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Analysis of secondary metabolites in Xinjiang Morus nigra leaves using different extraction methods with UPLC-Q/TOF-MS/MS technology

  • Xue-Li Guo , Lu Yang EMAIL logo , Si-Lin Yu , Ke Zhang , Jin-Hui Wang and Hang-Yu Wang EMAIL logo
Published/Copyright: September 25, 2023
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Open Chemistry
From the journal Open Chemistry Volume 21 Issue 1

Abstract

Objective

The differences in the chemical composition of Morus nigra (M. nigra) extracts from four different extraction methods, ultrasound-assisted extraction with pure water (WU), pure water decoction extraction (WD), ultrasonic-assisted extraction with formic acid water (FAU), and pure water heat reflux extraction (WHR), were identified using ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) technology.

Method

A Waters ACQUITY BEH C18 (1.7 μm, 2.1 mm × 100 mm) was used, with a column temperature of 45°C, mobile phase of methanol and 0.1% formic acid aqueous solution, and gradient elution with a flow rate of 0.4 mL/min. Detection was performed in positive and negative ion modes, and compounds were identified using Progenesis QI software and mass spectrometry data reported according to the literature and laboratory self-built databases of the Mulberry genus. Multivariate statistical techniques, such as principal component analysis and orthogonal partial least squares-discriminant analysis, were applied to differential cluster metabolic profiles and chemical components and to screen the differential chemical components of M. nigra leaves.

Results

There were significant differences in the chemical composition between WD and the other extraction methods of M. nigra leaves. A total of 13 differential metabolites (4 flavonoids, 3 organic acids, 3 phenylpropanoids, 2 alkaloids, and 1 trisaccharide) were identified.

Conclusion

The multivariate statistical analysis and UPLC-Q-TOF-MS/MS method established in this study identified the differential chemical constituents of Xinjiang M. nigra leaves using different extraction methods, which provides a basis for the quality control of M. nigra leaves, and provides basic data for revealing the influence of extraction methods on the synthesis and accumulation of M. nigra leaf metabolites, which has certain reference significance.

Keywords: Morus nigra L. leaves; different extraction methods; secondary metabolites; UPLC-Q-TOF-MS/MS

Abbreviations

DNJ

1-deoxynojirimycin

FAU

ultrasonic-assisted extraction with formic acid water

OPLS-DA

orthogonal partial least squares-discrimination analysis

PCA

principal component analysis

UPLC-Q/TOF-MS

ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry

WD

pure water decoction extraction

WHR

pure water heat to reflux extraction

WU

ultrasound-assisted extraction with pure water

1 Introduction

Mulberry leaves are dried leaves of Morus alba L. or Morus nigra L. (M. nigra), members of the Moraceae family. Mulberry leaves have a wide range of pharmacological effects, including antibacterial [1], hypolipidemic [2], hypoglycemic [2,3], anti-inflammatory [4], anti-aging [5], and anti-tumor [6] effects. Modern studies have found that mulberry leaves contain flavonoids, steroids, alkaloids, amino acids, organic acids, phenylpropanoids, and other components [7]. As a widely planted mulberry variety in Xinjiang, China, M. nigra is an important supplement and the source of medicinal mulberry leaves.

At present, the research on M. nigra leaves mainly focuses on the pharmacological effects of single chemical components [8] or pharmacodynamic evaluation of crude extracts [4,6,9,10], and there is no in-depth study on the components and contents of mulberry leaf extracts in different extraction solvents. The study of chemical constituents of traditional Chinese medicine is premised on clarifying pharmacological actions, mechanism of actions, and clinical efficacy, and it is of great reference value to analyze and compare the differential chemical constituents of M. nigra leaves using different extraction methods based on plant metabolomic techniques (Table 1).

Table 1

Metabolomics analysis

Methods Advantages Disadvantages References
Gas chromatography–mass spectrometry High resolution, high sensitivity, a large number of reference databases Pretreatment such as derivatization is required; compounds with poor thermal stability and high molecular weight are not suitable [11,12,13]
Liquid chromatography–mass spectrometry High resolution, fast analysis speed, and high sensitivity, suitable for the detection of high boiling point, thermally unstable, and high molecular weight compounds — [14,15,16]
Capillary electrophoresis–mass spectrometry Analysis of ionic compounds; samples do not need to be derivatized Separation analysis and micro samples and specific uses; not well-used in plant research [17,18]
Nuclear magnetic resonance Simple pre-processing; low sample usage; the amount of information that can be measured is large The detection sensitivity is low and the detection range is narrow, which cannot realize the detection of trace substances [19,20]

Extensive untargeted metabolomics analysis based on ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) is a rapid and reliable method for the detection of metabolites in a variety of plant medicinal herbs [21]. In this study, UPLC-Q-TOF-MS/MS technology was used for non-targeted analysis to more comprehensively evaluate M. nigra leaves using different extraction methods. In addition, statistical analysis was performed to identify and quantify signature compounds and to provide the basis for quality control (QC) of M. nigra leaves.

2 Materials and methods

2.1 Materials and reagents

Sample: M. nigra leaves, collected in 2021 from Xinhe County, Aksu City, Xinjiang, China, were naturally dried and stored in the laboratory for later use.

Reagents: Purified water was prepared by purification with Milli-Q. Methanol, acetonitrile, and formic acid were of MS grade (Thermo Fisher Scientific China Ltd). The purity of the 1-deoxynojirimycin (DNJ) standard (lot number ZZS-20-052-A4, specification 25 mg) and fagomine standard (lot number 21A029-A1, specification 2 mg) was >98%, and the standards were purchased from Shanghai ZZBIO Co., Ltd.

2.2 Instruments and equipment

The instruments and equipment used in this study included ultra-high performance phase quadrupole time-of-flight mass spectrometer (Waters, USA); SW23 water bath (Julabo, Germany); desktop low-temperature high-speed centrifuge (Thermo, USA); ultrapure water system (Millipore, USA); KQ-600E CNC ultrasonic cleaner (Jiangsu Kunshan Ultrasonic Instrument Co., Ltd); analytical balance (Mettler-Toledo Instruments Shanghai Co., Ltd); and a QL-866 rapid mixer (Haimen Kylin-Bell Lab Instruments).

2.3 Sample preparation procedures

2.3.1 Ultrasound-assisted extraction

Extractions with solvents of pure water, 40% ethanol, 70% ethanol, and 95% ethanol were investigated. Briefly, 1 g of M. nigra leaf powder was weighed and added to 20 mL of extract solvent in a 50 mL conical flask with a stopper. The bottle was placed into an ultrasonic cleaner for the extraction process carried out under the following experimental conditions: ultrasonic temperature 25°C, ultrasonic time 30 min, and material ratio 1:20 (w/v, sample powder/purified water). The sample was filtered after extraction, and 1 mL of filtrate was diluted to 10 mL with methanol in a volumetric flask. After shaking, the mixture was centrifuged at 10,000 rpm for 5 min. Each sample was extracted three times in parallel.

2.3.2 Pure water decoction extraction

This referred to the method of Deng et al. [22] with a slight modification. Briefly, 10 g M. nigra leaves were weighed in a round-bottomed flask. Then, 200 mL of purified water was added for a material ratio of 1:20 (w/v, sample/purified water), followed by decoction and extraction in a 95°C water bath for 2 h. Each extract was filtered and fixed to 150 mL, then pipetted into a 0.05 mL to 2 mL volumetric flask, and the volume was fixed with methanol. Each sample was extracted three times in parallel. This method was denoted as WD.

