Chemical profiling analysis of Maca using UHPLC-ESI-Orbitrap MS coupled. ingredients

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1 Chemical profiling analysis of Maca using UHPLC-ESI-Orbitrap MS coupled with UHPLC-ESI-QqQ MS and the neuroprotective study on its active ingredients Yanyan Zhou 1# Peng Li 2# Adelheid Brantner 3 Hongjie Wang 1 Xinbin Shu 4 Jian Yang 1 Nan Si 1 Lingyu Han 1 Haiyu Zhao 1 & Baolin Bian 1 1 Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China 2 Institute of Chinese Medical Sciences, University of Macau, Macau, China 3 Institute of Pharmaceutical Sciences Pharmacognosy, University of Graz, Graz, Austria 4 Shandong Rosemed Biopharm LTC, Yanzhou, Shandong province, China #: These two authors contributed equally to the paper Correspondence author at: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences m3 #6823 RT: AV: 1 NL: 1.07E7 T: FTMS + c ESI d Full ms @cid35.00 [ ] O N H O Relative Abundance (CH 2 ) n successively C 18 H 30 ON C 8 H 11 NO C 7 H 8 O H 2 O OH m/z Figure S1 The MS/MS spectrum of N-(3-methoxybenzyl)-(9Z,12Z,15Z)-octadecatrienamide (No. A 32 )

2 m3 #7798 RT: AV: 1 NL: 2.63E7 T: FTMS + c ESI d Full ms @cid35.00 [ ] Relative Abundance O N H C 15 H C 7 H 8 N (CH 2 ) n successively H 2 O C 7 H 8 C 6 H m/z Figure S2 The MS/MS spectrum of N-benzylhexadecanamide (No. A 54 ) m2 #8038 RT: AV: 1 NL: 6.41E6 T: FTMS + c ESI d Full ms @cid35.00 [ ] N H O C 16 H Relative Abundance C 6 H (CH 2 ) n successively H 2 O C 7 H 8 C 7 H 8 N m/z Figure S3 Proposed structures and MS/MS spectrum for N-benzylheptadecanamide (No. A 58 )

3 m2 #7839 RT: AV: 1 NL: 1.13E6 T: FTMS + c ESI d Full ms @cid35.00 [ ] O N H O Relative Abundance C 18 H 32 O NH 3 H2 O C 8 H 11 NO C 7 H 8 O m/z Figure S4 Proposed structures and MS/MS spectrum for N-(3-methoxybenzyl)-(9Z)-octadecenamide (No.A 55 ) m2 #3054 RT: AV: 1 NL: 1.37E7 T: FTMS + c ESI d Full ms @cid35.00 [ ] OCH 3 Relative Abundance N OH -C 6 H 9 NO C 6 H 5 H2O m/z Figure S5 Proposed structures and MS/MS spectrum for 3-benzyl-1,2-dihydro-N-hydroxypyridine-4-methoxy (No. A 65 )

4 m2 #3634 RT: AV: 1 NL: 5.45E6 T: FTMS + c ESI d Full ms @cid35.00 [ ] N N Relative Abundance C 14 H 15 N 2 CH2 CH C 7 H m/z Figure S6 Proposed structures and MS/MS spectrum for 1,3-dibenzyl-2(R)-ethyl-4,5-dimethylimidazilium (No. A 84 ) m2 #2284 RT: AV: 1 NL: 2.60E5 T: FTMS + c ESI d Full ms @cid35.00 [ ] O C OH NH Relative Abundance N H C 7 H 9 N 2 O 2 C 7 H C 6 H H 2 O m/z Figure S7 Proposed structures and MS/MS spectrum for (1R,3S)-1-ethyltetrahydro-β-5,6-carboline-3-carboxylic acid (No. A 111 )

