Skip to main content
NCBI home page
RSC Advances logoLink to RSC Advances
. 2026 Jul 14;16(37):40318–40331. doi: 10.1039/d6ra03310e

LC-QTOF-MS metabolic profiling and anti-MRSA activity of Asparagus officinalis L. spear methanolic extract in a skin infection mouse model

Passent M Abdel-Baki a, Mariam Hassan b,c, Noha A Ahmed b, Dina Gamal d, Amal E Khaleel a, Meselhy R Meselhy a, Riham A El-Shiekh a,✉
PMCID: PMC13367281  PMID: 42453954

Abstract

Asparagus officinalis L. spears offer substantial potential as a functional food due to their rich nutritional profile and bioactive compounds. The emergence and spread of multidrug-resistant (MDR) pathogens, particularly in low- and medium-income countries, have necessitated extensive research and concerted efforts to combat infections. Novel One Health approaches are being sought to swiftly identify antibiotics that retain efficacy against these MDR pathogens. In this study, we evaluated the antibacterial activity of the methanolic extract of A. officinalis L. spears against Staphylococcus aureus USA300 (MARSA USA300); a MDR bacterium in vitro and in vivo. The methanolic extract demonstrated significant in vitro inhibition of the biofilm formation (IC50 value of 3.415 mg mL−1) and enhanced biofilm detachment (EC50 value of 1.573 mg mL−1) against MRSA. Additionally, topical treatment of MRSA USA300-infected BALB/C mice open lesion with the extract (50 mg mL−1 in 25% DMSO, once for three days) significantly reduced the bacterial count by 2.877 and 2.583 logs lower than that of the negative control). LC-QTOF/MS tentatively identified 99 compounds in the methanolic extract of A. officinalis spears. The findings obtained here demonstrate the potential of A. officinalis L. spears for pharmaceutical, agricultural, and food industry applications.

Asparagus officinalis L. spears offer substantial potential as a functional food due to their rich nutritional profile and bioactive compounds.graphic file with name d6ra03310e-ga.jpg

1. Introduction

The emergence and continuous evolution of multidrug-resistant (MDR) pathogens pose a significant threat to humans, animals, and the environment, particularly in low- and middle-income countries (LMICs). In 2017, the Center for Disease Control (CDC) reported an estimated 119 247 cases of positive clinical cultures resulting in 19 832 associated deaths among hospitalized patients, including both community- and nosocomial-acquired infections.1 Among these, methicillin-resistant Staphylococcus aureus (MRSA) stands out as one of the most significant contributors. MRSA is accountable for a diverse array of infections extending from superficial skin lesions to invasive diseases involving soft-tissue infections, endocarditis, osteomyelitis, and pneumonia.2 The prevalence of MRSA among S. aureus infections in the Arab countries of the middle East and North Africa was found to be as high as 57% in Egypt and Tunisia, and 67% in Palestine, Jordan, Lebanon and Iraq.3 Such situation leads to increased healthcare costs and poorer patient outcomes.

Despite substantial antibiotic discoveries during the golden age (1960s through 1980s), subsequent advancements have stagnated dramatically.4 Multidrug resistance arises through a variety of mechanisms, either independently or through synergistic interactions including biofilm formation in S. aureus, which significantly facilitates the emergence of resistant strains.5 The emergence of strains resistant to multiple antibiotic classes including tetracyclines, fluoroquinolones, macrolides, aminoglycosides, and lincosamides, has made clinical management of MRSA infections challenging.2 Nevertheless, several antibiotics are currently in the clinical pipelines, but most represent derivatives of existing classes that do not target new molecular mechanisms.6 Accordingly, developing new antibacterial agents with unique chemical scaffolds and molecular target(s), effective against MRSA, remains a high priority.

Natural products, including plant metabolites, serve as essential sources for drug discovery, providing lead compounds that undergo structural optimization and biological refinement to become viable therapeutic agents. Many of these have contributed significantly to antibiotic development by targeting bacterial membranes, membrane components, membrane biosynthetic enzymes, or membrane-embedded proteins.7 Accordingly, plant-derived antimicrobials offer promising alternatives worth exploring.

Asparagus (Asparagus officinalis L., Family: Asparagaceae) is a perennial herb that holds considerable promise as a nontraditional horticultural crop in Egypt.8 It has gained significant importance as a vegetable crop in various regions across Asia, Africa, Europe, and America, with a cultivation history spanning over 2000 years.9 Asparagus spears are known for their unique texture, flavor, exceptional nutritional values, medicinal properties, and unique therapeutic effects. Asparagus has gained international recognition as one of the top ten dishes worldwide, earning the title of the King of Vegetables.10 The spears are rich in essential amino acids, fats, vitamins, and mineral elements such as potassium (K), calcium (Ca), and magnesium (Mg).11 The spears also possess a diverse array of bioactive phytochemicals including,12,13 organic acids, amino acids, peptides and derivatives, saponins, flavonoids, anthocyanins, bioactive oligosaccharides and polysaccharides. Studies have reported their antioxidant, antiepileptic, antitumor, hypolipidemic, decompression, anti-oxidation, anti-aging, hypotensive, hypoglycemic, immunomodulatory, and antifungal effects.12 Furthermore, extracts from asparagus spears have been traditionally used for their stimulating, diuretic, antitussive, and laxative properties.12

In this study, we evaluate the antibacterial activity of the methanolic extract of A. officinalis L. spears against methicillin-resistant S. aureus USA300 (MRSA USA300) both in vitro and in vivo. Additionally, LC-QTOF-MS was employed to tentatively identify the chemical metabolites in the extract.

2. Material and methods

2.1. Plant material and extraction

The spears of Asparagus officinalis L. were collected during the harvest season in May 2022 from a commercial farm in Al-Qanatir Al-Khayriyyah, city, Qalyubia, Egypt. The plant material was authenticated by Mrs Trease Labib, Consultant of Plant Taxonomy at the El-Orman Botanic Garden, Giza, Egypt, and a voucher specimen (No. 16.2.2023) was deposited at the museum of the department of Pharmacognosy, Faculty of Pharmacy, Cairo University, Cairo Egypt. The spears were sliced, lyophilized, and ground to fine powder. Part of the fine powder (1.0 kg) was extracted with methanol (2 L × 4) by maceration at room temperature. Solvents were evaporated under reduced pressure to give 55.0 g of dry extract.14

2.2. Antibacterial and anti-virulence activities

The antibacterial activity of the extract was tested against the highly virulent methicillin-resistant Staphylococcus aureus USA300 (MRSA USA300).15 The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined using the broth micro-dilution method.15,16 The anti-virulence activity of the extract was investigated by testing sub-MIC concentrations of the extract against MRSA USA300 biofilm. The biofilm inhibition and biofilm eradication activities were determined using crystal violet biofilm assay in 96-well flat bottom plates.17 The solvent of the extract (DMSO) was used as the control in the nontreated wells (100% reference values). The biofilm inhibition % and biofilm eradication % were calculated using the equation:182.2.

OD: optical density.

2.3. In vivo MRSA skin infection model

The in vivo experiment and all animal procedures were approved by the Research Ethics Committee of the Faculty of Pharmacy, Cairo University (Approval number: MI (3694)). MRSA skin infection model was performed as previously reported.19 Briefly, eighteen male BALB/C mice (seven weeks old) were housed with food and water. The dorsal backs of the mice were shaved before the experiment. The shaved area in the back of each mouse was injected intra-dermally with 50 µL MRSA USA300 suspended in phosphate buffered saline (11 × 109 colony-forming units CFU). Forty-eight hours post-infection and formation of open skin lesions, mice were randomly distributed into three groups (n = 6). The first group was used as a negative control (did not receive any treatment). The second group was treated with 25% DMSO (vehicle control). The third group was topically treated with 100 µL of the extract (50 mg mL−1 in 25% DMSO) once daily for three days. Twenty-four hours post the last treatment, the experiment was terminated, animals were euthanized, and the skin lesions were removed and homogenized in sterile saline. Samples were diluted and tested for the aerobic viable count of MRSA by culturing on mannitol salt agar (MSA). The microbial loads recovered from each group were calculated and analyzed. The 3 days evaluation window was strategically selected to capture the acute phase of the cutaneous infection. Because immunocompetent mice naturally exhibit progressive host-mediated clearance and spontaneous infection resolution over extended periods, an acute 3 days endpoint ensures maximum assay sensitivity and prevents data convergence across the experimental groups.

2.4. LC-QTOF-MS analysis

Ten mg of A. officinalis spears methanolic extract was dissolved in 1 mL of LC-MS grade methanol, followed by centrifugation at 13 000g for 10 minutes. The supernatant was then filtered through a 0.22 µm nylon syringe filter prior to analysis using LC- MS/MS operated in positive and negative ESI modes. LC-MS/MS analysis of A. officinalis spears methanolic extract was performed using ExionLC™ AD UPLC, coupled with TripleTOF 5600+ Time-of-Flight Tandem Mass Spectrometer (AB SCIEX, Agilant, Germany) in the positive and negative ionization modes. A sample (10 µL each) of the methanolic extract (1 µg mL−1) was used in each mode. A XSelect HSS T3 XP column (Waters, 2.5 µm, 2.1 mm × 150 mm) was used at 40 °C. In the positive ionization mode, a gradient elution was adopted at a flow rate of 0.3 mL min−1. Using a mobile phase composed of 5 mM ammonium formate buffer pH 3 containing 1% methanol (A) and acetonitrile (B) was used as follows: isocratic for 1.0 min with 10% B and 4.0 min with 90% B, gradient elution of B from 10% to 90% in 20 min, and then to 10% B in 3 min. For the negative ionization mode, the same technique was employed using a mobile phase composed of B and solvent C (5 mM ammonium formate buffer pH8 containing 1% methanol).

