Betulonic Acid Research Articles

Synthesis of Betulonic and Betulinic Acids Derivatives with Sulfonamide Fragment Linked to the Triterpene Core by Amidoalkane Spaser

A series of potentially biologically active sulfonamides of betulonic and betulinic acids with a sulfonamide fragment containing residues of dibutylamine or N-heterocycles, which is linked to a triterpene core by an amidoethane or amidopropane spacer, was synthesized. Betulonic acid sulfonamides were obtained by chlorohydride conjugation of acid with 2-aminoethane and 3-aminopropanesulfonamides and used as precursor compounds for the transition to similar betulinic acid derivatives by selective reduction of the 3-keto group with the action of NaBH4 in EtOH. The structure of all synthesized triterpene sulfonamides was confirmed by 1D and 2D NMR spectroscopy and mass spectrometry.

Determination of triterpenic acids in Ribes stenocarpum maxim by HPLC-APCI-MS with 2-(12,13-dihydro-7H-dibenzo[a,g]carbazol-7-yl)ethyl4-methylben-zenesulfonate labelling agent

The study utilized a fluorescent labeling reagent called 2-(12,13-dihydro-7H-dibenzo[a,g]carbaz-ol-7-yl)ethyl4-methylben-zenesulfonate (DDCETS) to label of triterpenic acids and the approach was used to measure the amount of triterpenic acids in different organs of the Ribes stenocarpum Maxim. (RSM) from several areas in Qinghai Province. The analytical potential of the reagent was evaluated using six triterpene acids, namely maslinic acid, corosolic acid, betulinic acid, betulonic acid, oleanolic acid and ursolic acid. The results showed that the method successfully determined that the main components of triterpenoids in RSM from different regions of Qinghai Province were oleanolic acid and ursolic acid. The highest content of oleanolic acid and ursolic acid was 2037.59 mg/Kg in branches and 2088.97 mg/Kg in fruits. The least distributed organs of oleanolic acid and ursolic acid in the RSM in each region were seeds, with the lowest content of 460.88 mg/Kg in Dongxia Township, Datong County. And when analyzed in relation to the geographical conditions of the diverse origins, the fruits and leaves of the RSM were more abundant in both types of triterpene acids in areas with lower altitude, temperature and precipitation.

Betulonic Acid Inhibits Type-2 Porcine Reproductive and Respiratory Syndrome Virus Replication by Downregulating Cellular ATP Production.

Porcine reproductive and respiratory syndrome (PRRS), caused by PRRS virus (PRRSV) infection, has been a serious threat to the pork industry worldwide and continues to bring significant economic loss. Current vaccination strategies offer limited protection against PRRSV transmission, highlighting the urgent need for novel antiviral approaches. In the present study, we reported for the first time that betulonic acid (BA), a widely available pentacyclic triterpenoids throughout the plant kingdom, exhibited potent inhibition on PRRSV infections in both Marc-145 cells and primary porcine alveolar macrophages (PAMs), with IC50 values ranging from 3.3 µM to 3.7 µM against three different type-2 PRRSV strains. Mechanistically, we showed that PRRSV replication relies on energy supply from cellular ATP production, and BA inhibits PRRSV infection by reducing cellular ATP production. Our findings indicate that controlling host ATP production could be a potential strategy to combat PRRSV infections, and that BA might be a promising therapeutic agent against PRRSV epidemics.

New Type of Ferrocene‐Betulonic Acid Hybrids. Bioactivity, Synthesis, Structure, IR and CD versus DFT Calculations

AbstractFerrocene‐modified natural compounds are of interest for the development of new pharmaceuticals due to improved bioavailability, increased bioactivity, and also as redox markers or vectors. Betulin is a natural plant‐derived product with a wide range of pharmacological properties. However, it is limited in its bioavailability and efficacy. In order to enhance its pharmacological profile, the ferrocene modification of betulin was used in this study. A hybrid of the betulin core (up to 30 % betulin in birch bark) with a ferrocene moiety combined through a piperazine linker (compound 12) was prepared in good yield from readily available ferrocene carboxylic acid and betulonic acid, a natural betulin derivative, via coupling reactions. Spectroscopic techniques, including 1H and 13C NMR, IR and UV‐vis spectroscopy, as well as circular dichroism (CD), were applied to characterize the physicochemical and chiroptical properties. The cytotoxicity of the synthesized hybrid 12 and its ferrocene precursor 13 on MCF‐7 human cancer cell lines were in vitro evaluated by the MTT assay. The IC50 cytotoxicity index of 12 was found to be 26 μM. The theoretically calculated energies of the HOMO‐LUMO orbitals and electronic characteristics of betulin 10, betulonic acid 11, hybrid 12 and ferrocenyl piperazine 13 were determined by the DFT method with hybrid functionals ωB97XD, LC‐ωPBE and B3LYP‐D3 and pure M06 L functionals, with a 6‐311+G(d), 6‐31G(d) or TZV basis set.

