Supplement
Detecting Admixture to Mango Purées of the Alphonso Cultivar (Mangifera indica L. cv. Alphonso) by
1H-NMR Spectroscopy
Claas Strecker*a and Victor Araa
a Chelab Dr. V. Ara GmbH & Co. KG, Carl-Zeiss-Straße 16, 30966 Hemmingen, Germany
* Correspondence: c.strecker@chelab.de; Tel.: +49-511-414443
This file contains:
Fig. S1: Exemplary 1H-NMR spectrum of an Alphonso sample (segment from 9.5 to 5.5 ppm).
Fig. S2: Exemplary 1H-NMR spectrum of an Alphonso sample (segment from 5.5 to 2.6 ppm).
Fig. S3: Exemplary 1H-NMR spectrum of an Alphonso sample (segment from 2.6 to 0.6 ppm).
Fig. S4: 1H-NMR spectrum of an Alphonso sample and spiking experiment with niacin.
Fig. S5: 1H-NMR spectrum of an Alphonso sample and spiking experiment with trigonelline.
Tab. S1: Information gains for assessing discriminative capabilities (Alphonso vs. non-Alphonso).
Fig. S6: Scatterplots of metabolite levels.
Tab. S2: Metabolite levels and LOFs of Alphonso samples in the training set.
Tab. S3: Metabolite levels and LOFs of Alphonso samples in the test set.
Tab. S4: Metabolite levels and LOFs of pure non-Alphonso samples.
Tab. S5: Metabolite levels and LOFs of self-mixed admixtures in the test set.
Tab. S6: Information gains for assessing discriminative capabilities (Alphonso vs. admixtures).
1
Fig. S1 Exemplary 1H-NMR spectrum of an Alphonso sample (segment from 9.5 to 5.5 ppm). In this study, peak areas of ten metabolites were retrieved. The signals that were used for this are labeled in the spectrum.
Fig. S2 Exemplary 1H-NMR spectrum of an Alphonso sample (segment from 5.5 to 2.6 ppm, same spectrum as in Fig. S1). In this study, peak areas of ten metabolites were retrieved. The signals that were used for this are labeled in the spectrum.
Fig. S3 Exemplary 1H-NMR spectrum of an Alphonso sample (segment from 2.6 to 0.6 ppm, same spectrum as in Fig. S1). In this study, peak areas of ten metabolites were retrieved. The signals that were used for this are labeled in the spectrum.
Fig. S4 1H-NMR spectrum of an Alphonso sample (black) and the same sample after spiking of niacin (red). With regard to the 1H-NMR signals of trigonelline (see Fig. S5) only the H-6 proton is clearly separated and usable for quantification of niacin.
3
Fig. S5 1H-NMR spectrum of an Alphonso sample (black) and the same sample after spiking of trigonelline hydrochloride (red).
Tab. S1 Information gains for assessing each metabolites ability to discriminate Alphonso and non- Alphonso cultivars. A higher information gain indicates a higher discriminative capability. For details on calculation, see subparagraph “Feature Importance” in the experimental section of the manuscript
Metabolite Information Gain Metabolite Information Gain
Niacin 0.685 Adenosine 0.269
Histidine 0.539 Alanine 0.246
Formic acid 0.450 Shikimic acid 0.224
Trigonelline 0.450 p-Digallic acid 0.000
Choline 0.379 Succinic acid 0.000
Fig. S6 Scatterplots of metabolite levels of a) choline and adenosine, b) alanine and shikimic acid, c) p-digallic and succinic acid (ordered by decreasing information gain (see Tab. S1)). d): Legend
5
Tab. S2 Metabolite levels and LOFs of Alphonso samples in the training set. Key for metabolites: A - Niacin, B - Trigonelline, C - Histidine, D - Adenosine, E - Formic acid, F - Shikimic acid, G - Choline, H - Alanine. LOFs were calculated as described in subparagraph “Model” in the experimental section of the manuscript. Sample ID 06 is a significant outlier with a LOF of 1.590. Therefore, it was excluded from the training set. After this, LOFs were recalculated and are reported in the “LOF adjusted” column
Metabolite
ID Cultivar A B C D E F G H LOF LOF
adjusted 0
1 Alphonso 51.6 116.7 71.7 92.2 232.9 1412.
5 3016.
