation ELSD Application

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1. Rhamnose 2. Fructose 3. Glucose 4. Sucrose 5. Raffinose 6. Lactose

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Sample; 5ug each Column; SHODEX ASAHIPAK NH2P50 (4.6x150mmL) Mobile phase; 75%CH3CN/H2O Flow rate; 1ml/min SC/DT/FLT; 30/45/4

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