A sensitive HPLC–MS/MS method for the simultaneous determination of anemoside B4, anemoside A3 and 23‐hydroxybetulinic acid: Application to the pharmacokinetics and liver distribution of Pulsatilla chinensis saponins

Pulsatilla chinensis saponins, the major active components in the herb, have drawn great attention as potential hepatitis B virus infection and hepatoma treatments. Here, a sensitive and accurate HPLC–MS/MS method was established for simultaneous determination of three saponins – anemoside B4, anemoside A3 and 23‐hydroxybetulinic acid – in rat plasma and liver, and fully validated. The method was successfully applied to a pharmacokinetics and liver distribution study of P. chinensis saponins. Consequently, 23‐hydroxybetulinic acid, with an extremely low content in the P. chinensis saponins, exhibited the highest exposure in the liver and in sites before and after hepatic disposition, namely, in the portal vein plasma and systemic plasma, followed by anemoside B4, which showed the highest content in the herb, whereas anemoside A3 displayed quite limited exposure. The hepatic first‐pass effects were 71% for 23‐hydroxybetulinic acid, 27% for anemoside B4 and 37% for anemoside A3, corresponding to their different extents of liver distribution. To our knowledge, this is the first investigation on the liver first‐pass effect and distribution of P. chinensis saponins to date. These results also provide valuable information for the understanding of the pharmacological effect of P. chinensis saponins on liver diseases.     

Synthesis,Formulation, and Adjuvanticity of Monodesmosidic Saponins with Olenanolic Acid,Hederagenin and Gypsogenin Aglycones,and some C‐28 Ester Derivatives

In an attempt to discover a new synthetic vaccine adjuvant, the glycosylation of hederagenin, gypsogenin, and oleanolic acid acceptors with di‐ and trisaccharide donors to generate a range of mimics of natural product QS‐21 was carried out. The saponins were formulated with phosphatidylcholine and cholesterol, and the structures analyzed by transmission electron microscopy. 3‐O‐(Manp(1→3)Glcp)hederagenin was found to produce numerous ring‐like micelles when formulated, while C‐28 choline ester derivatives preferred self‐assembly and did not interact with the liposomes. When alone and in the presence of cholesterol and phospholipid, the choline ester derivatives produced nanocrystalline rods or helical micelles. The effects of modifying sugar stereochemistry and the aglycone on the immunostimulatory effects of the saponins was then evaluated using the activation markers MHC class II and CD86 in murine bone marrow dendritic cells. The most active saponin, 3‐O‐(Manp(1→3)Glcp)hederagenin, was stimulatory at high concentrations in cell culture, but this did not translate to strong responses in vivo.     

Facile Synthesis of Four Natural Triterpene Saponins with Important Antitumor Activity

The first synthesis of four natural triterpene saponins, which exhibit significant antitumor activities, was concisely achieved by adopting a stepwise glycosylation. The key intermediate 13 was afforded via Bu₂SnO-mediated regioseletive benzoylation. During the preparation of the target compounds, it was found that the α-L-arabinopyranosyl unit in intermediates 17 and 20 existed in the unusual ¹ C ₄ conformation, and after removing the benzoyl groups, the α-L-arabinopyranosyl unit was normal in a typical ⁴ C ₁ form.     

Quantification and identification of bioactive metabolites from Kalopanacis Cortex by HPLC with evaporative light scattering detection and ESI quadrupole TOF MS

A method based on HPLC coupled with an evaporative light scattering detection and ESI quadrupole TOF MS was established for the quantification and identification of phenolics and triterpene saponins in Kalopanacis Cortex using a gradient elution of acetonitrile with 0.1% formic acid and water with 0.1% formic acid on an RP C₁₈ column (4.6 × 250 mm, 5 μm). Diverse validation parameters, such as the linearity, LOD and LOQ, accuracy, precision, repeatability, and stability, were successfully obtained. Additionally, the efficiencies of different extraction methods were compared. The developed method was applied for the quantitative analysis of twelve representative metabolites in 61 Kalopanacis Cortex samples. The quantitation results showed that coniferin, kalopanaxsaponin C, septemlosides II, III, C, and D exhibited distinct regional patterns in Kalopanacis Cortex samples. These six compounds including one new triterpene saponin show potential as marker compounds for evaluating the quality of Kalopanacis Cortex and the geographical variation in its chemical composition.     

毛冬青总皂苷对AHNP诱导肝损伤大鼠骨中7种微量元素的影响

目的探讨了毛冬青总皂苷对AHNP诱导肝损伤大鼠骨中微量元素的影响。方法应用逆行胰胆管术注射牛磺胆酸钠（TAC）复制急性出血坏死性胰腺炎（AHNP）大鼠实验动物模型,进行了毛冬青总皂苷（高、低剂量）的治疗。观察了5组（假手术对照组、AHNP模型组、毛冬青总皂苷高剂量组、毛冬青总皂苷低剂量组、尼尔雌醇组、）大鼠股骨中Cu、Mn、Fe、Zn、Pb、Mg和ca等微量元素的变化。结果统计学结果显示与AHNP模型组比较,毛冬青总皂苷能提高AHNP大鼠骨质中cu、Fe、Mn、Mg、Ca等元素的含量,尼尔雌醇组中的zn含量提高,毛冬青总皂苷组中的zn含量反而下降;尼尔雌醇组与毛冬青总皂苷组中Pb含量下降明显。结论说明毛冬青总皂苷的治疗作用并不是单一的,可以认为中药毛冬青总皂苷是通过发挥对人体整体的调节作用来调节骨骼中微量元素的变化,改变骨质的代谢情况,促进骨密度的提高。     