6 1965.
6 1.16
9 1.072
0
2 Alphonso 95.7 113.9 76.1 92.2 145.0 2415.
7 3800.
2 2014.
6 1.03
3 0.990
0
3 Alphonso 116.3 109.3 87.3 80.2 157.8 1470.
4 4149.
3 1736.
1 0.97
1 0.985
0
4 Alphonso 56.7 104.8 57.8 89.6 116.3 1803.
3 2886.
8 2057.
4 1.07
4 1.101
0
5 Alphonso 83.9 120.6 92.8 81.6 189.3 1759.
4 3145.
9 2222.
3 1.00
5 0.988
0
6 Alphonso 107.2 138.2 171.0 87.4 147.8 2081.
8 3980.
4 3066.
5 1.59
0 -
0
7 Alphonso 100.6 111.4 93.1 77.0 161.8 1509.
2 3276.
3 1890.
2 1.05
8 1.022
0
8 Alphonso 89.8 128.7 58.9 95.7 146.1 1694.
0 2793.
5 2183.
3 0.99
5 1.024
0
9 Alphonso 95.1 117.1 99.9 82.8 158.4 854.6 3745.
6 1888.
2 0.97
6 0.969
1
0 Alphonso 72.9 107.4 82.6 65.9 169.0 1405.
0 3215.
5 1857.
0 0.92
7 0.969
Tab. S3 Metabolite levels and LOFs of Alphonso samples in the test set. Key for metabolites: A - Niacin, B - Trigonelline, C - Histidine, D - Adenosine, E - Formic acid, F - Shikimic acid, G - Choline, H - Alanine.
LOFs were calculated as described in subparagraph “Model” in the experimental section of the manuscript. Samples were evaluated not to be Alphonso samples if their LOF was higher than 1.194 (see
“Inlier?” column)
Metabolite
ID Cultivar A B C D E F G H LOF Inlier?
1
1 Alphonso 93.4 124.5 62.7 97.9 147.9 1655.
8 2735.
8 2266.
3 1.01
5 yes
1
2 Alphonso 69.2 133.4 80.5 83.0 174.7 3054.
8
3139.
1
2097.
2
1.14
2 yes
1
3 Alphonso 106.
6 120.7 100.5 84.5 240.8 2013.
1 3903.
7 2412.
3 1.18
5 yes
1
4 Alphonso 57.7 124.8 77.0 80.7 243.6 1649.
7
2870.
2
1304.
2
1.49
6 no
1
5 Alphonso 62.9 129.5 87.4 84.7 125.8 2358.
2 3093.
5 1740.
8 1.11
0 yes
1
6 Alphonso 60.4 121.3 70.0 75.1 101.0 2780.
3
2932.
3
2021.
8
1.08
3 yes
1
7 Alphonso 82.3 112.0 76.4 76.4 143.2 1504.
2 3606.
7 1655.
6 1.01
4 yes
7
Tab. S4 Metabolite levels and LOFs of pure non-Alphonso samples. Key for metabolites: A - Niacin, B - Trigonelline, C - Histidine, D - Adenosine, E - Formic acid, F - Shikimic acid, G - Choline, H - Alanine.
LOFs were calculated as described in subparagraph “Model” in the experimental section of the manuscript. Samples were evaluated not to be Alphonso samples if their LOF was higher than 1.194 (see
“Inlier?” column)
Metabolite
ID Cultivar A B C D E F G H LOF Inlier?
1
8 Totapuri 7.4 43.1 24.0 59.3 110.2 762.8 1858.
0 3821.