A complete scan (50–1000 m/z) and up to 15 information-dependent MS/MS scans were automatically switched between when acquiring mass spectra in negative-ion mode. In both ionization modes, automatic switch was used to alternate between complete scans (50.0000–1000.0000 m/z) and up to 15 information-dependent MS/MS scans. The MS/MS scan started at 104 ppm, the intensity threshold. The following parameters were established: nebulizer gas pressure of 45 PSI, heater gas pressure of 45 PSI, ion spray voltage of 4500 V, normalized collision energy of 35 V, and declustering potential of 80 V. The dynamic exclusion setting was set to 10 seconds. Raw data files were loaded into MS-DIAL 3.52 (https://prime.psc.riken.jp/)20 for data-independent MS/MS deconvolution. The software Analyst (TF 1.7.1, Sciex) was used for instrument control, data acquisition, and processing of the MS/MS spectra. By comparing experimental extract spectra against library retention times (RT), precise precursor masses (m/z), and MS/MS spectra, as well as taking into account similar acquisition parameters (e.g., resolution, collision energy, ionization, MS level) to ensure the validity of the match and decision criteria, compound identifications with the high level of confidence (MSI level 2) were obtained. The diagnostic MS/MS fragments, ionization behavior, parent compound information, and review of Asparagus chemical constituents that have been published provided additional evidence for compound annotations.21–27

3. Results and discussion

3.1. Antibacterial and anti-virulence activities

Various opportunistic pathogens can cause skin infections,28,29 with MRSA being one of the most challenging due to its resistance to antibiotics.30 In this study, the extract from the spears of A. officinalis L. demonstrated significant antibacterial and anti-virulence activities against the highly virulent MRSA USA300 strain, as reflected by MIC of 8.33 ± 1.67 and MBC of the same value. Although numerous medicinal plants have been investigated for their antimicrobial activity against MRSA,31 to the best of our knowledge, this is the first study to show the antimicrobial activity of the extract of A. officinalis spears against MRSA.32

Biofilm formation plays a critical role in increasing bacterial resistance, making it a key target for antibacterial therapies. The antibiofilm activity of the extract was tested at sub-MIC concentrations (5–0.15625 mg mL−1). The tested extract significantly inhibited biofilm formation of MRSA USA300 with IC50 of 3.415 mg mL−1 (Fig. 1). The previously established MRSA USA300 biofilm was significantly eradicated by the tested extract with EC50 value of 1.573 mg mL−1 (Fig. 1). The recorded biofilm eradication activity was higher than the biofilm inhibition activity with significant difference at a concentration range of 1.25–0.15625 mg mL−1 (two-way ANOVA, Sidak's post-hoc test, p < 0.05) (Fig. 1). In our study, the IC50 for biofilm inhibition (3.415 mg mL−1) was higher than the EC50 for biofilm eradication (1.573 mg mL−1). The significant differences in biofilm inhibition and eradication activities at a lower concentration range (1.25–0.15625 mg mL−1) suggest a concentration-dependent mechanism, with the extract being more effective in disrupting established biofilms than in preventing their formation. This indicates that the active components of the extract may target specific elements of biofilm integrity, potentially interfering with the extracellular matrix or disrupting signaling pathways that maintain biofilm structure. In vitro assessments have demonstrated that the extract of A. officinalis spears exhibit a considerable antibacterial activity against MRSA. The mechanism of action is thought to involve the disruption of bacterial cell membranes by saponins and other phytochemicals, leading to increased permeability and eventual cell lysis.33 When compared with previous studies, the antibacterial and biofilm-disrupting properties of A. officinalis are consistent with the known activities of other plant-derived extracts, which have shown efficacy against various Gram-positive bacteria, including MRSA.34–36 However, the precise bioactive compounds responsible for these effects remain unidentified. Future research should focus on isolating and characterizing these compounds and investigating their specific mechanisms of action at the molecular level.

Fig. 1. Anti-biofilm activity. Effect of different sub-minimum inhibitory concentrations (MIC) (5–0.15625 mg mL−1) of the extract of the spears of A. officinalis L. on MRSA USA300 biofilm. Results are expressed as mean biofilm inhibition % ± standard error and mean biofilm detachment % ± standard error. *, **& **** indicate that the difference is significant at p < 0.05, 0.01 & 0.0001, respectively, (two-way ANOVA, Sidak's post-hoc test).

Fig. 1

Open in a new tab

3.2. In vivo MRSA skin infection model

The in vivo antibacterial activity of the spears of A. officinalis L. extract was tested against MRSA using skin infection model. Three groups of male BALB/C mice (n = 6) were injected intradermally with MRSA USA300 suspension. Forty-eight hr. post-infection, an abscess wound developed at the site of infection. The tested extract significantly decreased the bacterial count of MRSA USA300 compared to the negative control and the DMSO groups (One-way ANOVA, Tukey's post-hoc test, P < 0.0001) (Fig. 2). The bacterial count retrieved from the extract-treated group was 2.877 and 2.583 logs lower than that of both the negative control and the DMSO groups, respectively (Fig. 2).

Fig. 2. Efficacy of the extract of the spears of A. officinalis L. in an in vivo murine model of MRSA skin infection. Eighteen BALB/C mice were divided into three groups (n = 6). (A) Photo image of the efficacy of different treatment groups on MRSA skin infection in the posterior backs of mice at the end of the experiment. (B) Efficacy of different treatment groups on the bacterial load in murine model MRSA skin infection. Each data point in the figure represents a mouse. Results are expressed as mean ± standard error. **** indicate that the difference is significant at p < 0.0001, respectively, (one-way ANOVA, Tukey's post-hoc test).

Fig. 2

Open in a new tab

Many medicinal plants have been investigated for their antimicrobial activity against wound and skin infections,29 but relatively little research has been conducted on Asparagus spears.37 The findings from the MRSA skin infection model further support the antibacterial potential of the extract of A. officinalis spears against MRSA USA300. The significant reduction in bacterial load in the extract-treated group compared to both the negative control and DMSO groups highlights its efficacy in treating MRSA-induced skin infections. These results are particularly significant given the virulence of the MRSA USA300 strain, which is commonly associated with skin and soft tissue infections that are notoriously difficult to treat.38 Results indicated that treatment with asparagus extract significantly reduced bacterial load and improved healing outcomes compared to untreated controls. This suggests dual mechanisms: direct antibacterial activity plus host immune modulation and anti-adhesion/anti-invasion effects.39

When compared with previous in vivo studies on plant extracts, A. officinalis showed considerable promise as a therapeutic agent. Plant-based treatments are gaining attention due to their lower likelihood of promoting resistance and their multifaceted modes of action, which may include disrupting bacterial cell membranes, inhibiting virulence factor production, and modulating host defenses.40–42 Overall, the significant reduction in bacterial load in the extract-treated group provides a strong rationale for further exploration of A. officinalis spears as a natural antibacterial agent, particularly for infections caused by antibiotic-resistant pathogens such as MRSA USA300. A limitation of the current in vivo study is the absence of a standard commercial antibiotic control group, such as vancomycin. Crucially, because vancomycin is a purified, mono-molecular pharmaceutical entity, direct numerical benchmarking against a crude, multi-component botanical extract represents an unequal comparison. Furthermore, in strict accordance with international animal welfare ethics and the 3Rs principle (Reduction), an independent positive control cohort was intentionally omitted to minimize the total number of animals sacrificed, given that the rapid staphylococcal clearance metrics of vancomycin in this specific infection model are already highly predictable and well-established in our previous work.43

3.3. Metabolic characterization of A. officinalis spears methanolic extract

With the aim of identifying the secondary metabolites of A. officinalis spears that could be correlated with the antibacterial and anti-virulence activities, we performed LC-QTOF-MS analysis of the spears methanolic extract. Ninety-nine compounds were tentatively identified in the positive and negative ionizations modes. These compounds belong to various chemical classes, including 7 organic acids, 17 amino acids, 6 sugars, 4 nucleosides, 19 phenolic acids and their derivatives, 12 flavonoids, 3 sulfur containing compounds, 6 steroidal saponins and 23 fatty acids and, lipids and 2 miscellaneous compounds. The details on their pseudomolecular ions, molecular formulas, retention time, tentative identification, and chemical class are listed in Table 1. The representative base peak chromatograms in the positive and negative ionization modes are represented in the SI (Fig. S1 and S2), respectively. The MS/MS spectra of representative compounds are represented in Fig. S3–S52.

Table 1. Compounds identified in the methanolic extract of A. officinalis spears by LC-QTOF-MS analysis in positive and negative ionization modesa.

Cpd RT (min) Name Mol. formula m/z (error/ppm) Ms/Ms fragments Rel. ab. Ref.
[M + H]+ [M–H]− Positive Negative
Organic acids
1 1.03 Citric acid C6H8O7 193.0347 (−2.19) 191.0200 (−1.43) 139, 111, 87 173, 111, 85 + 21
2 1.09 Furoic acid C5H4O3 — 111.0081 (5.96) — 67 + 21
3 1.17 Quinic acid C7H12O6 — 191.0549 (6.31) — 173, 129, 111, 87 ++ 27
4 1.187 Fumaric acid C4H4O4 — 115.0032 (4.6) — 71 + 21
5 1.20 Malic acid C4H6O5 — 133.0140 (1.84) — 115, 71 ++ 23
6 1.52 Pantothenic acid C9H17NO5 220.1182 (−1.15) 218.1019 (6.83) 202 146 + 22
7 5.35 Abscisic acid C15H20O4 265.1432 (0.89) 263.1292 (−1.2) 247, 229, 205 219, 204, 152 + 23
 
Amino acids and derivatives
8 1.21 l-Glutamine C5H10N2O3 147.0761 (2.18) 145.0617 (1.13) 130, 102, 84 127 + 23
9 1.18 l-Glutamic acid C5H9NO4 — 146.0453 (3.95) — 128 + 23
10 1.23 Glutamine hexose C11H20N2O8 309.1296 (−1.16) 307.1148 (−0.36) — 145 + 21
11 1.250 Pyroglutamic acid C5H7NO3 130.0501 (−1.79) 128.0351 (1.68) 84, 56 115, 111 ++ 27
12 1.26 Aspartic acid C4H7NO4 134.0451 (−2.37) 132.0299 116, 88, 74 115, 88, 71 ++ 27
13 1.263 Proline C5H9NO2 116.0711 (−4.3) 114.0555 (4.8) 98, 70 84, 76 ++ 27
14 1.39 l-Asparagine C4H8N2O3 133.061 (−1.75) 131.0461 (0.88) 116, 87, 74 114, 113 + 23
15 1.3901 Histidine C6H9N3O2 156.077 (−1.59) 154.0622 (0) 110, 83 — + 25
16 1.44 (Iso)Leucine C6H13NO2 132.1016 (2.33) 130.0870 (2.69) 86 128 + 22
17 1.364 Valine C5H11NO2 118.0855 (6.45) 116.0710 (−3.43) 58 — +
18 1.544 Lysine C6H14N2O2 147.1136 (−5.45) 145.098 (1.72) 130 127, 101, 84 + 21
19 1.67 Methionine C5H11NO2S 150.0587 (−2.51) 148.0443 (−3.54) 133, 104 128, 102 ++ 25
20 1.80 l-Tyrosine C9H11NO3 182.0812 (−0.17) 180.0663 (1.75) 165, 136, 91 163, 134, 119 + 22
21 1.84 l-(Iso)leucine-hexose C12H23NO7 294.1550 (−0.93) 292.1411 (−2.81) 276, 258, 132 130 + 22
22 2.19 l-Phenylalanine C9H11NO2 166.0864 (−0.88) 164.0718 (−0.59) 120, 103 147 ++ 22
23 2.23 l-Phenylalanine-hexose C15H21NO7 328.1397 (0.75) 326.1236 (2.83) 310, 292, 264, 166 164 + 22
24 3.10 l-Tryptophan C11H12N2O2 205.0973 (−0.71) 203.0826 (0.01) 188, 146, 118 142 116 + 22
 