Synthesis and Investigation of Biological Activity of New Betulonic Acid Derivatives Containing 1,2,3-Triazole Fragments.

The results of this study showed that the compounds synthesized by the authors have significant potential due to their antibacterial and cytotoxic properties. The apparent antibacterial activity demonstrated by the compounds suggests that they are active antimicrobial agents against common microbial pathogens that cause various socially significant infectious diseases. Compound 6 showed pronounced antimicrobial activity against the Gram-positive test strain Staphylococcus aureus ATCC 6538, and compound 7 demonstrated pronounced antimicrobial activity against the Gram-negative test strain Escherichia coli ATCC 25922 (MIC = 6.3 µg/mL). This allowed us to consider these compounds to have great potential.

Synthesis and anticancer evaluation of new lupane triterpenoid derivatives containing various substituent at the 2 or 3 position

Betulonic acid benzyl ester 1 has been subjected to a series of structural modifications for the purpose of new triterpenoid synthesis and evaluating for anticancer activity. The one-pot two step synthesis of 2α-(aminomethyl)betulinic acid benzyl ester derivatives 3a–f (yield 46–69 %) was achieved by the Mannich reaction of compound 1 with methyleneiminium salts, generated in situ from N,N-disubstituted bis(amino)methanes 2a–f by the action of acetyl chloride in dichloromethane, and subsequent reduction of aminomethylation products with sodium borohydride. Minor 2β-(aminomethyl) triterpenoids 4c,d,f were also isolated (yield 6–15 %). We found, that the stereoselective reaction of triterpenoid 1 with acetylides, generated at –78 °C from alkynes in the presence of n-BuLi, has been useful and noteworthy as the key step in providing of new alkyne substituted triterpenoids – benzyl 3-alkynyl-3-deoxy-2(3),20(29)-lupadiene-28-oates or 3-deoxy-2(3)-dehydro-28-oxoallobetulin derivative. The new compounds were examined for anticancer activity against the human cell lines (MTT assay). All tested derivatives were non-toxic on human fibroblasts. The 3‐(phenylethynyl)lupa-2(3),20(29)-diene 9 showed selective cytotoxicity on cervical cancer cell lines. Tumor cells death trigged by the most active compound 4f resulted from apoptotic processes. These data make the series of synthesized 2 or 3 substituted lupane derivatives as promising compounds with anticancer potential.

Studying immunotherapeutic properties of the conjugate based on BCG antigens with betulonic acid in guinea pigs infected with <i>Mycobacterium scrofulaceum</i>

Studying immunotherapeutic properties of the conjugate based on BCG antigens with betulonic acid in guinea pigs infected with <i>Mycobacterium scrofulaceum</i>

Propranolol chiral analysis using voltammetric sensor based on triterpenoid-graphene oxide hybrid material

Enantioselective sensors are potentially in demand at all stages of the production, storage and use of drugs, including identifying impurities of undesirable isomers and improving stereoselective synthesis methods. A key aspect for enantioselective sensors design is the development of appropriate molecular architectures that make it possible to create recognition sites with different affinities for enantiomers. This experimental work proposes a new chiral sensor based on glassy carbon electrode (GCE) modified by graphene oxide (GO) covalent functionalized with pentacyclic triterpenoid betulonic acid (BetA) for voltammetric recognition and determination of propranolol (Prop) enantiomers. Characterization of the materials was carried out using FTIR spectroscopy, scanning electron microscopy and cyclic voltammetry. Differential pulse voltammetry was used for Prop enantiomers recognition and quantification. The potential difference reached 30 mV along with enanitoselectivity coefficient ipS/ipR equal to 1.30. The calculated binding energy for S-Prop/GO-BetA complex is higher by 4.801 kcal moll−1 compared to R-Prop/GO-BetA indicating a more favorable mutual arrangement of molecules. The linear determination range was established as 5.0 × 10-6 ∼ 1.0 × 10-4 mol·L-1 and 1.0 × 10-4 ∼ 4.0 × 10-4 mol·L-1 for both enantiomers. The detection limits were determined to be 3.9⋅10-7 mol·L-1 and 5.0⋅10-7 mol·L-1 for S-Prop and R-Prop, respectively. The standard spike-recovery tests were carried out in human biological fluids with recoveries 97.3 % − 106.9 % for both enantiomers. The proposed sensor was used to determine the ratio of Prop enantiomers in mixture by regression analysis with projection to latent structures (PLS). The purpose was to build a PLS-model based on a calibration set containing a known number of Prop enantiomers and recognize a test set prepared independently. All samples of test set are correctly identified and the RSD did not exceed 8.7 %. GCE modified by covalent functionalized GO with BetA showed better stability compared with noncovalent functionalization in terms of electrode degradation.

