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6 protocols using z 3 hexenal

1

Chemical Compounds for Behavioral Assays

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Authentic samples of the compounds tentatively identified by GC‐MS were purchased for structure confirmation by GC peak enhancement and for behavioural assays. (Z)‐3‐Hexenal (50% solution in triacetin), (E)‐2‐hexenal (99%), p‐cymene (99%), 1,8‐cineole (99%), γ‐terpinene (95%), linalool oxide [2‐(5‐methyl‐5‐vinyltetrahydro‐1‐furyl)‐2‐propanol] (>97%) and (R/S)‐camphor (96%) were obtained from Sigma‐Aldrich (Gillingham, Dorset, UK). (Z)‐3‐Hexen‐1‐ol (98%) and (Z)‐3‐hexenyl acetate (99%) were purchased from Alfa Aesar (Heysham, Lancs, UK), and (Z)‐ocimene (90%) was obtained from Bush Boake Allen (London). 3‐Ethylacetophenone (95%) was obtained from Maybridge (Tintagel, Cornwall, UK). (E)‐Ocimene (95%) was synthesised according to Chou et al.13 (E)‐4,8‐Dimethyl‐1,3,7‐nonatriene (95.6%) was synthesised from geraniol by oxidation to the corresponding aldehyde, followed by Wittig methylenation.14
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2

GC-MS Analysis of Volatile Compounds

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After sampling for 10 min the fiber was desorbed for 1 min in the injection port which was constantly kept at 250°C. Compounds were separated on HP-5 ms column (30 m × 250 μm, 0.25 μm film thickness; Agilent) in an Agilent 7890A gas chromatograph with a temperature program set to 40°C for 5 min, increasing to 140°C at a rate of 5°C per min, followed by increasing temperature to 250°C at a rate of 15°C per min and an additional 5 min at 250°C. Helium was used as the carrier gas with the transfer column flow set to 3 mL per minute and a flow rate of 1 mL per min thereafter. Mass spectra were generated by an Agilent 7200 accurate-mass quadrupole time-of-flight mass spectrometer, operating in electron ionization mode (70 eV) at 230°C and collected with an acquisition rate of 20 scans per second. volatiles were identified and quantified using standard solutions of Z-3-hexenal, Z-3-hexenol, E-2-hexenal, E-2-hexenol and (E,Z)-2,6-nonadienal (Sigma-Aldrich). (Z,Z)-3,6-nonadienal was synthesized from (Z,Z)-3,6-nonadienol (Ventos) as described below.
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3

Analytical Reagents for Chemical Protocols

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Unless otherwise stated, all the reagents used in this study were analytically pure. The water used for various experiments was purified with a Milli-Q purification system. Tween 80, glucose anhydrous, cupric sulfate, potassium sodium tartrate tetrahydrate, sodium hydroxide, magnesium chloride, acetyl-CoA, linoleic acid sodium salt, nicotinamide adenine dinucleotide, dithiothreitol, Triton X-100, 4-morpholineethanesulfonic acid, polyvinylpolypyrrolidone (PVPP), dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium acetate, acetaldehyde, and butanol were obtained from Yuanye Co., Ltd. (Shanghai, China). BTH, E-2-hexenal, hexanal, Z-3-hexenal, E-2-hexenol, Z-3-hexenol, hexanol, Z-3-hexenyl acetate, hexyl acetate, 2-octanol, linolenic acid, and linoleic acid were HPLC grade and obtained from Sigma-Aldrich (Shanghai, China).
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4