2.3.3 Ultrasonic-assisted extraction with formic acid water

The laboratory established methods were as follows. Briefly, 1 g of M. nigra leaf powder was weighed, 10 mL of 70% methanol and 0.05% formic acid water was added to a 25 mL stoppered Erlenmeyer flask with the powder, and then the bottle was placed into an ultrasonic cleaner for the extraction process. This was carried out under the following experimental conditions: the ultrasonic temperature was 25°C, the ultrasonic processing time was 10 min, and the material ratio was 1:10 (w/v, sample powder/70% methanol + 0.05% formic acid water). Next the extracted sample was moved to a centrifuge tube and centrifuged at 10,000 rpm for 5 min, and 0.2 mL of the supernatant was placed in a 2 mL volumetric flask. The volume was fixed with methanol and mixed well. Each sample was extracted three times in parallel. This method was denoted as FAU.

2.3.4 Heat reflux extraction

According to the existing methods of the laboratory, the extraction of solvent pure water and 90% ethanol was investigated. First, 100 g M. nigra leaves were weighed in a 2,000 mL round-bottomed flask, and 1,000 mL of extract solvent was added for a material ratio of 1:10 (w/v, sample/purified water). Reflux extraction was performed with an electric heating jacket for 2 h, the filtrate was filtered with gauze, and 1,000 mL of purified water was added to a round-bottomed flask containing the filter residue. Reflux extraction was performed for 2 h, during which the filtrate was combined with the first filtrate. The filtrate was concentrated using a pressure reducing device to generate an extract. The extract was heated to remove the remaining solvent to obtain 39 g of water-extracted extract. Then, weigh 0.1 g of extract into a 2 mL volumetric flask, and fix the volume to the scale with 70% methanol, mixed, and centrifuged at 10,000 rpm for 5 min. Then, 0.05 mL of the supernatant was placed in a 2 mL volumetric flask, and the volume was set with methanol and mixed. Each sample was extracted three times in parallel.

2.3.5 QC sample

Briefly, 50 μL of each prepared sample was mixed well to prepare the QC sample. The QC sample was used to balance the system by continuously injecting the sample. Inject one QC sample for every four samples when checking the stability of the instrument [23].

2.3.6 Preparation of standard solution

For the standard solution, 2 mg of the reference substance DNJ was accurately weighed and dissolved with 2 mL of 70% acetonitrile. A standard solution of 1 mg/mL was obtained. The standard solution was diluted with mass spectrometry methanol to obtain a 500 ng/mL standard solution.

Next 0.8 mg of the control buckwheat base was weighed and dissolved in 2 mL of 60% acetonitrile. A standard solution of 0.4 mg/mL was obtained. The standard was then diluted with MS-grade methanol to 500 ng/mL of the standard solution.

2.3.7 Chromatographic conditions

The chromatographic column was a Waters ACQUITY BEH C18 (1.7 μm, 2.1 mm × 100 mm). The mobile phase was methanol (A) and a 0.1% formic acid aqueous solution (B). Gradient elution conditions are shown in Table 2. The flow rate was 0.4 mL/min, the column temperature was 45°C, the pool temperature was 4°C, and the injection volume was 2 μL.

Table 2

Mobile phase gradient elution conditions for samples

Time (min) A (%) B (%)
0 5 95
3 40 60
10 95 5
12 95 5
13 5 95
15 5 95

2.3.8 Mass spectrometry conditions

The ion source was an electrospray ionization (ESI) source in both positive and negative ion modes with MSE and a source temperature of 120°C. The desolvation gas was nitrogen at 800 L/h and 450°C. The capillary voltage was 3 kV in positive mode and 2.5 kV in negative mode. The cone voltage was 50 V and the molecular weight scan range was 100–1,200 Da. The energy at the high energy scan was 15–40 eV. Accurate mass numbers were corrected with a leucine enkephalin (LE) solution at 200 pg/mL. Other parameters were m/z 556.2771 in positive ion mode, 554.2615 in negative ion mode, and acquisition mode was without correction. The operating software for the UPLC-Q-TOF-MS system was Masslynx4.1.

2.3.9 Data processing and analysis

A MassLynx 4.1 workstation controlled the instrument and acquired MS data. The data were processed with Progenesis QI, and the import data format was profile mass spectrum of the high-resolution mass spectrum. LockMass correction was selected. The positive ion was 556.2766, and the negative ion was 554.2620; the adduct ions in the positive ion mode were [M + H]+, [M + Na]+, and [M + H-H2O]+; and the adduct ions in the negative ion mode were [M-H]−, [M + HCOO]−, and [M-H-H2O]−. When data were aligned, the data file with common features among all data was selected as the reference file. At peak extraction, the chromatographic peaks of the buffer column were not analyzed based on the total ion current profile, and the default parameters were selected for the remaining parameters. When samples were grouped, they were grouped according to their experimental design. For compound identification, the compounds with fragment ions were selected first, the database built for mulberry was searched, and the mass error of parent ions and fragment ions was ≤5 ppm. Mapping software used SIMCA-P14.1 and GraphPad Prism 9.2.0, and data were calculated using Excel spreadsheets.

The data matrices of ESI+ and ESI− modes were synthesized and imported into SIMCA-P14.1 software for statistical analysis. The differences in the chemical composition of M. nigra leaf herbs with different extraction methods were visually expressed by preliminarily observing the aggregation of each sample. Each sample was classified via orthogonal partial least squares-discriminant analysis (OPLS-DA) based on principal component analysis (PCA). Specifically, differential indicator components were screened by combining the score scatter plot (S-plot) and variable importance projection (VIP) values from the OPLS-DA model. The software GraphPad Prism 9.2.0 was used to calculate and compare the contents in different extraction methods of M. nigra leaves.

3 Results and discussion

3.1 Optimization of sample preparation methods

Pure water, 40% ethanol, 70% ethanol, and 95% ethanol were selected for the extraction solvent of the ultrasound-assisted extraction method of M. nigra leaves. In the heating and reflux extraction method of M. nigra leaves, 90% ethanol and pure water were selected for the investigation of the extraction solvent, and the sample treatment method was determined through investigation.

For the ultrasound-assisted extraction method, the total ion chromatograms (TIC) of the four different extraction solvents are as shown in Figure 1. The peak shapes were similar with essentially no difference. Compared with the other extraction solvents, pure water ultrasound-assisted extraction (WU) had the highest response value, and pure water as the extraction solvent was more advantageous.

Figure 1 Ultrasound-assisted extraction TICs with different extraction solvents: pure water (a), 40% ethanol (b), 70% ethanol (c), and 95% ethanol (d).
Figure 1

Ultrasound-assisted extraction TICs with different extraction solvents: pure water (a), 40% ethanol (b), 70% ethanol (c), and 95% ethanol (d).

The TIC diagrams of the heat reflux extraction method employing 90% ethanol and water as solvents are compared in Figure 2. The peak shape of the two TIC diagrams was similar, and there is no obvious difference from the response value. The heat reflux extraction method with pure water (WHR) as the extraction solvent was higher and more advantageous.

Figure 2 Heat reflux extraction TICs using different extraction solvents: 90% ethanol (a) and pure water (b).
Figure 2

Heat reflux extraction TICs using different extraction solvents: 90% ethanol (a) and pure water (b).

Pure water was chosen as the extraction solvent for both methods for subsequent experiments.

3.2 QC analysis

Because of the large number of samples analyzed, the TIC of QC samples was visually compared (Figures 3 and 4) to rule out analytical interferences caused by differences in test results from instrument or method instability. The results showed that the response intensities of each peak and retention time were the same, indicating that the instrument was in a stable state and the analytical method repeatability was good.

Figure 3 TIC of M. nigra leaf QC sample in positive ion mode.
Figure 3

TIC of M. nigra leaf QC sample in positive ion mode.

Figure 4 TIC of M. nigra leaf QC sample in negative ion mode.
Figure 4

TIC of M. nigra leaf QC sample in negative ion mode.