5 ZIFU #2011 RT: AV: 1 NL: 8.35E3 T: FTMS - c ESI d Full ms @cid35.00 [ ] OH 100 O OH O - OSO 90 3 OH S N OH Relative Abundance H 2 O H 2 O C 8 H 12 O 5 S C 8 H 7 ON -Glc C 8 H 7 NS 10 0 C 8 H 12 O SO m/z Figure S8 Proposed structures and MS/MS spectrum for acetyl-benzylglucosinolate (No. G 11 ) m2 #6647 RT: AV: 1 NL: 6.10E6 T: FTMS + c ESI d Full ms @cid35.00 [ ] Relative Abundance OH O O CO C 10 H 18 C 9 H 16 CH 2 H 2 O m/z Figure S9 Proposed structures and MS/MS spectrum for 5-oxo-6E,8E-nineteencarbodienoic acid (No. M 9 )

6 RT: Relative Abundance Time (min) RT: Figure S10 The LC-MS chromatogram of Fr Relative Abundance Time (min) Figure S11 The LC-MS chromatogram of Fr 5

7 RT: Relative Abundance Time (min) Figure S12 The LC-MS chromatogram of Fr 6

8 G 5 -HPLC G 3 -HPLC G 5 -MS G 3 -MS G 5 -HNMR G 3 -HNMR G 8 -HPLC A 52 -HPLC G 8 -MS A 52 -MS G 8 -HNMR A 52 -HNMR

9 A 8 -HPLC A 54 -HPLC A 8 -MS A 54 -MS A 8 -HNMR A 54 -HNMR O 6 -HPLC A 66 -HPLC O 6 -MS A 66 -MS O 6 -HNMR A 66 -HNMR

10 A 67 -HPLC A 5 -HPLC A 67 -MS A 5 -MS A 67 -HNMR A 5 -HNMR A 4 -HPLC C 3 -HPLC A 4 -MS C 3 -MS A 4 -HNMR C 3 -HNMR

11 A 121 -HPLC C 5 -HPLC A 121 -MS C 5 -MS C 5 -HNMR A 121 -HNMR O 9 -HPLC O 9 -MS O 9 -HNMR Figure S13 The HPLC, MS and HNMR spectra of 15 reference standards

12 Table S1 Identification of Macamides and common amide alkaloids from Maca by UHPLC-ESI-Orbitrap MS NO name t R (min) Formula ESI(+)/expected ESI(+)/measured Delta (ppm) Fragamentor information A 1 A 2 benzylamine C 7 H 9 N N-methyl-3-hydroxy-Benzeneacetamide C 9 H 11 NO , , , A 3 N-benzyl-2E,4E-fivecarbondienamide C 12 H 11 NO , , , A 4 A 5 2-phenylacetamide C 8 H 9 NO , N-acetylbenzylamine C 9 H 11 NO , , A 6 N-(3-hydroxy-benzyl)-2Z-fivecarbon acrylamide C 10 H 13 NO , , , , A 7 N-(3-hydroxy-benzyl)-11Z,13Z,15Z,17Zoctadecatetrienamide C 25 H 35 NO , , , , A 8 N-benzylbenzamide C 14 H 13 NO , , A 9 N-benyl- Sevencarbamide C 13 H 19 NO , , , A 10 N-benzyl-2E,4E,6E-fourcarbonleukotrieneamide C 14 H 15 NO , , A 11 N-benzyl-2E,4E-sixcarbondienamide C 13 H 13 NO , , , A 12 N-(3-hydroxy-benzyl)-8E,11E,13E,15E,17E-octa decapentienamide C 25 H 33 NO ; , , , A 13 N-benzyl-octanamide C 15 H 23 ON , , , , A 14 N-(3,4-dihydroxy-benzyl)-hexadecanamide C 22 H 41 NO , , , , A 15 N-benzyl-(9Z,12Z,15Z)-octadecatetraenamide C 25 H 35 NO , , , , A 16 N-(3-hydroxy-benzyl)-11E,13E,15E,17E-octadec atetrienamide C 25 H 35 NO , , , A 17 N-benzyl-5-oxo-6E,8E-octadecadienamide [31] C 25 H 37 NO , , , , , ,