Sugars and nucleosides
25 1.239 Hexose C6H12O6 — 179.0562 (−0.49) — 161, 143, 71 + 44
26 1.19 Xylonic/arabionic/ribonic acid C5H10O6 — 165.0407 (−1.44) — 147, 95, 75 + 23
27 1.191 Sedoheptulose C7H14O7 — 209.0666 (0.36) — 191, 159, 129, 85 + 23
28 1.225 Cytidine C9H13N3O5 244.0933 (−2.07) 242.077 (5.12) 184, 112, 70 150 + 25
29 1.247 Disaccharide C12H22O11 — 341.1090 (−0.19) — 204, 179 + 44
30 1.303 Gluconic acid C6H12O7 — 195.0508 (1.15) — 129, 87, 75 + 45
31 1.34 Threonic acid C4H8O5 — 135.0301 (−1.49) — 117, 75 tr 23
32 1.56 Uridine C9H12N2O6 245.0762 (2.51) 243.0626 (−1.39) 113 200, 152, 110 + 21
33 1.83 Guanosine C10H13N5O5 284.1004 (−5.14) 282.0850 (−2.15) 152 150 tr 22
34 1.86 Adenosine C10H13N5O4 268.1041 (−0.26) 266.0902 (−2.71) 136 198, 130 tr 25
 
Phenolic acids and derivatives
35 1.323 Caffeoyl hexose C15H18O9 343.1053 (−8.6) 341.08750 (0.89) — 179, 161, 135, 89 + 24
36 1.457 Protocatechuic acid hexose C13H16O9 317.0854 (4.14) 315.0702 (6.19) — 153, 145 + 46
37 1.50 5-Caffeoylquinic acid (chlorogenic acid) C16H18O9 355.1029 (−1.53) 353.0877 (0.3) 163, 145, 135 191, 179, 173, 135 + 23
38 1.87 p-Coumaric acid C9H8O3 165.0548 (−1.09) 163.0400 (0.41) — 119 + 21
39 2.04 3-Caffeoylquinic acid (neochlorogenic acid) C16H18O9 355.1029 (−1.53) 353.0877 (0.3) 163, 145, 135 191, 179, 135 + 22
40 2.243 Benzyl-O-rutinoside C19H28O10 417.1752 (0.78) 415.1617 (−1.75) — 353, 164 ++ 47
41 2.46 3-p-Coumaroylquinic acid C16H18O8 339.1074 (0.13) 337.0928 (0.27) 147, 119 191, 173, 163, 119 + 21
42 2.461 4-Caffeoylquinic acid (cryptochlorogenic acid) C16H18O9 355.1029 (−1.53) 353.0877 (0.3) 163, 145, 135 191, 179, 173, 135 +++ 22
43 2.90 3-O-Feruloylquinic acid C17H20O9 369.1187 (−1.88) 367.1035 (−0.12) 177, 145 193, 191, 173, 134 ++ 48
44 3.61 4-p-Coumaroylquinic acid C16H18O8 339.1074 (0.13) 337.0928 (0.27) 147, 119 191, 173, 163, 119 ++ 21
45 4.05 5-p-Coumaroylquinic acid C16H18O8 339.1074 (0.13) 337.0929 (−0.03) 147, 119 191, 173, 163, 119 + 21
46 4.20 4-O-Feruloylquinic acid C17H20O9 369.1187 (−1.88) 367.1035 (−0.12) 177, 145 193, 173, 134 + 48
47 5.28 Vanillic acid hexose C14H18O9 — 329.08 (−1.19) — 167 + 22
48 6.81 Feruloyl glycerol C13H16O6 269.1032 (−4.6) 267.0872 (0.79) — 252, 175 + 21
49 6.91 Feruloyl hexose C16H20O9 357.1181 (−0.26) 355.1044 (−2.65) 195, 163 193 + 22
50 10.52 Ferulic acid C10H10O4 195.0667 (−7.81) 193.0491 (7.9) — 178, 134 ++ 21
51 11.02 Dicoumaroyl glycerol C21H20O7 385.128 (0.47) 383.1134 (0.59) 147, 119 219, 163, 119 + 21
52 11.28 Coumaroyl-feruloyl glycerol C22H22O8 415.1394 (1.58) 413.1240 (0.46) 177, 147 249, 193, 163, 177, 134 ++ 21
53 11.50 Diferuoloyl glycerol C23H24O9 445.1494 (−0.21) 443.1339 (1.39) 177, 145 249, 193, 134 ++ 21
 
Flavonoids
54 1.49 Catechin C15H14O6 291.0850 (4.53) 289.069 (9.52) 273 — ++ 25
55 4.45 Apigenin-6,8-di-C-glucoside (vicenin 2) C27H30O15 595.1658 (−0.09) 593.1501 (1.84) 577, 475, 457, 325 473, 383, 353 ++ 22
56 4.47 Quercetin hexosyl rutinoside C33H40O21 773.2142 (−0.93) 771.1981 (1.08) 627, 465, 303 609, 301 ++ 23
57 5.01 Kaempferol hexosyl rutinoside C33H40O20 757.2193 (−0.97) 755.2055 (−1.96) 611, 595, 449, 287 593, 285 ++ 21
58 5.15 Isorhamnetin hexosyl rutinoside C34H42O21 787.2296 (−0.59) 785.2135 (1.38) 641, 625, 479, 317 623, 315 ++ 21
59 6.35 Quercetin dihexoside C27H30O17 627.1560 (−0.68) 625.1427 (−2.68) — 301, 299 +++ 22
60 6.38 Rutin C27H30O16 611.1608 (−0.23) 609.1459 (0.34) 465, 303 301 +++ 23
61 6.60 Quercetin hexoside C21H20O12 465.1035 (−1.61) 463.0868 (3.02) 303 301, 286 +++ 23
62 6.80 Kaempferol rutinoside (nicotiflorin) C27H30O15 595.1663 (−0.39) 593.1510 (0.33) 449, 287 285 ++ 23
63 6.97 Isorhamnetin rutinoside (narcissin) C28H32O16 625.1766 (−0.46) 623.1618 (−0.07) 479, 317 315 ++ 23
64 7.17 Kaempferol hexoside C21H20O11 449.1082 (−0.81) 447.0930 (0.64) 287 285 ++ 21
65 7.29 Isorhamnetin hexoside C22H22O12 479.1203 (−3.97) 477.1034 (0.94) 317 315 +++ 23
 
S-containing compounds
66 2.03 Asparagusic acid C4H6O2S2 — 148.9735 (0.96) — 104, 63 + 22
67 3.55 Asparaptine C10H18N4O3S2 307.0898 (−1.61) 305.0746 (0.51) 248, 229, 212, 170, 150 259, 241, 173 + 25
68 8.37 Thiamine C12H16N4OS 265.1092 (9.69) — 219, 144, 122 — + 25
 
Steroidal saponins
69 7.35 Aspacochinoside M C45H74O19 901.4797bb (−0.62) 917.4746 (0.6) 739, 433 — + 26
70 8.69 Furostane-triol glycoside C45H76O19 903.4951bb (−0.34) 919.4914 (−0.65) 741, 579 417, 399 — ++ 26
71 9.14 Aspacochioside D C45H74O18 885.4839bb (0.37) 901.4791 (1.26) 723, 415 — + 26
72 9.56 Aspacochioside A C45H76O18 887.5019bb (−2.28) 903.4978 (−2.11) 741, 417 — + 26
73 10.45 Yamogenin II C45H72O17 867.4739bb (−0.37) 883.4696 (0.08) 721, 413 — +++ 26
74 16.28 Dioscin C45H72O16 869.4897 (−0.45) — 723, 577, 415 — ++ 26
 
