# Identification of Quality Markers in Viola kunawaresis Royel Using HPLC Fingerprints, Chemometric Analysis, and Network Pharmacology

**Authors:** Haifeng Liu, Rongmei Zhao, Hong Xu, Tabusi Manaer, Le Pan, Lu Jin, Amatjan Ayupbek

PMC · DOI: 10.1002/bmc.70410 · Biomedical Chromatography · 2026-03-06

## TL;DR

This study identifies key chemical markers in a medicinal plant used in Uyghur medicine to improve quality control and understand its antiasthmatic effects.

## Contribution

The study introduces a novel approach combining HPLC, chemometrics, and network pharmacology to identify quality markers in VTH.

## Key findings

- Six representative quality markers were identified for VTH using HPLC and chemometric analysis.
- Network pharmacology and molecular docking revealed potential antiasthmatic effects of the identified markers.
- The study provides a foundation for the chemical basis and mechanism of action of VTH.

## Abstract

Violae tianshanicae herba (VTH), a widely used crude drug in Uyghur medicine in China, is renowned for its immunomodulatory, anti‐inflammatory, antibacterial, antiviral, and antidiabetic activities, with notable efficacy in treating asthma. However, systematic research on its quality markers (Q‐markers) remains limited, underscoring the need to analyze its chemical composition to enable effective quality control. A high‐performance liquid chromatography (HPLC) fingerprint method was developed for 21 VTH batches in order to identify the common peaks of 23 flavonoids and esculetin, which were verified using liquid chromatography‐mass spectrometry (LC–MS) and reference standards. In this study, chemometric methods were employed to differentiate samples from four geographical origins and to screen for differential components. In addition, a thorough examination of virtual target prediction in network pharmacology, in conjunction with molecular docking validation, has identified Q‐markers with potential antiasthmatic effects. A group of six representative Q‐markers (including kaempferol 3‐O‐sophorosyl‐7‐O‐glucoside, esculetin, kaempferol 3‐O‐rutinosyl‐7‐O‐glucoside, nicotiflorin, narcissoside, and astragalin) was selected for the purpose of developing a method to quantitatively analyze these components simultaneously in VTH samples. This study provides important references for the comprehensive quality control of VTH and establishes the foundation for researchers to explore the chemical basis and mechanisms of action of VTH.

## Linked entities

- **Chemicals:** esculetin (PubChem CID 5281416), nicotiflorin (PubChem CID 5318767), narcissoside (PubChem CID 5481663), astragalin (PubChem CID 5282102)
- **Diseases:** asthma (MONDO:0004979)