13 A 18 N-benzyl-13-oxo-9E,11E-octadecadienamide C 25 H 37 NO A 19 N-benzyl-9-oxo-12E,15Z-Octadecadienamide C 25 H 37 NO A 20 N-benzyl-9-oxo-12E,15E-Octadecadienamide C 25 H 37 NO , , , , , , , , , , , , A 21 N-butyl-14,16-dihydroxy-9E, 11E, 13Eoctadecatrieneamide C 22 H 39 NO , , , , A 22 N-benzyl-16-hydroxy-9-oxo-3E,5E,7E-octadecatr ieneamide C 25 H 35 NO , , , , A 23 N-benzyl-9-oxo-12E-octadecenamide C 25 H 39 NO , , , , A 24 A 25 N-benzyl-16-hydroxy-9-oxo-15E,17E-octadecatri eneamide N-eyhyl-12-(2-4 -)-cyclohexadiene-dodeceneam ide C 25 H 35 NO , , , C 20 H 35 NO , , , , A hydroxyl-octadecanamide C 18 H 37 NO , , , A 27 N-cyclohexane-(3 -hydroxyl)-tridecane amide C 20 H 39 NO , , , , A 28 N-(3,4-dihydroxybenzyl)-(9Z)-tetradecanamide C 20 H 39 NO , , , , A 29 N-(3,4-dihydroxy-benzyl)-(9E)-tetradecanamide C 20 H 39 NO , , , , A 30 N-benzyl-9-oxo-12N-octadecenamide C 25 H 39 NO , , , , A 31 N-benzyl-8E, 10E-hexadecadienamide C 23 H 35 NO , , , , A 32 N-(3-methoxybenzyl)-(9Z,12Z,15Z)-octadecatrie namide C 26 H 39 NO , , , , A 33 N-ethyl-(9E,11E)-octadecadienamide C 20 H 37 NO , , , , A 34 N-benzyl-(9Z,12Z,15Z)-octadecatrienamide C 25 H 37 NO , , , ,

14 A 35 N-benzyl-9Z,12Z-hexadecadienamide C 24 H 37 NO , , , , A 36 N-benzyl-tetradeceneamide C 21 H 35 NO , , , , A 37 N-(3-methoxybenzyl)-(9Z)-benzylhexadecanamid e C 23 H 39 NO , , , , , A 38 N-benzyl)-13-oxo-11E-heptadecenamide C 24 H 39 NO , , , , A 39 N- benzyl-9e,12e- hexadecadienamide C 24 H 37 NO , , , , , , A 40 N-benzyl-16-hydroxy-9-oxo-13E,15E-octadecadi enamide C 25 H 41 NO , , , , , A 41 N-benzyl-4E-hexadecenamide C 23 H 37 NO , , , , A 42 N-benzyl-4Z-hexadecenamide C 23 H 37 NO , , , , A 43 N-(3-methoxybenzyl)-(9Z,12Z)-octadecadienami de C 26 H 41 NO , , , , A 44 N-octadecanamide C 18 H 37 NO , , , , A 45 N-benzyl-(9Z,12Z)-octadecadienamide [29] C 25 H 39 NO , , , , , A 46 N-benzylpentadecanamide C 22 H 37 NO , , , , A 47 N-ethyl-(2E,16E)-octadecadienamide C 20 H 39 NO , , , A 48 N-(3,4-dimethoxybenzyl)-hexadecanamide C 25 H 43 NO , , , A 49 N-benzyl-14E,8E-eicosenoicdienamide C 22 H 39 NO , , , , , A 50 N-benzyl-9Z, 12Z-heptadecadienamide C 24 H 39 NO , , , , A 51 N-benzyl-9Z,12Z-nonadecadienamide C 26 H 41 NO , , , , A 52 N-(3-methoxybenzyl)-hexadecanamide C 24 H 41 NO , , , , A 53 N-(3-methoxybenzyl)-(9E)-octadecenamide C 26 H 43 NO , , , , A 54 N-benzylhexadecanamide [29] C 23 H 39 NO , , , ,

