Rapid Determination of Major Compounds in the Ethanol Extract of Geopropolis from Malaysian Stingless Bees, Heterotrigona itama, by UHPLC-Q-TOF/MS and NMR
Abstract
:1. Introduction
2. Results and Discussion
2.1. Identification of Compounds from EEGP by UHPLC-Q-TOF/MS
2.1.1. Identification of Compounds in EEGP Based on Known Authentic Standards
2.1.2. Identification of Compounds in EEGP using METLIN and MSC Software
2.2. Identification of Unknown Compounds using Preparative HPLC (PHPLC) and NMR
3. Materials and Methods
3.1. Chemicals
3.2. UHPLC-Q-TOF/MS Analysis of EEGP
3.2.1. Sample Preparation
3.2.2. UHPLC System and Mass Spectrometry
3.3. PHPLC Purification and NMR Analysis for Identification of Unknown Compounds in EEGP
3.3.1. PHPLC
3.3.2. NMR
4. Conclusions
Supplementary Materials
Supplementary File 1Acknowledgments
Author Contributions
Conflicts of Interest
References
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Sample Availability: Samples of the compounds are available from the authors. |
Peak | Rt | m/z (−) | Error (ppm) | m/z (+) | Error (ppm) | Formula | MS/MS (m/z) (−) | MS/MS (m/z) (+) | Identification | Confirmation |
---|---|---|---|---|---|---|---|---|---|---|
1 | 2.2 | 169.0144 | 1.18 | 171.0283 | −2.92 | C7H6O5 | 125.0234, 107.0128 | / | Gallic acid [17,30] | MSC and authentic standard |
2 | 3.1 | 179.0342 | −4.47 | 181.0489 | −3.31 | C9H8O4 | 135.0399,109.0301 | / | Caffeic acid [18] | MSC and authentic standard |
3 | 3.9 | 353.0876 | −0.56 | 355.1019 | −1.41 | C16H18O9 | 191.0129, 179.0488, 173.0004, 161.0535, 154.9881 | / | Caffeoylquinic acid [31] | MSC |
4 | 4.6 | 331.1031 | −1.21 | 333.1170 | −3.00 | C14H20O9 | 211.0224, 169.0133, 125.0232, 107.0127 | / | Gallic acid-hexose [32] | MSC |
5 | 5.5 | 167.0362 | 1.20 | 169.0491 | −2.37 | C8H8O4 | 153.0206, 108.0213 | / | Vanillic acid [33] | MSC |
6 | 7.9 | 167.0354 | 2.39 | / | C8H8O4 | 153.0211, 108.0214 | / | Isomer of vanillic acid [33] | MSC | |
7 | 9.3 | 195.0877 | 0 | C8H10N4O2 | / | 150.0888, 138.0668, 135.0477, 110. 0713 | Caffeine [34] | MSC | ||
8 | 12.8 | 151.0402 | 0.66 | 153.0543 | −1.96 | C8H8O3 | 137.0233, 123.0463, 107.0120 | / | Vanillin [33] | MSC |
9 | 16.5 | 197.0452 | −1.52 | 199.0611 | 5.02 | C9H10O5 | 153.0481, 124.0161, 107.0477, 106.0062 | / | Syringic acid [33] | MSC and authentic standard |
10 | 21.8 | 153.0192 | −0.65 | 155.0344 | 3.23 | C7H6O4 | 123.0416, 109.0285 | / | Protocatechuic acid [33] | MSC |
11 | 25.2 | 125.0244 | 0 | 127.0391 | 0.79 | C6H6O3 | 107.0130 | / | Pyrogallol [33] | MSC |
12 | 31.8 | 121.0293 | −1.65 | 123.0440 | −0.81 | C7H6O2 | 105.0349 | / | Benzoic acid | authentic standard |
13 | 33.1 | 583.1118 | 4.29 | / | C28H24O14 | 431.0997, 313.0578, 269.0467,169.0143, 125.0242 | / | Vitexin-O-gallate [35] | MSC | |
14 | 34.0 | 583.1120 | 4.63 | C28H24O14 | 431.0970, 313.0577, 269.0461, 169.0142, 125.0234 | / | Isomer of vitexin- O-gallate [35] | MSC | ||
15 | 34.9 | 205.0863 | −3.41 | 207.1014 | −0.97 | C12H14O3 | / | 189.0526, 150.0297, 149.0232, 122.0335 | Acetyleugenol [36] | MSC |
