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.

Immediate-release Mechanism of Dehydrotrametenolic Acid and Dehydroeburicoic Acid in Poriae Cutis Total Triterpenoids Tablets Based on Liquid-solid Compact Technique

[Objectives]To study the immediate-release mechanism of dehydrotrametenolic acid and dehydroeburicoic acid in Poriae Cutis total triterpenoids tablets by liquid-solid compacts technique.[Methods] Taking dehydrotrametenolic acid and dehydroeburicoic acid as indicators,differences in dissolution were compared between liquid-solid compressed tablets and crude drug of total triterpenoids in Poriae Cutis;crude drug,powder of liquid-solid compressed tablets and excipients of liquid-solid compressed tablets were characterized by differential scanning calorimetry( DSC).[Results]Liquid-solid compact technique could significantly improve dissolution rate of dehydrotrametenolic acid and dehydroeburicoic acid. The total dissolution rate of dehydrotrametenolic acid and dehydroeburicoic acid in Poriae Cutis total triterpenoids tablets was 92%,t50( time for 50% total dissolution rate) and t_D( time for 63. 2% total dissolution rate) were 11. 18 min and 22. 71 min; the total dissolution rate of dehydrotrametenolic acid and dehydroeburicoic acid in crude drug of total triterpenoids was 29%,and t50 and t_D were231. 06 min and 359. 23 min. DSC showed that there was no mutual interaction between excipients and total triterpenoids in Poriae Cutis; on the DSC curve for powder of liquid-solid compressed tablets,the absorption peak vanished completely,indicating that the drug exists in the amorphous form in the liquid-solid powder.[Conclusions] Liquid-solid compact technique can increase the dissolution rate of total triterpenoids in Poriae Cutis and allow rapid release of poorly water soluble drugs.

Optimization of Extraction Process and Determination of Total Triterpenoids from Semen Trichosanthis by Response Surface Methodology

[Objectives] To optimize the extraction process and determine the total triterpenoids from Semen Trichosanthis.[Methods]The Box-behnken response surface methodology was applied to optimize the extraction conditions of total triterpenoids in Semen Trichosanthis.And the spectrophotometry was used to determine the content of total triterpenoids in Semen Trichosanthis.[Results] The optimal extraction conditions were solid to liquid ratio of 40∶ 1,ultrasonic time of 20 min,and ultrasonic power of 100 W.Under these conditions,the content of total triterpenoids in Semen Trichosanthis was significantly different among the 10 production areas,among which the No.9 production area(Anhui 1) had the highest total triterpenoids content of 45.71 mg/g,while No.7 production area(Shandong) had the lowest total triterpenoids content of 15.22 mg/g.[Conclusions]The Box-behnken response surface methodology is reliable for the extraction of total triterpenoids in Semen Trichosanthis,and the spectrophotometry is proper for determining the content of total triterpenoids in Semen Trichosanthis.There are large differences in the content of total triterpenoids in Semen Trichosanthis produced in different areas.

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.

Optimization of Extraction Method of Triterpenoids from Artemisia selengnesis Turcz

[Objectives]The research aimed to optimize the extraction method of triterpenoids from A. selengnesis. [Methods]The ultrasonic extraction of triterpenoids in A. selengnesis was optimized by single factor analysis( including infusion time,ethanol concentration,ultrasonic power,extraction time,and liquid-solid ratio) and response surface method. [Results]The optimum extracting process of triterpenoids was as follows: infusion time of 65 min,ethanol concentration of 80%,ultrasonic power of 275 W,extraction time of 30 min,and liquid-solid ratio of 34 m L/g. Under the optimized condition,the extraction content of triterpenoids reached( 18. 26 ± 0. 10) mg/g. [Conclusions] The optimized extraction had a good modal on practice in developing and utilization.

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.

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.

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.

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 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).

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.