15 A 55 N-(3-methoxybenzyl)-(9Z)-octadecenamide C 26 H 43 NO , , , , A 56 N-benzyl-9Z-octadecenamide [29] C 25 H 41 NO , , , , A 57 N-benzyl-10E,12E-twentycarbondienamide C 27 H 43 NO , , , , A 58 N-benzylheptadecanamide [29] C 24 H 41 NO , , , , A 59 N-benzyl-12Z-nonadecenamide C 26 H 43 NO , , , , , A 60 N-benzylheptadecanamide C 24 H 41 NO , , , , A 61 N-benzyl-12E-nonadecenamide C 26 H 43 NO , , , , A 62 N-benzyloctadecanamide C 25 H 43 NO , , , , A 63 N-benzyloctadecanamide C 25 H 43 NO , , , , A 64 N -benzyltwentycarboxamide C 27 H 47 NO , , , , Structures confimed by comparison with reference standards. Bold characters: the base peaks in MS n spectra Table S2 Identification of Macaridines from Maca by UHPLC-ESI-Orbitrap MS NO name t R (min) Formula ESI(+)/expected d ESI(+)/measure Delta (ppm) Fragamentor information A 65 A 66 3-benzyl-1,2-dihydro-N-hydroxypyridine-4-met hoxy 3-benzyl-1,2-dihydro-N-hydroxypyridine-4-car baldehyde C 13 H 15 NO , , , , C 13 H 13 NO , , , , A 67 (3-methoxybenzyl) N-pyridine-4-carbaldehyde C 14 H 13 NO , , , Structures confimed by comparison with reference standards. Bold characters: the base peaks in MS n spectra Table S3 Identification of Imidazole alkaloids from Maca by UHPLC-ESI-Orbitrap MS

16 NO name t R (min) Formula ESI(+)/expected ESI(+)/measured Delta (ppm) Fragamentor information A 68 1-dibenzyllimidazilium C 8 H 12 N , , , A 69 1-dibenzyl-4,5-dimethylimidazilium C 12 H 14 N , , A 70 1-dibenzyl-trimethylimidazilium C 13 H 16 N , , , A 71 1-dibenzyl-2-ethyl-4,5-dimethylimidazilium C 14 H 18 N , , , , A 72 1-dibenzyl-2-propane-4,5-dimethylimidazilium C 15 H 18 N , A 73 1-dibenzyl-2-propyne-4,5-dimethylimidazilium C 15 H 16 N , , , A 74 1-dibenzyl-2-propane-4,5-dimethylimidazilium C 15 H 20 N , , , A 75 1-dibenzyl-2-butene-4,5-dimethylimidaziliumc C 16 H 20 N , , A 76 A 77 1-dibenzyl-2-isopropane-4,5-dimethylimidazili um 1-dibenzyl-2-(1,3-butadiene)-4,5-dimethylimid azilium C 15 H 20 N , , , , C 16 H 18 N , , , A 78 1-dibenzyl-2-butyl-4,5-dimethylimidazilium C 16 H 22 N , , , A 79 1-dibenzyl-2-phenyl-4,5-dimethylimidazilium C 18 H 18 N , , , , A 80 1-dibenzyl-2-isobutyl-4,5-dimethylimidazilium C 16 H 22 N , , , A 81 1-dibenzyl-2-(2-pentynylalkenyl)-4,5-dimethyli midazilium C 17 H 18 N , , , , A 82 1,3-dibenzyl-4,5-dimethylimidazolium C 19 H 20 N , , , , A 83 1,3-dibenzyl-2,4,5-trimethylimidazilium C 20 H 22 N , , , , A 84 A 85 1,3-dibenzyl-2(R)-ethyl-4,5-dimethylimidaziliu m 1,3-dibenzyl-2-(1,3-diacetylene)-4,5-dimethyli midazilium C 21 H 24 N , , , , C 23 H 20 N , , , , A 86 1,3-dibenzyl-2-vinyl-4,5-dimethylimidazilium C 21 H 22 N , , , ,