Fatty acids and lipids
75 7.77 Trihydroxy-octadecadienoic acid C18H32O5 — 327.2178 (−0.31) — 229, 211, 171 + 22
76 8.46 Trihydroxy-octadecaecaenoic acid C18H34O5 — 329.2331 (0.75) — 229, 211, 171 ++ 22
77 11.56 9-Hydroperoxy-octadecadienoic acid C18H32O4 — 311.2222 (1.87) — 293 171, 139 + 22
78 11.87 12,13-Dihydroxy-octadecenoic acid C18H34O4 — 313.2380 (1.38) — 295, 183, 129 + 22
79 14.11 LPI (18 : 3) C27H47O12P 595.2881 (−0.52) 593.2728 (0.74) 577, 559, 335, 317 277, 241 ++ 21
80 14.69 9-Hydroxy-octadecadienoic acid C18H32O3 297.2429 (−1.62) 295.2272 (2.26) 279 277, 171 tr 22
81 14.90 13-Hydroxy-octadecadienoic acid C18H32O3 297.2429 (−1.62) 295.2272 (2.26) 279 277, 195 tr 22
82 15.12 LPI (18 : 2) C27H49O12P 597.3039 (−0.77) 595.2900 (−1.87) 579, 561, 337, 317 415, 315, 279, 241 + 49
83 15.48 LPI (16 : 0) C25H49O12P 573.3031 (0.59) 571.2901 (−2.12) 555, 537, 313, 155 315, 255, 241, 214, 225, 191, 153 + 25
84 15.94 Linoleoyldigalactopyranosyl glycerol C33H56O14 677.3749 (−0.91) 675.3586 (1.67) 353, 261 415, 397, 277, 235 ++ 22
85 16.08 Phytosphingosine C18H39NO3 318.3002 (0.22) — 282 — + 25
86 16.20 LPG (16 : 0) C22H45O9P 485.2879 (−1.04) 483.2723 (1.12) 467, 449, 313, 155 255, 227, 153 + 25
87 16.43 LPA (16 : 0) C19H39O7P 411.2515 (−2.15) 409.2355 (1.37) — 171, 153, 79 + 25
88 17.20 LPE (18 : 2) C23H44NO7P 478.2933 (−1.01) 476.2785 (−0.5) 337, 306 279, 196, 140 + 50
89 17.29 LPC (18 : 2) C26H50NO7P 520.3405 (−1.41) 564.3309cc (−0.37) 502, 184 504, 279, 242, 224 + 50
90 17.65 Linoleoylgalactopyranosyl glycerol C27H46O9 515.3217 (−0.47) 513.3064 (0.99) 497, 353, 261 277, 253, 235 +++ 22
91 17.96 LPE (16 : 0) C21H44NO7P 454.2933 (−1.07) 452.2777 (1.24) 313, 282 255, 196, 140 tr 25
92 18.11 LPC (16 : 0) C24H50NO7P 496.3402 (−0.88) 540.3305cc (0.36) 478, 184, 104 480, 255, 224 tr 25
93 20.53 12-Oxo-phytodienoic acid C18H28O3 293.2117 (−1.98) 291.1966 (−0.11) 275, 219 247 +++ 22
94 20.65 Linolenic acid C18H29O2 — 277.2175 (−0.71) — 227 ++ 22
95 22.47 Linoleic acid C18H32O2 — 279.2335 (−1.95) — 261, 227 ++ 47
96 23.55 Palmitic acid C16H32O2 — 255.2328 (0.6) — 237, 227 ++ 25
97 24.11 Oleic acid C16H34O2 — 281.2486 (0.01) — 227 ++ 47
 
Miscellaneous compounds
98 5.63 Trimethoxystilbene hexose C23H28O9 449.1827 (−4.67) 447.1652 (1.91) — 285, 225 ++ 22
99 20.14 Di-tert-butylphenol C14H22O — 205.1598 (−0.05) — 206, 189 tr 22
Open in a new tab
a

LPA: lysophosphatidic acid; LPC: lysophosphatidycholine; LPE: lysophosphatidylethanolamine; LPG: lysophosphatidylglycerol; LPI: lysophosphatidylinositol; Rel. ab., relative abundancy; Rt, retention time;

b

[M + H–H2O]+.

c

[M + HCOO]−, tr, traces (<5 thousands); +, present (5–500 thousands); ++, abundant (>500 thousands – 50 × 106), +++, highly abundant (50 × 106). All values are presented as signal intensity

3.3.1. Identification of organic acids

Organic acids content in food contribute to its organoleptic qualities and act as preservatives, improving quality and safety.46 Seven organic acids were detected in the methanolic extract of A. officinalis spears. The negative ionization 1mode provided better response than the positive mode.51 Decarboxylation and/or water elimination gave the most intense fragments for most of the organic acids.46 For example, malic acid (5) showed a base peak at m/z 115 corresponding to [M–H–H2O]− and fragment ion at m/z 71 corresponding to [M–H–H2O–CO2]− (Fig. S3). Major fragments in the positive ionization mode were characterized by the loss of water.52 For instance, abscisic acid (7) exhibited a fragment ion at m/z 247 [M + H–H2O]+ and a base peak at m/z 229 [M + H–2H2O]+ (Fig. S4).

3.3.2. Identification of amino acids and derivatives

Plants often contain free amino acids, which can indicate fruit growth, maturation, and ripening.53 The amino acid scaffold serves as the basis for the structures of many dozen recognized antibacterial, antifungal, or antiprotozoal medicines.54 The amino acid skeleton plays a critical role in most of them, as they are frequently structural analogues of amino acid intermediates from various microbial metabolic pathways. Other amino acids act as effective inhibitors of various bacterial target enzymes. Seventeen amino acids were tentatively identified, including l-glutamine (8), l-(iso)leucine (16), l-phenylalanine (22), alongside glycosides as glutamine hexose (10), l-(iso)leucine-hexose (21), l-phenylalanine-hexose (23). Amino acids showed a common loss of NH3 (17 m/z), followed by loss of H2O (18 m/z) and CO and H2O (46 m/z).55 For example, tyrosine (20) fragmentation showed ions at m/z 165 and 136 and at m/z 163 and 134 in the positive (Fig. S5) and negative (Fig. S6) ionization modes, respectively. In the negative ionization mode, the glycosides showed loss of the attached sugar (−162 Da) and base peak formation at m/z 145, 130 and 164 for compounds 10, 21 and 23, respectively (Fig. S7–S9).46 Amino acids were previously detected in A. officinalis spears.22,23

3.3.3. Identification of sugars and nucleosides

In the negative ionization mode, a hexose (25), sedoheptulose (27) and 3 sugar acids (26, 30 and 31) were identified, exhibiting loss of water molecules (−18 Da) (Fig. S10 and S11). A disaccharide (29) was detected showing a fragment ion at m/z 179 due to loss of hexose molecule (−162 Da) (Fig. S12)56.

Nucleosides were recognized for their striking antibacterial potential.57 In the positive ionization mode, four nucleosides were identified as cytidine (28), uridine (32), guanosine (33) and adenosine (34)]. Each detected nucleosides exhibited loss of ribose moiety (−132 Da) yielding base peak ions at m/z 112, 113, 152 and 136 for compounds 28, 32, 33 and 34, respectively (Fig. S13 and S14).

3.3.4. Identification of phenolic acids and their derivatives

Phenolic acids and their derivatives, known for their antioxidant and health-promoting properties, represented a significant group among the identified compounds. Nineteen compounds (35–53) belonging to this class were tentatively identified, underscoring the potential bioactive properties associated with the sample. Phenolic compounds are found in an abundance of vegetables, fruits, and cereals. The high consumption of foods rich in phenolic compounds could prevent a series of diseases as they exhibit various pharmacological potentials, including antiviral, antioxidant, antitumor, antiallergic, anti-inflammatory, fungal, and antibacterial activities.58

Compounds 35, 36, 47 and 49 both showed loss of hexose moiety (−162 Da) yielding fragment ions at m/z 179 [caffeic acid–H]−, m/z 153 [protocatechuic acid–H]−, m/z 167 [vanillic acid–H]− and m/z 193 [ferulic acid–H]−, respectively and were identified as caffeoyl hexose (35), protocatechuic acid hexose (36), vanillic acid hexose (47) and ferulic hexose (51) (Fig. S14).21,22

Compounds 37, 39 and 42 exhibited the same [M–H]− at m/z 353, yielding fragments at m/z 191[quinic acid–H]−, 179 [caffeoyl–H]−, 173 [quinic acid–H–H2O]−, and 135 [caffeoyl–H–COO]−which are specific for caffeoylquinic acids.59. Compounds 37 and 39 both yielded base peak at m/z 191 and were identified as 5-O-caffeoylquinic acid (chlorogenic) and 3-O-caffeoylquinic acid (neochlorogenic acid), respectively based on the relative abundance of the peak at m/z 179 (Fig. S15 and S16). Compound 42 was identified as 4-O-caffeoylquinic acid (cryptochlorogenic acid) as it displayed a base peak at m/z 173 (Fig. S17).

Compound (38) exhibited [M–H]− at m/z 163.0404 and fragment ion at m/z 119.1, showing the loss of COO group (Fig. S18). Therefore, the compound was tentatively identified as coumaric acid.21

Regarding compounds 41, 44 and 45, they showed the same [M–H]− at m/z 337, yielding diagnostic fragment ion for p-coumaroyl moiety at m/z 163 and 119.60 Compound 41 yielded base peak at m/z 163 and was identified as 3-p-coumaroylquinic acid (Fig. S19). Compounds 44 and 45 were identified as 4-p-coumaroylquinic acid and 5-p-coumaroylquinic acid due to the distinctive base peak at m/z 173 and the relative abundance of fragment ion at m/z 191 (Fig. S20 and 21).61

Compounds 43 and 46 showed pseudomolecular ion [M–H]− at m/z 367 that gave rise to fragment ions at m/z 193.049 [ferulic acid–H]− and 134 [ferulic acid–H–CO2–CH3]− corresponding to feruloyl moiety and m/z at 191.055 [quinic acid–H]−, and 173.044 [quinate–H2O]− characteristic to quinic moiety (Fig. S22 and S23). They were identified relative to their MS/MS spectra as 3-O-feuloylquinic acid (43) and 4-O-feuloylquinic acid (46).48 Four compounds corresponding to coumaric and/or ferulic acid esterified with glycerol were recognized as feruloyl glycerol (48), dicoumaroyl glycerol (51), coumaroyl-feruloyl glycerol (52) and diferuloyl glycerol (53). Compound 48 showed fragment ions at m/z 252 and 175 that corresponded to the loss of methyl group from the molecular ion [M–H–CH3]− and loss of water molecule from the feruloyl residue [ferulic acid–H-18]−, respectively (Fig. S24). Regarding compounds 51–53, fragments ions at m/z 193 and/or163 indicated the presence of feruloyl and/or coumaroyl moiety (Fig. S25–S27). Coumaroyl and feruloyl glycerols esters were previously detected in A. officinalis spears.21

Phenolic acids were reported to have anti-staphylococcal properties by influencing the adhesion properties of S. aureus, inhibiting the production of α-hemolysin by this microorganism, and interfering with the stability of the cell membrane and with the metabolic activity of the cells of S. aureus.62 The potential of Sortase A (SrtA) as a drug target for treating S. aureus infections has garnered considerable attention, given its role in mediating the virulence factors crucial for bacterial adhesion, invasion, and evasion of host immunity. In this perspective, inhibiting SrtA activity has emerged as a promising strategy for combating S. aureus infections. While chlorogenic acid does not exhibit significant direct antimicrobial activity against S. aureus, it has been found to inhibit the activity of SrtA in vitro and impede the binding of S. aureus to fibrinogen (Fg), a crucial step in the infection process. Using molecular dynamics simulations and mutagenesis assays, it has been established that chlorogenic acid interacts with the binding sites of C184 and G192 within SrtA. These findings provide valuable insights into the molecular interactions underlying the inhibitory effects of chlorogenic acid on SrtA activity. Moreover, in vivo studies have demonstrated that chlorogenic acid effectively prevents the formation of S. aureus-induced renal abscess in mice, resulting in a significant survival advantage.63 Additionally, evidence that caffeoylquinic acids derivatives have a preferential binding to major facilitator super family efflux systems, a key MDR determinant in Gram-positive bacteria.64 It is worthy to mention that the combination of protocatechuic acid and chlorogenic acid with antibiotics against several common MDR bacteria enhance the antimicrobial activity of most of the tested antibiotics against the bacterial strains of Escherichia coli, S. aureus, Streptococcus iniae, and Proteus mirabilis.65 Moreover, chlorogenic acid, ferulic acid, and p-coumaric acid showed antimicrobial, antibiofilm, and antivirulence potentials against several MDR pathogens.66

3.3.5. Identification of flavonoids

Flavonoids, renowned for their diverse physiological effects and potential health benefits, accounted for twelve of the identified compounds (54–65). Compound 54 with characteristic pseudomolecular ions at m/z 291.0850 and m/z 289.069 in the positive and negative ionization modes, respectively was identified as catechin 25, and compound 55 showed typical fragmentation pattern of vicenin 2, the di-C-glucoside of apigenin (Fig. S28 and S29)45.

