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LAAN-C-XX011A Lifescience

O HO O MS 1 MS 2 C 14 H 16 N 10 HO HO O MS 3

Application Note No.10

Application Note No.10

Application Note No.10 Intensity (x100,000,000) 7.5 5 5.0 2.5 1 2 3 4 6 7 8 9 10 +:TIC -:TIC +:181.0707 (20) +:195.0882 (2.00) +:307.0800 (2.00) +:459.0927 (2.00) +:291.0848 (2.00) +:443.0973 (2.00) -:305.0686 (2.00) -:457.0791 (2.00) -:289.0725 (2.00) -:441.0829 (2.00) 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 1 min Peaks 1: (-)-Gallocatechin, 2: Theophylline, 3: (+)-Catechin, 4: (-)-Epigallocatechin, 5: Caffeine, 6: (-)-Epigallocatechin gallate, 7: (-)-Epicatechin, 8: (-)-Gallocatechin gallate,9: (-)-Epicatechin gallate, 10: (-)-Catechin gallate Intensity (x100,000,000) 2.00 1.75 1.50 1.25 1.00 A B C D E %RSD Area R.T. A 2.087 0.201 B 3.174 0.216 C 2.213 0.182 D 2.861 0.149 0.75 E 0.821 0.129 0.50 0.25 0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 min

( 10,000,000) TIC 1.25 1.00 0.75 0.50 0.25 0 5.0 7.5 1 12.5 15.0 17.5 2 22.5 25.0 ID Compound Name RT ID Compound Name RT 1 L-Alanine-2TMS 4.82 37 Lyxose methyloxime-tms 1 10.11 2 Oxalic acid-2tms 5.15 38 Asaparagine-3TMS 10.11 3 Malonic acid-2tms 5.93 39 Lyxose methyloxime-tms 2 10.22 4 Urea-2TMS 6.27 Ribitol-5TMS (IS) 10.58 5 Serine-2TMS 6.48 40 L-Glutamine-4TMS 10.58 6 Methyl 5-oxo-2-pyrrolidinecarboxylate 6.56 41 Xylonic acid-3tms 10.66 7 Ethanolamine-3TMS 6.58 42 Theanine 10.83 8 L-Leucine-2TMS 6.63 43 L-Glutamine-3TMS 10.87 9 Phosphoric acid-3tms 6.67 44 Shikimic acid-4tms 11.11 10 L-Isoluecine-2TMS 6.85 45 Citric acid-4tms 11.23 11 L-Threonine-2TMS 6.86 46 Caffeine 11.51 12 L-Proline-2TMS 6.89 47 Quinic Acid- TMS 11.57 13 Glycine-3TMS 6.98 48 Fructose-Methyloxime-5TMS 1 11.65 14 Succinic acid-2tms 7.01 49 Fructose-Methyloxime-5TMS 2 11.72 15 Glyceric acid-3tms 7.22 50 Glucose-Methyloxime-5TMS 11.83 16 Fumaric acid-2tms 7.31 51 L-Lysine-4TMS 11.89 17 L-Serine-3TMS 7.50 52 Mannose-Methyloxime-5TMS 11.96 18 3,4-Bis[(trimethylsilyl)oxy]dihydro-2(3H)-furanone-2TMS 7.65 53 L-Tyrosine-3TMS 12.02 19 L-threonine-3TMS 7.75 54 Benzoic acid-4tms 12.14 20 L-Aspartic acid-2tms 8.05 55 Gluconic acid-5tms 1 12.47 21 4-Ketoglucose methyloxime-3tms 8.50 56 Palmitic acid-tms 12.60 22 2-Methylmaric acid-3tms 8.52 57 Gluconic acid-5tms 2 12.62 23 Arabino-Hexos-2-ulose-4TMS 8.61 58 Inositol-6TMS 13.08 24 Malic acid-3tms 8.64 59 Galactose-Methyloxime-5TMS 13.32 25 L-Aspartic acid-3tms 8.91 60 Phosphoric acid propylester-4tms 13.65 26 Pyroglutamic acid-2tms 8.95 61 Stearic acid-tms 13.79 27 4-aminobutyric acid-3tms 8.98 62 L-Tryptophan-3TMS 13.82 28 L-Norvaline-3TMS 9.13 63 Xylopyranose-4TMS 14.37 29 L-Threonic acid-4tms 9.28 64 Glucose-6-phosphate-Methyloxime-5TMS 1 14.40 30 Isopropylmalic acid-3tms 9.40 65 Glucose-6-phosphate-Methyloxime-5TMS 2 14.47 31 Glycerol-3TMS 9.54 66 Glucoheptulose-Methyloxime-TMS 14.74 32 Ornithine-3TMS 9.65 67 Di-n-octyl phthalate 15.53 33 L-Glutamic acid-3tms 9.70 68 Maltose-8TMS 15.79 34 Phenylalanine-2TMS 9.79 69 Sucrose-8TMS 16.16 35 Asparagine-4TMS 9.81 70 Arabinopyranose-4TMS 16.33 36 2,3,4,5-Tetrahydroxypentanoic acid-1,4-lactone-3tms 11 71 Raffinose-8TMS 19.92

Application Note No.10 a) PCA Score plot 10 Low Rank 30 High Rank 5 40 0 35 20 15 25 10 05-5 45-10 R 2 X(PC1) : 0.37 R 2 X(PC2) : 0.16 Q 2 :0.39-15 -10-5 0 5 10 15 b) Loading plot of PC1 Peak X (Unknown) 0.15 High Rank 0.10 5 p[1] -0-5 -0.10-0.15 Low Rank Apparatus GC/MS LC/MS Analyte type organic acids amino acids saccharides sugar alcohol catechines xantines

Inten.( 1,000,000) 4.0 3.0 345.0818 MS 1 2.0 1.0 223.0650 297.0826 265.2011 367.0695 443.9989 100 200 300 400 m/z Inten.( 1,000,000) 2.0 153.0178 MS 2 1.0 153.0203 327.0712 100 200 300 400 m/z 1.5 1.0 Inten.( 100,000) MS 3 0.5 327.0672 139.0469 309.0658 100 200 300 400 m/z Predicted formula of Peak1C 14 H 16 O 10

Application Note No.10 Theogallin C 14 H 16 O 10 Exact Mass : 344.0743 HO O O HO O HO m/z 153 C 7 H 5 O 4 + m/z 309 C 14 H 12 O 8 + m/z 327 C 14 H 14 O 9 + 4.0 3.0 m/z calc. 345.0822 0.4 mda 153.0178 m/z calc. 153.0188 1.0 mda 153.0203 m/z calc. 153.0188 1.5 mda 327.0712 345.0818 2.0 1.0 223.0650 297.0826 265.2011 367.0695 443.9989 100 200 300 400 m/z 2.0 1.0 Inten.(x1,000,000) m/z calc. 327.0716 0.4 mda 100 200 300 400 m/z Inten.(x1,000,000) 1.5 1.0 Inten.(x1,000,000) m/z calc. 327.0716 0.4 mda 0.5 m/z calc. 309.0610 1.8 mda 327.0712 139.0469 309.0628 100 200 300 400 m/z MS 1 MS 2 MS 3

3218-06006-30A-IK 2010.6