Intestinal Absorption of Ergostane and Lanostane Triterpenoids from Antrodia cinnamomea Using Caco-2 Cell Monolayer Model

Antrodia cinnamomea is a precious medicinal mushroom.It exhibits promising therapeutic effects on cancer,intoxication,hypertension,hepatitis,and inflammation.Its major bioactive constituents are ergostane and lanostane triterpenoids.In this study,we used intestinal Caco-2 cell monolayer model to reveal the intestinal absorption property of 14 representative triterpenoids from A.cinnamomea.The bidirectional transport through the monolayer at different time points was monitored by a fully validated LC/MS/MS method.In the case of pure compounds,ergostanes 5(25R-antcin H),6(25Santcin H)and 10(25R-antcin B)could readily pass through the Caco-2 cell layer,whereas lanostanes 13(dehydroeburicoic acid)and 14(eburicoic acid)could hardly pass through.When the cells were treated with A.cinnamomea extract,antcins A,B,C,H and K(1–6 and 9–11)were absorbed via passive transcellular diffusion,and showed high PAB and PBA values(>2.5×10^(-5) cm/s).Meanwhile,the lanostanes dehydrosulphurenic acid(8),15a-acetyldehydrosulphurenic acid(12),13 and 14 exhibited poor permeability.Transport features of these compounds were consistent with their pharmacokinetic behaviors in rats.This study could also be helpful in predicting the intestinal absorption of A.cinnamomea in human.

A Review of Triterpenoids and Their Pharmacological Activities from Genus Kadsura

Plants in genus Kadsura,belong to the Schisandraceae family including genus Schisandra and genus Kadsura,revealed folk efficacy and medicinal application in the treatment of rheumatoid arthritis and gastroenteric disorders for a long history.Previous study indicated that plants from genus Kadsura mainly contain lignans,triterpenoids and essential oils,and showed the active compounds were lignans and triterpenoids.Among all compounds from Schisandraceae family,the triterpenoids between the genus Schisandra and genus Kadsura had obvious differences.In this paper,we reviewed214triterpenoids and their pharmacological activities reported from genus Kadsura in the past30years(from1987to2017),aiming to provide reference for further study.

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.

Limonoids and triterpenoids from the twigs and leaves of Dysoxylum hainanense

Four new limonoids,dysohainanins A-D(1-4),and two new triterpenoids,dysohainanins E and F(5 and 6),together with seven known ones were isolated from the twigs and leaves of Dysoxylum hainanense Merr.The structures of the new compounds were determined by a variety of spectroscopic methods.The cytotoxic activities of these compounds were evaluated,and the known compound ent-19-nor-4,16,18-trihydroxy-8(14)-pomaren-15-one(13)showed in vitro cytotoxicity against HL-60,A-549,MCF-7,and SW480 cells,with IC50 values of 24.3,28.1,30.7,and 22.5mM,respectively.Compounds 2 and 3 were tested their insecticidal activities using brine shrimp and both of them were inactive.

Triterpenoids and Semisynthetic Derivatives with Antimicrobial Activities from the Leaves of <i>Caloncoba glauca</i>(Flacourtiaceae)

Chemical investigation of the MeOH extract from the leaves of C. glauca yielded nine known triterpenoids (1-9) belonging to the cycloartane and friedelane series. Two of these compounds namely glaucartanoic acid A (1) and 3β,21β-dihydroxy-30-nor-(D:A)-friedoolean-20(29)-en-27-oic acid (5) were subjected to chemical derivatizations and afforded five new derivatives: diacetylglaucartanoic acid A (1a), 24-acetylglaucartanoic acid A (1b), glaucartanoic acid A methyl ester (1c), 24-methoxyglaucartanoic acid A methyl ester (1d), and 3β,21β-diacetoxy-30-nor-(D:A)-friedoolean-20(29)-en-27-oic acid (5a). Their structures were assigned based on their NMR and MS data and by comparison with literature values. The MeOH extract, isolated compounds and some new semi-synthetic derivatives were subjected to in vitro antimicrobial assays against a panel of pathogenic microorganisms, including Gram-positive and Gram-negative bacteria, and fungi using broth microdilution method. The MeOH extract displayed activity towards all the tested pathogenic bacterial and fungal strains with good activity (MIC < 100 μg/mL) against Staphylococcus aureus ATCC25923 and Shigella flexneri SDINT. Compounds 3 and 5 showed the most potent antimicrobial effect.

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.

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.

Lactam Triterpenoids from the Bark of Toona sinensis

Three new limonoid-type triterpenoids,namely toonasins A–C(1–3)with a rare lactam E ring,along with six known compounds(4–9)were isolated from the barks of Toona sinensis.The structures of new compounds were elucidated by interpretation of spectroscopic data,and the relative configuration of compound 1 was further characterized by X-ray crystallographic analyses.The isolated compounds were evaluated for their cytotoxic activities against five human tumor cell lines(HL-60,SMMC-7721,A-549,MCF-7 and SW480),and compounds 3 and 5 showed weak cytotoxicities.