3.3 TIC comparison

UPLC-Q-TOF-MS analysis of M. nigra leaves using different extraction methods showed that the total ion current maps of the samples from the four different extraction methods were generally similar, but there were some differences, indicating that the types and contents of the main components of M. nigra leaves were different to varying degrees (Figures 5 and 6).

Figure 5 TIC of M. nigra leaves extracted using different methods in positive ion mode. Notes: A: FAU; B: WD; C: WU; and D: WHR.
Figure 5

TIC of M. nigra leaves extracted using different methods in positive ion mode. Notes: A: FAU; B: WD; C: WU; and D: WHR.

Figure 6 TIC of M. nigra leaves extracted using different methods in negative ion mode. Notes: A: FAU; B: WD; C: WU; and D: WHR.
Figure 6

TIC of M. nigra leaves extracted using different methods in negative ion mode. Notes: A: FAU; B: WD; C: WU; and D: WHR.

3.4 Analysis of compounds in different extraction methods of M. nigra leaves

Substance peaks detected in the ESI+ and ESI− patterns were abundant, and 46 and 80 secondary metabolites were identified in the mulberry self-built library based on fragment score screening. Table 3 shows information on the identified compounds.

Table 3

Compounds identified in M. nigra leaves using four different extraction methods

No. Identification Rt (min) m/z Adducts Formula Mass error (ppm) Compound type
1 Guangsangon I 0.55 625.170 M + H C35H28O11 −0.09 Flavone
2 DNJ 0.56 164.092 M + H C6H13NO4 2.50 Alkaloid
3 Fagomine 0.57 148.097 M + H C6H13NO3 2.50 Alkaloid
4 Macrourin B 0.59 523.102 M + Na C28H20O9 4.14 Flavonoid analog
5 Lactic Acid 0.61 135.030 M + FA-H C3H6O3 −2.62 Organic acid
6 Galacturonic acid 0.61 193.035 M-H C6H10O7 −0.09 Organic acid
7 Melezitose 0.63 503.162 M-H, M + FA-H C18H32O16 −0.38 Trisaccharide
8 2′,3′-Dihydro-4′,6-dihydroxy-2′-(1-hydroxy-1-methylethyl)-3,6′-bibenzofuran 0.63 371.113 M + FA-H C19H18O5 −3.55 Flavonoid analog
9 Quinic acid 0.64 191.056 M-H2O-H, M-H, M + FA-H C7H12O6 −2.98 Organic acid
10 Maltose 0.65 323.098 M-H2O-H C12H22O11 −2.67 Disaccharide
11 Citric acid 0.7 191.019 M-H2O-H, M-H C6H8O7 −3.08 Organic acid
12 3-Epi-fagomine 1.07 170.079 M + Na C6H13NO3 1.81 Alkaloid
13 Protocatechuic acid methyl ester 1.42 149.024 M-H2O-H C8H8O4 −3.89 Phenol
14 Cyanidin-3-glucoside 1.52 488.072 M + K C21H21O11 + 1.05 Flavone
15 Syringic acid 1.7 197.045 M-H C9H10O5 −4.14 Organic acid
16 Vanillic acid 1.71 167.034 M-H C8H8O4 −3.68 Organic acid
17 Cryptochlorogenic acid 1.84 353.087 M-H C16H18O9 −2.05 Phenylpropin
18 Caffeic acid 1.84 179.034 M-H2O-H, M-H C9H8O4 −2.15 Phenylpropin
19 Cycloartomunin 1.94 471.139 M + Na C26H24O7 −4.99 Flavone
20 Moracin R 1.95 389.125 M + FA-H C19H20O6 2.91 Flavonoid analog
21 2′,4′,7-Trihydroxy-8-(2-hydroxyethyl)flavone 1.97 313.072 M-H C17H14O6 0.54 Flavone
22 m-Coumaric acid 2.03 163.039 M-H C9H8O3 −3.96 Phenylpropin
23 Oxyresveratrol 2.07 243.065 M-H C14H12O4 −3.71 Phenylpropin
24 Dauroside D 2.07 339.071 M-H C15H16O9 −2.89 Phenylpropin
25 Plantagoside 2.13 465.103 M-H C21H22O12 −2.60 Flavone
26 Cis-Mulberroside A 2.16 567.171 M-H C26H32O14 −2.49 Phenylpropin
27 4-[(2R)-8-(2-Hydroxyethyl)-7-methoxy-3,4-dihydro-2H-chromen-2-yl]-1,3-benzenediol 2.18 355.093 M + K C18H20O5 −2.51 Flavonoid
28 Cyanidin-3,5-diglucoside 2.22 609.145 M-H C27H30O16 −1.24 Flavone
29 Dihydrooxyresveratrol 2.3 227.072 M-H2O-H C14H14O4 1.84 Other
30 Ferulic acid 4-O-β-d-glucuronide 2.34 351.072 M-H2O-H, M-H C16H18O10 −1.29 Phenylpropin
31 7-Hydroxycoumarin 2.34 161.024 M-H C9H6O3 −2.44 Phenylpropin
32 Mongolicin G 2.37 681.270 M + H C40H40O10 1.38 Phenylpropin
33 Neochlorogenic acid 2.46 353.087 M-H C16H18O9 −1.87 Phenylpropin
34 Eriodictyol 2.48 287.058 M-H C15H12O6 4.93 Flavone
35 Delphinidin-3,5-diglucoside 2.51 625.141 M-H C27H30O17 −0.71 Flavone
36 Morusimic acid D 2.53 312.254 M + H-H2O C18H35NO4 1.36 Organic acid
37 Kaempferol-7-O-β-d-glucopyranoside 2.54 487.065 M + K C21H20O11 2.57 Flavone
38 p-Coumaric acid 2.54 163.039 M-H C9H8O3 −4.32 Phenylpropin
39 Luteolin-4′-glucoside 2.54 447.095 M-H C21H20O11 3.57 Flavone
40 Morusignin G 2.58 379.154 M + H-H2O C23H24O6 −0.76 Flavone
41 Mesozygin A 2.66 589.152 M + H-H2O C35H26O10 4.99 Other
42 Mulberroside F. 6-glc 2.66 403.103 M-H C20H20O9 −1.11 Flavonoid analog
43 Mulberroside F 2.66 565.155 M-H, M + FA-H C26H30O14 −1.81 Flavonoid analog
44 Oxyresveratrol 2-O-β-d-glucopyranoside 2.68 405.119 M-H C20H22O9 0.02 Phenylpropin
45 d-Glucopyranose 2.71 161.045 M-H2O-H C6H12O6 −4.49 Monosaccharide
46 Mulberrofuran E 2.79 671.205 M + K C39H36O8 1.34 Flavonoid analog
47 3-(3,4-Dihydroxyphenyl)prop-2-enoic acid 2.8 179.035 M-H2O-H, M-H C9H8O4 0.41 Phenylpropin
48 Spiraeoside 2.8 463.088 M-H C21H20O12 −1.23 Flavone
49 6,7-Dihydroxycoumarin 2.81 177.019 M-H C9H6O4 −3.74 Phenylpropin
50 dl-Arginine 2.82 175.119 M + H C6H14N4O2 2.43 Amino acid
51 Morusyunnansin F 2.98 309.149 M + H-H2O C20H22O4 1.31 Flavonoid
52 Wittifuran H 2.98 475.214 M-H2O-H C29H34O7 2.33 Flavonoid analog
53 Cathafuran C 2.98 421.164 M + FA-H C24H24O4 −3.93 Flavonoid analog
54 Astragalin 3.03 487.065 M+K C21H20O11 1.73 Flavone
55 Sinapic acid 3.03 223.061 M-H C11H12O5 −3.17 Phenylpropin
56 5-Hydroxy Apiosylskimmin 3.03 453.105 M-H2O-H C20H24O13 2.46 Phenylpropin
57 Isoquercitrin 3.05 463.088 M-H C21H20O12 −0.27 Flavone