17 A 87 1,3-dibenzyl-2(S)-ethyl-4,5-dimethylimidaziliu m C 21 H 24 N , , , A 88 1-dibenzyl-2-heptyl-4,5-dimethylimidazilium C 19 H 28 N , , , A 89 1,3-dibenzyl-2-propyl-4,5-dimethylimidazilium C 22 H 26 N , , , , , A 90 A 91 A 92 A 93 1,3-dibenzyl-2(1,3-glutaricalkynyl)-dimethylim idazilium 1,3-dibenzyl-2-isopropyl-4,5-dimethylimidazili umchloride 1-dibenzyl-2-isoheptyl-4,5-dimethylimidaziliu m 1,3-dibenzyl-2-propenyl-4,5-dimethylimidaziliu m C 24 H 22 N , , , , C 22 H 26 N , , , , C 19 H 28 N , , , , C 22 H 24 N , , , , , , A 94 1,3-dibenzyl-2-phenyl-4,5-dimethylimidazilium C 25 H 24 N , , , , , A 95 1,3-dibenzyl-2-butyl-4,5-dimethylimidazilium C 23 H 28 N , , , , , , A 96 A 97 A 98 1,3-dibenzyl-2-isobutyl-4,5-dimethylimidaziliu m 1-dibenzyl-2-(5-en-1,3-heptadiyne)-4,5-dimeth ylimidazilium 1,3-dibenzyl-2-(5-en-1,3-propylene alkyne)-4,5-dimethylimidazilium C 23 H 28 N , , , , , , C 26 H 24 N , , , C 27 H 26 N , , , , A 99 1,3-dibenzyl-2-pentyl-4,5-dimethylimidazilium C 24 H 30 N , , , , , A 100 A 101 A 102 A 103 1,3-dibenzyl-2-(3,5,7-trienepropyne) -4,5-dimethylimidazilium 1-(3,5-cyclohexadiene)-cyclohexyl-2-hexyl-4,5 -dimethyhydridelimidazilium 1-dibenzyl-3-cyclohexyl-2-isopropenyl-4,5-dim ethylimidazilium 1-(1,3-cyclohexadiene)-3-cyclohexyl-2-hexyl-4,5-dimethyhydridelimidazilium C 27 H 26 N , , , , C 21 H 35 N , , , , C 21 H 35 N , , , C 21 H 35 N , , , , A dibenzyl-3-cyclohexyl-2-propenyl-4,5-dimeth C 21 H 35 N , , , ,

18 ylimidazilium A 105 1,3-dibenzyl-2-hexyl-4,5-dimethylimidazilium C 25 H 32 N , , , , , , , Structures confimed by comparison with reference standards. Bold characters: the base peaks in MS n spectra Table S4 Identification of β-carboline alkaloids from Maca by UHPLC-ESI-Orbitrp MS NO name t R (min) Formula ESI(+)/expected ESI(+)/measured Delta (ppm) Fragamentor information A 106 A 107 A 108 (1R,3S)-1-methyltetrahydro-β-5,6-hydride carboline-3-carboxylic acid (3S)-tetrahydro-β-4,5-hydridecarboline-2-carbo xylic acid (1S,3S)-1-methyltetrahydro-β-5,6-hydride carboline-3-carboxylic acid C 13 H 16 N 2 O , , , C 11 H 12 N 2 O , , , , C 13 H 16 N 2 O , , , , A 109 A 110 A 111 A 112 A 113 A 114 A 115 A 116 (1R,3S)-tetrahydro-β-5,6-hydridecarboline-3-ca rboxylic acid (1R,3S)-1-methyltetrahydro-β-carboline-3-carb oxylic acid (1R,3S)-1-ethyltetrahydro-β-5,6-carboline-3-car boxylic acid (1R,3S)-1-butanetetrahydro-β-5,6-hydride carboline-3-carboxylic acid (1R,3S)-1-pentanetetrahydro-β-5,6-carboline-3- carboxylic acid (1R,3S)-1-butanetetrahydro-β-5,6-hydride carboline-3-carboxylic acid (1R,3S)-1-pentanetetrahydro-β-5,6-hydride carboline-3-carboxylic acid (1R,3S)-1-(2-methylbutane)tetrahydro-β-5,6-hy dride carboline-3-carboxylic acid C 12 H 12 N 2 O , , , , C 13 H 14 N 2 O , , , , C 14 H 16 N 2 O , , , , , , C 16 H 18 N 2 O , , , C 17 H 20 N 2 O , , , , C 16 H 20 N 2 O , , , , C 17 H 22 N 2 O , , , , C 17 H 22 N 2 O , , , ,