Compounds 56–65, belonged to the flavonol subclass, were assigned as quercetin, kaempferol, or isorhamnetin glycosides. Their MS–MS showed cleavage at the glycosidic O-linkage with elimination of the sugar residues. The glycosides were annotated with respect to the difference between the m/z values of the protonated/deprotonated pseudomolecular ions and the fragment ions detected in the positive and negative ionization modes. Mass differences of 162 Da, 146 Da, 308 Da and 470 Da are assigned to glucosyl, rhamnosyl, rutinosyl, and glucosyl rutinosyl loss respectively. In the positive/negative ionization modes, the aglycone ions corresponding to quercetin (m/z 303/301), kaempferol (m/z 287/285), and isorhamnetin (m/z 317/315) were observed as major peaks of these compounds in the MS/MS spectra. For example, the positive and negative MS/MS spectra of compound 56 (Fig. S30 and S31). Accordingly, these compounds were tentatively identified as: quercetin glucosyl rutinoside (56), kaempferol glucosyl rutinoside (57), isorhamnetin glucosyl rutinoside (58), quercetin dihexoside (59), quercetin rutinoside (rutin) (60), quercetin hexoside (61), kaempferol rutinoside (nicotiflorin) (62), isorhamnetin rutinoside (63), kaempferol hexoside (64) and isorhamnetin hexoside (65).23

These compounds are recognized for their effects against resistant bacteria, antioxidant, anti-inflammatory, and anticancer activities, among others. The effects of flavonoids in conjunction with antibiotics in MRSA were investigated. Where flavonoids, alone or in combination, destroyed the bacterial cell membrane. When flavonoids were combined with antibiotics, they were found to increase each other's antibacterial activity. In most cases, the flavonoids and antibiotics produced additive effects. However, synergism was discovered in selected combinations.67 Additionally, they present antibacterial activities against MRSA strains, such as quercetin and its derivatives which exhibit remarkable antibacterial activities against MRSA.68 Rutin's multifaceted mechanisms of action, including antibiofilm activity against Pseudomonas aeruginosa and MRSA urinary catheter biofilms (reduction of biofilm biomass, reduction of cell viability, exopolysaccharide, and extracellular DNA levels), moderate reduction of bacterial adhesion to human keratinocytes, suppression of virulence factors included an impact on P. aeruginosa protease, pyocyanin, rhamnolipid, and elastase production, modulation of gene expression (lasI, lasR, rhlI, rhlR, pqsA and mvfR), and interference with membrane permeability, make it a promising compound for the development of novel antimicrobial therapies.66 PBP2a, a cell wall synthesizing protein, causes MRSA resistance to β-lactam antibiotics but is susceptible to ceftobiprole, a 5th-generation cephalosporin. To develop natural alternatives, in silico-based docking analysis with 37 quercetin derivatives was performed. Quercetin 3-O-rutinoside interacted efficiently with PBP2a, achieving the highest LibDock score. Its structural stability and molecular dynamics simulations confirmed its effectiveness. Quercetin 3-O-rutinoside also showed antibacterial activity at different concentrations, suggesting that the quercetin 3-O-rutinoside could be used as an alternative or in combination with other drugs in treating MRSA infection.69

3.3.6. Identification of S-containing compounds

Three sulfur-containing compounds were detected, suggesting the involvement of sulfur metabolism within the biological system under investigation. Compound 66 ([M–H]−, m/z 148) was assigned as asparagusic acid showing a fragment ion at m/z 104 due to loss of CO2 (44 Da) (Fig. S32).70 In the positive ionization mode, compound 67 [[M + H]+, m/z 307] showed fragment ion peaks at m/z 248 [loss of 59 Da (CH5N3)] and m/z 150 (loss of 157 Da (C6H11N3O2)] suggesting the existence of arginine and asparagusic acid moieties (Fig. S33). Thus compound 67 was assigned as asparaptin.71 Also, in the positive ionization mode, compound 68 showed fragment ions at m/z 144 and m/z 122 corresponding to pyrimidinic and thiazolic rings, respectively (Fig. S34) and was tentatively identified as thiamine.72 These compounds often possess unique biological activities and have been found to be effective anti-MRSA agents.73

3.3.7. Identification of steroidal saponins

Six steroidal saponins, which are bioactive compounds with potential therapeutic applications, were identified as aspacochinoside M (69), furostane-triol glycoside (70), aspacochioside D (71), aspacochioside A (72), yamogenin II (73) and dioscin (74).26 In the positive ionization mode, Compounds 69–73 displayed precursor ions [M + H–H2O]+ and no [M + H]+ was detected, followed by loss of glycosyl moieties. For example, compound 70 showed precursor ion at m/z 903.4951, fragment ions at m/z 741, 579, 417 due to successive loss of glucosyl moieties (−162 Da) (Fig. S35). On the other hand, compound 74 showed a protonated pseudomolecular ion [M + H]+ at m/z 869.4897 and fragment ions were detected m/z 723, m/z577, m/z 415 (loss of glucose) and m/z 397 due to successive loss of 2 rhamnose moieties, 1 glucose and H2O) (Fig. S36). Their presence underscores the potential pharmacological relevance of the sample. A combination between saponin fractions of Zygophyllum album and penicillin showed a potential agent against MRSA that can serve as possible model for new antibacterial drug.74 Saponin has shown to be very promising against MDR bacteria in association with aminoglycoside, fluoroquinolones and beta-lactam, however additional studies should be carried out to better elucidate the mechanism of actions.75 Diosgenin inhibits MRSA biofilm formation by activating sigB expression, increasing sarA, agrA, and RNAIII expression, which decreases spa and icaA expression, thereby inhibiting the synthesis of polysaccharide intercellular adhesins and reducing biofilm formation.76 Antibacterial saponins from A. racemosus, commonly known as Shatavari, have been the focus of several studies due to their significant medicinal properties. The roots of A. racemosus are rich in steroidal saponins, isoflavones, and other bioactive compounds. These constituents contribute to its antibacterial properties against various pathogens, including E. coli and Bacillus subtilis.77,78 Research indicates that extracts from A. racemosus exhibit notable antimicrobial activity. For instance, a study highlighted the effectiveness of its root extract against a range of bacteria, suggesting potential applications in treating infections. The antibacterial potency is attributed to the presence of saponins, which disrupt bacterial membranes.77,79 Recent phytochemical analyses have identified new furostanol saponins with promising antibacterial activity. These findings underscore the therapeutic potential of A. racemosus beyond traditional uses.80 Finally, this study of Asparagus officinalis L. (asparagus) through HR-LC/MS-based metabolite profiling has revealed significant insights into its antibacterial properties, particularly against Methicillin-Resistant Staphylococcus aureus (MRSA) in both in vitro and in vivo models. The findings underscore the potential of Asparagus officinalis as a natural alternative or adjunct therapy for treating MRSA infections. Given the rising concerns over antibiotic resistance, incorporating plant-based therapies could provide a multifaceted approach to infection management. Moreover, the rich profile of bioactive compounds in asparagus opens avenues for developing novel antimicrobial agents through further pharmacological studies and clinical trials.

3.3.8. Identification of fatty acids and lipids

In the negative ionization mode, seven oxylipins were identified as trihydroxy-octadecadienoic acid (75), trihydroxy-octadecaecaenoic acid (76), 9-hydroperoxy-octadecadienoic acid (77), 12,13-dihydroxy-octadecenoic acid (78) and 12-oxo-phytodienoic acid (93).22 Furthermore, compounds 80 and 81 exhibited precursor ions [M–H]− at m/z 295.2272 and they were identified as 9-hydroxy-octadecadienoic and 13-hydroxy-octadecadienoic acid owing to the presence of fragment ion at m/z 171 and 195, respectively (Fig. S37 and S38).22 In addition, four free fatty acids showed precursor ions [M–H]− at m/z 277.2175, 279.2335, 255.2328 and 281.2486 and were identified as linolenic acid (93), linoleic acid (94), palmitic acid (95) and oleic acid (96), respectively.

Twelve lipids were identified belonging to different chemical classes (2 glyceroglycolipids 9 lysophospholipids and 1 sphingolipid).