0 4.39
9 no
1
9 Totapuri 11.1 37.2 24.3 65.1 46.7 379.1 1353.
0
4480.
5
5.46
3 no
2
0 Totapuri 16.8 67.2 48.8 101.5 196.7 1405.
0 2662.
1 7455.
8 9.28
0 no
2
1 Kesar 74.6 15.0 43.1 83.0 66.4 748.2 2915.
2
3065.
7
4.27
6 no
2
2 Kesar 92.9 11.0 33.9 78.6 39.3 702.9 3330.
3 5989.
1 7.73
2 no
2
3 Kesar 27.0 8.1 20.2 47.3 21.1 377.9 1116.
1
2135.
0
4.66
1 no
2
4 Sindura 21.0 83.8 57.8 52.7 45.5 2800.
0 2986.
2 4643.
3 4.77
3 no
2
5 Sindura 11.5 94.2 39.0 67.1 55.6 1460.
4
2695.
2
5112.
3
5.37
2 no
2
6 Sindura 19.5 87.9 56.3 86.1 53.1 1484.
5 2714.
1 5077.
8 5.28
2 no
2
7 Chato de Ica 8.8 35.8 19.2 40.6 23.7 968.4 1117.
6
1031.
6
4.15
5 no
2
8 Chato de Ica 10.0 24.3 16.3 16.9 129.3 3473.
0 767.5 705.4 5.02
1 no
2
9 Yai 6.6 28.1 27.6 46.7 23.3 593.0 1161.
9
2941.
7
4.31
9 no
3
0 Nam Dok Mai 21.0 3.3 26.9 46.1 3.0 836.3 806.0 2988.
5 5.11
3 no
Tab. S5 Metabolite levels and LOFs of self-mixed admixtures in the test set. The “Admixture Ratio”
column allows to track how these admixtures were generated. Each admixture was generated by mixing 35% of a pure non-Alphonso sample (C1 (component 1), see Tab. S4) and 65% of an Alphonso sample of the test set (C2 (component 2), see Tab. S3). Key for metabolites: A - Niacin, B - Trigonelline, C - Histidine, D - Adenosine, E - Formic acid, F - Shikimic acid, G - Choline, H - Alanine. LOFs were calculated as described in subparagraph “Model” in the experimental section of the manuscript. Samples were evaluated not to be Alphonso samples if their LOF was higher than 1.194 (see “Inlier?” column)
Admixture ratio Metabolite
ID C1 C2 A B C D E F G H LOF Inlier?
3
1 ID 18
(35%) ID 17
(65%) 59.7 86.3 58.7 72.7 151.
5 1256.
2 3029.
4 2352.
6 1.34
8 no
3
2 ID 18
(35%) ID 14
(65%) 38.7 98.8 62.0 82.5 200.
0 1343.
7 2572.
6 2246.
1 1.11
5 yes
3
3 ID 19
(35%) ID 15
(65%) 54.8 89.3 54.9 78.9 95.2 1700.
1 2488.
3 2451.
2 1.50
0 no
3
4 ID 19
(35%) ID 11
(65%) 67.9 91.2 56.2 91.3 139.
8 1238.
2 2301.
1 2888.
4 1.80
7 no
3
5 ID 20
(35%) ID 16
(65%) 44.0 92.2 56.7 69.5 115.
1 2142.
8 2434.
3 2738.
4 1.77
8 no
3
6 ID 20
(35%) ID 15
(65%) 49.8 96.6 76.8 72.5 119.
8 1860.
1 2554.
1 2587.
0 1.45
0 no
3
7 ID 21
(35%) ID 14
(65%) 64.1 87.7 57.6 83.2 205.
6 1373.
2 2956.
3 1985.
0 1.14
5 yes
9
Tab. S5 Continued
Admixture ratio Metabolite
ID C1 C2 A B C D E F G H LOF Inlier?
3
8 ID 21
(35%) ID 13
(65%) 92.8 90.7 72.0 85.5 169.
