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An Affordable Evaporative Light Scattering Detector for Every HPLC 蒸发光散射检测器 Melissa J. Wilcox Alltech Associates, Inc. 2051 Waukegan Road Deerfield, IL.

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Presentation on theme: "An Affordable Evaporative Light Scattering Detector for Every HPLC 蒸发光散射检测器 Melissa J. Wilcox Alltech Associates, Inc. 2051 Waukegan Road Deerfield, IL."— Presentation transcript:

1 An Affordable Evaporative Light Scattering Detector for Every HPLC 蒸发光散射检测器 Melissa J. Wilcox Alltech Associates, Inc Waukegan Road Deerfield, IL Phone: ALLTECH Web Site: PP083

2 The New ELSD 800 – The First High-Performance, Low-Cost ELSD

3 Small and Stackable

4 Unique Method of Detection Three steps:  Nebulization  Mobile Phase Evaporation  Detection

5 Step 1: Nebulization  Column effluent passes through nebulizer needle  Mixes with nitrogen gas  Forms dispersion of droplets

6 Step 2: Mobile Phase Evaporation  Droplets pass through a heated zone  Mobile phase evaporates from the sample particle  Dried sample particles remain

7 Step 3: Detection  Sample particles pass through an optical cell  Sample particles interrupt laser beam and scatter light  Photodiode detects the scattered light

8 The ELSD Improves Baseline Stability and Detection Sensitivity Compared to RI Column:Prevail™ Carbohydrate ES, 5µm, 53 x 7mm (Part No ) Mobile Phase: Acetonitrile:Water (75:25) Flowrate: 2.0mL/min ELSD RI 1. Fructose 2. Glucose 3. Sucrose

9 The ELSD’s Response Accurately Reflects Equal Concentrations of Two Brominated Flame Retardants Hexabromocyclododecane (500ppm) 2. Tetrabromobisphenol A (allyl ether) (500ppm) Column:Alltima™ C18, 5µm, 250 x 4.6mm (Part No ) Mobile Phase: Acetonitrile:Water (95:5) Flowrate: 1.0mL/min UV at 254nm ELSD

10 The ELSD Responds to All Components in this Antihistamine Formulation Diphenhydramine 2. PEG, Gelatin 3. Sorbitan 4. Glycerol 5. Sorbitol UV ELSD Column:Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm (Part No ) Mobile Phase:Acetonitrile:0.1% Ammonium Carbonate, pH 6.5 (80:20) Flowrate: 1.0mL/min

11 Using Acetone in a Mobile Phase Gradient Results in a 50% Reduction in Analysis Time Fructose 2. Glucose 3. Maltose 4. Maltotriose 5. Maltopentaose 6. Maltohexaose 7. Maltoheptaose Column:Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm (Part No ) Mobile Phase:A: ACN B: Water Gradient:(Time,%B),(0,20),(50,35) Flowrate: 1.0mL/min Detector:Alltech ELSD Column:Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm (Part No ) Mobile Phase: A: ACN:Acetone (75:25) B: Water Gradient:(Time,%B),(0,24),(18,50) Flowrate: 1.0mL/min Detector:Alltech ELSD Unmodified Mobile Phase Modified Mobile Phase

12 Detect Difficult Samples like Triglycerides without Derivatizing 1. LLO 2. LLP 3. OOL 4. POL 5. PPL 6. OOO 7. OOP 8. PPO 9. OOS Column: Alltima™ C18, 3µm, 150 x 4.6mm Mobile Phase: A: Dichloromethane B: Acetonitrile Gradient:(Time,%B),(0,70),(10,55), (18,70),(20,70) Flowrate: 1.5mL/min Detector:Alltech ELSD 9515

13 Use ELSD in Parallel with MS to Obtain Maximum Structural and Concentration Information 1. FMOC-Ala-OH 2. FMOC-Trp-OH 3. FMOC-Arg(Pmc)-OH Column:YMC ODS-A, 5µm, 50 x 4.6mm Mobile Phase:A: 0.05% TFA in Water B: 0.035% TFA in Acetonitrile Gradient:(Time,%B),(0,30),(3.7,95),(4.2,95) Flowrate: 3.5mL/min MS ELSD