Synthesis, Pharmacokinetic Profile, Anticancer Activity and Toxicity of the New Amides of Betulonic Acid-In Silico and In Vitro Study.

Betulonic acid (B(O)A) is a pentacyclic lupane-type triterpenoid that widely exists in plants. There are scientific reports indicating anticancer activity of B(O)A, as well as the amides and esters of this triterpenoid. In the first step of the study, the synthesis of novel amide derivatives of B(O)A containing an acetylenic moiety was developed. Subsequently, the medium-soluble compounds (EB171 and EB173) and the parent compound, i.e., B(O)A, were investigated for potential cytotoxic activity against breast cancer (MCF-7 and MDA-MB-231) and melanoma (C32, COLO 829 and A375) cell lines, as well as normal human fibroblasts. Screening analysis using the WST-1 test was applied. Moreover, the lipophilicity and ADME parameters of the obtained derivatives were determined using experimental and in silico methods. The toxicity assay using zebrafish embryos and larvae was also performed. The study showed that the compound EB171 exhibited a significant cytotoxic effect on cancer cell lines: MCF-7, A-375 and COLO 829, while it did not affect the survival of normal cells. Moreover, studies on embryos and larvae showed no toxicity of EB171 in an animal model. Compared to EB171, the compound EB173 had a weaker effect on all tested cancer cell lines and produced less desirable effects against normal cells. The results of the WST-1 assay obtained for B(O)A revealed its strong cytotoxic activity on the examined cancer cell lines, but also on normal cells. In conclusion, this article describes new derivatives of betulonic acid-from synthesis to biological properties. The results allowed to indicate a promising direction for the functionalization of B(O)A to obtain derivatives with selective anticancer activity and low toxicity.

Qualitative and semiquantitative analyses of the chemical components of the seed coat and kernel of Ziziphi Spinosae Semen

Ziziphi Spinosae Semen refers to the dried seed of Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chou. The seed is composed of a reddish brown coat and a yellow kernel. A comparative study was conducted to investigate differences in the chemical composition and their relative contents between the seed coat and kernel of Ziziphi Spinosae Semen. First, the chemical compounds found in the seed coat and kernel were characterized and identified using ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). The analytical results tentatively identified 57 chemical compounds based on reference-compound comparison, literature retrieval, and chemical-database (e. g., MassBank) searches; these compounds included 14 triterpenes, 23 flavonoids, 7 alkaloids, 6 carboxylic acids, and 7 other types of compounds. The mass error of the identified compounds was within the mass deviation range of 5×10-6 (5 ppm). Next, two methods of multivariate statistical analysis, namely, principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), were used to compare the differential compounds between the two seed parts. A total of 17 differential compounds were screened out via OPLS-DA based on a variable importance in projection (VIP) value of >5. The results revealed that betulinic acid, betulonic acid, alphitolic acid, and jujuboside Ⅰ mainly existed in the seed coat whereas the 13 other compounds, such as spinosin, jujuboside A, and 6‴-feruloylspinosin, mainly existed in the seed kernel. Therefore, these 17 differential compounds can be used to distinguish between the two seed parts. Finally, a semiquantitative method was established using UPLC and a charged aerosol detector (CAD) with inverse gradient compensation in the mobile phase. Six representative compounds with different types were selected to examine the CAD response consistency: magnoflorine (alkaloid), spinosin (flavone), 6‴-feruloylspinosin (flavone), jujuboside A (triterpenoid saponin), jujuboside B (triterpenoid saponin), and betulinic acid (triterpenoid acid). The results showed that the relative standard deviation (RSD) of the average response factors at different levels of these six compounds was 7.04% and that their response intensities were similar. Moreover, each compound in the fingerprint demonstrated good response consistency, and the peak areas obtained directly reflected the contents of each compound. Based on the semiquantitative fingerprints obtained, betulinic acid and oleic acid were considered the main components of the seed coat. The betulinic acid content in the seed coat was approximately 7 times higher than that in the seed kernel. Spinosin, jujuboside A, linoleic acid, betulinic acid, and oleic acid were the main components of the seed kernel. The spinosin content in the seed kernel was 18 times higher than that in the seed coat. In addition, the jujuboside A content in the seed kernel was 24 times higher than that in the seed coat. The proposed method can accurately determine the main components and compare the relative contents of these components in different seed parts. In summary, this study identified the differences in chemical components between the seed coat and kernel of Ziziphi Spinosae Semen and clarified the main components and their relative contents in these parts. The findings can not only provide a basis for the identification of chemical compounds and quality research on different parts of Ziziphi Spinosae Semen but also promote the development and utilization of this traditional Chinese medicine.