58 (2S)-8-Hydroxyethyl-7,4′-dimethoxyflavane-2′-O-β-d-glucopyranoside 3.07 537.196 M + FA-H C25H32O10 −2.67 Flavonoid
59 (2R,3S)-Guibourtinidol-3-0-α-d-apiofuranosyl-(1→6)-O-β-d-glucopyranoside 3.09 551.178 M-H C26H32O13 1.69 Flavonoid
60 Resveratrol 3.15 229.086 M + H C14H12O3 −0.19 Phenylpropin
61 Kaempferol-3-O-β-d-sophoroside 3.15 655.153 M + FA-H C27H30O16 2.82 Flavone
62 Kuwanol D 3.21 409.201 M + H C25H28O5 1.20 Phenylpropin
63 Umbelliferone 3.24 161.024 M-H C9H6O3 −2.83 Phenylpropin
64 Austrafuran A 3.25 449.123 M + H-H2O C25H22O9 −0.82 Flavonoid analog
65 Wittifuran E 3.25 303.051 M + FA-H C14H10O5 −1.55 Flavonoid analog
66 Chlorogenic acid 3.57 353.089 M-H C16H18O9 2.26 Phenylpropin
67 Lariciresinol 3.6 359.150 M-H C20H24O6 −0.92 Phenylpropin
68 Rutin 3.71 609.146 M-H C27H30O16 −0.02 Flavone
69 Morin 3.74 301.034 M-H C15H10O7 −3.99 Flavone
70 Isoquercetin 3.74 463.088 M-H C21H20O12 −0.93 Flavone
71 Gossypin 3.74 461.072 M-H2O-H, M-H C21H20O13 −1.37 Flavone
72 Norartocarpetin 7-glucoside 3.75 493.098 M + FA-H C21H20O11 −0.84 Flavone
73 Pinoresinol 3.76 403.161 M-H2O-H, M + FA-H C20H22O6 −1.62 Phenylpropin
74 Moracinflavan B 3.81 313.145 M + H-H2O C19H22O5 4.98 Flavone
75 Quercetin-7-O-β-d-glucopyranoside 3.84 445.077 M-H2O-H C21H20O12 −1.65 Flavone
76 Morin-3-O-β-d-glucopyranoside 3.84 341.030 M-H2O-H C17H12O9 −1.39 Flavone
77 2′,4′,5,7-Tetrahydroxy-3-methoxyflavone 3.84 297.040 M-H2O-H C16H12O7 −1.59 Flavone
78 Oxyresveratrol 3-glc 3.86 387.110 M-H2O-H C20H22O9 4.41 Phenylpropin
79 2′,7-Dihydroxy-4′-methoxy-8-prenylflavan 2,7-diglc 3.94 645.255 M-H2O-H C33H44O14 −0.41 Flavonoid
80 Syringaresinol 4 417.155 M-H C22H26O8 −1.43 Phenylpropin
81 Stearyltriarabinoferulate 4 901.443 M + FA-H C43H68O17 −1.19 Phenylpropin
82 14-Methoxy-dihydromorusin 4.01 435.180 M + H-H2O C26H28O7 −0.58 Flavone
83 Morusignin D 4.01 387.109 M + FA-H C19H18O6 1.24 Xanthones
84 Moracinflavan C 4.13 331.154 M + H C19H22O5 −0.85 Flavone
85 Kaempferol-3-O-glucoside 4.13 447.093 M-H C21H20O11 −1.25 Flavone
86 (E)-4-Isopentenyl-3,5,2′,4′-tetrahydroxystilbene 4.16 357.135 M + FA-H C19H20O4 1.59 Phenylpropin
87 Kuwanon O 4.18 717.228 M + Na C40H38O11 −4.56 Flavone
88 (2S)-2′,4′-Dihydroxyl-7-methoxyl-8-butyricflavane 4.18 339.123 M-H2O-H C20H22O6 −2.17 Flavonoid
89 Sanggenol G 4.21 529.219 M + Na C30H34O7 −2.25 Flavone
90 Kuwanon N 4.25 799.249 M + K C45H44O11 −4.03 Flavone
91 6-Methoxy-5,7,4′-trihydroxyisoflavone 4.3 339.026 M + K C16H12O6 −0.84 Flavone
92 Taxifolin 4.3 285.040 M-H2O-H C15H12O7 −1.97 Flavone
93 Moracenin C 4.39 799.250 M + K C45H44O11 −2.50 Flavone
94 Kuwanon U 4.39 483.204 M + FA-H C26H30O6 3.99 Flavone
95 7-O-β-d-Glucopyranosid Benzofuran 4.56 339.072 M-H C15H16O9 −0.39 Phenylpropin
96 Linoleic acid 4.68 303.230 M + Na C18H32O2 1.40 Organic acid
97 Alboctalol 4.79 471.142 M + H-H2O C28H24O8 −4.42 Other
98 Notabilisin B 4.87 547.207 M + K C30H36O7 −3.84 Flavone
99 Morusignin A 4.96 329.103 M + H C18H16O6 3.15 Xanthones
100 Sanggenol E 5.59 559.306 M + H-H2O C35H44O7 1.19 Flavone
101 Mulberroside A. 2-glc 5.62 405.118 M-H C20H22O9 −1.99 Phenylpropin
102 Wittifuran U 5.7 393.171 M + H-H2O C24H26O6 3.96 Flavonoid analog
103 Yunanensol A 5.74 441.189 M + H C25H28O7 −4.39 Flavone
104 2-(5-Hydroxymethyl-2-formylpyrrol-1-yl)isovaleric acid lactone 5.97 246.052 M + K C11H13NO3 −2.46 Other
105 Albafuran C 6.37 563.172 M + H-H2O, M + K C34H28O9 2.83 Flavonoid analog
106 Mornigrol G 6.37 477.131 M + K C25H26O7 −0.24 Flavone
107 Moracin C 6.37 309.113 M-H C19H18O4 −0.23 Flavonoid analog
108 2′,4′,7-Trihydroxy-(2S)-flavone 6.37 253.050 M-H2O-H C15H12O5 −0.90 Flavone
109 Leachianone G 6.38 355.120 M-H C20H20O6 4.18 Flavone
110 Wittifuran D. 6.4 293.117 M + H-H2O C19H18O4 −1.15 Flavonoid analog
111 Sanggenol K 6.51 509.251 M + H C30H36O7 −3.99 Flavone
112 2′,4′,5,7-Tetrahydroxyflavanone 6.77 271.060 M + H-H2O C15H12O6 −0.49 Flavone
113 Oleic acid 7.22 265.251 M + H-H2O C18H34O2 −4.81 Organic acid
114 Nigrasin J 7.36 451.177 M + H-H2O C26H28O8 4.56 Flavone
115 Wittifuran R 7.85 457.203 M-H2O-H C29H32O6 1.63 Flavonoid analog
116 Mortatarin D 7.96 521.252 M + FA-H C30H36O5 −4.30 Flavone
117 Mornigrol E 8.16 437.160 M-H C25H26O7 −2.02 Flavone
118 Notabilisin A 8.19 451.175 M-H C26H28O7 −2.66 Flavone
119 Chaminic acid 8.22 167.106 M + H C10H14O2 −4.67 Organic acid
120 2′,4′-Dihydroxy-7-methoxy-8-prenylflavan 8.43 339.161 M-H C21H24O4 1.32 Flavonoid
121 Notabilisin C 8.51 507.236 M + H C30H34O7 −4.41 Flavone
122 Wittifuran P 8.69 505.225 M + FA-H C29H32O5 3.10 Flavonoid analog
123 Nonadecanoic acid 9.46 343.285 M + FA-H C19H38O2 −3.06 Organic acid
124 Wittifuran T 9.51 505.223 M + FA-H C29H32O5 −0.99 Flavonoid analog
125 Behenic acid 9.67 323.330 M + H-H2O C22H44O2 −3.21 Organic acid
126 Stearic acid 9.96 283.263 M-H C18H36O2 −3.29 Organic acid

In both models, mass spectrometry information was identified using software, secondary metabolites of M. nigra leaves were identified through the mulberry plant database, and a total of 126 compounds were identified by referring to the relevant literature. The compounds were assigned to 54 flavonoids, including flavonoids and flavanone, 17 stilbenoids, including benzofuran, 3 alkaloids, 28 phenylpropanoids, 13 organic acids, 1 amino acid, 1 sugar, 2 xanthones, 1 phenol, and 4 other compounds.