Comprehensive Volatile Compound Analysis Protocol

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Methanol (HPLC grade) was purchased from Merck (Darmstadt, Germany) NaCl (analytical grade) was purchased from Sinopharm Chemical Reagent Co., Ltd (Beijing, China); and pure water was obtained from the Milli-Q purification system (Millipore, Bedford, MA).
The chemical standards were purchased as follows: (Z)-3-hexenal, 2-octanol, hexanal, (E)-2-hexenal, geranic acid, phellandrene, β-myrcene, β-damascenone, D-limonene, citronellol, benzyl alcohol, neral, α-terpineol, geranial, geraniol, rose oxide II (cis), rose oxide I (trans), phenylethyl alcohol from Sigma (St. Louis, MO, USA); octanoic acid, pentanal, octanal, nonanal, benzaldehyde, 3-methylbutanal, hexanoic acid, (Z)-3-hexenol, 1-octen-3-ol, heptanol, ethyl acetate, ethyl butyrate, ethyl isobutyrate, butyl acetate, ethyl pentanoate, methyl salicylate, hexyl acetate, (E)-2-hexenoic acid, P-cymene, terpinolene, linalool, 4-terpineol, geranyl acetone from Dr. Ehrenstorfer (Germany); β-ionone from Fluka (Buchs, Switzerland); ethyl hexanoate from Nu-chek (USA); octanol, hexanol, (E)-2-hexenol from Chem Service (USA), and n-alkanes (C7-C27) from Supelco (Bellefonte, PA).
The automatic solid-phase microextraction (SPME) fibres of 50/30 μm divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) were purchased from Supelco (Bellefonte, PA, USA).
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5

Volatile Compound Analysis of Plant Samples

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Dichloromethane, acetone, methanol, diethyl ether, ethyl acetate, n-butanol, sodium sulphate (anhydrous), and n-alkane mix (C8–C26) were acquired from Merck (Darmstadt, Germany). methanol and acetonitrile were HPLC-MS grade also from Merck (Darmstadt, Germany) for HPLC-MS analyses. Dichloromethane was distilled prior to use during volatile analysis. Pure reference standards of methyl butanoate, hexanal, (Z)-3-hexenal, 1,8-cineole, hexanol, (Z)-3-hexen-1-ol, and ethyl 3-hydroxy-butanoate were purchased from Sigma-Aldrich (Milwaukee, WI, USA). Pure reference standards of 4-hydroxy-4-methyl-2-pentanone, ethyl butanoate, 2,3-butanediol, and terpinen-4-ol were generously supplied by Disaromas S.A. (Bogotá, Colombia).
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6

Comprehensive Volatile Compound Analysis

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Methanol HPLC grade from Merck (Darmstadt, Germany), NaCl (analytical grade) was obtained from Sigma–Aldrich (Millipore, Bedford, MA, USA) and pure water was obtained from the Milli-Q purification system (Millipore, Bedford, MA, USA). The standards (purity > 95%) used for the quantification and identification of the volatile compounds and the chemical standards were obtained as follows: (E)-2-hexenal, geranic acid, (Z)-3-hexenal, 2-octanol, hexanal, phellandrene, β-damascenone, β-myrcene, D-limonene, benzyl alcohol, citronellol, neral, geranial, α-terpineol, (Z)-rose oxide II, geraniol, (E)-rose oxide I and phenylethyl alcohol from Sigma (St. Louis, MO, USA); hexanoic acid, benzaldehyde, octanoic acid, octanal, nonanal, pentanal, 3-methylbutanal, 1-octen-3-ol, (Z)-3-hexenol, heptanol, ethyl butyrate, ethyl acetate, butyl acetate, ethyl pentanoate, ethyl isobutyrate, methyl salicylate, (E)-2-hexenoic acid, hexyl acetate, terpinolene, P-cymene, linalool, geranyl acetone, 4-terpineol, β-ionone and ethyl hexanoate was obtained from Nu-Chek Prep (Elysian, MN, USA); hexanol, octanol, (E)-2-hexenol from Chem Service Inc. (West Chester, PA, USA). The automatic solid phase microextraction (SPME) fibers of 50/30 μm divinylbenzene/polydimethylsiloxane/carboxen (DVB/PDMS/CAR) were obtained from Supelco (Bellefonte, PA, USA).
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