Table 1.
RRLC gradienta elution program for the separation of fat- and water-soluble vitamins on Agilent Poroshell 120 EC-C18 reverse phase column.
Table 2.
Gradient program employed for the separation of PITC derivatized amino acidsa.
Figure 1.
RRLC-MS/MS TIC chromatogram of a. Fat-soluble vitamin standards, b. Phytococktail sample under the optimum analysis conditions.
r: retinol, e: ergocalciferol, t: tocopherol, p: phylloquinone.
Figure 2.
RRLC-MS/MS TIC chromatogram of a. Water-soluble vitamin standards, b. Phytococktail sample under the optimum analysis conditions.
t: thiamine, na: nicotinic acid, p: pyridoxine, nd: nicotinamide, pa: pantothenic acid, f: folic acid, c: cyanocobalamin, b: biotin, r: riboflavin.
Figure 3.
Extracted RRLC-MRM ion chromatogram of the analytes.
The quantifier, qualifier and MRM transitions of extracted ions for blank, four fat-soluble standards and phytococktail sample have been shown for 1A–1I: vit A, 2A–2I: vit D2, 3A–3I: vit E and 4A–4I: vit K1.
Figure 4.
Extracted RRLC-MRM ion chromatogram of the analytes.
The quantifier, qualifier and MRM transitions of extracted ions for blank, nine water-soluble standards and phytococktail sample have been shown for 1A–1I: vit B1, 2A–2I: vit B2, 3A–3I: vit B3 (nicotinic acid), 4A–4I: vit B3 (nicotinamide), 5A–5I: vit B5, 6A–6I: vit B6, 7A–7I: B7, 8A–8I: vit B9 and 9A–9I: vit B12.
Table 3.
Optimized MS/MS transition parameters.
Table 4.
Intra- and inter-day precision of retention time, recovery and linearity (n = 5).
Table 5.
Intra- and inter-day precision of response.
Table 6.
Validation data for determination of fat- and water-soluble vitamins (n = 3).
Figure 5.
Recoveries of a. water- and b. fat-soluble vitamins (%) obtained from phytococktail spiked before extraction (SBE) and spiked after extraction (SAE).
Figure 6.
Mean content of water- and fat-soluble vitamins in phytococktail sample before extraction (SBE) and spiked after extraction (SAE).
Table 7.
Content of fat- and water-soluble vitamins in phytococktail (n = 5).
Figure 7.
RP-HPLC chromatogram of A. 21 amino acid standards, B. Amino acid profile of the phytococktail.
a: L-Arginine; b: L-Aspartic Acid; c: L-Glutamic Acid; d: L-Serine; e: Glycine; f: L-Histidine; g: L-Threonine; h: L-Alanine; i: L-Proline; j: L-2-amino-n-butyric acid; k: L-Valine; l: L-Methionine; m: L-Cystine HCl; n: L-Cystine; o: L-Isoleucine; p: L-Leucine; q: L-Nor Leucine; r: L-Tryptophan; s: L-Phenylalanine; t: L-Ornithine; u: L-Lysine.
Table 8.
Content, type of amino acid, retention time (RT), peak area as quantitated by RP-HPLC.
Figure 8.
Different amino acids identified in the phytococktail by RP-HPLC.
1: L-Arginine; 2: L-Aspartic Acid; 3: L-Glutamic Acid; 4: L-Serine; 5: Glycine; 6: L-Histidine; 7: L-Threonine; 8: L-Alanine; 9: L-Proline; 10: L-Valine; 11: L-Methionine; 12: L-Cystine HCl; 13: L-Cystine; 14: L-Isoleucine; 15: L-Leucine; 16: L-Nor Leucine; 17: L-Phenylalanine; 18: L-Ornithine; 19: L-Lysine.
Figure 9.
Different fatty acids identified in the phytococktail by GC-FID.
Figure 10.
GC-FID chromatogram of a. 37 FAMEs standards, b. FAMEs of the phytococktail.
1: Butyric acid methyl ester (C4∶0); 2: Caproic acid methyl ester (C6∶0); 3: Caprylic acid methyl ester (C8∶0); 4: Capric acid methyl ester (C10∶0); 5: Undecanoic acid methyl ester (C11∶0); 6: Lauric acid methyl ester (C12∶0); 7: Tridecanoic acid methyl ester (C13∶0); 8: Myristic acid methyl ester (C14∶0); 9: Myristoleic acid methyl ester (C14∶1); 10: Pentadecanoic acid methyl ester (C15∶0); 11: cis-10-Pentadecenoic acid methyl ester (C15∶1); 12: Palmitic acid methyl ester (C16∶0); 13: Palmitoleic acid methyl ester (C16∶1); 14: Heptadecanoic acid methyl ester (C17∶0); 15: cis-10-Heptadecenoic acid methyl ester (C17∶1); 16: Stearic acid methyl ester (C18∶0); 17: Elaidic acid methyl ester (C18∶1n9t); 18: Oleic acid methyl ester (C18∶1n9c); 19: Linolelaidic acid methyl ester (C18∶2n6t); 20: Linoleic acid methyl ester (C18∶2n6c); 21: Arachidic acid methyl ester (C20∶0); 22: cis-11-Eicosenoic acid methyl ester (C20∶1); 23: α-Linolenic acid methyl ester (C18∶3n3); 24: Heneicosanoic acid methyl ester (C21∶0); 25: cis-11,14-Eicosadienoic acid methyl ester (C20∶2); 26: Behenic acid methyl ester (C22∶0); 27: cis-8,11,14-Eicosatrienoic acid methyl ester (C20∶3n6); 28: Erucic acid methyl ester (C22∶1n9); 29: cis-11,14,17-Eicosatrienoic acid methyl ester (C20∶3n3); 30: Arachidonic acid methyl ester (C20∶4n6); 31: Tricosanoic acid methyl ester (C23∶0); 32: cis-13,16-Docosadienoic acid methyl ester (C22∶2); 33: Lignoceric acid methyl ester (C24∶0); 34: Nervonic acid methyl ester (C24∶1); 35: cis-4,7,10,13,16,19-Docosahexaenoic acid methyl ester (C22∶6n3); 36: γ-Linolenic acid methyl ester (C18∶3n6); 37: cis-5,8,11,14,17-Eicosapentaenoic acid methyl ester (C20∶5n3).
Table 9.
Fatty acid composition of the phytococktail.
Table 10.
Major nutritional composition and mineral content of the phytococktail.