An improved qualitative method for detecting and isolating shark-repelling semiochemicals Eric M. Stroud Michael M. Herrmann Dr. Samuel Gruber AES July.

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Presentation transcript:

An improved qualitative method for detecting and isolating shark-repelling semiochemicals Eric M. Stroud Michael M. Herrmann Dr. Samuel Gruber AES July 09, 2005

Introduction Analytical method Analytical method Observe decay process Observe decay process Field testing Field testing

HPLC-UV/Vis Solvents: Methanol + 0.1% acetic acid Water + 0.1% acetic acid

HPLC-UV/Vis Ternary HPLC Pump Gradient control: 0-10 minutes: 100% methanol/acetic acid minutes: Linear gradient to 100% water/acetic acid minutes: 100% water/acetic acid minutes: Linear gradient to methanol/acetic acid minutes: 100% methanol/acetic acid

HPLC-UV/Vis Injector with 50uL loop

HPLC-UV/Vis Guard column

HPLC-UV/Vis Column heater: Maintain temperature at 25C Column: Waters Novapak C18 RP 3.9x150 mm

HPLC-UV/Vis UV / Vis Detectors Monitor: 240nm-340nm - semios 440nm – secondary amines 570nm – primary amines

HPLC-UV/Vis Integrator or PC

Why ??? November 2004 Field Test November 2004 Field Test Visual/odor was OK Visual/odor was OK Delivery method same Delivery method same Same species Same species What happened? What happened?

Good Batch – Early Eluting

~7 minutes ~6 minutes ~5 minutes

Bad Batch – Early Eluting

Good Batch – Late Eluting

Bad Batch – Late Eluting

GWS Head – Early Eluting

GWS Head – Late Eluting

GWS Field Trials 6 C. perezii, 2 C. acronotus 6 C. perezii, 2 C. acronotus 500mL GWS, 2 minute span 500mL GWS, 2 minute span 4 C. perezii, 2 C. acronotus gone 4 C. perezii, 2 C. acronotus gone 2 C. perezii disinterested for 10 mins with constant attractant 2 C. perezii disinterested for 10 mins with constant attractant

A2, good batch, 1 yr A13N, 1 yr SQ1, 1 yr

GWS Head Tiger shark carcass Nurse shark carcass Lemon shark carcass, 2yr. C. limbatus carcass GWS carcass Lemon shark carcass, 1yr. Southern ray GWS Tail

0 days

7 days

0 days 7 days 21 days

0 days 7 days 21 days 40 days

Secondary 440nm

Primary 570nm

Conclusions HPLC is suitable for degradation studies HPLC is suitable for degradation studies GWS head currently has highest yield of target compounds GWS head currently has highest yield of target compounds Storage procedures / shelf life with extracts Storage procedures / shelf life with extracts