## Full-text entities

- **Genes:** Tnf (tumor necrosis factor) [NCBI Gene 21926] {aka DIF, TNF-a, TNF-alpha, TNFSF2, TNFalpha, Tnfa}, ST2 (suppression of tumorigenicity 2) [NCBI Gene 6761], AKT1 (AKT serine/threonine kinase 1) [NCBI Gene 207] {aka AKT, PKB, PKB-ALPHA, PRKBA, RAC, RAC-ALPHA}, PTGS2 (prostaglandin-endoperoxide synthase 2) [NCBI Gene 5743] {aka COX-2, COX2, GRIPGHS, PGG/HS, PGHS-2, PHS-2}, Nfkb1 (nuclear factor of kappa light polypeptide gene enhancer in B cells 1, p105) [NCBI Gene 18033] {aka NF-KB1, NF-kappaB, NF-kappaB1, p105, p50, p50/p105}, Il17a (interleukin 17A) [NCBI Gene 16171] {aka Ctla-8, Ctla8, IL-17, IL-17A, Il17}, MTOR (mechanistic target of rapamycin kinase) [NCBI Gene 2475] {aka FRAP, FRAP1, FRAP2, RAFT1, RAPT1, SKS}, IGHE (immunoglobulin heavy constant epsilon) [NCBI Gene 3497] {aka IgE}, Il1b (interleukin 1 beta) [NCBI Gene 16176] {aka IL-1beta, Il-1b}, TNF (tumor necrosis factor) [NCBI Gene 7124] {aka DIF, IMD127, TNF-alpha, TNFA, TNFSF2, TNLG1F}, COX2 (cytochrome c oxidase subunit II) [NCBI Gene 4513] {aka COII, MTCO2}, KRT16 (keratin 16) [NCBI Gene 3868] {aka CK16, FNEPPK, K16, K1CP, KRT16A, NEPPK}, CDK7 (cyclin dependent kinase 7) [NCBI Gene 1022] {aka CAK, CAK1, CDKN7, HCAK, MO15, STK1}, NFE2L2 (NFE2 like bZIP transcription factor 2) [NCBI Gene 4780] {aka IMDDHH, NRF2, Nrf-2}, AHR (aryl hydrocarbon receptor) [NCBI Gene 196] {aka FVH3, RP85, bHLHe76}, Hmox1 (heme oxygenase 1) [NCBI Gene 15368] {aka D8Wsu38e, HO-1, HO1, Hemox, Hmox, Hsp32}, NFKB1 (nuclear factor kappa B subunit 1) [NCBI Gene 4790] {aka CVID12, EBP-1, KBF1, NF-kB, NF-kB1, NF-kappa-B1}, MUC5AC (mucin 5AC, oligomeric mucus/gel-forming) [NCBI Gene 4586] {aka MUC5, TBM, leB, mucin}, Akt1 (Akt serine/threonine kinase 1) [NCBI Gene 11651] {aka Akt, LTR-akt, PKB, PKB/Akt, PKBalpha, Rac}, EGFR (epidermal growth factor receptor) [NCBI Gene 1956] {aka ERBB, ERBB1, ERRP, HER1, NISBD2, NNCIS}, IL5 (interleukin 5) [NCBI Gene 3567] {aka EDF, IL-5, TRF}, GSK3B (glycogen synthase kinase 3 beta) [NCBI Gene 2932], PIK3CB (phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit beta) [NCBI Gene 5291] {aka P110BETA, PI3K, PI3KBETA, PIK3C1}, Hsp90aa1 (heat shock protein 90, alpha (cytosolic), class A member 1) [NCBI Gene 15519] {aka 86kDa, 89kDa, Hsp86-1, Hsp89, Hsp90, Hspca}, VIP (vasoactive intestinal peptide) [NCBI Gene 7432] {aka PHM27}, Egfr (epidermal growth factor receptor) [NCBI Gene 13649] {aka 9030024J15Rik, Erbb, Errb1, Errp, Wa5, wa-2}, HSP90AA1 (heat shock protein 90 alpha family class A member 1) [NCBI Gene 3320] {aka EL52, HEL-S-65p, HSP86, HSP89A, HSP90A, HSP90N}, Mmp9 (matrix metallopeptidase 9) [NCBI Gene 17395] {aka B/MMP9, Clg4b, Gel B, MMP-9, pro-MMP-9}, Ephb2 (Eph receptor B2) [NCBI Gene 13844] {aka Cek5, Drt, ETECK, Erk, Hek5, Nuk}, HIF1A (hypoxia inducible factor 1 subunit alpha) [NCBI Gene 3091] {aka HIF-1-alpha, HIF-1A, HIF-1alpha, HIF1, HIF1-ALPHA, MOP1}, Ptgs2 (prostaglandin-endoperoxide synthase 2) [NCBI Gene 19225] {aka COX2, Cox-2, PES-2, PGHS-2, PHS II, PHS-2}, Gsk3b (glycogen synthase kinase 3 beta) [NCBI Gene 56637] {aka 7330414F15Rik, 8430431H08Rik, GSK-3, GSK-3beta, GSK3}, VEGFA (vascular endothelial growth factor A) [NCBI Gene 7422] {aka L-VEGF, MVCD1, VEGF, VPF}, KRT6A (keratin 6A) [NCBI Gene 3853] {aka CK-6C, CK-6E, CK6A, CK6C, CK6D, K6A}, IL33 (interleukin 33) [NCBI Gene 90865] {aka C9orf26, DVS27, IL1F11, NF-HEV, NFEHEV}, IL4 (interleukin 4) [NCBI Gene 3565] {aka BCGF-1, BCGF1, BSF-1, BSF1, IL-4}, Il6 (interleukin 6) [NCBI Gene 16193] {aka Il-6}
- **Diseases:** airway inflammation (MESH:D007249), airway hyperresponsiveness (MESH:D012130), prostate cancer (MESH:D011471), atherosclerosis (MESH:D050197), PDB (MESH:D011488), cancer (MESH:D009369), ALI (MESH:D055371), cough (MESH:D003371), lung injury (MESH:D055370), asthma (MESH:D001249), HCA (MESH:D003027), microbial infections (MESH:D015163), hypoxic (MESH:D002534), V. kunawarensis (MESH:D015419), RSD (MESH:D012019), PCA (MESH:C566443), Kaposi sarcoma-associated herpesvirus infection (MESH:D012514)
- **Chemicals:** Kaempferol 3-O-rhamnoside-7-O-glucoside (-), Methanol (MESH:D000432), narcissoside (MESH:C031062), formic acid (MESH:C030544), kaempferol 3-O-sophoroside (MESH:C064309), isorhamnetin 3-O-glucoside (MESH:C432511), megastigmanes (MESH:D045792), nicotiflorin (MESH:C513882), quercetin (MESH:D011794), phosphoric acid (MESH:C030242), nictoflorin (MESH:C492687), prostaglandin (MESH:D011453), Astragalin (MESH:C001579), acetonitrile (MESH:C032159), Esculetin (MESH:C007628), saccharides (MESH:D002241), MDA (MESH:D008315), rutin (MESH:D012431), flavonols (MESH:D044948), water (MESH:D014867), kaempferol (MESH:C006552), kaempferol 3-O-sophoroside 7-O-glucoside (MESH:C509452), lipid (MESH:D008055), LPS (MESH:D008070), Kaempferol 3-O-glucoside (MESH:C511963), PGE2 (MESH:D015232), apigenin (MESH:D047310), hyperoside (MESH:C021304), coumarins (MESH:D003374), Compound 21 (MESH:C000711730), isorhamnetin 3-O-rutinoside (MESH:C515815), hydrogen (MESH:D006859), isorhamnetin (MESH:C047368), alkaloids (MESH:D000470), NO (MESH:D009569), Flavonoids (MESH:D005419), ethanol (MESH:D000431), ROS (MESH:D017382), Compound 20 (MESH:C028342), glycosides (MESH:D006027)
- **Species:** Mus musculus (house mouse, species) [taxon 10090], Homo sapiens (human, species) [taxon 9606], Human betaherpesvirus 5 (no rank) [taxon 10359]
- **Mutations:** serine/threonine, M10A
- **Cell lines:** RAW 264.7 — Mus musculus (Mouse), Mouse leukemia, Cancer cell line (CVCL_0493)

## Full text

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## Figures

13 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12964187/full.md

## References

74 references — full list in the complete paper: https://tomesphere.com/paper/PMC12964187/full.md

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Source: https://tomesphere.com/paper/PMC12964187