Different chemical classes of lysophospholipids were identified including lysophosphatidylinositols (LPIs), lysophosphatidylglycerols (LPGs), lysophosphatidic acid (LPAs), lysophosphatidylethanol amines (LPEs) and lysophosphatidylcholines (LPCs). Compounds 79, 82 and 83 were identified as LPIs and were annotated as LPI (18 : 3), LPI (18 : 2) and LPI (16 : 0), respectively. They were tentatively identified based on their characteristic fragmentation in the negative ionization mode. For example, LPI (18 : 2) exhibited precursor ion [M–H]− at m/z 595.2900. Moreover, diagnostic fragment ions at m/z 279 related to the carboxylate 18 : 2 anion and at m/z 241 corresponding to inositol-1,2-cyclic phosphate were detected (Fig. S39)49. Compound 86 was annotated as lysophosphatidylglycerol [LPG (16 : 0)] showing precursor ion at m/z 483.2723 [M–H]− as well as fragment ions at m/z 255 and 153 characteristic to the carboxylate anion and the dehydrated glycerophosphate (C3H6PO5) after loss of water, respectively (Fig. S40).25 One LPA (compound 87) was detected in the negative ionization mode at m/z 409.2355 [M − H]− and was identified as LPA (16 : 0) owing to the presence of characteristic fragment ions at m/z 153 (phosphoglycerol) and 79 (phosphonate group) related to the head group (Fig. S41).25 Compounds 88 and 91 were identified as LPE (18 : 2) and LPE (16 : 0), respectively. In the positive ionization mode, their protonated precursor ions [M + H]+ were detected at m/z 478.2933 and 454.2933, respectively. In addition, characteristic fragment ions were observed at m/z 337 and 313, respectively due to loss of phosphoethanolamine polar head [M + H-141]+ (Fig. S42).50 In the negative ionization mode, fragment ions were detected at m/z 279 and 255, respectively corresponding to the carboxylate anion. Diagnostic fragment ions for LPEs were detected at m/z 196 (due to neutral loss of acyl chain) and 140 (deprotonated phosphoethanolamine) (Fig. S43).50 Two LPCs were identified as LPC (18 : 2) (compounds 89) and LPC (16 : 0) (compound 92). In the positive ionization mode, they showed protonated precursor ions [M + H]+ at m/z 520.3405 and m/z 496.3402, respectively. Fragment ions appeared at m/z 502 and m/z 478 in compound 89 and 92, respectively due to loss of water molecule [M + H–H2O]+. In addition to diagnostic ions at m/z 184 and m/z 104 assigned to the phosphocholine head group and the choline ion, respectively (Fig. S44). In the negative ionization mode, the LPCs were identified as formate adduct [M + HCOO]− at m/z 564.3309 and 540.3305, respectively. In both compounds, the predominant fragment ions occur at m/z 279 and 255 which are the carboxylate 18 : 2 and 16 : 0 anions, respectively (Fig. S45)50.

Two glyceroglycolipids were identified as linoleoyldigalactopyranosyl glycerol (84) and linoleoylgalactopyranosyl glycerol (90). In the positive ionization mode, the spectrum of compounds 84 and 90 showed the same fragment ion at m/z 353 due to loss of 2 hexoses (−324 Da) and 1 hexose moiety (−162 Da) respectively (Fig. S46 and S47). While in the negative ionization mode, fragment ion at m/z 277 was observed in the spectrum of both compounds corresponding to the fatty acyl chain (Fig. S48 and S49). For example, compound 84 showed fragment ion at m/z 415 and 397 due to neutral loss of the fatty acyl group as fatty aldehyde (−260 Da) and free fatty acid (−278 Da), respectively. In addition to fragment ion at m/z 235 resulting from neutral loss of fatty acid (−278 Da) and hexoside moiety (−162 Da) (Fig. S48).81

Compound 85 was annotated as phytosphingosine showing protonated molecular ion at m/z 318.3002 and fragment ions at m/z 300 [M + H–H2O]+ and 282 [M + H–2H2O]+ due to successive loss of water molecules (Fig. S49).25

Lipids have critical roles in the body, such as providing energy, forming cell membranes, and signalling.82

3.3.9. Miscellaneous compounds

Two compounds that were not classified in the previous chemical classes were described in this section. One stilbene; trimethoxystilbene hexoside (98) and one alkyphenol di-ter-butylphenol (99) were tentatively identified. Compound 98 showed precursor ion at m/z 447.1652 and fragment ion at m/z 285 due to loss of hexosyl moiety (Fig. S50).22 Compound 99 exhibited [M–H]− at m/z 205.1598 and a fragment ion at m/z 189 suggesting neutral loss of methane (−16 Da) (Fig. S51)22.

Among the investigated constituents, several phenolic acids, flavonoids and fatty acids have been previously reported to exhibit anti-MRSA activity, including protocatechuic acid, p-coumaric acid, chlorogenic acid, quercetin, apigenin, catechin, rutin, linoleic acid, and oleic acid.68,83–85 For example, linoleic acid, and oleic acid inhibited efflux pumps,83 phenolic acids including protocatechuic acid, p-coumaric acid, and vanillic acid inhibited the growth of MRS.84 Additionally, the antibacterial mechanism of flavonoids such as quercetin and rutin against MRSA is mainly achieved by disruption of membrane structures, inhibition of efflux pumps, and inhibition of β-lactamases and bacterial virulence factors.86,87

4. Conclusion

Globally, the number of fatal infections caused by MDR bacteria has increased particularly in LMICs. More than 130 000 deaths due to MRSA were indicated in 2021, more than doubling from 57 200 in 1990.88 This study demonstrates that A. officinalis L. spears methanolic extract effectively combats MRSA USA300 through multiple mechanisms, addressing the escalating global MDR crisis. Asparagus represents a valuable agricultural crop, offering numerous health benefits and culinary appeal. The identification of ninety-nine compounds provides valuable insights into the chemical diversity and complexity of asparagus spears. These findings contribute to our understanding of the chemical profile and offer potential leads as MRSA inhibitors. Incorporating Asparagus-based antimicrobial agents into food products can help prevent or inhibit the growth of bacterial pathogens, leading to increased shelf life and improved safety. Further research and development, including toxicity and safety evaluations, are necessary to explore the therapeutic potential of A. officinalis L. Nonetheless, these initial findings reveal promising applications in pharmaceutical, agricultural, and food industries.

5. Limitation of the study

Semi-quantitative and targeted LC-MS/MS analyses are recommended in the future study to report relative abundances for major constituents, implement bioactivity-guided chromatographic fractionation followed by purification and structural elucidation (LC-MS/MS, NMR) of active components, and compare profiles across independent extract batches to establish marker compounds and quality-control assays for lot-to-lot consistency. These measures will facilitate identification of active principle(s), support reproducibility, and inform downstream preclinical development.

Ethical approval

This experimental design was conducted and approved following the instructions of the institutional animal care and use committee of Cairo University (Approval number: MI (3694)). All animal experiments were conducted in accordance with the regulations of Directive 2010/63/EU in Europe and this study is reported in accordance with ARRIVE guidelines (https://arriveguidelines.org).

Conflicts of interest

There is no conflicts of interest among authors.

Supplementary Material

RA-016-D6RA03310E-s001

Data availability

All data are available within the manuscript and its supplementary information (SI). Supplementary information is available. See DOI: https://doi.org/10.1039/d6ra03310e.