19 A 117 A 118 A 119 A 120 (1R,3S)-1-hexyltetrahydro-β-5,6-hydride carboline-3-carboxylic acid (1R,3S)-1-(3-methylpentane)tetrahydro-β-5,6-h ydridecarboline-3-carboxylic acid (1R,3S)-1-(1,2-dimethylbutane)tetrahydro-β-5, 6-hydride carboline-3-carboxylic acid (1R,3S)-1-isobutanetetrahydro-β-5,6-hydride carboline-3-carboxylic acid C 18 H 24 N 2 O , , , , C 18 H 24 N 2 O , , , , C 18 H 24 N 2 O , , , , C 16 H 18 N 2 O , , , , A 121 (1R,3S)-1-methyl-β-carboline-3-carbaldehyde C 13 H 10 N 2 O , , Structures confimed by comparison with reference standards. Bold characters: the base peaks in MS n spectra Table S5 Identification of Acids from Maca by UHPLC-ESI-Orbitrap MS NO name t R (min) Formula ESI(-)/expected ESI(-)/measured Delta (ppm) Fragamentor information C 1 C 2 C 3 C 4 C 5 3-methoxyphenylacetic acid 2.17 C 9 H 10 O , , ,4-dihydroxy-3,5-cyclopentyl dienoic acid 2.33 C 6 H 7 O Nicotinic acid 3.98 C 6 H 5 NO , , , hydroxy-3-methoxybenzoicacid 5.31 C 8 H 8 O , , , Succinic acid 6.80 C 4 H 6 O , Table S6 Identification of Glucosinolates from Maca by UHPLC-ESI-Orbitrap MS NO name t R (min) Formula ESI(-)/expected ESI(-)/measured Delta (ppm) Fragamentor information G 1 indolyl-3-methylglucosinolate C 15 H 29 O 10 NS , , , , G 2 indolyl-5-methylglucosinolate C 15 H 29 O 10 NS , , , , G 3 m-methoxybenzylglucosinolate C 15 H 20 O 10 NS , , , , , ,

20 G 4 p-methoxybenzylglucosinolate C 15 H 20 O 10 NS , , , , , , G 5 benzylglucosinolate C 14 H 19 O 9 NS , , , , , , , G 6 benzylglucosinolate C 14 H 19 O 9 NS , , , , , , , G 7 th-hydroxybenzylglucosinolate C 14 H 19 O 10 NS , , , , , G 8 p-hydroxybenzylglucosinolate C 14 H 19 O 10 NS , , , , , , G 9 4-methoxyindolyl-3-methyloxyglucosinolate C 17 H 22 O 10 N 2 S G 10 4-methoxyindolyl-3-hexylhydroxyglucosinolate C 22 H 36 O 10 N 2 S , , , , , , , , , , G 11 acetyl-benzylglucosinolate C 16 H 21 O 10 NS , , , , , , G 12 acetyl-benzylglucosinolate C 16 H 21 O 10 NS , , , , , , G 13 Pent-4-enylglucosinolate C 12 H 24 O 7 N 2 S , , , , , G 14 indolyl3-hexyl-4-methyl-cyclohexaneglucosino late C 22 H 38 O 9 N 2 S , , , , Structures confimed by comparison with reference standards. Bold characters: the base peaks in MS n spectra Table S7 Identification of Macaenes from Maca by UHPLC-ESI-Orbitrap MS NO name t R (min) Formula ESI(+)/expected ESI(+)/measured Delta Fragamentor information (ppm) M 1 5-oxo-6E,8E,10E,12E-octadecatetraenoic acid C 18 H 26 O , , , , M 2 5-oxo-6E,8E,10E,12E,14E-octadecapentaenoic acid C 18 H 24 O , , , ,