The mass spectrometry data detected in positive and negative ion modes were analyzed using SIMCA-P14.1. PCA was used to cluster the data, and the score plot and the degree of convergence of M. nigra leaves with different extraction methods were obtained, as shown in Figure 7. PCA of the four extraction methods showed that the first two principal components explained 78.50% of the original variable information (PC1: 56.30% and PC2: 22.20%). It can be seen from the figure that the PCA model established in this experiment was well clustered. The sample sites of M. nigra leaves with different extraction methods were completely separated, and the samples of medicinal materials of the same extraction method were well gathered in the same area. In addition, as seen in Figure 7, the sample points of WU, FAU, and WHR were close in the first principal component, indicating that the chemical composition structure between the extraction methods was similar. As shown in Figure 7, the sample point aggregation was divided into two areas, which were far apart, indicating that the difference was large.

Figure 7 PCA score plots of M. nigra leaf samples using different extraction methods.
Figure 7

PCA score plots of M. nigra leaf samples using different extraction methods.

3.5 Analysis of differential metabolites using different extraction methods of M. nigra leaves

Due to different extraction methods, the chemical constituents in M. nigra leaf extracts were different, and 126 secondary metabolites identified were stoichiometrically analyzed using the map cloud mapping platform to find the number of differential compounds under the different extraction methods. The results are shown in Figure 8. From the Wayne diagram, it can be seen that there are 55 common components in the four extractions. There were six common components from the other three extraction methods but not the reflux extraction, which showed that the number of metabolites in the reflux extraction was less than that from the other three extraction methods. Among them, there were four signature differential metabolites extracted by formic acid and one signature differential metabolite extracted by water decoction. Some differences in chemical composition were observed among the four groups, and the results were consistent with the PCA results.

Figure 8 Wayne diagram of metabolites from M. nigra leaf using different extraction methods.
Figure 8

Wayne diagram of metabolites from M. nigra leaf using different extraction methods.

PCA analysis showed that there were some differences in the chemical composition of M. nigra leaves extracted by different extraction methods, and the chemical compositions of M. nigra leaves extracted by UA, FAU, and WHR showed more similarities compared with mulberry leaves extracted by WD. To understand the differences of M. nigra leaf extracts from different extraction methods, according to the PCA model results of the different extraction methods, OPLS-DA modeling was adopted for different groups of M. nigra leaves. The results showed that almost all of the M. nigra leaves extracted by WD and other groups could be divided into two groups, indicating that the compounds in the two groups of M. nigra leaves were significantly different (Figure 8). The model was validated by setting the number of tests to 200. The R 2 and Q 2 points on the left side (Figure 9b) were lower than those on the right side. The regression line of the Q 2 points intersected the longitudinal axis below the origin, and the model validation results (R 2 X = 0.951, R 2 Y = 0.991, Q 2 = 0.988) suggested that no overfitting occurred and the model was reliable (Figure 9). To further elucidate the differential compounds in the M. nigra leaves extracted by WD and other extraction methods, S-plots were plotted according to the established OPLS-DA model, and the compounds were screened according to a VIP value ≥1. According to the m/z value and retention time of differential metabolites, these data were matched with the component identification results or database, and a total of 13 differential components were screened and identified. These included four organic acids, three flavonoids, three phenylpropanoids, two alkaloids, and one trisaccharide. These differential metabolites were quinic acid, DNJ, delphinidin-3,5-diglucoside, melezitose, ferulic acid, 4-O-β-d-glucuronide, isoquercetin, rutin, stearic acid, fagomine, citric acid, neochlorogenic acid, cryptochlorogenic acid, and caffeic acid. Among them, DNJ and fagomine were confirmed by comparison with the control. Tables 3 and 4 shows information on the differential compounds.

Figure 9 OPLS-DA diagram (a), substitution test diagram (b) of M. nigra leaves using different extraction methods, and S-plot results (c).
Figure 9

OPLS-DA diagram (a), substitution test diagram (b) of M. nigra leaves using different extraction methods, and S-plot results (c).

Table 4

Differential metabolites of M. nigra leaves using four different extraction methods

No. Identification VIP Compound type MS/MS fragmentation
1 Quinic Acid 7.60 Organic acid 191.056, 179.056, 175.025, 173.046, 161.046, 149.046, 133.014, 101.024
2 DNJ 4.20 Alkaloid 146.147, 128.136, 110.122, 96.106, 82.135, 69.091
3 Delphinidin-3,5-diglucoside 3.99 Flavone 625.141, 463.088, 462.080, 461.073
4 Melezitose 2.79 Trisaccharide 503.162, 341.109, 323.098, 267.072, 237.062, 191.056, 143.035
5 Ferulic acid 4-O-β-d-glucuronide 1.61 Phenylpropanoids 369.083, 353.088, 341.088, 307.082, 287.056, 199.040, 191.020
6 Isoquercetin 1.52 Flavone 463.088, 300.028, 283.025, 271.025, 255.030, 254.022,2 43.030
7 Rutin 1.44 Flavone 609.146, 463.082, 461.073, 343.046, 300.028, 271.025, 255.030, 243.030, 227.035, 151.004
8 Stearic acid 1.42 Organic acid 283.264, 255.233
9 Fagomine 1.37 Alkaloid 130.152, 112.139, 94.127, 86.121, 80.109, 68.107
10 Citric acid 1.33 Organic acid 175.025, 173.009, 157.014, 147.030, 133.014, 129.019, 115.004, 111.009
11 Neochlorogenic acid 1.22 Phenylpropanoids 343.285, 295.228, 269.212, 199.170, 171.103
12 Cryptochlorogenic acid 1.22 Phenylpropanoids 353.088, 307.082, 191.056, 179.035, 177.019, 173.046, 135.045, 134.037, 133.030, 127.040
13 Caffeic acid 1.07 Organic acid 179.035, 135.045, 134.037, 133.030

3.6 Analysis of a differential metabolite cleavage rule under different extraction methods of medicinal M. nigra leaves

3.6.1 Flavonoid assay identification

Flavonoids are a class of characteristic compounds in M. nigra leaves, which have pharmacological effects, such as anti-oxidation and anti-cancer, and improve cardiovascular function [24]. Mass spectrometric fragmentation of flavonoids was investigated using rutin as an example, which formed a [M-H]− quasi-molecular ion peak at m/z 609.1461, and a fragment from lost rutinose at m/z 300.0275 upon collision-induced dissociation. Fragment m/z 300.0275 formed fragment m/z 151.0037 through reverse Diels–Alder (RDA) cleavage [25]. Possible mass spectrometric fragmentation pathways of rutin are shown in Figure 10.

Figure 10 Possible cleavage pathways for rutin.
Figure 10

Possible cleavage pathways for rutin.