Revised structures of Arillatanosides A–C from Polygala arillata

Arillatanosides A–C are three triterpenoid saponins from Polygala arillata Buch–Ham that have been reported previously, but with partially incorrect structures. Further investigation of their NMR data led to the conclusion that the terminal α‐L ‐arabinopyranosyl unit originally proposed for Arillatanosides A–C ( I – III ) is actually a β‐D ‐xylopyranosyl unit. Thus, the correct structures of Arillatanosides A–C are represented by 1 – 3 . Complete NMR assignments of Arillatanosides A–C ( 1 – 3 ) and the related polygalasaponin XXXV ( 4 ) were achieved using modern 2D NMR techniques, such as DQF H–H COSY, HMQC, HMBC, TOCSY, 2D HMQC–TOCSY. Copyright © 2002 John Wiley & Sons, Ltd.     

Three new steroidal saponins from Helleborus thibetanus

Three new steroidal saponins including two spirostanol glycosides (1–2) and one furostanol glycoside 1-sulphate (3) were isolated from the dried roots and rhizomes of Helleborus thibetanus. Structures of the compounds were determined on the basis of extensive use of 1-D and 2-D NMR experiments, together with HR–ESI–MS and IR measurements, as well as the results of acid hydrolysis. Compounds 1–2 represented steroidal saponins with an unusual substitution pattern, which possessed a double bond at C-25 and were glycosylated at 1-OH.     

Three new triterpenoid saponins from the roots of Ardisia crenata and their cytotoxic activities

Three new triterpenoid saponins, ardisicrenoside O (1), ardisicrenoside P (2) and ardisicrenoside Q (3) together with three known compounds, 3β,16α-dihydroxy-30-methoxy-28, 30-epoxy-olean-12-en, cyclamiretin A 3-O-β-d-glucopyranosyl-(1→2) -α-l-arabinopyranoside and cyclamiretin A 3-O-β-d-glucopyranosyl-(1→4) -α-l-arabinopyranoside were isolated from the roots of Ardisia crenata Sims. Their structures were determined by one- and two-dimensional NMR techniques, including HSQC, HMBC and TOCSY experiments, as well as acid hydrolysis and GC analysis. All isolates were evaluated for the cytotoxic activities on two human cancer cell lines and compounds 3, 5 and 6 showed significant cytotoxicity.     

Efficient method for large-scale preparation of two components H and I of Sg-6 saponins from whole seeds of wild soybean (Glycine soja Sieb. and Zucc.)

New saponin components, Sg-6 saponins, have been recently reported from the seeds of wild soybean (Glycine soja) which may have specific health benefits. To evaluate the possible health benefits, a large amount of Sg-6 saponins are needed, but general group A acetyl saponins and new Sg-6 saponins are eluted in overlapping peaks by ordinal preparative high-performance liquid chromatography and/or open column methods. A new method is proposed in this report. This method includes (1) deacetylation of group A acetyl saponins in alkali condition with KOH, (2) precipitation of Sg-6 saponins in acid condition with HCl, (3) recovery of Sg-6 saponins with aqueous methanol from the precipitate, and (4) elution of Sg-6 saponins by preparative reverse-phase open column. With this method, from 450?g of wild soybean whole seed powder, about 1?g of Sg-6 saponins (mixture of six components) was clearly separated from other saponins with 61% recovery.     

Three Escin‐Like Triterpene Saponins: Assamicins VI,VII, and VIII from the Seeds of Aesculus assamica Griff

Three new escin‐like triterpene saponins, assamicins VI ( 1 ), VII ( 2 ), and VIII ( 3 ), were isolated from the seeds of A. assamica, together with a known saponin, isoescin Ib ( 4 ). Their structures were established as 28‐O‐acetyl‐21‐O‐(3,4‐di‐O‐angeloyl‐6‐deoxy‐β‐glucopyranosyl)‐3‐O‐{O‐β‐glucopyranosyl‐(1→2)‐O‐[β‐glucopyranosyl‐(1→4)]‐β‐glucopyranuronosyl}protoaescigenin ( 1 ), 21‐O‐angeloyl‐3‐O‐{O‐α‐rhamnopyranosyl‐(1→2)‐O‐[β‐glucopyranosyl‐(1→3)]‐β‐glucopyranuronosyl}protoaescigenin ( 2 ), and 21‐O‐angeloyl‐3‐O‐{O‐[β‐glucopyranosyl‐(1→3)]‐β‐glucopyranuronosyl}protoaescigenin ( 3 ) on the basis of spectroscopic analysis (protoaescigenin=(3β,4β,16α,21β,22α)‐olean‐12‐ene‐3,16,21,22,23,28‐hexol; angelic acid=(2Z)‐2‐methylbut‐2‐enoic acid).