Betulonic and Platanic Acids as a Basis for the Synthesis of New Therapeutically Effective Substances

Natural substances produced by plants represent an important source of new medicinal products. This article is focused on the description of the antimicrobial, antiviral, and cytotoxic activity of the synthesized derivatives of betulonic and platanic acids, especially their oxime-based derivatives. It is also aimed at providing overview of the possible use of these derivatives in the treatment of infectious or cancer diseases.

Chemical constituents of Syzygium cerasiforme leaves and their nitric oxide inhibitory activity in LPS‐activated RAW264.7 cells

AbstractA phytochemical study of the methanolic extract of Syzygium cerasiforme leaves led to the isolation of seven compounds (1–7). Their chemical structures were identified as benzyl‐6ʹ‐O‐galloyl‐β‐d‐glucopyranoside (1), 3‐methoxy‐4‐hydroxyphenol 1‐O‐β‐D‐(6ʹ‐O‐galloyl)‐glucopyranoside (2), betulonic acid (3), 3‐epi‐betulinic acid (4), 7α‐methoxy‐stigmast‐5‐en‐3β‐ol (5), dihydrodehydrodiconiferyl alcohol (6), and secoisolariciresinol (7), by HR‐ESI‐MS, 1D‐ and 2D NMR spectra, and by comparison with the previously published data. Compounds 1–7 exhibited NO production inhibitory activity in LPS‐activated RAW264.7 cells with IC50 values of 33.17, 25.51, 51.09, 54.17, 39.54, 6.98, and 58.12 µm, respectively, compared to that of the positive control compound, L‐NMMA, which showed an IC50 of 32.5 µm. This is the first report on these compounds from S. cerasiforme.

Oxime derivatives of betulonic acid and platanic acid as novel cytotoxic or antiviral agents

Less frequently studied plant triterpenoids betulonic acid and platanic acid were selected to design, synthesize and investigate their oxime derivatives as novel and potentially effective cytotoxic and/or antiviral agents.

Hepatotropic activity of a betulonic acid based compound.

Using the model of cyclophosphamide (CP)-induced immunosuppression in C57BL/6 mice, the hepatotropic effects of a conjugate of betulonic acid with 9-(4-methylpiperazin-1-ylmethyl)-2-(1,2,3-triazolyl) oreozelone (BABC) have been studied. In the liver of treated animals the expression of genes for cytochromes (CYP 1A1, CYP 1A2, CYP 3A44, CYP 2B10, CYP 2C29, CYP 17A1), PPARA, and cytokines (TNF-α, IL-1β, IL-12α, IL-10) and the relative levels of NF-κB p65, GST-π, and NAT-1 proteins were determined. On day six after administration of the compound and CP to animals a significant (3.2-fold) increase in the expression of the CYP 2B10 as compared to the control group was observed. Treatment of mice with the compound and CP also caused a 2.4-fold increase in the mRNA level of the pro-inflammatory TNF-α gene as compared to the group of animals receiving CP. Administration of the studied compound to intact animals was accompanied by a 2.5-fold increase in the IL-1β expression and a 1.8-fold decrease in the IL-10 expression as compared to the control group. An increase in the expression of pro-inflammatory cytokine genes in the liver of animals treated with the compound was accompanied by an increase in the content of NF-κB p65 (by 1.6 times), as well as an increase in the relative amount of NAT-1 protein (by 2.7 times) as compared to control animals.

