HPLC

。。。the supernatant was filtered with a microporous membrane (0.45mm) for highperformance liquid chromatography (HPLC) analysis, and Comatex C18 column (250 mm x 4.6 mm, 5 mm), column temperature 30 ℃. The elution system consisted of a mobile phase A (ultra-pure water) with a gradient elution procedure at a flow rate of 1.0 mL·min-1, injection volume of 10.0 mL, column temperature of 30 ℃, and detection wavelength of 520 nm.

来源文献：Yao L, Liang D, Xia H, Pang Y, Xiao Q, Huang Y, Zhang W, Pu C, Wang J, Lv X. Biostimulants promote the accumulation of carbohydrates and biosynthesis of anthocyanins in 'Yinhongli' plum. Front Plant Sci. 2023 Jan 6;13:1074965. doi: 10.3389/fpls.2022.1074965. PMID: 36684717; PMCID: PMC9854126.

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.

来源文献：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.

UPLC-MS/MS

The analytical conditions were as follows, UPLC: column, C18 (1.8 μm, 2.1 mm × 100 mm); the mobile phase was consisted of solvent A, pure water with 0.1% formic acid, and solvent B, acetonitrile. Sample measurements were performed with a gradient program that employed the starting conditions of 95% A, 5% B. Within 9 min, a linear gradient to 5% A, 95% B was programmed, and a composition of 5% A, 95% B was kept for 1min. Subsequently, a composition of 95% A, 5.0% B was adjusted within 1.10 min and kept for 2.9 min. The column oven was set to 40 ℃. The injection volume was 4 μL.

The ESI source operation parameters were as follows: ion source, turbo spray; source temperature 550 ℃; ion spray voltage (IS) 5500 V (positive ion mode)/-4500 V (negative ion mode); ion source gas I (GSI), gas II(GSII), curtain gas (CUR) were set at 50, 60, and 30.0 psi, respectively; the collision gas (CAD) was high. Instrument tuning and mass calibration were performed with 10 and 100 μM polypropylene glycol solutions in QQQ and LIT modes, respectively. QQQ scans were acquired as MRM experiments with collision gas (nitrogen) set to 5 psi. DP and CE for individual MRM transitions were done with further DP and CE optimization. A specific set of MRM transitions were monitored for each period according to the metabolites eluted within this period.

来源文献：[1] Xiao Y , Li K , Zhang H ,et al.The profile of buckwheat tannins based on widely targeted metabolome analysis and pharmacokinetic study of ellagitannin metabolite urolithin A[J].LWT, 2022, 156:113069-.DOI:10.1016/j.lwt.2022.113069.

HPLC

Anthocyanins were separated in an HPLC system . A C18 column (5 μm, 4.6 mm × 150 mm) was used; the loading volume was 20 μL, mobile phase A was 10% formic acid, mobile phase B was acetonitrile. The linear gradient elution design was: 0–13 min – acetonitrile 0–20%, 20 min –acetonitrile 40%, 25 min – acetonitrile 0%; column temperature was 25 ℃, flow rate was 1 mL/min, and detection wavelength was 520 nm. A standard curve was prepared using cyanidin 3-O-galactoside (Beijing Solarbio Science & Technology Co. Ltd., Beijing, China).

来源文献：Wang Z, Song M, Li Y, Chen S, Ma H. Differential color development and response to light deprivation of fig (Ficus carica L.) syconia peel and female flower tissues: transcriptome elucidation. BMC Plant Biol. 2019 May 23;19(1):217. doi: 10.1186/s12870-019-1816-9. PMID: 31122203; PMCID: PMC6533723.

HPLC

The phenolic acid contents were also measured usingHigh Performance Liquid Chromatography.Chromatographic separation was conductedwith an C18 column (4.6 mm × 250 mm,5 mm) and a binary solvent system of (A) methanol (0.1%H3PO4) and (B) water (0.1% H3PO4). The elution wasperformed at a flow rate of 1.0 mL/min and the columntemperature was maintained at 30℃. The contents of phenolicacids [CGA, neochlorogenic acid (NCGA), cryptochlorogenicacid (CCGA), p-coumaryl quinic acid, and caffeoyl shikimicacid] were detected at 320 nm and the contents of anthocyanin(cyanidin-3-O-glucoside chloride and cyanidin 3-O-rutinosidechloride) were quantified at 525 nm.

来源文献：Su Z, Jia H, Sun M, Cai Z, Shen Z, Zhao B, Li J, Ma R, Yu M, Yan J. Integrative analysis of the metabolome and transcriptome reveals the molecular mechanism of chlorogenic acid synthesis in peach fruit. Front Nutr. 2022 Jul 19;9:961626. doi: 10.3389/fnut.2022.961626. PMID: 35928835; PMCID: PMC9344011.

HPLC

HPLC analysis was carried out using Waters e2695 (Waters Corp., Milford, MA, USA) equipped with 2998PDA detector. The separation was performed using a Symmetry? C18 column (4.6 × 250 mm; i.d., 5 μm; Waters Corporation, Milford, MA). HPLC setting conditions followed the previously reported protocol with some modifications (Liu, Chang, Liu, & Shen, 2016). The mobile phase consisted of acetonitrile (A) and 2% formic acid aqueous solution (B). The column flow rate was 0.7 mL/min with gradient elution program of 0–10 min, 8–10% A; 10–12 min, 10–15% A; 12–15 min, 15–30% A; 15–18 min, 30–60% A; 18–20 min, 60–80% A; 20–22 min, 80–8%A; 22–25 min, 8%A. The injection volume was 10 μL with column temperature of 30 ℃, and the detection wavelength was set at 520 nm.

来源文献：[1] A M H , B J D A , A L D ,et al.Anti-fatigue activity of purified anthocyanins prepared from purple passion fruit ( P. edulis Sim ) epicarp in mice[J].Journal of Functional Foods, 65[2024-01-05].DOI:10.1016/j.jff.2019.103725.