jack c. cappozzo separation science webinar march 12, 2013 · lc-qtof sample prep and...

Jack C. Cappozzo Separation Science Webinar

March 12, 2013

Research/Analytical Purpose Adulteration (economical/food safety)

Authenticity (economical/quality)

Quality

Sensory…Analytical Instrumentation

Adulteration Nineteen Arrested Over Alleged Olive Oil Fraud

(Spain, Olive Oil Times)

Spanish Police Say Palm, Avocado, Sunflower Was Passed Off as Olive Oil (Spain, Olive Oil Times)

Two Plead Guilty in Olive Oil Fraud Scheme, Sentenced to Two Years in Jail (Spain, 2011, Olive Oil Times)

Toxic Oil Syndrome (TOS), Spain 1981

Adulteration Quality

Health

Nutritional Loss

Illness

Economics

Economics At 10% adulteration (soybean oil), the producer saves

about $0.20/Kg……commodity pricing

Metric Ton of Olive Oil….@$2900

Metric Ton….$200

3 M Metric Tons in Spain

10% = 300,000 Metric Tons @ $60 million (in Savings)

Retail…10% oil (100 Kg) savings…$5000-10,000

Analytical Tests Needed for EVOO/OO 23 tests (reduce that to about 20 tests)

$$$$$$$$

Olive Oil (Extra Virgin) (Standards of Identity) Sterol composition and

content Fatty acid composition Saturated fatty acid content

in position 2 of the triglyceride

Unsaponifiable material Wax content Erythrodiol and uvaol ECN 42 content UV Light Absorbency Moisture and Volatiles Metal Traces

Hydrocarbons: stigmastadiene, stigmasta-3, 5 diene, and campesta-3, 5 diene

Trans fatty acid isomer Total aliphatic alcohols DNA characterization Color Aspect Free Acidity Peroxide Value Insoluble Impurities Flash Point

Olive Oil (Standards of Identity)

Halogenated Solvents

Additives

Heavy Metals & Pesticide Residues

Sensory

Olive Oil (IFSH Focus) Technology for Olive Oil

Authenticity & Adulteration

Rapid Technology for Olive Oil Quality (Sensory)

Collaborators

Agilent

Alpha Mos

AOCS

LC-MS/MS and QTOF GC-MS

ICP-MS

Electronic Nose (FID)

Statistical Analysis Software

Goal Rapid Techniques to relate to

sensory quality

(Rapid) Technology to confirm Authenticity and Adulteration

Technology LC-Q-TOF

Electronic Nose/GC-MS PCA

Statistical Mapping

Mass Spectroscopy LC-MS-TOF (Q-

TOF)

LC-MS/MS

LC-QTOF Sample Prep and Chromatographic Parameters Method: 5 mL oil diluted with 5 mL hexane extracted twice with 10 mL

methanol, evaporated and reconstituted in 50% methanol in water. (Polar Fraction)

Chromatographic separation achieved on a Poroshell 120 EC-C18 2.7 µm column kept at 45oC

Gas temp was 325oC with a flow of 10 L/min

Nebulizer was 45 psig

Analysis was conducted in positive ionization mode

Injection volume 5 µL

The mobile phase was a gradient of 0.1% acetic acid in water and acetonitrile starting at 5% ACN and increasing to 100% ACN over 25 min with a post time of 3 min.

Italian Olive Oil/Soybean Oil

Overlay

Oil Analysis Using QTOF Technology Chromatograms Produced Using MFE (Molecular Feature Extraction) and Contain Several Thousands of Compounds

Olive Oil from Palestine

Soybean Oil

Mass Profiler Professional MPP software allows multivariance statistical analysis of MS data

Data can be filtered based on set parameters and organized into entity lists.

Mass, Retention Time

PCA Italian Olive Oil

Olive Oil Varieties

PCA Analysis of Olive Oils and Olive Oils Combined with Vegetable Oils

Venn Diagram of Olive Oils and Olive Oils Overlap with Soybean

Specific entity lists can be displayed in many different ways. An example is A Venn diagram representing similarities and differences among various groups of samples.

ID Browser for Identification of Compounds included in Entity Lists

An entity list can be also identified based on available databases and libraries using the ID browser

Table of representative 38 compounds identified in EVOO. Over 200 compounds identified.

