Hplc grade dichloromethane

Chromatography vials were filled with 10 μl of each sample as well as with 1 ml of HPLC grade dichloromethane (Fisher Scientific). Analyses were performed using an Agilent 7890B GC with a 5977A MS. The DB-624, 30 m × 0.320 mm × 0.32 mm capillary column with flow rate of helium of 7 ml/min was used. Temperature of injector, mass transfer line and ion source was 280 °C, column temperature increased from 110 to 250 °C (10 °C/min) with a hold for 1 min. The injection volume was 1 μl with a splitless injection. The full scan examined masses between 30 and 300 amu. Qualitative analyses of the flavored liquids were carried out using the NIST 14 MS library as well as the FFNSC 3 flavouring library. All samples were run in triplicate.
Following this analysis, each chemical was screened, by CAS number, using the Good Scents Company's chemical inventory (http://www.thegoodscentscompany.com /). This database identified if the detected compound was used as a flavouring and helped to identify hazardous chemicals (e.g., harmful, irritant) from the compounds safety data sheets.

HPLC grade dichloromethane, methanol, and isopropanol were purchased from Fisher Scientific (Pittsburgh, PA, USA). The internal standard (IS), C₁₃H₂₈ (n-tridecane), was obtained from the Polyscience Corporation (Niles, IL, USA). The reference standards of α-humulene, β-caryophyllene, caryophyllene oxide, β-elemene, and soybean oil were purchased from Sigma-Aldrich (St. Louis, MO, USA). LC-MS grade ammonium acetate, ammonium formate, and formic acid were also purchased from Sigma-Aldrich. Beverage grade CO₂ was purchased from nexAir (Memphis, TN, USA).

Epicoprostanol, 5β-cholestan-3α-ol, heptane (99%), anhydride pyridine (99.9%), and N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) (99.9%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). HPLC-grade n-hexane, HPLC-grade methanol, potassium hydroxide (85%) (KOH), acetone, and HPLC-grade dichloromethane were purchased from Thermo Fisher (Waltham, MA, USA).