21 M 3 5-oxo-6E,8E,10E-octadecatrienoic acid C 18 H 28 O , , , , M 4 5-oxo-6E,8E,10E,12E,14E,16E,18E,20E,22Etwentysixcarbonnonaenoic acid C 26 H 32 O , , , M 5 5-oxo-6E,8E,10E,12E-eicosatetraenoic acid C 20 H 30 O , , , , , , M 6 5-oxo-6E,8E,10E,12E,14E,16E,18E,20E,22E,2 4E,26E -octacosundecenoic acid C 28 H 32 O , , , , M 7 5-oxo-6E,8E-octadecadienoic acid C 18 H 30 O , , , , M 8 5-oxo-6E,8E,10E-eicosatrienoic acid C 20 H 32 O M 9 5-oxo-6E,8E-nineteencarbodienoic acid C 19 H 32 O , , , , , , , , , M 10 5-oxo-6E,8E,10E,12E,14E,16E,18E-eicosatana batienoic acid C 20 H 24 O , , , , M 11 5-oxo-6E- eicosanoic acid C 20 H 36 O , , , , , Structures confimed by comparison with reference standards. Bold characters: the base peaks in MS n spectra Table S8 Idetification of Other compounds from Maca by UHPLC-ESI-Orbitrap MS NO name t R (min) Formula ESI(+)/expected ESI(+)/measured Delta (ppm) Fragamentor information O 1 O 2 2-furoate methyl ester 2.98 C 6 H 6 O , formaldehyde-2-hydroxy-5-methoxypyridine C 7 H 7 NO , , O 3 N-(3-hydroxy-5-methyl-benzyl)-2Z-2-amino-de catrienamide C 11 H 12 N 2 O , , , , O 4 O 5 3-hydroxybenzylcyanide C 8 H 7 NO catechin C 15 H 14 O , , , ,

22 O 6 Licochalcone A C 21 H 22 O , , , , O 7 N-ethyl-tetradecene ester C 16 H 35 NO , , , O 8 2,3-dihydroxy-acetic acid methyl ester C 9 H 9 O , , , O 9 dibutyl phthalate C 16 H 22 O , , Structures confimed by comparison with reference standards. Bold characters: the base peaks in MS n spectra Table S9 Precision, stability and repeatability of 15 compounds Compounds Precision (RSD%, n=3) Stability (RSD%, n=6) Repeatibility (RSD%, n=6) Intra day Inter day G G G A A A O A A A A C

23 C A O Table S10 Recoveries of 15 constituents (n=9) Compound Addition level Initial / μg Added/ μg Measured/ μg Recovery rate /% Average/% RSD/% 1: G 5 G 3 G 8 A 52 A 8 A 54 1: : : : : : : : : : : : : : : :

24 O 6 A 66 A 67 A 5 A 4 C 3 C 5 1: : : : : : : : : : : : : : : : : : : : : : A 121 1:

25 O 9 1: : : : : Table S11 Samples of Maca from different places NO. Places Weight(g) 1 Xingjiang Xinjiang Xizang Xizang Peru Peru Yunnan Yunnan Yunnan Yunnan Yunnan Yunnan Yunnan Yunnan Yunnan Yunnan Yunnan Leaves of Maca (12) Yunnan