Name Formula Mass RT

12-amino-octadecanoic acid C18 H37 N O2 299.282 21.911

2-Oxo-4E-hexenoic acid C6 H8 O3 128.0478 1.657

Vanillin C8H8O3 152.0473 1.298

3,4-bis(methylene)-Hexanedioic acid C8 H10 O4 170.0577 1.657

2,3-Dihydroxy-3-methylvaleric acid C6 H12 O4 148.0733 4.995

Oleamide C18 H35 N O 281.2714 21.911

2,2-dimethyl-4-pentenoic acid C7 H12 O2 128.0831 8.676

(R)-3-methyl-2-oxo-Pentanoic acid C6 H10 O3 130.0628 3.276

2,2-dimethyl-3-butenoic acid C6 H10 O2 114.0691 3.919

2-Hydroxycinnamic acid C9 H8 O3 164.0465 10.404

3-oxo-4-pentenoic acid C5 H6 O3 114.031 3.61

(5R)-5-Hydroxyhexanoic acid C6 H12 O3 132.079 16.458


2-Octenedioic acid C8 H12 O4 172.0729 2.709


2-keto-n-caprylic acid C8 H14 O3 158.0938 1.716


Isovalerylglucuronide C11 H18 O8 278.1018 4.495

2E,4E,6E,8E-Decatetraenedioic acid C10 H10 O4 194.0573 14.721

11-amino-undecanoic acid C11 H23 N O2 201.1724 1.248

2-ethyl-3E-hexenoic acid C8 H14 O2 142.0984 1.949

12-amino-dodecanoic acid C12 H25 N O2 215.1878 2.166

6,8-dihydroxy-octanoic acid C8 H16 O4 176.1056 9.803


13-methyl-pentadecanoic acid C16 H32 O2 256.2393 0.931

10,12,14-octadecatrienoic acid C18 H30 O2 278.2254 20.984

3,4,5-Trimethoxycinnamic acid C12 H14 O5 238.0826 15.314

10-oxo-5,8-decadienoic acid C10 H14 O3 182.093 10.629

3,6-dioxo-decanoic acid C10 H16 O4 200.1035 10.629

2S-amino-pentanoic acid C5 H11 N O2 117.0795 6.372

5-oxo-7-octenoic acid C8 H12 O3 156.0798 11.306

(+)-catechin C15 H14 O6 290.0809 8.108

Tyrosol C8H10O2 138.0681 11.306

(+)-pinoresinol C20H22O6 358.1416 12.684

Syringic acid C9H10O5 198.0536 10.195

Caffeic acid C9H8O4 180.0423 9.752

Elenolic acid C11 H14 O6 242.0788 4.578

p-coumaric acid C9H8O3 164.0465 10.404

Name Formula Mass RT

12-amino-octadecanoic acid C18 H37 N O2 299.282 21.911

2-Oxo-4E-hexenoic acid C6 H8 O3 128.0478 1.657

Vanillin C8H8O3 152.0473 1.298

3,4-bis(methylene)-Hexanedioic acid C8 H10 O4 170.0577 1.657

2,3-Dihydroxy-3-methylvaleric acid C6 H12 O4 148.0733 4.995

Oleamide C18 H35 N O 281.2714 21.911



2,2-dimethyl-3-butenoic acid C6 H10 O2 114.0691 3.919

2-Hydroxycinnamic acid C9 H8 O3 164.0465 10.404

3-oxo-4-pentenoic acid C5 H6 O3 114.031 3.61



2-Octenedioic acid C8 H12 O4 172.0729 2.709


2-keto-n-caprylic acid C8 H14 O3 158.0938 1.716


Isovalerylglucuronide C11 H18 O8 278.1018 4.495

2E,4E,6E,8E-Decatetraenedioic acid C10 H10 O4 194.0573 14.721

11-amino-undecanoic acid C11 H23 N O2 201.1724 1.248

2-ethyl-3E-hexenoic acid C8 H14 O2 142.0984 1.949

12-amino-dodecanoic acid C12 H25 N O2 215.1878 2.166

6,8-dihydroxy-octanoic acid C8 H16 O4 176.1056 9.803


13-methyl-pentadecanoic acid C16 H32 O2 256.2393 0.931

10,12,14-octadecatrienoic acid C18 H30 O2 278.2254 20.984

3,4,5-Trimethoxycinnamic acid C12 H14 O5 238.0826 15.314

10-oxo-5,8-decadienoic acid C10 H14 O3 182.093 10.629

3,6-dioxo-decanoic acid C10 H16 O4 200.1035 10.629

2S-amino-pentanoic acid C5 H11 N O2 117.0795 6.372

5-oxo-7-octenoic acid C8 H12 O3 156.0798 11.306

(+)-catechin C15 H14 O6 290.0809 8.108

Tyrosol C8H10O2 138.0681 11.306

(+)-pinoresinol C20H22O6 358.1416 12.684

Syringic acid C9H10O5 198.0536 10.195

Caffeic acid C9H8O4 180.0423 9.752

Elenolic acid C11 H14 O6 242.0788 4.578

p-coumaric acid C9H8O3 164.0465 10.404

Blue=phenolic compounds Green=fatty acids

3-D PCA

PCA Analysis of Italian (P) Greek (G) Olive Oils and Soybean Oil(B).

Cluster analysis based on five sensors with the highest discrimination power.

Palestinian Olive Oil Adulterated with 2%, 5% and 10% Soybean Oil

IFSH Direction Oxidation correlated to Analytical Methods

PV, FFA, …

Sensory correlated to Analytical Methods

Continue with Adulteration and Authenticity

ICP-MS

HS-GC-MS

Statistical Analysis

PCA

Mass Profiler Professional

IFSH Institute for Food Safety & Health (IFSH) Bedford Park, IL

National Center for Food Safety and Technology (NCFST)

At the Illinois Institute of Technology (IIT) Chicago

50:50 IIT:FDA

Collaborative Model of Industry:Academia:FDA

Food Safety…Chemistry, Process Engineering, Microbiology (molecular biology)

Nutrition (biological/clinical)

In existence for 23 years

Thank you

Questions?

jack c. cappozzo separation science webinar march 12, 2013 · lc-qtof sample prep and...

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