Asponchimides A-E: new enantiomeric N-acetyldopamine trimers from Aspongopus chinensis / 中国天然药物

Five new racemic N-acetyldopamine (NADA) trimers, asponchimides A-E (1-5), were isolated from Aspongopus chinensis, a prominent traditional Chinese medicinal insect employed for alleviating pain, treating indigestion, and addressing kidney ailments. Compounds 1-5 were successfully resolved by chiral high-performance liquid chromatography (HPLC), yielding five pairs of enantiomers: (+)- and (-)-asponchimides A-E (1a/1b-5a/5b). Their structural identities were discerned by extensive spectroscopic analyses, including high-resolution mass spectrometry (HRMS), ultraviolet-visible (UV-Vis) spectroscopy, infrared (IR) spectroscopy, and nuclear magnetic resonance (NMR), and their absolute configurations were determined by electronic circular dichroism (ECD) calculations. Compounds 1-5 are pioneering instances of NADA trimers featuring a Δ7 double bond. When subjected to a series of bioassays, a majority of the compounds exhibited weak inhibitory activity against nitric oxide (NO) production in LPS-induced RAW 264.7 cells.

Targeted trace ingredients coupled with chemometric analysis for consistency evaluation of Panax notoginseng saponins injectable formulations / 中国天然药物

Evaluating the consistency of herb injectable formulations could improve their product quality and clinical safety, particularly concerning the composition and content levels of trace ingredients. Panax notoginseng Saponins Injection (PNSI), widely used in China for treating acute cardiovascular diseases, contains low-abundance (10%-25%) and trace saponins in addition to its five main constituents (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, and ginsenoside Rd). This study aimed to establish a robust analytical method and assess the variability in trace saponin levels within PNSI from different vendors and formulation types. To achieve this, a liquid chromatography-triple quadrupole mass spectrometry (LC-MS/MS) method employing multiple ions monitoring (MIM) was developed. A "post-column valve switching" strategy was implemented to eliminate highly abundant peaks (NR1, Rg1, and Re) at 26 min. A total of 51 saponins in PNSI were quantified or relatively quantified using 18 saponin standards, with digoxin as the internal standard. This study evaluated 119 batches of PNSI from seven vendors, revealing significant variability in trace saponin levels among different vendors and formulation types. These findings highlight the importance of consistent content in low-abundance and trace saponins to ensure product control and clinical safety. Standardization of these ingredients is crucial for maintaining the quality and effectiveness of PNSI in treating acute cardiovascular diseases.

Offline two-dimensional liquid chromatography coupled with ion mobility-quadrupole time-of-flight mass spectrometry enabling four-dimensional separation and characterization of the multicomponents from white ginseng and red ginseng / 药物分析学报

Inherent complexity of plant metabolites necessitates the use of multi-dimensional information to accomplish comprehensive profiling and confirmative identification. A dimension-enhanced strategy, by offline two-dimensional liquid chromatography/ion mobility-quadrupole time-of-flight mass spec-trometry (2D-LC/IM-QTOF-MS) enabling four-dimensional separations (2D-LC, IM, and MS), is proposed. In combination with in-house database-driven automated peak annotation, this strategy was utilized to characterize ginsenosides simultaneously from white ginseng (WG) and red ginseng (RG). An offline 2D-LC system configuring an Xbridge Amide column and an HSS T3 column showed orthogonality 0.76 in the resolution of ginsenosides. Ginsenoside analysis was performed by data-independent high-definition MSE (HDMSE) in the negative ESI mode on a Vion TM IMS-QTOF hybrid high-resolution mass spectrometer, which could better resolve ginsenosides than MSE and directly give the CCS information. An in-house ginsenoside database recording 504 known ginsenosides and 58 reference compounds, was estab-lished to assist the identification of ginsenosides. Streamlined workflows, by applying UNIFI TM to auto-matedly annotate the HDMSE data, were proposed. We could separate and characterize 323 ginsenosides (including 286 from WG and 306 from RG), and 125 thereof may have not been isolated from the Panax genus. The established 2D-LC/IM-QTOF-HDMSE approach could also act as a magnifier to probe differ-entiated components between WG and RG. Compared with conventional approaches, this dimension-enhanced strategy could better resolve coeluting herbal components and more efficiently, more reli-ably identify the multicomponents, which, we believe, offers more possibilities for the systematic exposure and confirmative identification of plant metabolites.