14 Replace Fluorescence Detectors with ELSD to Simply Methods by Eliminating Pre- or Post-column Derivatization 1. Glycine (Gly) 2. Serine / Asparagine (Ser/Asn) 3. Aspartic Acid (Asp) 4. Glutamine (Gln) 5. Alanine / Threonine (Ala/Thr) 6. Glutamic Acid (Glu) 7. Cysteine / Lysine (Cys/Lys) 8. Histidine (His) 9. Proline (Pro) 10. Arginine (Arg) 11. Valine (Val) 12. Methionine (Met) 13. Tyrosine (Tyr) 14. Isoleucine (Ile) 15. Leucine (Leu) 16. Phenylalanine (Phe) 17. Tryptophan (Trp) Column:Prevail™ C18, 5µm, 250 x 4.6mm (Part No ) Mobile Phase:A:5mM Heptafluorobutyric Acid, pH 1.0 w/0.7% TFA B:Acetonitrile Gradient:(Time,%B),(0,0),(6,0),(8,15),(25,35) Flowrate: 1.0mL/min Detector:Alltech ELSD 9413

15 Fat Soluble Vitamins — Micro Scale Column:Alltima™ C18, 3µm, 150 x 1mm (Part No ) Mobile Phase: Methanol:Chloroform (90:10) Flowrate: 50µL/min Detector:Alltech ELSD Vitamin A (100ng on-column) 2. Vitamin E (160ng on-column) 3. Vitamin K1 (130ng on-column)

16 Catechins in Green Tea — Prep Scale 1. Caffeine 2. Epicatechin 3. Epigallocatechin Gallate Column:Alltima™ C18, 5µm, 150 x 22mm (Part No ) Mobile Phase: (87:13) 0.05% TFA:Acetonitrile Flowrate: 23mL/min Detector:Alltech ELSD 10140

17 The ELSD 800 does not Respond to Changes in Mobile Phase Composition, Allowing Rapid Separations using Extreme Gradients Aspartame (0.9mg/mL) 2. Cortisone (0.3mg/mL) 3. Reserpine (0.3mg/mL) 4. Dioctyl Phthalate (0.3mg/mL) Column:Platinum™ C18, 3µm, 20 x 4.6mm (Part No ) Mobile Phase: A: 0.05% Formic Acid in Water B: Acetonitrile with 0.05% Formic Acid Gradient:(Time,%B),(0,5),(3,90),(7,90),(10,5) Flowrate: 1.0mL/min Detector:Alltech ELSD 800

18 Near Ambient Temperature Evaporation Maximizes Sensitivity for Semi-Volatiles Column:Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm Mobile Phase: Acetonitrile: 0.04% Ammonium Hydroxide (80:20) Flowrate: 1.0mL/min Detector:Alltech ELSD Glycerol

19 iso-Erythritol 2. Fructose 3. Sorbitol 4. Mannitol 5. Glucose (DP1) 6. Inositol 7. Sucrose 8. Maltitol 9. Maltose (DP2) 10. Raffinose 11. Maltotriose (DP3) 12. Maltotetraose (DP4) 13. Maltopentaose (DP5) 14. Maltohexaose (DP6) 15. Maltoheptaose (DP7) 16. Maltooctaose (DP8) 17. Maltononaose (DP9) 18. Maltodecaose (DP10) 19. Maltoundecaose (DP11) 20. Maltododecaose (DP12) 21. Maltotridecaose (DP13) 22. Maltotetradecaose (DP14) Mono-, Di-, Tri-, Oligosaccharides Column:Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm (Part No ) Mobile Phase:A: Acetonitrile B: 0.04% Ammonium Hydroxide in Water Gradient:(Time,%B)(0,17),(25,27),(40,45),(60,55) Flowrate: 1.0mL/min Detector:Alltech ELSD

20 Column:Allsphere™ Silica, 3µm, 100 x 4.6mm (Part No ) Mobile Phase: A: Isopropanol:Hexane:Water (58:40:2) B: Isopropanol:Hexane:Water (52:40:8) Gradient:(Time,%B),(0,0),(7,100),(19,100),(20,0) Flowrate: 1.25mL/min Detector:Alltech ELSD Egg Yolk Phospholipid 1. Phosphatidylethanolamine 2. Phosphatidylcholine 10214

21 10142 Ginsenosides in Korean Ginseng 1. Rh 2 2. Rh 1 3. Rg 2 4. Rg 3 5. Rg 1 6. Rf Column:Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm (Part No ) Mobile Phase: A: Acetonitrile:Water:Isopropyl Alcohol (80:5:15) B: Acetonitrile:Water:Isopropyl Alcohol (80:25:15) Gradient:(Time,%B),(0,25,),(28,85),(35,80),(45,75), (50,90),(51,100),(57,25) Flowrate: 0.8mL/min Detector:Alltech ELSD 7. Re 8. Rd 9. Rc 10. Rb Rb 1

22 % Tween 80 Column:Alltima™ C18, 5µm, 250 x 4.6mm (Part No ) Mobile Phase: Water:Methanol (60:40) Flowrate: 1.0mL/min Detector:Alltech ELSD


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