Chemical derivatization strategies for enhancing the HPLC analytical performance of natural active triterpenoids

Triterpenoids widely exist in nature,displaying a variety of pharmacological activities.Determining triterpenoids in different matrices,especially in biological samples holds great significance.High-performance liquid chromatography(HPLC)has become the predominant method for triterpenoids analysis due to its exceptional analytical performance.However,due to the structural similarities among botanical samples,achieving effective separation of each triterpenoid proves challenging,necessitating significant improvements in analytical methods.Additionally,triterpenoids are characterized by a lack of ultraviolet(UV)absorption groups and chromophores,along with low ionization efficiency in mass spectrometry.Consequently,routine HPLC analysis suffers from poor sensitivity.Chemical derivatization emerges as an indispensable technique in HPLC analysis to enhance its performance.Considering the structural characteristics of triterpenoids,various derivatization reagents such as acid chlorides,rhodamines,isocyanates,sulfonic esters,and amines have been employed for the derivatization analysis of triterpenoids.This review comprehensively summarized the research progress made in derivatization strategies for HPLC detection of triterpenoids.Moreover,the limitations and challenges encountered in previous studies are discussed,and future research directions are proposed to develop more effective derivatization methods.

Four new lanostane triterpenoids featuring extendedπ-conjugated systems from the stems of Kadsura coccinea

Four new 14(13→12)-abeolanostane triterpenoids featuring extendedπ-conjugated systems,kadcoccitanes E-H(1-4),were obtained from the stems of Kadsura coccinea through using a HPLC−UV-guided approach.Their structural and configurational determination was accomplished through extensive spectroscopic analysis coupled with quantum chemical calculations.Kadcoccitanes E-H were tested for their cytotoxic activities against five human tumor cell lines(HL-60,A-549,SMMC-7721,MDA-MB-231,SW-480)but none of them exhibited activities at the concentration 40μM.

Triterpenoids from the Dai Medicinal Plant Winchia calophylla

A new compound (1) named as winchic acid and eight known ones were isolated from the stem barks of Winchia colophylla A. DC. (collected in Xishuangbanna, Yunnan Province of China). The new compound 1 was identified as 3beta-hydroxy-27-(4-hydroxy-3-methoxy-E-cinnamoyloxy)-lup-20(29)-en-28-oic acid on the basis of 1D and 2D NMR experiments. The eight known compounds were determined to be lupenone, lupenyl acetate, betulinic acid, alpha-amyrin acetate, ursolic acid, ptiloepoxide, beta-amyrin and cycloeucalenol, respectively.

Triterpenoids from Rubus buergeri

A new triterpenoid, methyl buergericate (12a), which was elucidated as 2 alpha, 3 alpha, 19 alpha, 24-tetrahydroxyl olean-12-en-28-oic acid methyl ester by spectroscopic methods, together with eleven known triterpenoids was isolated from Rubus buergeri Miq. The isolation of the above chemical constituents from this plant was reported for the first time.

Triterpenoids from the Fruiting Body of Ganoderma lucidum

A new triterpenoid, named ganolactone, was isolated from the CH 2Cl 2 soluble fraction of the fruiting body of Ganoderma lucidum. Its structure was deduced as 7β hydroxy 3,11,15 trioxo lanosta 8 en 24→20s lactone (1) on the basis of spectral analysis (UV, IR, MS. 1H NMR, 13 C NMR and 2D NMR) and was confirmed by X ray diffraction. In addition, three known compounds, ganoderiol A (2), ganoderiol B (3) and ganodermatriol (4), were obtained.

Determination of ganoderic acids in rat plasma after oral administration of total triterpenoids from Ganoderma lucidum

To investigate the absorption of total triterpenoids from Ganoderma lucidum in rats. HPLC-DAD and LC-MS methods were used to identify ganoderic acids in rat plasma after oral administration of total triterpenoids from G. lucidum by comparing their HPLC retention behaviors, UV absorption spectra, and mass spectra with authentic samples. Five ganoderic acids, ganoderic acid C2, C6, G, B and A were simultaneously detected in rat plasma. Ganoderic acids can be directly absorbed into circulation after oral administration of total triterpenoids from G. lucidum in rats.

Two New Lanostane-type Triterpenoids from Mulgedium tataricum

Two new triterpenoids, lanost-9（11）, 23Z（24）-dicnc-3β, 25-diol （1） and lanost-9（11）, 25- diene-3β, 24-diol （2） were isolated from Mulgedium tataricum L. Their structures wcrc elucidated by spectral methods.