3.6.2 Organic acids

Organic acid compounds are important compounds in M. nigra leaves, which have the effects of eliminating soreness and freckles and enhancing skin beauty. Using citric acid as an example, possible fragmentation patterns of organic acid compounds were analyzed, as shown in Figure 11. Shown in negative mode [M-H]−, a quasi-molecular ion peak was found at m/z 190, and fragmentation by secondary scanning yielded m/z 147 [M-H-H2O-CO2]− and m/z 111 [M-H-CO2-2H2O]− fragments [26].

Figure 11 Possible cleavage pathways of citric acid.
Figure 11

Possible cleavage pathways of citric acid.

3.6.3 Alkaloids

Alkaloids are important components in M. nigra leaves. Modern pharmacological studies have shown that mulberry leaves can inhibit increased blood glucose, with the effect of preventing and treating diabetes. The main active component of lowering blood glucose in mulberry leaves is DNJ-based polyhydroxyalkaloids [27]. Using DNJ as an example to investigate the possible mass spectrometric fragmentation of alkaloids, molecular ion peaks were obtained from DNJ in the positive ion mode at 164.1580 [M + H]+, and two characteristic fragment ions were obtained by energy collision, which were m/z 96.1063 [M + H-(OH)4]+ and m/z 82.1253 [M + H-CH6O4]+. The possible mass spectrometric fragmentation pathway of DNJ is shown in Figure 12 [28].

Figure 12 DNJ possible cleavage pathways.
Figure 12

DNJ possible cleavage pathways.

3.6.4 Phenylpropanoids

Phenylpropanoid compounds are important compounds in M. nigra leaves and have antioxidant activity [29]. The possible mass spectrometric fragmentation of phenylpropanoid compounds was analyzed using cryptochlorogenic acid as an example, as shown in Figure 13. The cryptochlorogenic acid parent ion in negative mode was 353.0878 [M-H]−, and a secondary fragment ion m/z 191.0561 was produced by the loss of one molecule of caffeoyl from the parent ion [M-H-caffeoyl]−. Then, m/z 179.0349 was produced by the loss of one molecule of caffeoyl from the parent ion [caffeoyl-H]−. On this basis, a portion of CO2 was removed to produce m/z 135.0444 [caffeoyl-H-CO2]− [30,31].

Figure 13 Possible cleavage pathway of cryptochlorogenic acid.
Figure 13

Possible cleavage pathway of cryptochlorogenic acid.

3.6.5 Trisaccharides

Trisaccharides are important active components in M. nigra leaves and can be used to repair islet cells with a significant hypoglycemic effect. Melezitose was used as an example to elaborate on the possible mass spectrometric behavior of trisaccharides. In the negative ion mode, the matrix parent ion was 503.1617 [M-H]−, and the fragment ion m/z 341.1089 was obtained by losing one molecule of glucose (the matrix ion), and the fragment ion m/z 323.0983 was obtained by losing one molecule of water.

3.7 Content analysis of the main compounds in M. nigra leaves using different extraction methods

A total of 13 different chemical components were identified by analysis of M. nigra leaf samples from four different extraction methods, which can serve as specific markers to distinguish different extraction methods. The flavonoid rutin is the main active ingredient against cancer and diabetes [32,33]. The alkaloid DNJ is a natural glycosidase inhibitor with strong postprandial glucose inhibition [34]. The phenylpropanoid compound neochlorogenic acid is an important component in the treatment of inflammation [35]. According to the standard DNJ, the corresponding contents of the differential components were calculated from the corresponding abundance values between the samples of each group under each extraction method. The standard deviations of the mean values of the content of each substance between the samples of M. nigra leaves from different extraction methods were calculated to obtain the content changes in the differential components between the M. nigra leaves from different extraction methods (Figure 14). Rutin and isoquercitrin were obtained at a higher yield by WHR. The remaining 11 differential compounds were extracted by WD, and the content in M. nigra leaves was higher than that of the other extraction methods, indicating that the WD method was more efficient compared with the other three extraction methods. This indicated that the WD extraction method of M. nigra leaves had higher medicinal value. Among them, quinic acid, delphinidin-3,5-diglucoside, ferulic acid 4-O-β-d-glucuronide, neochlorogenic acid, cryptochlorogenic acid, isoquercetin, rutin, and caffeic acid were not found in the reflux extraction method, which is important identification information for distinguishing the extraction of M. nigra leaves using different extraction methods.

Figure 14 Content of differential chemical constituents in M. nigra leaves extracted using different methods.
Figure 14

Content of differential chemical constituents in M. nigra leaves extracted using different methods.

4 Conclusion

In this study, UPLC-Q-TOF-MS/MS and multivariate statistical analysis were used to analyze different extraction methods, WU, FAU, WHR, and WD, of M. nigra leaves. A total of 128 chemical constituents were identified and classified. The results showed that WD was different from the other three methods. Compared to other methods evaluated in this study, rutin and isoquercitrin in the differential metabolites were more abundant in WHR. The differential metabolites quinic acid, DNJ, delphinidin-3,5-diglucoside, melezitose, ferulic acid 4-O-β-d-glucuronide, isoquercetin, rutin, stearic acid, fagomine, citric acid, neochlorogenic acid, cryptochlorogenic acid, and caffeic acid were more abundant with WD extractions. This result laid a foundation for the selection of extraction methods for M. nigra leaves in certain pharmacological applications, but further studies are still needed. The results can provide a reference for further optimization of the extraction method of metabolites and provide a basis for the QC of M. nigra leaves.

Acknowledgements

We thank LetPub (www.letpub.com) for its linguistic assistance during the preparation of this manuscript.

  1. Funding information: This research was funded by the Application for Open Project of the Key Laboratory of Xinjiang Autonomous Region (Analysis of Genetic Variation in Morus nigra L. and Research on Key Genes of 1-DNJ Alkaloid Synthesis) and Innovation project of germplasm resources of main forest and fruit tree species (lgxy202108).

  2. Author contributions: L.Y. and H.Y.W.: conceived and supervised the study; L.Y., W.J.H., and K.Z.: designed the experiments; X.L.G. and S.L.Y.: performed the experiments; and X.L.G.: analyzed the data and wrote the manuscript. All authors reviewed the results and approved the final version of the manuscript.

  3. Conflict of interest: Authors state no conflict of interest.

  4. Ethical approval: The conducted research is not related to either human or animal use.

  5. Data availability statement: The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

[1] Yiğit D, Yiğit N. Antibacterial activty of black mulberry (morus nigra) fruits and leaves. Erzincan Univ J Sci Technol. 2008;1:39–48.Search in Google Scholar

[2] Yi-heng LIU, Ling Y. Hypoglycemic and hypolipidemic effects of Morus nigra leaves on type 2-diabetic mice. Nat Prod Res Dev. 2015;27:1411.Search in Google Scholar

[3] Oryan S, Eidi M, Yazdi E, Eidi A, Solati J. Hypoglycaemic effect of alcoholic extract of Morus nigra L. leaves in normal and diabetic rats. J Med Plants. 2003;2:27–32.Search in Google Scholar

[4] Padilha MM, Vilela FC, Rocha CQ, Dias MJ, Soncini R, dos Santos MH, et al. Antiinflammatory properties of Morus nigra leaves. Phytother Res. 2010;24:1496–500.10.1002/ptr.3134Search in Google Scholar PubMed

[5] Turgut NH, Mert DG, Kara H, Egilmez HR, Arslanbas E, Tepe B, et al. Effect of black mulberry (Morus nigra) extract treatment on cognitive impairment and oxidative stress status of d-galactose-induced aging mice. Pharm Biol. 2016;54:1052–64.10.3109/13880209.2015.1101476Search in Google Scholar PubMed