7 1587.
0 3602.
3 2633.
6 1.47
4 no
3
9 ID 22
(35%) ID 13
(65%) 100.
0 78.7 76.0 80.2 176.
2 1536.
6 3631.
5 3426.
6 2.79
5 no
4
0 ID 22
(35%) ID 15
(65%) 82.4 77.5 71.1 89.1 101.
5 1832.
6 3153.
3 3229.
2 2.46
4 no
4
1 ID 23
(35%) ID 16
(65%) 59.6 89.2 63.4 79.0 113.
1 2157.
0 2772.
0 2741.
6 1.72
8 no
4
2 ID 23
(35%) ID 13
(65%) 87.1 86.7 80.2 85.4 171.
6 1578.
5 3372.
0 3092.
6 2.09
9 no
4
3 ID 24
(35%) ID 17
(65%) 61.1 100.
9 71.2 70.3 111.
3 1998.
5 3442.
4 2633.
1 1.41
5 no
4
4 ID 24
(35%) ID 16
(65%) 48.4 115.
1 80.8 70.5 85.6 2960.
8 3127.
2 2874.
3 1.79
4 no
4
5 ID 25
(35%) ID 14
(65%) 44.0 113.
6 70.8 77.9 214.
2 1631.
8 2911.
8 2818.
5 1.45
6 no
4
6 ID 25
(35%) ID 13
(65%) 74.4 110.
0 86.8 80.6 168.
3 1821.
2 3508.
3 3223.
1 2.02
7 no
4
7 ID 26
(35%) ID 11
(65%) 66.0 115.
9 52.1 93.8 121.
2 1607.
2 2761.
7 3174.
0 1.95
9 no
4
8 ID 26
(35%) ID 15
(65%) 53.3 109.
5 75.2 85.3 99.3 2056.
3 2906.
0 2824.
9 1.48
4 no
4
9 ID 27
(35%) ID 13
(65%) 67.2 94.2 71.1 64.4 126.
6 1667.
5 2982.
2 1808.
9 1.08
6 yes
5
0 ID 27
(35%) ID 14
(65%) 39.4 100.
5 64.4 67.6 179.
5 1441.
9 2325.
1 1254.
0 1.58
3 no
5
1 ID 28
(35%) ID 15
(65%) 49.3 79.2 58.6 58.0 135.
6 2737.
5 2122.
2 1281.
5 2.18
9 no
5
2 ID 28
(35%) ID 12
(65%) 43.8 89.3 57.4 57.1 160.
5 3212.
4 2243.
4 1529.
4 1.92
5 no
5
3 ID 29
(35%) ID 16
(65%) 41.4 90.7 55.7 69.8 57.4 2029.
6 2345.
1 2536.
8 1.79
8 no
5
4 ID 29
(35%) ID 11
(65%) 63.1 88.0 59.6 89.4 72.2 1302.
6 2233.
5 2761.
5 1.87
9 no
5
5 ID 30
(35%) ID 13
(65%) 85.1 79.1 78.6 90.1 116.
8 1914.
2 3141.
7 3602.
4 2.93
5 no
5
6 ID 30
(35%) ID 14
(65%) 50.8 86.0 77.6 90.9 144.
9 1685.
7 2558.
2 3103.
3 2.16
3 no
Tab. S6 Information gain for assessing each metabolites ability to discriminate Alphonso and self-mixed admixtures (in that 35% of non-Alphonso cultivars are admixed to Alphonso samples). A higher information gain indicates a higher discriminativ capability. For details on calculation, see subparagraph
“Feature Importance” in the experimental section of the manuscript
Metabolite Information Gain Metabolite Information Gain
Niacin 0.000 Adenosine 0.000
Histidine 0.210 Alanine 0.224
Formic acid 0.000 Shikimic acid 0.000
Trigonelline 0.326 p-Digallic acid 0.000
Choline 0.162 Succinic acid 0.000
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