Fig. 1

Extracted ion chromatograms of amino acids (leucine, phenylalanine, tyrosine, and threonine) which were separated by the PFP column with mobile phase additives (A) 0.1% formic acid; (B) 0.1% acetic acid; (C) 5 mM ammonium acetate (pH = 6.5) in positive ionization mode.

Fig. 2

Extracted chromatograms of metabolites. (a) nucleotides: cytidine, adenosine, and 2-deoxyguanosine; (b) amines: serotonin, thiamine, and tyramine; (c) amino acids: glutamate, lysine, and tyrosine; (d) others: tryptophan, carnitine, and pantothenic acid; obtained on the HILIC column with solvent additives: 10 mM NH4HCO2 (pH = 3) (left); 5 mM NH4HCO2 (pH = 3) (middle); 10 mM NH4Ac (pH = 6.5) (right) in positive ionization mode.

Fig. 3

MS response of metabolite standards analyzed by the PFP column with various solvent additives: 0.1% formic acid (blue), 0.1% acetic acid (red), and 0.02% acetic acid (green) in negative ionization mode. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 4

Extracted ion chromatograms of metabolite standards: hydrocortisone (1), tyramine (2), serotonin (3), 2′-deoxyguanosine (4), tryptophan (5), 3,4-dihydroxyphenylacetic acid (6), biotin (7), pantothenic acid (8), mannose (9), lauric acid (10), N-acetyl-L-aspartic acid (11), caprylic acid (12), maleic acid (13), and taurocholic acid (14); analyzed by the HILIC column with different solvent additives in negative ionization mode. Under different condition groups (a) and (b) include metabolites with relatively low and high ESI intensity, respectively.

Fig. 5

(a) Base Peak Chromatograms (BPCs) of zebrafish embryos reconstituted in water/methanol (10/90, v/v) and analyzed by the HILIC with optimal solvent additives; (b) the number of metabolites detected in the same zebrafish samples under the same chromatographic conditions in positive ion mode.

Fig. 6

(a) Base Peak Chromatograms (BPCs) of zebrafish extracts reconstituted in 100% (or 10%) water in methanol obtained with optimal chromatographic conditions, and (b) the number of metabolites detected in the same zebrafish samples under the same chromatographic conditions in negative ion mode.

Acknowledgments
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