© 2009 Perkin Elmer As Speciation in Apple Juice Charles Schneider, Presenter Kenneth Neubauer, Co-Author PerkinElmer.

Slides:



Advertisements
Similar presentations
Dr. Birgit Schmauser, BfArM, Bonn
Advertisements

Analytical Method Development and Validation
Instrumental Analysis
Cruces-Blanco, C., Gamiz-Gracia, L., Garcia-Campana A.M., Applications of Capillary Electrophoresis in Forensic Analytical Chemistry Trends in Analytical.
Chromatographic Process Provides the analyte transport. Immobile phase. Mixture of components dispersed in the mobile phase.
Microspectrophotometry Validation. Reasons for Changing Instruments Reduced reliability. Limited efficiency. Limited availability and cost of replacement.
1 1. Introduction H: High P : Performance (Pressure) L : Liquid C : Chromatography GC : Gas chromatography TLC: Thin layer chromatography IC : Ion chromatography.
Ion-Pair Chromatography In addition to the aqueous buffer and an organic solvent that is typical for reversed-phase, the mobile phase contains a counter.
Applications of Analytical Chemistry at RTI International  Argon plasma instruments Atomic Emission (ICP-AES) – 3 Quadrupole Mass Spectrometers (ICP-MS)
World Health Organization
Figure 1. Influence of sample solvent on peak shape. The sample is dissolved in buffer with (a) 0%, (b) 30%, (c) 50%, and (d) 70% acetonitrile. Sample.
Results Initial chromatographic conditions [Figure 2a caption] for the separation of the degradation products of aspirin were problematic due to the long.
Chem. 133 – 5/14 Lecture. Announcements I Lab Stuff Pass out Peer Review Assignments See Term Project Handout for Format of Poster Today should be check.
1 April 7, 2009 C. Lu Monitoring the RPC Gas Mixture with a Gas Chromatograph C. Lu Princeton University.
 Amino acid analysis refers to the methodology used to determine the amino acid composition or content of proteins, peptides, and other pharmaceutical.
Evaluating Robustness of Signal Timings for Conditions of Varying Traffic Flows 2013 Mid-Continent Transportation Research Symposium – August 16, 2013.
Created with MindGenius Business 2005® Performance parameters (Advanced) Theoretical Plate Model Performance parameters (Advanced) Theoretical Plate Model.
The following minimum specified ranges should be considered: Drug substance or a finished (drug) product 80 to 120 % of the test concentration Content.
Chromatographic separations Chapter 26 The “stuff” you do before you analyze a “complex” sample.
1.1 General description - Sample dissolved in and transported by a mobile phase - Some components in sample interact more strongly with stationary phase.
LECTURE 9 CHROMATOGRAPHIC SEPARATIONS The “stuff” you do before you analyze a “complex” sample.
Analytical considerations
Arsenic Speciation - Year 4/5 Bernine Khan University of Miami Dept. of Civil, Architectural & Environmental Engineering July 9th, 2001.
Chem. 231 – 1/28 Lecture. Introduction Goals of Course Discussion of More Practical Aspects of Separation Science Provide Specific Lab Training in Use.
INTERNATIONAL LABOUR ORGANIZATION Conditions of Work and Employment Programme (TRAVAIL) 2012 Module 13: Assessing Maternity Protection in practice Maternity.
Bias and Errors. Some Terms Used to Describe Analytical Methods Accuracy Precision LOD RDL LOQ Selectivity Sensitivity Linearity Ruggedness.
Higher Grade / Intermediate 2 Physical Education Preparation of the Body October Break Homework.
Simple Scale-up on a 940-LC Analytical to Preparative HPLC
Preparation of the Body Lesson 6 Int 2 / Higher Grade Physical Education.
CHROMATOGRAPHY Chromatography basically involves the separation of mixtures due to differences in the distribution coefficient.
Title: Lesson 3 Rate Law and Reaction Order Learning Objectives: – Know that rate law can only be derived from experimental data – Understand the concept.
Chem. 231 – 2/18 Lecture. Announcements Set 2 Homework – Due Wednesday Quiz 2 – Next Monday Set 1 Labs –should be switching instruments today (or after.
Quality Assurance How do you know your results are correct? How confident are you?
7000A GC-QQQ Applications Pesticides in Foods (1).
Advanced Analytical Chemistry – CHM 6157® Y. CAIFlorida International University Updated on 9/26/2006Chapter 3ICPMS Interference equations Isobaric.
By: Arafath and Nick. What is it  Chromatography is a technique that is used to separate the substances present in a mixture.  It is widely used to.
1/16/2016The GC-in-a-PC 1 Pneumatic Focusing Gas Chromatography PFGC Robert O’Brien, Portland State Univ. & VOC Technologies The GC-in-a-PC Development.
Exam next week: Chapter 4?
HPLC.
Why HPLC? Almost universal applicability Remarkable precision Highly commercially available (competition)
Introduction Arsenic is a metalloid that can be found in the environment in rocks, soil, and water. It occurs naturally as the minerals arsenopyrite (FeAsS),
Selling Restek LC Solutions for Analytical Scale Laboratories Rick Lake Product Line Manager Liquid Chromatography x 2379
Lecture 10 ANALYTICAL METHOD DEVELOPMENT AND VALIDATION IN HPLC AND GC. Lecture 10 – Chromatography, Dr. Rasha Hanafi 1© Dr. Rasha Hanafi, GUC.
FAME in AVTUR using GC Heart-Cut and refocusing – IP PM EL Piet Koppen – AC Analytical Controls EI Fame in Jet workshop March 1, 2012.
Lecture 8 Peak Parameters and Quantitative chromatography
Determination of metformin in urine (by Liquid Chromatography LC)
SEMINAR ON PRESENTED BY BRAHMABHATT BANSARI K. M. PHARM PART DEPARTMENT OF PHARMACEUTICS AND PHARMACEUTICAL TECHNOLGY L. M. COLLEGE OF PHARMACY.
Instrumental Analysis (I) HPLC Tutorial 8. Graded presentation Students in groups of 4-5 individuals are asked to prepare a presentation (weight=5% of.
PREPARED BY VAISHNANI BHAVIN M.PHARMA-I (Q.A.). Qualitative error sources in HPLC: Poor separation power. Some special substances from the samples can.
Forensic Toxicology Use Only Analysis of ETG, ETS using the Thermo Scientific Exactive Mass Spectrometer Kent Johnson Fortes lab, Portland Oregon.
이 장 우. 1. Introduction  HPLC-MS/MS methodology achieved its preferred status -Highly selective and effectively eliminated interference -Without.
Plasma Free Metanephrines Analysis using LC-MS/MS with Porous Graphitic Carbon Column Xiang He (Kevin) and Marta Kozak Thermo Fisher Scientific.
Indranil Sen, Wei Zou, Ryszard Gajek, Jianwen She*
Table 1. Quality Parameters Being Considered for Evaluation
Multi-Analyte LC-MS/MS Methods – Best Practice.
James Byrd, Marta Kozak 28 Apr 2011
Chem. 133 – 5/11 Lecture.
A sensitive and repeatable method for characterization of sulfonamides and trimethoprim in honey using QuEChERS extracts with Liquid-Chromatography-Tandem.
Troubleshooting Method Development – a case study. Diclofenac in milk.
Automation of Sample Preparation for Trace Analysis Haibin Wan
Dr. Birgit Schmauser, BfArM, Bonn
High Performance Liquid Chromatography (HPLC)
EPA Method Equivalency
Concentrations of arsenic in human urine: A correlation between total arsenic by ICP-MS and speciated arsenic by HPLC-ICP-MS Indranil Sen, Wei Zou, Josephine.
Hydride Generation–AFS for Arsenic Speciation in Food
Strategies for Eliminating Interferences in Optical Emission Spectroscopy Best practices to optimize your method and correct for interferences to produce.
The Analysis of Soils and Waters in Accordance with U. S
Topic Osmosis Level GCSE (or any course for students aged 11-16)
Best Practices for Identification and Quantitation
Darrell Clinton and H. Dorota Inerowicz Purdue University
Presentation transcript:

© 2009 Perkin Elmer As Speciation in Apple Juice Charles Schneider, Presenter Kenneth Neubauer, Co-Author PerkinElmer

… we explain that some scientific studies have shown that two forms of organic arsenic found in apple juice could also be harmful, and for that reason, the FDA counts these two forms of organic arsenic in with the overall content for inorganic arsenic.

Initial Goals Develop an Improved Method for As Speciation in Urine  As(III), As(V), MMA, DMA, AsB Increase the Speed of Analysis  Current methods require 10 minutes or longer  Some methods use of LC gradients  Shorter chromatograms but long sample-to-sample times  i.e. 8 minute chromatogram, but 20 minutes between samples Attain Baseline Resolution of All Species

As5 As3 MMA DMA AsB Improving the Separation 1 ppb Mixed As Standard Faster Separation, Better Resolution  Reversed-Phase Ion Pairing Chromatography

As5 As3 MMA DMA AsB Separation in Urine 2 Consecutive Injections of a Urine Sample Diluted 2x Urine Matrix Does Not Hinder the Separation  All 5 species are baseline resolved  Good Injection-to-Injection Stability

Other Matrices: Apple Juice Recent Media Focus on As in Apple Juice  What forms? Will the Newly-Developed Separation Work in Apple Juice?  Urine: High salt content  Juice: High sugar content FDA Method in Development for As Speciation in Fruit Juices