16 | 36.8 | 271.0613 | 0.37 | 273.0754 | −1.10 | C15H12O5 | 229.0477, 211.0348, 187.0375, 151.0012 | / | Pinobanksin | MSC and authentic standard |
17 | 38.4 | 161.0243 | −0.62 | 163.0396 | 3.68 | C9H6O3 | / | 135.0447, 133.0286, 107.0512, 105.0347 | Umbelliferone [37] | MSC |
18 | 39.7 | 285.0417 | 4.21 | 287.0557 | 2.44 | C15H10O6 | 255.0341, 239.0376, 227.0384, 211.0421, 199.0574, 124.0143, 107.0135 | / | Kaempferol [33] | MSC and authentic standard |
19 | 40.6 | 247.0972 | −1.62 | 249.1123 | 0.80 | C14H16O4 | 217.0884, 172.0888, 144.0939 | / | Prenyl caffeate [38] | MSC |
20 | 41.3 | 439.3583 | 0.23 | 441.3725 | −0.45 | C30H48O2 | / | / | 24(E)-cycloart-24-ene-26-ol-3-one [23] | NMR |
21 | 42.4 | 241.0874 | 1.66 | 243.1017 | 0.41 | C15H14O3 | / | 225.0560, 183.0806, 149.0597, 133.0665 | Lapachol [39] | MSC |
22 | 43.2 | 441.3736 | −0.45 | 443.3882 | −0.45 | C30H50O2 | / | / | 20-hydroxy-24-dammaren-3-one [24] | NMR |
23 | 44.9 | 407.1362 | 3.44 | 409.1488 | −1.22 | C20H24O9 | 245.0832, 230.0594, 202.0612, 187.0403, 173.0605, 137.0224 | / | Torachrysone-O-hexose [30] | MSC |
24 | 45.7 | 449.1462 | 2.00 | 451.1506 | −0.66 | C22H26O10 | 407.1322, 245.0482, 230.0065, 202.0516 | / | Torachrysone-O-(acetyl)-hexose [30] | MSC |
25 | 46.7 | 559.1467 | 1.79 | 561.1554 | −1.60 | C27H28O13 | 313.0575, 287.0938, 245.0827, 230.0596, 215.0367, 169.0143, 125.0232 | / | Torachrysone-O-(galloyl)-hexose [30] | MSC |
26 | 47.8 | 409.1656 | −0.24 | 411.1801 | −0.24 | C24H26O6 | / | 393.1688, 355.1167, 341.1009, 299.0541, 195.1013 | Mangostin [40] | MSC |
27 | 48.7 | 437.3424 | −0.23 | 439.3570 | −0.23 | C30H46O2 | / | 421.3449, 249.1849, 235.1698, 167.1074 | Ganoderol A [41] | MSC |
28 | 50.0 | 427.3582 | 0 | 429.3712 | −3.93 | C29H48O2 | / | 411.3611, 193.1608 | Saringosterol [42] | MSC |
29 | 52.3 | 411.3629 | −0.73 | 413.3757 | −5.08 | C29H48O | / | 395.3651, 135.1171 | Stigmasterol [43] | MSC |
30 | 56.0 | 423.3630 | −0.47 | 425.3777 | −0.24 | C30H48O | / | 407.3648, 271.2086, 135.1169, 109.1019 | Taraxerone [44] | MSC |
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Zhao, L.; Yu, M.; Sun, M.; Xue, X.; Wang, T.; Cao, W.; Sun, L. Rapid Determination of Major Compounds in the Ethanol Extract of Geopropolis from Malaysian Stingless Bees, Heterotrigona itama, by UHPLC-Q-TOF/MS and NMR. Molecules 2017, 22, 1935. https://doi.org/10.3390/molecules22111935
Zhao L, Yu M, Sun M, Xue X, Wang T, Cao W, Sun L. Rapid Determination of Major Compounds in the Ethanol Extract of Geopropolis from Malaysian Stingless Bees, Heterotrigona itama, by UHPLC-Q-TOF/MS and NMR. Molecules. 2017; 22(11):1935. https://doi.org/10.3390/molecules22111935
Chicago/Turabian StyleZhao, Lingling, Mengjiao Yu, Minghui Sun, Xiaofeng Xue, Tongtong Wang, Wei Cao, and Liping Sun. 2017. "Rapid Determination of Major Compounds in the Ethanol Extract of Geopropolis from Malaysian Stingless Bees, Heterotrigona itama, by UHPLC-Q-TOF/MS and NMR" Molecules 22, no. 11: 1935. https://doi.org/10.3390/molecules22111935