Natural products from the culture broth of an actinobacteria Streptomyces sp. M5205 and evaluation of their antifungal activity

One previously undescribed sesquiterpene analog, M5205 (1), together with four known analogs, betulinic acid (2), betulin (3), messagenic acid D (4), and betulonic acid (5), were isolated from the EtOAc extract of the culture broth of the actinomycete Streptomyces sp. M5205. The structures of all isolates were elucidated on the basis of extensive analyses of spectroscopic data and comparison with literature data. Their antifungal activity was also evaluated. Our results showed that 4 constituents (Compounds 1 &3-5) possessed mild antifungal activity against Fusarium sp. LC8, Neopestalotiopsis sp. BCRC 35002, and Colletotrichum gloeosporioides BCRC 35178. It is worth mentioning that the chemical composition of the type strain Streptomyces sp. M5205 has never been studied.

A SYSTEMATIC REVIEW ON CHEMICAL AND PHARMACOLOGICAL ACTIVITIES OF EXCOECARIA AGALLOCHA: A POTENTIAL SUNDARBAN PLANT

Among various plants in the Mangrove Forest of the tropical and subtropical region, E. agallocha is notable for possessing multifarious pharmacological activity. As E. agallocha belongs to the Euphoriaceae family, this plant is rich in many classes of phytochemicals that are therapeutically potent such as Diterpenoids, Triterpenoids, Alkaloids, Polyphenol, Sterol, Flavonoids, Tanine, Flavanone, etc. Its ancient use in the treatment of disorders such as epilepsy, rheumatism, ulcer, leprosy, and others has attracted the interest of researchers, who are now studying different parts of the plant. Plant parts have been studied for reducing inflammation, anti-microbial, anti-epileptic, and cytotoxic effects in several research. Phytoconstituents responsible for pharmacological activities have been distinguished and their possible mechanism of action along with IC₅₀ value has been presented. From wood, roots, and stems of the plant physiologically active constituents such as Agallochin, ent-kauran-16â-ol-3-one, Chebulagic acid, Betulinic acid, Betulonic acid, Corilagin, Geraniin, Excoecarin, Agallochaexcoerin, Vanillic acid are described as pharmacologically potent molecules as they interact with certain biomolecules and precipitate changes in the physiological state of the body. This review is elucidating the likely mechanism of action of each of these plant compounds that provide particular biological activity.

Exploration of 1,2,3-triazolo fused triterpenoids as inhibitors of human coronavirus 229E targeting the viral nsp15 protein.

The coronavirus disease-19 (COVID-19) pandemic has raised major interest in innovative drug concepts to suppress human coronavirus (HCoV) infections. We previously reported on a class of 1,2,3-triazolo fused betulonic acid derivatives causing strong inhibition of HCoV-229E replication via the viral nsp15 protein, which is proposedly related to compound binding at an intermonomer interface in hexameric nsp15. In the present study, we further explored the structure-activity relationship (SAR), by varying the substituent at the 1,2,3-triazolo ring as well as the triterpenoid skeleton. The 1,2,3-triazolo fused triterpenoids were synthesized by a multicomponent triazolization reaction, which has been developed in-house. Several analogs possessing a betulin, oleanolic acid, or ursolic acid core displayed favorable activity and selectivity (EC50 values for HCoV-229E: 1.6-3.5 μM), but neither of them proved as effective as the lead compound containing betulonic acid. The 18β-glycyrrhetinic acid-containing analogs had low selectivity. The antiviral findings were rationalized by in silico docking in the available structure of the HCoV-229E nsp15 protein. The new SAR insights will aid the further development of these 1,2,3-triazolo fused triterpenoid compounds as a unique type of coronavirus inhibitors.

