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.

Slides:

Advertisements

Similar presentations

Proteins: Structure reflects function….. Fig. 5-UN1 Amino group Carboxyl group carbon.

Advertisements

Amino Acids PHC 211.  Characteristics and Structures of amino acids  Classification of Amino Acids  Essential and Nonessential Amino Acids  Levels.

A Ala Alanine Alanine is a small, hydrophobic

Review of Basic Principles of Chemistry, Amino Acids and Proteins Brian Kuhlman: The material presented here is available on the.

Aminoácidos Unidad de las proteínas. Isómeros Clasificación Esenciales Valina (Val) Leucina (Leu) Isoleucina (Ile) Fenilalanina (Phe) Metionina (Met)

BASIC CELL BIOLOGY I CHEMISTRY of LIFE CLASSIFICATION OF THE ORGANIC COMPOUNDS, FUNCTIONAL GROUPS.

Metabolic fuels and Dietary components Lecture - 2 By Dr. Abdulrahman Al-Ajlan.

• Exam II Tuesday 5/10 – Bring a scantron with you!

5’ C 3’ OH (free) 1’ C 5’ PO4 (free) DNA is a linear polymer of nucleotide subunits joined together by phosphodiester bonds - covalent bonds between.

The Organic Molecules of Living Organisms

Lectures on Computational Biology HC Lee Computational Biology Lab Center for Complex Systems & Biophysics National Central University EFSS II National.

Amino Acids, Peptides, Protein Primary Structure Chapter 3.

Amino Acids, Peptides, Protein Primary Structure

Amino Acids, Peptides, Protein Primary Structure

FIGURE (part 2) Urea cycle and reactions that feed amino groups into the cycle. The enzymes catalyzing these reactions (named in the text) are distributed.

Molecular Techniques in Molecular Systematics. DNA-DNA hybridisation -Measures the degree of genetic similarity between pools of DNA sequences. -Normally.

©CMBI 2001 A Ala Alanine Alanine is a small, hydrophobic residue. Its side chain, R, is just a methyl group. Alanine likes to sit in an alpha helix,it.

You Must Know How the sequence and subcomponents of proteins determine their properties. The cellular functions of proteins. (Brief – we will come back.

Amino Acids, Peptides, and Proteins.. Classification of Amino Acids.

1 Biological Macromolecules Much larger than other particles found in cells Made up of smaller subunits Found in all cells Great diversity of functions.

Lesson today Protein Teacher : Isroli Laboratory : Animal Physiology and Biochemistry Faculty : Animal Agriculture Diponegoro University.

Help Memorizing Amino Acids General rule, side chains have a maximum of four C atoms, with other structures attached onto up to 4 carbon atoms.

Chapter 27 Amino Acids, Peptides, and Proteins. Nucleic Acids.

Proteins and Enzymes Nestor T. Hilvano, M.D., M.P.H. (Images Copyright Discover Biology, 5 th ed., Singh-Cundy and Cain, Textbook, 2012.)

Proteins account for more than 50% of the dry mass of most cells

1.What makes an enzyme specific to one type of reaction (in other words, what determines the function of a protein)? –SHAPE determines the function of.

How does DNA work? What is a gene?

Protein Synthesis. DNA RNA Proteins (Transcription) (Translation) DNA (genetic information stored in genes) RNA (working copies of genes) Proteins (functional.

©CMBI 2006 Amino Acids “ When you understand the amino acids, you understand everything ”

How Proteins Are Made Mrs. Wolfe. DNA: instructions for making proteins Proteins are built by the cell according to your DNA What kinds of proteins are.

The Use of Evaporative Light Scattering Detection in Pharmaceutical Analysis 1.

. Sequence Alignment. Sequences Much of bioinformatics involves sequences u DNA sequences u RNA sequences u Protein sequences We can think of these sequences.

Macromolecules Chapter 5. Macromolecules Large complex molecules Carbohydrates, proteins, lipids & nucleic acids.

LESSON 4: Using Bioinformatics to Analyze Protein Sequences PowerPoint slides to accompany Using Bioinformatics : Genetic Research.

Proteins – Amides from Amino Acids

Amino Acids are the building units of proteins

Learning Targets “I Can...” -State how many nucleotides make up a codon. -Use a codon chart to find the corresponding amino acid.

Fig Second mRNA base First mRNA base (5 end of codon) Third mRNA base (3 end of codon)

Welcome Back! February 27, 2012 Sit in any seat for today. You will have assigned seats tomorrow Were you absent before the break? Plan on coming to tutorial.

NOTES: 2.3 part 2 Nucleic Acids & Proteins. So far, we’ve covered… the following MACROMOLECULES: ● CARBOHYDRATES… ● LIPIDS… Let’s review…

intro-VIRUSES Virus NamePDB ID HUMAN PAPILLOMAVIRUS 161DZL BACTERIOPHAGE GA1GAV L-A virus1M1C SATELLITE PANICUM MOSAIC VIRUS1STM SATELLITE TOBACCO NECROSIS2BUK.

Macromolecules of Life Proteins and Nucleic Acids

UNIT 8 THE AMINO ACIDS. LEARNING OBJECTIVES  State the composition and describe the structure of amino acids.  Classify amino acids.  List essential,

AMA 108 Nutrition. Essentials of Nutrition Nutrients – essential elements found in food: Carbohydrates – provide body with energy - simple: sugars, not.

Transcription and Translation

Amino Acids ©CMBI 2001 “ When you understand the amino acids, you understand everything ”

Chapter 3 Proteins.

Amino Acids  Amino Acids are the building units of proteins. Proteins are polymers of amino acids linked together by what is called “ Peptide bond” (see.

Parts is parts…. AMINO ACID building block of proteins contain an amino or NH 2 group and a carboxyl (acid) or COOH group PEPTIDE BOND covalent bond link.

Amino acids Common structure of 19 AAs H3N+H3N+ COO - R H C Proline.

Genomics Lecture 3 By Ms. Shumaila Azam. Proteins Proteins: large molecules composed of one or more chains of amino acids, polypeptides. Proteins are.

Proteins Tertiary Protein Structure of Enzyme Lactasevideo Video 2.

Amino acids Proof. Dr. Abdulhussien Aljebory College of pharmacy

Biochemistry: The Chemistry of Life

Transport proteins Transport protein Cell membrane

Chapter 2 part 2: Biochemistry

Concept 5.3: Lipids are a diverse group of hydrophobic molecules

Fig. 5-UN1  carbon Amino group Carboxyl group.

A Ala Alanine Alanine is a small, hydrophobic

Proteins Genetic information in DNA codes specifically for the production of proteins Cells have thousands of different proteins, each with a specific.

The 20 amino acids.

The 20 amino acids.

Example of regression by RBF-ANN

Proteins Proteins have many structures, resulting in a wide range of functions Proteins do most of the work in cells and act as enzymes 2. Proteins are.

“When you understand the amino acids,

Presentation transcript:

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

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

Small and Stackable

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

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

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

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

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

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

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

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

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

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

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

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)

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

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

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

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

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

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

% 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