[6] Qadir MI, Ali M, Ibrahim Z. Anti-cancer activity of Morus nigra leaves extract. Bangladesh J Pharmacol. 2014;9:496–97.10.3329/bjp.v9i4.19783Search in Google Scholar

[7] Song W, Wang HJ, Bucheli P, Zhang PF, Wei DZ, Lu YH. Phytochemical profiles of different mulberry (Morus sp.) species from China. J Agric Food Chem. 2009;57:9133–40.10.1021/jf9022228Search in Google Scholar PubMed

[8] Li B. Study on the chemical constituents and bioactivities of leaves of Morus nigra L. Xinjiang: Tarim University; 2019.Search in Google Scholar

[9] Zeni A, Moreira TD, Dalmagro AP, Camargo A, Bini LA, Simionatto EL, et al. Evaluation of phenolic compounds and lipid-lowering effect of Morus nigra leaves extract. An Acad Bras Ciênc. 2017;89:2805–15.10.1590/0001-3765201720160660Search in Google Scholar PubMed

[10] Araujo CM, Lúcio Kde P, Silva ME, Isoldi MC, de Souza GH, Brandão GC, et al. Morus nigra leaf extract improves glycemic response and redox profile in the liver of diabetic rats. Food Funct. 2015;6:3490–9.10.1039/C5FO00474HSearch in Google Scholar PubMed

[11] Dobson G, Shepherd T, Verrall SR, Conner S, McNicol JW, Ramsay G, et al. Phytochemical diversity in tubers of potato cultivars and landraces using a GC-MS metabolomics approach. J Agric Food Chem. 2008;56:10280–91.10.1021/jf801370bSearch in Google Scholar PubMed

[12] Styczynski MP, Moxley JF, Tong LV, Walther JL, Jensen KL, Stephanopoulos GN. Systematic identification of conserved metabolites in GC/MS data for metabolomics and biomarker discovery. Anal Chem. 2007;79:966–73.10.1021/ac0614846Search in Google Scholar PubMed

[13] Wiklund S, Johansson E, Sjöström L, Mellerowicz EJ, Edlund U, Shockcor JP, et al. Visualization of GC/TOF-MS-based metabolomics data for identification of biochemically interesting compounds using OPLS class models. Anal Chem. 2008;80:115–22.10.1021/ac0713510Search in Google Scholar PubMed

[14] Yoshida H, Yamazaki J, Ozawa S, Mizukoshi T, Miyano H. Advantage of LC-MS metabolomics methodology targeting hydrophilic compounds in the studies of fermented food samples. J Agric Food Chem. 2009;57:1119–26.10.1021/jf803235mSearch in Google Scholar PubMed

[15] Anacardio R, Mullins FG, Hannam S, Sheikh MS, O'Shea K, Aramini A, et al. Development and validation of an LC–MS/MS method for determination of methanesulfonamide in human urine. J Chromatogr B. 2009;877:2087–92.10.1016/j.jchromb.2009.05.051Search in Google Scholar PubMed

[16] Helander A, Zheng Y. Molecular species of the alcohol biomarker phosphatidylethanol in human blood measured by LC-MS. Clin Chem. 2009;55:1395–405.10.1373/clinchem.2008.120923Search in Google Scholar PubMed

[17] López-Gonzálvez Á, Godzien J, García A, Barbas C. Capillary electrophoresis mass spectrometry as a tool for untargeted metabolomics. In: D’Alessandro A, editor. High-Throughput Metabolomics. Vol. 1978. New York: Springer; 2019. p. 55–77.10.1007/978-1-4939-9236-2_5Search in Google Scholar PubMed

[18] Villate A, San Nicolas M, Gallastegi M, Aulas PA, Olivares M, Usobiaga A, et al. Review: Metabolomics as a prediction tool for plants performance under environmental stress. Plant Sci. 2021;303:110789.10.1016/j.plantsci.2020.110789Search in Google Scholar PubMed

[19] McKay RT. Metabolomics and NMR. In Springer Berlin Heidelberg; 2022.10.1007/164_2022_616Search in Google Scholar PubMed

[20] Reo NV. NMR-based metabolomics. Drug Chem Toxicol. 2002;25:375–82.10.1081/DCT-120014789Search in Google Scholar PubMed

[21] Wang Z-Y, Xiong H, Duan LY, Wang CF, Du YL, Hong X, et al. UPLC-Q-TOF-MS based metabolomics study of hawthorn leaves in different geographical regions. Anal Methods. 2021;13:5458–66.10.1039/D1AY01150BSearch in Google Scholar

[22] Deng W, Sun Z, Luo X, Wu X. Optimization of extraction technology of total alkaloids from mulberry leaves by orthogonal design. Chin Arch Tradit Chin Med. 2012;30:608–9.Search in Google Scholar

[23] Arapitsas P, Ugliano M, Marangon M, Piombino P, Rolle L, Gerbi V, et al. Use of untargeted liquid chromatography–mass spectrometry metabolome to discriminate Italian monovarietal red wines produced in their different terroirs. J Agric Food Chem. 2020;14:13353–66.10.1021/acs.jafc.0c00879Search in Google Scholar PubMed PubMed Central

[24] Luo W-T, Feng JH, Wang LX, He SN, Sun KH, Liu Q, et al. Research progress on flavonoids from mulberry leaves. Chem Bioeng. 2021;38:8–13.Search in Google Scholar

[25] Li Z, Wei Y, Fan Y, Zhu J, Zhao T. Analysis of rutin by electrospray iontrap mass spectrometry. Chin J Anal Lab. 2015;34:186–9.Search in Google Scholar

[26] Qiao X, Wu S, Qi X, Feng J, Xiao X. Identification of chemical constituents in Crataegus pinnatifida var. major by UPLC/ESI-TOF/MS. Drugs Clin. 2014;29:120–4.Search in Google Scholar

[27] Jakobsen P, Lundbeck JM, Kristiansen M, Breinholt J, Demuth H, Pawlas J, et al. Iminosugars: potential inhibitors of liver glycogen phosphorylase. Bioorg Med Chem. 2001;9:733–44.10.1016/S0968-0896(00)00291-1Search in Google Scholar PubMed

[28] Xiong Z-Q, Li Y, Xu Y, Li SY, Cai JY, Lv Q, et al. Study on the difference of principal components different processed sericulture by UPLC-Q/TOF-MS. J Jiangxi Univ Chin Med. 2021;33:89–94.Search in Google Scholar

[29] Zhang W, Han F, He J. HPLC-DAD-ESI-MS/MS analysis and antioxidant activities of nonanthocyanin phenolics in mulberry (Morus alba L.). J Food Sci. 2008;73(6):C512–8.10.1111/j.1750-3841.2008.00854.xSearch in Google Scholar PubMed

[30] Zhao G-L, Miao XL, Song ZH, Gao W, Zhou GR, Li P. Analysis of chemical composition of Kun Yining Granules by UPLC-Q-TOF/MS method. Chin Tradit Pat Med. 2020;42:1074–82.Search in Google Scholar

[31] Lan X-Y, Zhu LB, Huang XZ, Liu DH, Hao QQ, Zhou L, et al. Study on identification and quantitation of main components in Artemisiae Argyi Folium. Chin Tradit Herb Drugs. 2021;52:7630–7.Search in Google Scholar

[32] Saha S, Prajapati DG, Ratrey P, Mishra A. Co-delivery nanosystem of Epigallocatechin Gallate and Rutin for anticancer and antibacterial activities. J Drug Deliv Sci Technol. 2022;70:103191.10.1016/j.jddst.2022.103191Search in Google Scholar