GC-MS and LC-MS/MS Analysis of Secondary Metabolites in the Methanolic Extract of Uncaria callophylla Blume ex Korth. Stems

Uncaria callophylla Blume ex Korth. is a wild plant species belonging to the Rubiaceae family. These plants thrive in the peat swamp forest of Kalimantan, Indonesia. The stems of Uncaria callophylla have been used by the Dayak Ngaju tribe to treat various diseases and to restore stamina while they work in the forest. The present study was conducted to identify the secondary metabolites in the methanolic extract of Uncaria callohylla stems to increase the knowledge about this plant and its potential in medicinal applications and drug discovery. The analysis was conducted using GC-MS and LC-MS/MS. GC-MS analysis was performed with the Agilent Technologies 7890 Gas Chromatography equipped with an Auto Sampler and 5975 Mass Selective Detector. The LC-MS analysis was carried out in the C18 column of the Waters Acquity UPLC system. MS analysis was performed with an electrospray ionization source (ESI) in positive and negative ion modes, and the compounds were identified using a UNIFI data processor with a mass spectrum library of natural active substances from the Waters Traditional Medical Scientific Library database based on UPLC/QTof MSE Data Acquisition. GC-MS analysis revealed the presence of 24 peaks, and nine of them showed quality 90% or higher, namely n-Hexadecanoic acid (16.63%), 1,3,7-Trimethyl-3,7-Dihydro-1H-Purine-2,6-Dione (13.07%), Hexadecanoic Acid, Methyl Ester; 1,2-Benzenedicarboxylic Acid, Diethyl Ester; 9,12-Octadecadienoic Acid, Methyl Ester; Methyl 14-Methylheptadecanoate; (9E,12E)-9,12-Octadecadienoic Acid; Z, E-3,13-Octadecadien-1-o1; and 2,6,10,14,18,22-Tetracosahexaene, 2,6,10,15,19,23-hexanethyl-, (all-E)-. LC-MS/MS analysis revealed the presence of five compounds, namely Arecatannin A1, Sweroside_2, Uncarine A, Epianhydrobelachinal, and Betulonic acid. All the secondary metabolites identified from GC-MS and LC-MS/MS analysis have biological activity, which indicates their medicinal potencies. From the result, it can be concluded that Uncaria callophylla stem methanolic extract contains various bioactive compounds justifies its traditional use, and therefore it can be developed further for new drug discovery.

Effective Three‐Step Construction of Betulonic Acid Hybrids with Heterocycle‐Containing Peptidomimetic Fragments

AbstractSynthesis of hybrid molecules containing a natural betulonic acid core and a peptidomimetic moiety was carried out by a three‐step transformation: e. g. (i) Ugi four‐component reaction involving heterocyclic amines to obtain some new azido‐substituted peptidomimetics; (ii) isolation of the compounds from the natural raw materials and obtaining betulonic acid propargyl esters; (iii) Cu‐catalyzed cycloaddition of peptide‐structured azides to betulonic acid propargyl esters, resulting in the construction of target molecules. The microbiological properties of the hybrid compounds and their precursors were tested against two Gram‐positive and two Gram‐negative bacterial strains.

Separation of betulin and its derivatives by high-performance liquid chromatography and their determination in extracts of some plants and chaga mushroom

The influence of conditions on the separation of betulin (BT), betulinic (BIA), and betulonic (BOA) acids by reversed-phase high-performance liquid chromatography (RP-HPLC) with isocratic elution was studied. It was shown that the order of peaks in chromatograms changed with varying the acetonitrile (ACN) content in the mobile phase, and a poor separation under certain conditions was observed. The highest peak resolution with minimal retention times was at a column temperature of 20 °C, flow rate of 0.25 ml/min, and 92.5% ACN in the mobile phase. The extracts from jujube (Ziziphus jujuba) dried fruit, chaga mushroom (Inonotus obliquus), and white birch bark (Betula pendula) were studied using the obtained conditions. For extracts from the first two sources, it was found that peaks of the compound studied interfered with unknown peaks. By varying the ACN content in the mobile phase with a small step from run to run and tracking the peaks, a baseline separation was achieved. The optimal % ACN in the mobile phase was 87 and 89 for the extracts from jujube and chaga mushroom, respectively. Jujube dried fruit was found to contain, in terms of dry weight of the jujube, 0.223 ± 0.008 mg/g of BIA and 0.044 ± 0.006 mg/g of BOA. Chaga mushroom studied contains 0.022 ± 0.004 mg/g of BT and 0.062 ± 0.009 mg/g of BIA. White birch bark contains 50.9 ± 0.7 mg/g of BT, 11.2 ± 0.3 mg/g of BIA, and 2.5 ± 0.3 mg/g of BOA.