References

  1. Bai C. Bai B. Strength use and workers' job burnout in the Centers for Disease Control and Prevention: The mediating role of psychological capital. J. Adv. Nurs. 2023;79(6):2328–2336. doi: 10.1111/jan.15586. [DOI] [PubMed] [Google Scholar]
  2. Elsebaei M. M. et al., Alkynyl-containing phenylthiazoles: Systemically active antibacterial agents effective against methicillin-resistant Staphylococcus aureus (MRSA) Eur. J. Med. Chem. 2018;148:195–209. doi: 10.1016/j.ejmech.2018.02.031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Tabaja H. Hindy J.-R. Kanj S. S. Epidemiology of methicillin-resistant Staphylococcus aureus in arab countries of the middle east and north African (MENA) region. Mediterr. J. Hematol. Infect. Dis. 2021;13(1):e2021050. doi: 10.4084/MJHID.2021.050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bhavna K., et al., Antibiotics role in past and present: A challenge to find new possibilites in development, in Antibiotics-Therapeutic Spectrum and Limitations, Elsevier, 2023, pp. 39–64 [Google Scholar]
  5. Abebe A. A. Birhanu A. G. Methicillin resistant Staphylococcus aureus: molecular mechanisms underlying drug resistance development and novel strategies to combat. Infect. Drug Resist. 2023:7641–7662. doi: 10.2147/IDR.S428103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Xu T. et al., Development of membrane-targeting osthole derivatives containing pyridinium quaternary ammonium moieties with potent anti-methicillin-resistant Staphylococcus aureus properties. J. Med. Chem. 2025;68(7):7459–7475. doi: 10.1021/acs.jmedchem.4c03167. [DOI] [PubMed] [Google Scholar]
  7. Kumar G. Engle K. Natural products acting against S. aureus through membrane and cell wall disruption. Nat. Prod. Rep. 2023;40(10):1608–1646. doi: 10.1039/D2NP00084A. [DOI] [PubMed] [Google Scholar]
  8. Desoukey S. et al., Antimicrobial effect of Asparagus officinalis L. extracts. Plant Arch. 2020;20(2):9253–9264. [Google Scholar]
  9. Moreno-Pinel R., et al., Asparagus (Asparagus officinalis L.) breeding, in Advances in Plant Breeding Strategies: Vegetable Crops: Volume 9: Fruits and Young Shoots, Springer, 2021, p. 425–469 [Google Scholar]
  10. Chitrakar B. et al., Protocols for extraction and purification of rutin from leafy by-products of asparagus (Asparagus officinalis) and characterization of the purified product. Food Chem. 2023;418:136014. doi: 10.1016/j.foodchem.2023.136014. [DOI] [PubMed] [Google Scholar]
  11. Chitrakar B. Zhang M. Adhikari B. Asparagus (Asparagus officinalis): Processing effect on nutritional and phytochemical composition of spear and hard-stem byproducts. Trends Food Sci. Technol. 2019;93:1–11. doi: 10.1016/j.tifs.2019.08.020. [DOI] [Google Scholar]
  12. Guo Q. et al., The bioactive compounds and biological functions of Asparagus officinalis L.–A review. J. Funct.Foods. 2020;65:103727. doi: 10.1016/j.jff.2019.103727. [DOI] [Google Scholar]
  13. Kohli D. Champawat P. S. Mudgal V. D. Asparagus (Asparagus racemosus L.) roots: nutritional profile, medicinal profile, preservation, and value addition. J. Sci. Food Agric. 2023;103(5):2239–2250. doi: 10.1002/jsfa.12358. [DOI] [PubMed] [Google Scholar]
  14. Çomaklı V. et al., Enzyme inhibition and antioxidant activities of Asparagus officinalis L. and analysis of its phytochemical content by LC/MS/MS. Chem. Biodivers. 2023;20(5):e202201231. doi: 10.1002/cbdv.202201231. [DOI] [PubMed] [Google Scholar]
  15. Ali N. B. et al., In Vitro and In Vivo Antibiofilm Activity of Red Onion Scales: An Agro-Food Waste. Molecules. 2023;28(1):355. doi: 10.3390/molecules28010355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Humphries R. M. et al., CLSI Methods Development and Standardization Working Group Best Practices for Evaluation of Antimicrobial Susceptibility Tests. J. Clin. Microbiol. 2018;56(4):e0193417. doi: 10.1128/JCM.01934-17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ismail M. M. et al., Exploring the Antivirulence Activity of Pulverulentone A, a Phloroglucinol-Derivative from Callistemon citrinus Leaf Extract, against Multi-Drug Resistant Pseudomonas aeruginosa. Antibiotics. 2021;10(8):907. doi: 10.3390/antibiotics10080907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. El-Naggar M. M. et al., Integration of terpesomes loaded Levocetrizine dihydrochloride gel as a repurposed cure for Methicillin-Resistant Staphylococcus aureus (MRSA)-Induced skin infection; D-optimal optimization, ex vivo, in silico, and in vivo studies. Int. J. Pharm. 2023;633:122621. doi: 10.1016/j.ijpharm.2023.122621. [DOI] [PubMed] [Google Scholar]
  19. Eltabeeb M. A. et al., Nanocomposite alginate hydrogel loaded with propranolol hydrochloride kolliphor® based cerosomes as a repurposed platform for Methicillin-Resistant Staphylococcus aureus-(MRSA)-induced skin infection; in vitro, ex vivo, in silico, and in vivo evaluation. Drug Deliv. Transl. Res. 15:556–576. doi: 10.1007/s13346-024-01611-z. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tsugawa H. et al., MS-DIAL: data-independent MS/MS deconvolution for comprehensive metabolome analysis. Nat. Methods. 2015;12(6):523–526. doi: 10.1038/nmeth.3393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Zhang M., et al., A Targeted Analysis of Flavonoids in asparagus Using the UPLC-MS Technique. 2020 [Google Scholar]
  22. Jiménez-Sánchez C. et al., Comprehensive, untargeted, and qualitative RP-HPLC-ESI-QTOF/MS2 metabolite profiling of green Asparagus (Asparagus officinalis) J. Food Compos. Anal. 2016;46:78–87. doi: 10.1016/j.jfca.2015.11.004. [DOI] [Google Scholar]
  23. Jiménez-Sánchez C. et al., Identification of bioactive compounds of Asparagus officinalis L.: Permutation test allows differentiation among “triguero” and hybrid green varieties. Molecules. 2021;26(6):1640. doi: 10.3390/molecules26061640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Li Y. et al., Profiles of phenolics and their synthetic pathways in Asparagus officinalis L. Food Chem.:Mol. Sci. 2024;8:100187. doi: 10.1016/j.fochms.2023.100187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Creydt M. et al., Plant metabolomics: evaluation of different extraction parameters for nontargeted UPLC-ESI-QTOF-mass spectrometry at the example of white Asparagus officinalis. J. Agric. Food Chem. 2018;66(48):12876–12887. doi: 10.1021/acs.jafc.8b06037. [DOI] [PubMed] [Google Scholar]
  26. Xue X. et al., Differentiation of three Asparagus species by UHPLC-MS/MS based molecular networking identification and chemical profile analysis. J. Pharm. Biomed. Anal. 2022;219:114863. doi: 10.1016/j.jpba.2022.114863. [DOI] [PubMed] [Google Scholar]
  27. Yang C. et al., Analysis of volatile organic compounds and metabolites of three cultivars of asparagus (Asparagus officinalis L.) using E-nose, GC-IMS, and LC-MS/MS. Bioengineered. 2022;13(4):8866–8880. doi: 10.1080/21655979.2022.2056318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ki V. Rotstein C. Bacterial Skin and Soft Tissue Infections in Adults: A Review of Their Epidemiology, Pathogenesis, Diagnosis, Treatment and Site Of Care. Can. J. Infect. Dis. Med. Microbiol. 2008;19(2):846453. doi: 10.1155/2008/846453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Bittner Fialová S. et al., Antibacterial Activity of Medicinal Plants and Their Constituents in the Context of Skin and Wound Infections, Considering European Legislation and Folk Medicine—A Review. Int. J. Mol. Sci. 2021;22(19):10746. doi: 10.3390/ijms221910746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Algammal A. M. et al., Methicillin-Resistant Staphylococcus aureus (MRSA): One Health Perspective Approach to the Bacterium Epidemiology, Virulence Factors, Antibiotic-Resistance, and Zoonotic Impact. Infect. Drug Resist. 2020;13(null):3255–3265. doi: 10.2147/IDR.S272733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Maureen U. O. et al., Methicillin-resistant Staphylococcus aureus (MRSA) and anti-MRSA activities of extracts of some medicinal plants: A brief review. AIMS Microbiol. 2019;5(2):117–137. doi: 10.3934/microbiol.2019.2.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Olas B. A Review of the Pro-Health Activity of Asparagus officinalis L. and Its Components. Foods. 2024;13(2):288. doi: 10.3390/foods13020288. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kumaran T. Citarasu T. Ethnopharmacological investigation and antibacterial evaluation of the methanolic extract of Asparagus racemosus (Shatavari) Trop. Plant Res. 2015;2(3):175–179. [Google Scholar]
  34. Quave C. L. et al., Effects of extracts from Italian medicinal plants on planktonic growth, biofilm formation and adherence of methicillin-resistant Staphylococcus aureus. J. Ethnopharmacol. 2008;118(3):418–428. doi: 10.1016/j.jep.2008.05.005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Panda S. K. et al., Indian medicinal plant extracts to control multidrug-resistant S. aureus, including in biofilms. South Afr. J. Bot. 2020;128:283–291. doi: 10.1016/j.sajb.2019.11.019. [DOI] [Google Scholar]
  36. El-Shiekh R. A. et al., Bioguided Isolation of Antibiofilm and Antibacterial Pregnane Glycosides from Caralluma quadrangula: Disarming Multidrug-Resistant Pathogens. Antibiotics. 2021;10(7):811. doi: 10.3390/antibiotics10070811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Guo Y. Literature analysis on asparagus roots and review of its functional characterizations. Front. Nutr. 2023;9:1024190. doi: 10.3389/fnut.2022.1024190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Johnson J. K. et al., Skin and soft tissue infections caused by methicillin-resistant Staphylococcus aureus USA300 clone. Emerging Infect. Dis. 2007;13(8):1195–1200. doi: 10.3201/eid1308.061575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Noumi E. et al., HR-LCMS-based metabolite profiling, antioxidant, and anticancer properties of Teucrium polium L. methanolic extract: Computational and in vitro study. Antioxidants. 2020;9(11):1089. doi: 10.3390/antiox9111089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tatiya-aphiradee N. Chatuphonprasert W. Jarukamjorn K. In vivo antibacterial activity of Garcinia mangostana pericarp extract against methicillin-resistant Staphylococcus aureus in a mouse superficial skin infection model. Pharm. Biol. 2016;54(11):2606–2615. doi: 10.3109/13880209.2016.1172321. [DOI] [PubMed] [Google Scholar]
  41. Tatiya-aphiradee N. Chatuphonprasert W. Jarukamjorn K. Anti-inflammatory effect of Garcinia mangostana Linn. pericarp extract in methicillin-resistant Staphylococcus aureus-induced superficial skin infection in mice. Biomed. Pharmacother. 2019;111:705–713. doi: 10.1016/j.biopha.2018.12.142. [DOI] [PubMed] [Google Scholar]
  42. Salem M. A. et al., In vivo Antibacterial Activity of Star Anise (Illicium verum Hook.) Extract Using Murine MRSA Skin Infection Model in Relation to Its Metabolite Profile. Infect. Drug Resist. 2021;14(null):33–48. doi: 10.2147/IDR.S285940. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. El-Hema H. S. et al., A multi-target nitrogen-fused azole drug platform derived from a pyrazoline-thiadiazole moiety: In vivo antimicrobial validation and comprehensive anticancer investigation supported by computational studies. Pharmaceutics. 2026;18(4):424. doi: 10.3390/pharmaceutics18040424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Saied D. B. Farag M. A. How does maturity stage affect seeds metabolome via UPLC/MS based molecular networking and chemometrics and in relation to antioxidant effect? a case study in 4 major cereals and legumes. Food Chem. 2023;426:136491. doi: 10.1016/j.foodchem.2023.136491. [DOI] [PubMed] [Google Scholar]
