The mixture of n-alkanes for the IT solution was prepared in cyclohexane at a concentration of 100 mg/L; internal standards (ISs) solutions were prepared in dichloromethane (GC grade) at a concentration of 4 mg/mL.
Pure standards for identity confirmation of pyruvic acid, malonic acid, phosphoric acid, succinic acid, glyceric acid, fumaric acid, malic acid, citric acid, Alanine-Ala, Aspartic acid-Asp, Glutamic acid-Glu, Glycine-Gln, Isoleucine-Ile, Leucine-Leu, Methionine-Met, Phenylalanine-Phe, Proline-Pro, Serine-Ser, Threonine-Thr, Tryptophan-Trp, Tyrosine-Tyr, Valine-Val, glycerol, xylitol, mannitol, myo-inositol, fructose, glucose, and the internal standards (ISs), 4-chlorophenyl alanine (quality control–QC for derivatization), and 1,4-dibromobenzene (QC for GC normalization) were purchased from Merck (Milan, Italy).
Derivatization reagents and HPLC grade solvents: O-methyl hydroxylamine hydrochloride (MOX), (N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) methanol, pyridine, n-hexane, cyclohexane, dichloromethane, and toluene were also obtained from Merck.