Novel C-17 spirost protostane-type triterpenoids from with anti-inflammatory activity in Caco-2 cells

Twenty-one protostane-type triterpenoids with diverse structures, including nine new compounds (-), were isolated from the of Linn. Structurally, alisolides A‒F (-), composed of an oxole group coupled to a five-membered ring, represent unusual C-17 spirost protostane-type triterpenoids. Alisolide H () is a novel triterpenoid with an unreported endoperoxide bridge. Alisolide I () represents the first example of 23,24-acetal triterpenoid. Their structures were elucidated based on spectroscopic analysis, wherein the absolute configurations of ‒, were further confirmed by the Mo(OAc)-induced ECD method. Furthermore, all isolates were evaluated for their inhibitory effects on LPS-induced NO production in Caco-2 cells, and all the compounds showed remarkable inhibitory activities, with IC values in the range of 0.76-38.20 μmol/L.

General Situations and Several Thoughts on Japanese Pharmacopoeia, Korean Pharmacopoeia and Taiwan Herbal Pharmacopoeia / 世界科学技术-中医药现代化

Through comparative analysis on the quality standards in Japanese Pharmacopoeia Seventeenth Edition,Korean Pharmacopoeia,Taiwan herbal Pharmacopoeia and pharmacopoeias of neighboring countries and regions,thoughts and suggestions are proposed on how to use the advantages of them,and research thoughts for available reference are provided for the construction of more scientific and feasible quality standards of traditional Chinese medicine.

Approaches to establish Q-markers for the quality standards of traditional Chinese medicines

Traditional Chinese medicine (TCM) has played a pivotal role in maintaining the health of Chinese people and is now gaining increasing acceptance around the global scope. However, TCM is confronting more and more concerns with respect to its quality. The intrinsic "multicomponent and multitarget" feature of TCM necessitates the establishment of a unique quality and bioactivity evaluation system, which is different from that of the Western medicine. However, TCM is investigated essentially as "herbal medicine" or "natural product", and the pharmacopoeia quality monographs are actually chemical-markers-based, which can ensure the consistency only in the assigned chemical markers, but, to some extent, have deviated from the basic TCM theory. A concept of "quality marker" (Q-marker), following the "property-effect-component" theory, is proposed. The establishment of Q-marker integrates multidisciplinary technologies like natural products chemistry, analytical chemistry, bionics, chemometrics, pharmacology, systems biology, and pharmacodynamics, etc. Q-marker-based fingerprint and multicomponent determination conduce to the construction of more scientific quality control system of TCM. This review delineates the background, definition, and properties of Q-marker, and the associated technologies applied for its establishment. Strategies and approaches for establishing Q-marker-based TCM quality control system are presented and highlighted with a few TCM examples.

Several Thoughts and Suggestions on International Quality Standard System Construction of Traditional Chinese Medicine / 世界科学技术-中医药现代化

Monographs of Chinese medicine into the United States Pharmacopeia and the European Pharmacopoeia is the prerequisite and foundation for the aim of Chinese medicine standards leading the international standard-set-ting. By comparative analysis of the key issues of quality standards among Chinese, American and European Phar-macopoeia, thoughts and suggestions are proposed on how to implement the construction of international quality stan-dard of traditional Chinese medicine. Meanwhile, under the pressure of the present international environment, the pa-per can also provide some reference and advices which can help to break down the difficult situation for the process of internationalization of traditional Chinese medicine.

Metabolic pathway and metabolites of total diterpene acid isolated from Pseudolarix kaempferi / 药学学报

The preliminary metabolic profile of total diterpene acid (TDA) isolated from Pseudolarix kaempferi was investigated by using in vivo and in vitro tests. Pseudolaric acid C2 (PC2) was identified as the predominant metabolite in plasma, urine, bile and feces after both oral and intravenous administrations to rats using HPLC-UV and HPLC-ESI/MS(n), and demethoxydeacetoxypseudolaric acid B (DDPB), a metabolite proposed to be the glucoside of PC2 (PC2G), as well as pseudolaric acid C (PC), pseudolaric acid A (PA), pseudolaric acid A O-beta-D glucopyranoside (PAG), pseudolaric acid B O-beta-D glucopyranoside (PBG) and deacetylpseudolaric acid A (DPA) originated from TDA could also be detected. It was demonstrated by tests that the metabolism of TDA is independent of intestinal microflora, and neither of pepsin and trypsin is in charge of metabolism of TDA, TDA is also stable in both pH environments of gastric tract and intestinal tract. The metabolites of TDA in whole blood in vitro incubation were found to be PC2, DDPB and PC2G, which demonstrated that the metabolic reaction of TDA in vivo is mainly occurred in blood and contributed to be the hydrolysis of plasma esterase to ester bond, as well as the glucosylation reaction. These results clarified the metabolic pathway of TDA for the first time, which is of great significance to the in vivo active form and acting mechanism research of P. kaempferi.