  45. Okba M. M. et al., UPLC-ESI-MS/MS profiling of the underground parts of common Iris species in relation to their anti-virulence activities against Staphylococcus aureus. J. Ethnopharmacol. 2022;282:114658. doi: 10.1016/j.jep.2021.114658. [DOI] [PubMed] [Google Scholar]
  46. Otify A. M. et al., Unveiling metabolome heterogeneity and new chemicals in 7 tomato varieties via multiplex approach of UHPLC-MS/MS, GC-MS, and UV-vis in relation to antioxidant effects as analyzed using molecular networking and chemometrics. Food Chem. 2023;417:135866. doi: 10.1016/j.foodchem.2023.135866. [DOI] [PubMed] [Google Scholar]
  47. Farag M. A. et al., Metabolomics reveals distinct methylation reaction in MeJA elicited Nigella sativa callus via UPLC-MS and chemometrics. Plant Cell, Tissue Organ Cult. 2015;122:453–463. doi: 10.1007/s11240-015-0782-7. [DOI] [Google Scholar]
  48. Kuhnert N. et al., How to distinguish between feruloyl quinic acids and isoferuloyl quinic acids by liquid chromatography/tandem mass spectrometry. Rapid Commun. Mass Spectrom. 2010;24(11):1575–1582. doi: 10.1002/rcm.4537. [DOI] [PubMed] [Google Scholar]
  49. Huang W. et al., Comprehensive profiling of Platycodonis radix in different growing regions using liquid chromatography coupled with mass spectrometry: from metabolome and lipidome aspects. RSC Adv. 2022;12(7):3897–3908. doi: 10.1039/D1RA08285J. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Silva A. C. R. et al., Lipid characterization of arabica and robusta coffee beans by liquid chromatography-ion mobility-mass spectrometry. J. Food Compos. Anal. 2022;111:104587. doi: 10.1016/j.jfca.2022.104587. [DOI] [Google Scholar]
  51. Flores P. Hellín P. Fenoll J. Determination of organic acids in fruits and vegetables by liquid chromatography with tandem-mass spectrometry. Food Chem. 2012;132(2):1049–1054. doi: 10.1016/j.foodchem.2011.10.064. [DOI] [Google Scholar]
  52. Levsen K. et al., Even-electron ions: a systematic study of the neutral species lost in the dissociation of quasi-molecular ions. J. Mass Spectrom. 2007;42(8):1024–1044. doi: 10.1002/jms.1234. [DOI] [PubMed] [Google Scholar]
  53. Oms-Oliu G. et al., Metabolic characterization of tomato fruit during preharvest development, ripening, and postharvest shelf-life. Postharvest Biol. Technol. 2011;62(1):7–16. doi: 10.1016/j.postharvbio.2011.04.010. [DOI] [Google Scholar]
  54. Nowak M. G. Skwarecki A. S. Milewska M. J. Amino acid based antimicrobial agents-synthesis and properties. ChemMedChem. 2021;16(23):3513–3544. doi: 10.1002/cmdc.202100503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Piraud M. et al., ESI-MS/MS analysis of underivatised amino acids: a new tool for the diagnosis of inherited disorders of amino acid metabolism. Fragmentation study of 79 molecules of biological interest in positive and negative ionisation mode. Rapid Commun. Mass Spectrom. 2003;17(12):1297–1311. doi: 10.1002/rcm.1054. [DOI] [PubMed] [Google Scholar]
  56. Abdel-Baki P. M. et al., Comprehensive identification of chemical fingerprint and potential quality markers of leaves and roots of two Aloe species via LC-MS/MS and computational analyses in relation to anti-virulence activity. Future J. Pharm. Sci. 2025;11(1):92. doi: 10.1186/s43094-025-00847-x. [DOI] [Google Scholar]
  57. Thomson J. M. Lamont I. L. Nucleoside analogues as antibacterial agents. Front. Microbiol. 2019;10:952. doi: 10.3389/fmicb.2019.00952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Kauffmann A. C. Castro V. S. Phenolic compounds in bacterial inactivation: a perspective from Brazil. Antibiotics. 2023;12(4):645. doi: 10.3390/antibiotics12040645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Willems J. L. et al., Analysis of a series of chlorogenic acid isomers using differential ion mobility and tandem mass spectrometry. Anal. Chim. Acta. 2016;933:164–174. doi: 10.1016/j.aca.2016.05.041. [DOI] [PubMed] [Google Scholar]
  60. Fang N. Yu S. Prior R. L. LC/MS/MS characterization of phenolic constituents in dried plums. J. Agric. Food Chem. 2002;50(12):3579–3585. doi: 10.1021/jf0201327. [DOI] [PubMed] [Google Scholar]
  61. Nagy Á. Abrankó L. Profiling of hydroxycinnamoylquinic acids in plant extracts using in-source CID fragmentation. J. Mass Spectrom. 2016;51(12):1130–1145. doi: 10.1002/jms.3847. [DOI] [PubMed] [Google Scholar]
  62. Luís Â. et al., Antistaphylococcal and biofilm inhibitory activities of gallic, caffeic, and chlorogenic acids. Biofouling. 2014;30(1):69–79. doi: 10.1080/08927014.2013.845878. [DOI] [PubMed] [Google Scholar]
  63. Wang L. et al., The therapeutic effect of chlorogenic acid against Staphylococcus aureus infection through sortase A inhibition. Front. Microbiol. 2015;6:1031. doi: 10.3389/fmicb.2015.01031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Fiamegos Y. C. et al., Antimicrobial and efflux pump inhibitory activity of caffeoylquinic acids from Artemisia absinthium against gram-positive pathogenic bacteria. PLoS One. 2011;6(4):e18127. doi: 10.1371/journal.pone.0018127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Chai B., et al., In Vitro Synergistic Interactions of Protocatechuic Acid and Chlorogenic Acid in Combination with Antibiotics against Animal Pathogens, Synergy, 2019, 9, p. 100055 [Google Scholar]
  66. Ivanov M. et al., Polyphenols as inhibitors of antibiotic resistant bacteria—mechanisms underlying rutin interference with bacterial virulence. Pharmaceuticals. 2022;15(3):385. doi: 10.3390/ph15030385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Amin M. U. et al., Antibiotic additive and synergistic action of rutin, morin and quercetin against methicillin resistant Staphylococcus aureus. BMC Compl. Alternative Med. 2015;15:1–12. doi: 10.1186/s12906-015-0580-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  68. Su Y. et al., Studies of the in vitro antibacterial activities of several polyphenols against clinical isolates of methicillin-resistant Staphylococcus aureus. Molecules. 2014;19(8):12630–12639. doi: 10.3390/molecules190812630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  69. Rani N. et al., Quercetin 3-O-rutinoside mediated inhibition of PBP2a: computational and experimental evidence to its anti-MRSA activity. Mol. BioSyst. 2014;10(12):3229–3237. doi: 10.1039/C4MB00319E. [DOI] [PubMed] [Google Scholar]
  70. Pegiou E. et al., Metabolomics reveals heterogeneity in the chemical composition of green and white spears of asparagus (A. officinalis) Metabolites. 2021;11(10):708. doi: 10.3390/metabo11100708. [DOI] [PMC free article] [PubMed] [Google Scholar]
  71. Nakabayashi R. et al., Top-down targeted metabolomics reveals a sulfur-containing metabolite with inhibitory activity against angiotensin-converting enzyme in Asparagus officinalis. J. Nat. Prod. 2015;78(5):1179–1183. doi: 10.1021/acs.jnatprod.5b00092. [DOI] [PubMed] [Google Scholar]
  72. Gentili A. Simultaneous determination of water-soluble vitamins in selected food matrices by liquid chromatography/electrospray ionization tandem mass spectrometry. Rapid Commun. Mass Spectrom. 2008;22(13):2029–2043. doi: 10.1002/rcm.3583. [DOI] [PubMed] [Google Scholar]
  73. Verma S. K. et al., Sulfur (SVI)-containing heterocyclic hybrids as antibacterial agents against methicillin-resistant Staphylococcus aureus (MRSA) and its SAR. Bioorg. Chem. 2024:107241. doi: 10.1016/j.bioorg.2024.107241. [DOI] [PubMed] [Google Scholar]
  74. Sharaf M. H. et al., New combination approaches to combat methicillin-resistant Staphylococcus aureus (MRSA) Sci. Rep. 2021;11(1):4240. doi: 10.1038/s41598-021-82550-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  75. do Socorro Costa M. et al., Evaluation of the antibacterial and inhibitory activity of NorA and MepA efflux pumps from Staphylococcus aureus by diosgenin. Life Sci. 2022;308:120978. doi: 10.1016/j.lfs.2022.120978. [DOI] [PubMed] [Google Scholar]
  76. Kim S.-Y. Kim M. Kim T.-J. Regulation of σB-Dependent Biofilm Formation in Staphylococcus aureus through Strain-Specific Signaling Induced by Diosgenin. Microorganisms. 2023;11(10):2376. doi: 10.3390/microorganisms11102376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  77. Shah M. A. et al., Antibacterial activity of chemical constituents isolated from Asparagus racemosus. Bangladesh J. Pharmacol. 2014;9(1):1–3. [Google Scholar]
  78. Pallela P. N. V. K. et al., Antibacterial activity assessment and characterization of green synthesized CuO nano rods using Asparagus racemosus roots extract. SN Appl. Sci. 2019;1:1–7. [Google Scholar]
  79. Battu G. Kumar B. Phytochemical and antimicrobial activity of leaf extract of Asparagus racemosus Willd. Pharmacogn. J. 2010;2(12):456–463. doi: 10.1016/S0975-3575(10)80031-8. [DOI] [Google Scholar]
  80. Pandey A. R. et al., Furostanol saponins from Asparagus racemosus as potential hypoglycemic agents. Phytochemistry. 2022;201:113286. doi: 10.1016/j.phytochem.2022.113286. [DOI] [PubMed] [Google Scholar]
  81. Masullo M. et al., Pouteria lucuma pulp and skin: In depth chemical profile and evaluation of antioxidant activity. Molecules. 2021;26(17):5236. doi: 10.3390/molecules26175236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  82. Ali S. et al., Food processing and challenges in the food production and quality: The foodomics approach. Food Biosci. 2023;56:103217. doi: 10.1016/j.fbio.2023.103217. [DOI] [Google Scholar]
  83. Liang M. et al., Phytochemicals with activity against methicillin-resistant Staphylococcus aureus. Phytomedicine. 2022;100:154073. doi: 10.1016/j.phymed.2022.154073. [DOI] [PubMed] [Google Scholar]
  84. Salaheen S. et al., Eradication and sensitization of methicillin resistant Staphylococcus aureus to methicillin with bioactive extracts of berry pomace. Front. Microbiol. 2017;8:253. doi: 10.3389/fmicb.2017.00253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  85. Amin M. U. et al., Antibiotic additive and synergistic action of rutin, morin and quercetin against methicillin resistant Staphylococcus aureus. BMC Compl. Alternative Med. 2015;15(1):59. doi: 10.1186/s12906-015-0580-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  86. Lin R.-D. et al., The effects of antibiotics combined with natural polyphenols against clinical methicillin-resistant Staphylococcus aureus (MRSA) Planta Med. 2008;74(08):840–846. doi: 10.1055/s-2008-1074559. [DOI] [PubMed] [Google Scholar]
  87. Xu S. et al., Plant flavonoids with antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) ACS Infect. Dis. 2024;10(9):3086–3097. doi: 10.1021/acsinfecdis.4c00292. [DOI] [PubMed] [Google Scholar]
  88. Naghavi M. et al., Global burden of bacterial antimicrobial resistance 1990–2021: a systematic analysis with forecasts to 2050. Lancet. 2024;404(10459):1199–1226. doi: 10.1016/S0140-6736(24)01867-1. [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

RA-016-D6RA03310E-s001
RA-016-D6RA03310E-s001.pdf (1.5MB, pdf)

Data Availability Statement

All data are available within the manuscript and its supplementary information (SI). Supplementary information is available. See DOI: https://doi.org/10.1039/d6ra03310e.

Articles from RSC Advances are provided here courtesy of Royal Society of Chemistry

RESOURCES