FDA Method Elemental Analysis Manuals: Section 4.10: HPLC-ICP-MS Determination of Four Arsenic Species in Fruit Juice  Version Draft 0.82 (August 2010)  Author: Sean D. Conklin Species:  As(III), As(V), MMA, DMA  Check for AsB interference Can this separation be improved?  Shorter time  Better resolution between peaks

Goals Focus on the Separation Chemistry of As Species in Apple Juice  Sample preparation  Chromatographic conditions  Affect of juice matrix  Variability among different apple juices Once the LC Conditions are Established, Focus on Quantitation  Optimal ICP-MS conditions  Calibration strategies Characterize Method

Following FDA LC Methodology Separation of 5 Species Slightly better separation and shorter chromatogram  Different version of ion-exchange column listed in the FDA Method

Following FDA LC Methodology Apple Juice – 5x Dilution Juice matrix does not affect retention times or separation/resolution of peaks Apple Juice - 5x Dilution + 5 ppb Spike Dilution in mobile phase

Applying Improved As in Urine LC Methodology Apple Juice +/- 1 ppb Spike Apple Juice Matrix Does Not Affect the Separation  No matrix-induced peak shift  Peak shape is unaffected  Dilution in mobile phase Black: Juice (5x dilution) Blue: Juice (5x) + 1ppb spike As5 As3 MMA DMA AsB ICP-MS in Standard Mode  No Interferences

Effect of Dilution Factor Lower Dilution Factor Doesn’t Affect the Chromatography  No change in retention time or peak shape  But some dilution is necessary  Matrix match mobile phase and sample Black: Juice – 5x dilution Blue: Juice – 2x dilution As5 As3 MMA DMA Benefit: Lower concentrations can be measured  How low is relevant?

As5 As3 MMA DMA AsB Effect of Dilution Factor Apple Juice Diluted 2x +/- 1 ppb Spike Lower Dilution Factor Does Not Affect the Chromatography  No matrix-induced peak shift  Peak shape is unaffected  No interferences Black: Juice (2x dilution) Blue: Juice (2x) + 1ppb spike

Stability 20 Injections over 100 Minutes of Apple Juice Diluted 2x Good Stability  Still need to evaluate longer-term stability As5 As3 MMA DMA

Effect of Injection Volume Benefit: Ability to See Lower Levels  How low is relevant? Drawbacks of Larger Injection Volumes  Overload the column for samples with high concentrations  Possible increased sample matrix effects on the chromatography Black = 20 µL Blue = 40 µL As5 As3 MMA DMA 0.1 ppb Standard

2 points/sec 4 points/sec Effect of Sampling Rate Higher Sampling Rate Gives Noisier Peaks  More difficult to integrate low levels  Can’t sample too slow or else peak integration is less accurate  How low is relevant? As5 As3 MMA DMA As5 As3 MMA DMA

Calibration MMA As5 As3 DMA As5 As3 DMA MMA Calibration Standards Blank, 0.1, 0.25, 0.5, 1.0 ppb (prepared in diluent)

Initial Quantitation Tests As5 As3 DMA MMA 5 Consecutive Injections of Apple Juice Diluted 2x

Initial Quantitation Results: Various Apple Juices All Apple Juice Diluted 2x and Run Against Aqueous External Calibration Curve

Initial Quantitation Results: Various Apple Juices Comparison: Total As vs. Sum of Species Total As  Measured directly by ICP-MS (i.e. no LC) on a different instrument in another lab by a different chemist  Summed As Species  Sum of individual species from previous slide  MMA and DMA standards made to final concentration of total molecule  As is ≈ 54% of the total molecule  Summed species are adjusted

Summary – Initial Results Four As species present in apple juice can be separated and measured in 4 minutes Various aspects of the chromatography have been characterized  Effects of sample matrix, dilution factor, sampling rate, injection volume Method has been applied to several apple juice samples  Separation is not affected by the different juices tested Initial results show good stability  More stability tests need to be performed Accurate quantitative results are obtained using aqueous calibration standards  Sum of species agrees with total As measurements All measurements can be made at As75 in Standard mode  No interferences observed, so cell mode is not needed

Going Forward Initial results are encouraging and suggest the method will work Test Method Ruggedness  Day–to-day results  Run same samples over several days  Stability over long runs  8-12 hours  Variation from column-to-column  Repeat tests on a 2 nd column of the same type  Matrix Effects  Can this method be applied to other matrices? Once these criteria have been met, the details of the method will be published

Questions That Need Answering Other juices need testing? Which ones? What levels need to be measured?  How low is biologically relevant? How will the test be implemented?  Measure juices for total As and only those about a certain level subject to speciation analysis? Are there SRMS? What else needs to be shown for the method to gain acceptance?

Other Matrices: Rice Run by Alternate Separation Method How Would the New Method Work?  Better separation of other As peaks in brown rice? Thai Rice As5 As3 DMA Brown Rice As5 As3 DMA ? ? ?