Characterization of flavonoids in Millettia nitida vat.hirsutissima by HPLC/DAD/ESI-MSn / 药物分析学报

Millettia nitida var.hirsutissima is a Chinese herbal medicine used for the treatment of gynecological diseases.An HPLC/DAD/ESI-MSn method was established for the rapid separation and characterization of bioactive flavonoids in M.nitida var.hirsutissima.A total of 32 flavonoids were detected,of which 14 compounds were unambiguously characterized by comparing their retention time,UV,and MS spectra with those of the reference standards,and the others were tentatively identified based on their tandem mass spectrometry fragmentation data obtained in the negative ionization mode on line.Nineteen of these compounds characterized were reported from this plant for the first time.

Study on proteomics of Hela cell apoptosis in bufalin-induced human cervical carcinoma / 中国中药杂志

<p><b>OBJECTIVE</b>To seek possible effect targets of bufalin in HeLa cells by studying the impact of bufalin on cell protein expression profile after treatment on human cervical carcinoma cell lines HeLa.</p><p><b>METHOD</b>Bufalin's ICs0was measured by MTr assay. The apoptosis of cells was observed by FCM (flow cytometry) and Hoechst 33342 staining assay. Differentiated expression protein spots were founded and identified using proteomic techniques, which could induce HeLa cell apoptosis.</p><p><b>RESULT</b>Bufalin showed remarkable cytotoxic effect on HeLa cells. IC50 (154 +/- 21.5) nmol X L(-1) indicated the possibility of inducing cell apoptosis. The protein expression profile showed 11 differentiated expression protein spots. Among the 11 proteins, nudix-type motif 5, vimentin, hnRNP C1/hnRNP C2 variant, HNRPK, HNRPK isoform a variant (two spots are the same protein), heat shock protein 27, macrophage-capping protein, SELENBP1 protein were down-regulated, while ribosomal protein, large, P0 and S-adenosylmethionine synthetase 2 were up-regulated by bufalin treatment. They may be effect targets of bufalin in HeLa cells. Western blotting showed consistent results in heat shock protein 27, vimentin and HNRPK between expression after treatment with bufalin and two-dimensional electrophoresis.</p><p><b>CONCLUSION</b>Bufa-Lin can induce apoptosis in human cervical carcinoma cells HeLa and the effect of bufalin may be related to the joint intervention with multiple protein targets.</p>

Metabolic pathway and metabolites of pseudolaric acid B / 药学学报

The metabolic profile of pseudolaric acid B (PB) was investigated by using in vivo and in vitro tests. Pseudolaric acid C2 (PC2) was identified as the specific metabolite of PB in plasma, urine, bile and feces using HPLC and HPLC-ESI/MS(n) after both oral and intravenous administration to rats, and almost no prototype was detected in all kinds of samples. The metabolic behaviors of PB orally administered in rats treated with antibiotics to eliminate intestinal microflora were identical with those in untreated rats, demonstrating that the metabolism of PB is independent of intestinal microflora. PB was stable in 48 h respective incubation with artificial gastric juice and artificial intestinal juice, suggesting that neither pepsin nor trypsin is in charge of metabolism of PB, and also demonstrating that PB is stable in both pH environments of gastric tract and intestinal tract. In vitro research on metabolism of PB in rat liver microsomes incubation revealed that little PB was metabolized and that the proposed metabolites were the demethoxy and demethoxydecarboxy products of the prototype. The amount of metabolites was extremely low compared with the prototype, indicating that liver microsomes are not responsible for the metabolism of PB either. PB was gradually metabolized into PC2 during 1 h in whole blood incubation in vitro, and the metabolic process showed dynamically dependent manner with incubation time. Once absorbed into blood, PB was quickly metabolized into PC2, accordingly, little prototype was detected in all kinds of samples. The metabolism was attributed to the rapid hydrolysis of C-19 ester bond by plasma esterase. These results clarified the metabolic pathway of PB for the first time, which was of great significance to identify the in vivo active form and interpret acting mechanism of the active compounds of P. kaempferi.

