UHPLC-Q-TOF

The analysis of metabolites was carried out using an UHPLC LC 30 for the chromatographic determination. The extract was injected into a C18 analytical column (2.0 mm I.D × 75 mm, 1.6 μm particle size). The mobile phase was 0.1% formic acid solution (solvent A) and acetonitrile (solvent B). The gradient elution programmer was as follows: 0~2 min, 2% B; 2~13 min, 2–90% B; 13~16 min, 100% B; 16~17 min, 100-2% B; 17–20min, 2% B. The flow rate was of 0.3 mL/min, and the column temperature was set at 40 ℃. The injection volume is 3 μL. The peak detection of the extract was used a X500R Q-TOF mass detector. Both primary mass and secondary fragmentation pattern (MS/ MS) spectra were recorded between m/z 50 and m/z 1000 Da.

References：

[1] Li W , Liu Y , Ye Y ,et al.Chemical profiling and metabolic mechanism of Pixian doubanjiang, a famous condiment in Chinese cuisine[J].LWT- Food Science and Technology, 2021(4):111274.DOI:10.1016/j.lwt.2021.111274.

LC-ESI-MS/MS

The chromatographic conditions were as follows: BEH C18 chromatographic column (150?mm?×?2.1?mm, 1.7 μm); mobile phase: A phase water, B phase acetonitrile, 0.1% (vol/vol) formic acid and 0.2 mM ammonium acetate were added t both phases. Step-stripping sequence: 0–1.0 min, 10% B; 0–9.0?min, 10% B–90% B; 9.0–11.0?min, 90% B–100% B; 11.0–11.1?min, 100% B–10% B; 11.1–13.0 min, 10% B. Column temperature: 30℃; injection volume: 2 μl; flow rate: 0.25?ml/min. Mass spectrum conditions: retention time 4.94?min?1; ion pair l: quantitative ion 433.1/271.1, collision energy ?22?V; ion pair 2: quantitative ion 433.1/150.9, collision energy ?42?V.

References：

Sun M, Li L, Wang C, Wang L, Lu D, Shen D, Wang J, Jiang C, Cheng L, Pan X, Yang A, Wang Y, Zhu X, Li B, Li Y, Zhang F. Naringenin confers defence against Phytophthora nicotianae through antimicrobial activity and induction of pathogen resistance in tobacco. Mol Plant Pathol. 2022 Dec;23(12):1737-1750. doi: 10.1111/mpp.13255. Epub 2022 Sep 12. PMID: 36094814; PMCID: PMC9644278.

UPLC-MS

Ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) was performed with liquid mass spectrometer for evaluation. The determination was performed on the SB-C18 column at the flow rate of 0.3 mL/min and the column temperature was 40 ℃. The flavonoid and anthocyanin components were detected at 210 nm.

References：

Shi M, Ali MM, He Y, Ma S, Rizwan HM, Yang Q, Li B, Lin Z, Chen F. Flavonoids Accumulation in Fruit Peel and Expression Profiling of Related Genes in Purple (Passiflora edulis f. edulis) and Yellow (Passiflora edulis f. flavicarpa) Passion Fruits. Plants (Basel). 2021 Oct 20;10(11):2240. doi: 10.3390/plants10112240. PMID: 34834602; PMCID: PMC8620868.

HPLC–MS

We used an HPLC system, which consisted of a binary solvent delivery system, an on-line degasser, an auto-sampler, a column temperature controller, and a diode array detector. Separation was conducted using a C18 column (5 μm, 150 mm × 4.6 mm), and the temperature was set to 40℃. The injection volume was 0.01 mL, and the flow rate was 1 mL/min. Gradient elution was conducted using a binary system consisting of 10 % formic acid in DW (A) and 10 % formic acid in acetonitrile (B). The utilized gradient was as follows: isocratic 1 % B for 6 min, 1% – 3% B for 4 min, 3 % – 15 % B for 20 min, 15 % – 40 % B for 20 min, 40 % – 90 % B for 5 min, 90 % B isocratic for 10 min, 90 % – 1 % B for 5 min, and 1 % B isocratic for 10 min. A re-equilibration operation followed. The UV detection wavelength was set to 280 nm.The voltages of the sample cone and the capillary were set to 17 and 3000 V for the positive ion mode and 22 and 2000 V for the negative ion mode.

References：

[1]Zhang,Junhong,Tian,et al.Isolation and identification of phenolic compounds in Chinese purple yam and evaluation of antioxidant activity[J].LWT-Food Science & Technology, 2018, 96:161-165.

UHPLC

Chromatographic separation of SPLFA and SPLF was done on an Agilent 1290 UHPLC system coupled to an auto sampler, a diode array detector (DAD) and an ACQUITY UHPLC? HSS T3 column (2.1 ×100 mm, 1.8 μm; Waters, Milford, MA, USA)maintained at 30 ℃. Eluent A was 0.05% formic acid in water and eluent B was 0.05% formic acid in acetonitrile. Elution of the SPLPA compounds was achieved using the following linear gradient elution (in %B): 0 min, 5%; 20 min, 60%; 20.1 min, 5%; 30 min, 5%, and for the SPLF compounds: 0 min, 20%, 25 min, 50%, 25.1 min, 20%, 35 min, 20%. The injection volume was 5 μL, with a flow rate of 0.3 mL/ min. All sample solutions were filtered through a 0.22 μm PTFE filter (Agilent Technologies) before analysis. The detection wavelength was 320 nm for SPLPA and 254 nm for SPLF.

References：

[1] Luo D , Mu T , Sun H .Profiling of phenolic acids and flavonoids in sweet potato (Ipomoea batatas L.) leaves and evaluation of their anti-oxidant and hypoglycemic activities[J].Food Bioscience, 2020, 39(44):100801.DOI:10.1016/j.fbio.2020.100801.