Analysis of three types of triterpenoids in tetraploid white birches(Betula platyphylla Suk.) and selection of plus trees

Betulin, oleanolic acid, and betulinic acid are naturally occurring pentacyclic triterpenoids that have significant medicinal value. Considerable amounts of these triterpenoids are available in the outer bark of white birch. In this study, we used ultrasound-assisted extraction （UAE） to extract triterpenoids from birch bark rapidly and with high efficiency. Using high performance liquid chro- matography （HPLC）, three types of triterpenoids were separated and detected. We examined the differences among triterpenoids extracted from diploid versus tetra- ploid white birch. Then, we used factor analysis to screen out tetraploid white birches with comprehensively excel- lent performance. The results indicate that the optimum conditions for extraction include the use of ethanol as an extraction solvent, a solid-to-liquid ratio of 0.1 g/10 ml, ultrasonic power set at 100 W, a temperature of 60 ℃ and an extraction time of 15 min. A reversed-phase C18 col- umn （4.6 mm × 250 mm × 5 μm） with a column tem- perature of 30 ℃ and the mobile phase composed of A （acetonitrile） and B （0.1% aqueous phosphoric acid, v/v） at a flow rate of 0.5 ml/min were used, and the detection wavelength was 195 nm. No significant difference wasobserved between diploid and tetraploid white birch in terms of the content of three types of triterpenoids （at a confidence level of 0.05）. As triterpenoid content, height, and DBH （diameter at breast height） are strongly interre- lated, we used factor analysis to evaluate all individuals, and we screened out six plus trees with excellent com- prehensive characters.

Advances in Research on Triterpenoids and Volatile Compositions of Kudingcha

Kudingcha, a commonly used traditional Chinese medicine, has multiple functions in anti-radiation, anti-aging, quickening the blood vessels, reducing blood lipid and blood pressure and so forth. Therefore, this article reviews the origins of Kudingcha, and analyzes the major triterpenoids and volatile chemicals, with an at- tempt to provide more references for the development and utilization of Kudingcha.

Cucurbitane triterpenoids from Hemsleya penxianensis

Two new cucurbitacins,jinfushanencins A(1)and B(2),seven new cucurbitane glycosides,jinfushanosides E-K(3-9),along with nine known analogues,were obtained from the tubers of Hemsleya penxianensis.Their structures were elucidated on the basis of extensive spectroscopic and chemical methods.Selected isolates were tested their anti-HIV-1 activities,and compound 5 showed weak anti-HIV-1 in C8166 cell(EC50=5.9μg/mL)with a selectivity index of 13.5.

Intercedenol A and B,Two New Triterpenoids from the Sea Cucumber Mensamaria intercedens

Two new triterpenoids named intercedenol A and B were isolated from the acid hydrolysate of the crude glycoside fraction. The structures were elucidated by ESI-MS and NMR spectrum.

Two New Triterpenoids from Saussurea petrovii

Two new taraxastane-type triterpenes. 3 beta. 21 beta -dihydroxyl-20(30)-en-taraxastane 1 and 3 beta. 30-dihydroxy-20(21)-en-taraxastane 2. were isolated from Saussurea petrovii. Their structures were elucidated by spectral methods (EIMS. IR. 1D and 2DNMR).

Natural variations in flavonoids and triterpenoids of Cyclocarya paliurus leaves

Cyclocarya paliurus is widely distributed in subtropical areas of China.Secondary metabolites in the leaves,including flavonoids and triterpenoids,provide protection against diseases such as hyperlipidemia,diabetes,hypertension,and hypoimmunity.In this study,343 C.paliurus leaves were collected from 32 distinct populations covering most of the main distribution,to deduce the response of flavonoids and triterpenoids to seven environmental factors.Principal component analysis was performed using seven flavonoid and six triterpenoid compounds.The results show that altitude,latitude,and longitude significantly contributed to variations in total flavonoid and triterpenoid contents.In addition,the interaction of these metabolites was extensively correlated with environment changes,and therefore should be considered during selection and cultivation.

Two new tirucallane triterpenoids from Aphanamixis grandifolia

Two new tirucallane triterpenoids,2a-ethoxy-2,3-secotirucalla-2,29-epoxy-7-ene-23-oxo-3-oic acid(1)and(23E)-2a-hydroxytirucalla-7,23,25-triene-3-one(2),along with the known 2,3-secotirucalla-2,3;2,29-diepoxy-7-ene-3,23-dione(3),were isolated from the leaves and twigs of Aphanamixis grandifolia.Their structures were elucidated by extensive NMR and MS data,and compound 3 was further confirmed by X-ray crystal diffraction analysis.Antimicrobial activities and insecticidal activities of these three compounds were also evaluated.Compound 1 showed good antimicrobial activity against Staphylococcus aureus with the MIC value of 1.56μg/mL,while compounds 1 and 2 showed insecticidal activity at 100 ppm,with the corrected mortality 79.1%and 60.6%,respectively.

Six New 9,19-Cycloartane Triterpenoids from Cimicifuga foetida L.