[33] Tian R, Yang W, Xue Q, Gao L, Huo J, Ren D, et al. Rutin ameliorates diabetic neuropathy by lowering plasma glucose and decreasing oxidative stress via Nrf2 signaling pathway in rats. Eur J Pharmacol. 2016;771:84–92.10.1016/j.ejphar.2015.12.021Search in Google Scholar PubMed

[34] Cipolla L, Cardona F, La Ferla B, Fusi P. 1-Deoxynojirimycin (DNJ) and pyrrolizidine derivatives as glycosidase inhibitors. Abstr Pap - Am Chem Soc; 2020.Search in Google Scholar

[35] Yu, M-H, Hung TW, Wang CC, Wu SW, Yang TW, Yang CY, et al. Neochlorogenic acid attenuates hepatic lipid accumulation and inflammation via regulating miR-34a in vitro. Int J Mol Sci. 2021;22:13163.10.3390/ijms222313163Search in Google Scholar PubMed PubMed Central

Received: 2022-12-20
Revised: 2023-03-04
Accepted: 2023-04-03
Published Online: 2023-09-25

© 2023 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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https://doi.org/10.1515/chem-2022-0320
Keywords for this article
Morus nigra L. leaves; different extraction methods; secondary metabolites; UPLC-Q-TOF-MS/MS
Creative Commons
BY 4.0

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  109. Important application value of injectable hydrogels loaded with omeprazole Schiff base complex in the treatment of pancreatitis
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  134. Treatment of gastric cancer by silver nanoparticles encapsulated by chitosan polymers mediated by Pistacia atlantica extract under ultrasound condition
  135. In vitro protective and anti-inflammatory effects of Capparis spinosa and its flavonoids profile
  136. Wear and corrosion behavior of TiC and WC coatings deposited on high-speed steels by electro-spark deposition
  137. Therapeutic effects of green-formulated gold nanoparticles by Origanum majorana on spinal cord injury in rats
  138. Melanin antibacterial activity of two new strains, SN1 and SN2, of Exophiala phaeomuriformis against five human pathogens
  139. Evaluation of the analgesic and anesthetic properties of silver nanoparticles supported over biodegradable acacia gum-modified magnetic nanoparticles
  140. Review Articles
  141. Role and mechanism of fruit waste polyphenols in diabetes management
  142. A comprehensive review of non-alkaloidal metabolites from the subfamily Amaryllidoideae (Amaryllidaceae)
  143. Discovery of the chemical constituents, structural characteristics, and pharmacological functions of Chinese caterpillar fungus
  144. Eco-friendly green approach of nickel oxide nanoparticles for biomedical applications
  145. Advances in the pharmaceutical research of curcumin for oral administration
  146. Rapid Communication
  147. Determination of the contents of bioactive compounds in St. John’s wort (Hypericum perforatum): Comparison of commercial and wild samples
  148. Retraction
  149. Retraction of “Two mixed-ligand coordination polymers based on 2,5-thiophenedicarboxylic acid and flexible N-donor ligands: The protective effect on periodontitis via reducing the release of IL-1β and TNF-α”
  150. Topical Issue on Phytochemicals, biological and toxicological analysis of aromatic medicinal plants
  151. Anti-plasmodial potential of selected medicinal plants and a compound Atropine isolated from Eucalyptus obliqua
  152. Anthocyanin extract from black rice attenuates chronic inflammation in DSS-induced colitis mouse model by modulating the gut microbiota
  153. Evaluation of antibiofilm and cytotoxicity effect of Rumex vesicarius methanol extract
  154. Chemical compositions of Litsea umbellata and inhibition activities
  155. Green synthesis, characterization of silver nanoparticles using Rhynchosia capitata leaf extract and their biological activities
  156. GC-MS analysis and antibacterial activities of some plants belonging to the genus Euphorbia on selected bacterial isolates
  157. The abrogative effect of propolis on acrylamide-induced toxicity in male albino rats: Histological study
  158. A phytoconstituent 6-aminoflavone ameliorates lipopolysaccharide-induced oxidative stress mediated synapse and memory dysfunction via p-Akt/NF-kB pathway in albino mice
  159. Anti-diabetic potentials of Sorbaria tomentosa Lindl. Rehder: Phytochemistry (GC-MS analysis), α-amylase, α-glucosidase inhibitory, in vivo hypoglycemic, and biochemical analysis
  160. Assessment of cytotoxic and apoptotic activities of the Cassia angustifolia aqueous extract against SW480 colon cancer
  161. Biochemical analysis, antioxidant, and antibacterial efficacy of the bee propolis extract (Hymenoptera: Apis mellifera) against Staphylococcus aureus-induced infection in BALB/c mice: In vitro and in vivo study
  162. Assessment of essential elements and heavy metals in Saudi Arabian rice samples underwent various processing methods
  163. Two new compounds from leaves of Capparis dongvanensis (Sy, B. H. Quang & D. V. Hai) and inhibition activities
  164. Hydroxyquinoline sulfanilamide ameliorates STZ-induced hyperglycemia-mediated amyleoid beta burden and memory impairment in adult mice
  165. An automated reading of semi-quantitative hemagglutination results in microplates: Micro-assay for plant lectins
  166. Inductively coupled plasma mass spectrometry assessment of essential and toxic trace elements in traditional spices consumed by the population of the Middle Eastern region in their recipes
  167. Phytochemical analysis and anticancer activity of the Pithecellobium dulce seed extract in colorectal cancer cells
  168. Impact of climatic disturbances on the chemical compositions and metabolites of Salvia officinalis
  169. Physicochemical characterization, antioxidant and antifungal activities of essential oils of Urginea maritima and Allium sativum
  170. Phytochemical analysis and antifungal efficiency of Origanum majorana extracts against some phytopathogenic fungi causing tomato damping-off diseases
  171. Special Issue on 4th IC3PE
  172. Graphene quantum dots: A comprehensive overview
  173. Studies on the intercalation of calcium–aluminium layered double hydroxide-MCPA and its controlled release mechanism as a potential green herbicide
  174. Synergetic effect of adsorption and photocatalysis by zinc ferrite-anchored graphitic carbon nitride nanosheet for the removal of ciprofloxacin under visible light irradiation
  175. Exploring anticancer activity of the Indonesian guava leaf (Psidium guajava L.) fraction on various human cancer cell lines in an in vitro cell-based approach
  176. The comparison of gold extraction methods from the rock using thiourea and thiosulfate
  177. Special Issue on Marine environmental sciences and significance of the multidisciplinary approaches
  178. Sorption of alkylphenols and estrogens on microplastics in marine conditions
  179. Cytotoxic ketosteroids from the Red Sea soft coral Dendronephthya sp.
  180. Antibacterial and biofilm prevention metabolites from Acanthophora spicifera
  181. Characteristics, source, and health risk assessment of aerosol polyaromatic hydrocarbons in the rural and urban regions of western Saudi Arabia
  182. Special Issue on Advanced Nanomaterials for Energy, Environmental and Biological Applications - Part II
  183. Green synthesis, characterization, and evaluation of antibacterial activities of cobalt nanoparticles produced by marine fungal species Periconia prolifica
  184. Combustion-mediated sol–gel preparation of cobalt-doped ZnO nanohybrids for the degradation of acid red and antibacterial performance
  185. Perinatal supplementation with selenium nanoparticles modified with ascorbic acid improves hepatotoxicity in rat gestational diabetes
  186. Evaluation and chemical characterization of bioactive secondary metabolites from endophytic fungi associated with the ethnomedicinal plant Bergenia ciliata
  187. Enhancing photovoltaic efficiency with SQI-Br and SQI-I sensitizers: A comparative analysis
  188. Nanostructured p-PbS/p-CuO sulfide/oxide bilayer heterojunction as a promising photoelectrode for hydrogen gas generation
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