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To analyze the fragmentation rule and pathway of pelargonidin, cyanidin, delphinidin, peonidin, petunidin and malvinidol under UHPLC-QTOF-MS positive mode electric spray, identify the characteristic product ions of six anthocyanins, and provide a theoretical basis for the establishment of mass spectrometry database and detection methods.
The chromatographic conditions were as follows: chromatographic column, Fusion-RPC18 (50 mm×2.0 mm, 4 μm), mobile phase 0.1% formic acid aqueous solution (A) and methanol (B), gradient elution (0-1 min, 95%A; 1-5 min, 95%A→10%A; 5-6 min, 1%A; 6-7 min, 95%A), flow rate 0.3 mL·min-1, column temperature 40 ℃, and injection volume 10 μL. The mass spectrometry conditions were as follows: TOF MS-IDA MS/MS, curtain gas, 0.20 MPa, collision gas CAD 7 MPa, IS voltage, 5 500 V/-4500 V, ion source temperature, 500 ℃, nebulizer gas, GAS1, 0.38 MPa, auxiliary, GAS2, 0.48 MPa, DP voltage, ±60 V, fragmentation voltage, (35±15) V, and time 0.2 s. Under the condition of positive mode of electrospray, the mass spectrometry data of six anthocyanins were measured. According to the pyrylium ions formed by the 2-phenylchromogenic structure of anthocyanins, and combined with the auxiliary analysis of the mass spectrometer database, the possible product ions were deduced.
The results showed that the six anthocyanins mainly undergo cleavage reactions on the pyranium ring, ultimately generating intermediate ions of pyrogallol and benzyl alcohol. On the other way, it occured α cracking, σ cracking causes the loss of functional groups on the ring, ultimately resulting in the formation of Chain hydrocarbons without functional groups.
The research results can provide support for the mass spectrometry characteristic ion data of six anthocyanins, which can be used to establish ion library data for anthocyanin products and also provide reference for the development and research of detection methods.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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