Characterization of polymeric phenolic acids and flavonoids in Clerodendranthi Spicati Herba using ultrahigh‐performance liquid chromatography coupled to quadrupole time‐of‐flight tandem mass spectrometry with target and nontarget data mining strategy

Abstract

Rationale Clerodendranthi Spicati Herba (CSH) is often used to treat urinary stones, urinary tract infections and nephritis in China. Much literature has reported that polymeric phenolic acids and flavonoids are the major bioactive components in CSH. Therefore, it is very meaningful to identify the polymeric phenolic acids and flavonoids in CSH. Methods Ultrahigh‐performance liquid chromatography coupled to quadrupole time‐of‐flight tandem mass spectrometry (UHPLC/QTOF‐MS/MS) analysis with target and nontarget data mining strategy was proposed to rapidly profile the polymeric phenolic acids and flavonoids in CSH. Diagnostic product ions and neutral loss filter were beneficial for identifying the polymeric phenolic acids and flavonoids from complex compounds in CSH. Results A total of 118 compounds, including 85 polymeric phenolic acids and 33 flavonoids, were reasonably identified in CSH by comparing their main fragmentation pathways with literature data, and 85 of them were discovered in CSH firstly by nontarget analysis. Nine potential compounds were characterized tentatively as new pentameric and hexameric phenolic acids in CSH. Six types of polymeric phenolic acids (monomer, dimer, trimer, tetramer, pentamer and hexamer) and four types of flavonoids (apigenin, kaempferol, luteolin and quercetin) were identified in CSH. Conclusions The results indicated that the UHPLC/QTOF‐MS/MS method coupled with target and nontarget data mining strategy was feasible and rational for identifying the polymeric phenolic acids and flavonoids in complex chemical constituents of CSH. The findings will be conducive to the discovery of the active ingredients of CSH and the establishment of quality standards.

Ultrasound-Assisted Extraction Optimization of α-Glucosidase Inhibitors from Ceratophyllum demersum L. and Identification of Phytochemical Profiling by HPLC-QTOF-MS/MS

“…Peak 3 (195.0514, C 6 H 12 O 7 ) was identified as gluconic acid according to reference [ 28 ]. Peaks 5, 10, 18, and 49 were individually identified as malic acid (133.0148, C 4 H 6 O 5 ), citric acid (191.0200, C 6 H 8 O 7 ), p -coumaric acid (163.0404, C 9 H 8 O 3 ), and azelaic acid (187.0984, C 9 H 16 O 4 ) due to the diagnostic MS/MS fragment ions at 115.002 [M − H − H 2 O] − , 111.0083 [M − H − CO 2 − 2H 2 O] − , 119.0487 [M − H − CO 2 ] − , and 125.0970 [M − H − C 2 H 2 O 2 ] − , respectively [ 29 , 30 ]. Peaks 19, 20, and 24 (C 15 H 18 O 8 ) showed the similar [M − H] − at 325.0938 and similar fragmentation pattern, suggesting they were isomers.…”

“…Peak 51 (287.0565, C 15 H 12 O 6 ) had two more hydrogen atoms compared with kaempferol; fragment ions at 259.0611 [M − H − CO] − , 151.0028 [ 1,3 A] − , 125.0239 [ 1,4 A] − allowed the assignment of dihydrokaempferol. Analogously, apigenin (peak 58) and its glycosides (peaks 21, 37, 43), luteolin (peak 54) and luteolin-7- O -hexoside (peak 34), naringenin (peak 59) and its glycosides (peaks 38, 45), myricetin-3- O -hexoside (peak 30), laricitrin-3- O -hexoside (peak 35), syringetin-3- O -hexoside (peak 40), chrysoeriol-7- O -hexoside (peak 44) were proposed by matching the MS and MS/MS data with those recorded in the literature and databases [ 29 , 35 , 41 , 42 ]. The detected fragmentation pattern of peak 58 is shown in Figure 5 e.…”

Comprehensive chemical and metabolic profiling of anti‐hyperglycemic active fraction from Clerodendranthi Spicati Herba

“…Guo et al have identified 21 phenolic acids, 11 MFs, and six diterpenoids in CSH [14]. In our previous study, 85 polymeric phenolic acids and 33 flavonoids were rapidly identified in CSH by UHPLC-QTOF-MS/MS with diagnostic product ions (DPIs) and neutral loss (NL) filtering strategy [15]. In summary, the chemical compositions of CSH include phenolic acids, flavonoids, MFs, diterpenoids, and triterpenoids.…”

“…The dried CSH (1.2 kg) was cut into pieces and exhaustively extracted twice under reflux with 10 L 80% ethanol (1 h each time). The solution was evaporated in vacuum to afford a residue (129 g) as the ethanol extract [15]. Then, 100 g of ethanol extract was distributed in 200 mL of waterethanol (4: 1, v/v) and loaded on a macroporous resin chromatography column, then eluted with 45 and 95% ethanol to afford two fractions, and after evaporation of the solvent in reduced pressure, the 45% EEF (phenolic acids and flavonoids fraction, 64 g) and 95% EEF (MFs and diterpenoids fraction, 2.9 g) were obtained.…”

“…Demethylation, sulfation, and glucuronidation were the main metabolic pathways of MFs. The metabolites were identified using the NL of 15 substituent were on ring B, and two hydroxyl substituents and one methoxy substituent were on ring A. Metabolite A6 had a precursor ion at m/z 317.0656, which was identified through filtration using DPI m/z 302.0421 via NL of 15 Da (⋅CH 3 ) as 5,7,3′,4′-tetrahydroxy-6-methoxyflavone. The DPIs m/z 168.0053 and m/z 135.0441 indicated two hydroxyl substituents were on ring B, and three hydroxyl substituents and one methoxy substituent were on ring A. Metabolites A5, A9, and A13 had a precursor ion at m/z 507.1133, which were identified using DPI m/z 331.0812 via NL of 176 Da (C 6 H 8 O 6 ) as 5,7,3′-trihydroxy-6,4′dimethoxyflavone-GluA.…”