Six new 9,19-cycloartane triterpene derivatives,as well as 3 known analogues(7–9),were isolated from the roots of Cimicifuga foetida L.Their structures were established on the basis of extensive spectroscopic analyses(IR,UV,ORD,HRESIMS,1D and 2D NMR).

Lanostane Triterpenoids from Fruiting Bodies of Ganoderma leucocontextum

Six new lanostane-type triterpenoids,namely leucocontextins S-X(1-6),together with twelve known compounds,were isolated from the fruiting bodies of Ganoderma leucocontextum.Their structures were established by MS and NMR data.

Protective effects of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja on STZ-stimulated INS-1 cells through regulating of autophagy and apoptosis

OBJECTIVE To explore the protective effects of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja on STZ-stimulated INS-1 cells through regulating of autophagy and apoptosis.METHODS INS-1cells were cultured in media containing 3n M STZ and different doses of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja.The proliferation of cells was examined by MTT assay,ROS content were detected by fluorescence enzyme label.The levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),hydrogen peroxidase(CAT),malondialdehyde(MDA)were also measured by colorimetry.The activities of caspase-3,9 were also observed by Caspase colorimetric assay kit in each group.The expressions of Bcl-2 m RNA and Bax m RNA were detected by Real time PCR,protein expression of LC3-Ⅱand PARP were detected by Western blotting.RESULTS Compared with the control group,total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja could promote the proliferation of INS-1 cells no matter with STZ or not when its concentration lower than 25μg·m L-1;but when its concentration higher than 100μg·m L-(1use individually)or 50μg·m L-1(combined use),total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja might significantly inhibite the growth of the cells whether STZ existed or not.Total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja(6.25,12.5,25μg·m L-1)might inhibit INS-1cell apoptosis,decrease intra-cellular ROS contents,improve the s upernatant liquid SOD,GSH-PX,CAT activities,decrease MDA level,promote INS-1 cell secreting insulin,decrease the protein expressions of autophagy protein LC3Ⅱand apoptosis regulating protein cleaved PARP protein,up-regulate the anti-apoptotic protein Bcl-2 m RNA expression,down-regulate the pro-apoptotic protein Bax m RNA expression and Bcl-2and Bax,reduce the activity of caspase-9 and caspase-3(P<0.05 or P<0.01).CONCLUSION Total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja has good protective effect on STZ-stimulated INS-1cells.It can inhibit STZ injured INS-1 cells to overproduce ROS production,enhance endogenous antioxidant enzymes(GSH-Px,SOD,CAT activities),reduce the expression of autophagy protein LC3Ⅱand apoptosisregulating protein cleaved PARP,up-reglute the antiapoptosis protein Bcl-2 expression,down-regulate the pro-apoptotic protein Bax expression,decrease the caspase-9 and caspase-3 activities,and improve the INS-1cell survival rate,and then play a protective effect on damaged INS-1 cells.

Four 14(13→12)‑Abeolanostane Triterpenoids with 6/6/5/6‑Fused Ring System from the Roots of Kadsura coccinea

Four new rearranged 6/6/5/6-fused lanostane-type triterpenoids,kadcoccitanes A-D(1-4),were isolated from the roots of Kadsura coccinea,and their structures were mainly elucidated by comprehensive analysis of their spectroscopic data.Additionally,the structure of 1 was ambiguously verifed by single-crystal X-ray difraction,while the structure of 2,which features a novel 8,16-epoxy motif,was validated by quantum chemical calculation of NMR parameters and ECD spectrum.Moreover,1 and 4 were found to exhibited anticoagulant activity,while 3 and 4 were found to possess anti-platelet aggregation activity.

New Dammarane Triterpenoids,Caffruones A–D,from the Cherries of Coffea arabica

In present study,four new dammarane-type triterpenoids,namely caffruones A–D(1–4),were isolated from the cherries of Coffea arabica.Their structures were elucidated by extensive spectroscopic analysis including 1D,2D NMR(HSQC,HMBC,^(1)H–^(1)H COSY,and ROESY),HRMS and IR spectra.This is the first time that tetracyclic triterpenes have been reported in genus Coffea.

Three new triterpenoids from Rubia schumanniana

Three new triterpenoids,3β-hydroxy-urs-30-p-Z-hydroxycinnamoyl-12-en-28-oic-acid(1),3β-hydroxy-olean-30-p-E-hydroxycinnamoyl-12-en-28-oic-acid(2)and 3β,6α-dihydroxy-urs-14-en-12-one(3),together with seven known triterpenoids,were isolated from the roots of Rubia schumanniana.Their structures were established by means of spectroscopic analysis.All compounds were evaluated for cytotoxic activity,and compounds 2-6 showed cytotoxicity with the IC50 values of 10.75~18.87μg/mL.