Characterization of diterpenoids in the bark of Pseudolarix kaempferi by HPLC-ESI/MSn / 药学学报

Fragmentation behavior of diterpenoids was investigated by ESI/MSn and the qualitative analysis of diterpenoids in the bark of Pseudolarix kaempferi was performed using high-performance liquid chromatography/ multi-stage mass spectrometry (HPLC-ESI/MSn). The characteristic fragmentation behaviors of the diterpenoids are the cleavages of the lactone ring and C4-O bond. Furthermore, the eliminations of substituent groups at C-18, C-7 and C-8 can also be observed in the MS" (n = 3-4) spectra. For C-4 acetoxy subsititued diterpenoids, [M+Na-60]+ and [M-H-104] are the base peaks of MS2 spectra in the positive and negative ionization modes, respectively. For C-4 hydroxyl subsititued diterpenoids, [M+Na-44]+ and [M-H-62] are the base peaks of MS2 in the positive and negative ionization modes, respectively. For C-18 glucosylated or esterized diterpenoids, [M+Na-44]+ is the base peak of MS2 spectra in positive ionization mode. These fragmentation rules were successfully exploited in the identification of diterpenoids in methanol/water (6:4) extract of P. kaempferi by LC-MS in positive ionization mode. A total of 9 diterpenoids were identified or tentatively characterized, and one of them is reported here for the first time. The described method could be utilized for the sensitive and rapid qualitative analysis of P. kaempferi.

Two new phenolic acids from drynariae rhizoma / 药学学报

To study the chemical constituents of Drynariae Rhizoma, nine phenolic acids were isolated from a 70% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, ODS, Sephadex LH-20, and semi-preparative HPLC. By spectroscopic techniques including 1H NMR, 13C NMR, 2D NMR, and HR-ESI-MS, these compounds were identified as 4, 4'-dihydroxy-3, 3'-imino-di-benzoic acid (1), protocatechuic acid (2), gallic acid (3), p-hydroxybenzoic acid (4), (E)-caffeic acid (5), ethyl trans-3, 4-dihydroxycinnamate (6), caffeic acid 4-O-beta-D-glucopyranoside (7), p-coumaric acid 4-O-beta-D-glucopyranoside (8), and 23(S)-12-O-caffeoyl-12-hydroxyllauric acid glycerol ester (9), separately. Among them, 1 and 9 are new compounds, and 3, 4, and 6 were isolated from Drynaria species for the first time.

Fingerprints of Tongmai Keli by HPLC-DAD-MS / 药学学报

In order to clarify the chemical composition of Tongmai Keli, a HPLC fingerprint was established, and the 22 peaks were characterized by LC-DAD-MS. The herbal sources of these peaks were assigned. The results implied that genistin 8-C-glucoside, puerarin, daidzein 8-C-apiosyl (1, 6)-O-glucoside, daidzin, and salvianolic acid B were the main constituents of in Tongmai Keli. Ten batches of Tongmai Keli produced by different pharmaceutical companies were analyzed. Although different batches contained similar compounds, the contents of major compounds were significantly different. The method established in this study could be used for the quality control of Tongmai Keli.

Determination of the binding rate of rat plasma protein with salvianolic acid B / 药学学报

This paper is aimed to report the development of a method for the determination of the binding rate of plasma protein with salvianolic acid B. In vitro, equilibrium dialysis method was used to imitate the binding process between salvianolic acid B and plasma protein, in vivo, ultrafiltration method was used and the binding rate with HPLC was determined. Plasma samples were treated with methanol to precipitate the protein, and the buffer solution was directly determined after filtering. The calibration curve of the buffer solution was linear in the range of 0.5-20 microg mL(-1). The calibration curve of the plasma was linear in the range of 2-200 microg mL(-1). The extract recovery was 68.6%-81.9%. RSDs of intra- and inter-day precisions were all less than 8.5%. The binding rates of plasma protein with salvianolic acid B in vitro was 75.2% and in vivo was 92.1%. This paper shows the high binding power of salvianolic acid B to plasma protein with high sensitivity, good reproduction, simple management and fulfilling the requirement.

Distribution study of two constituents in rat after oral administration of Smilax china extract / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the distribution in rat after oral administration of Smilax china extract.</p><p><b>METHOD</b>Oxyresveratrol and resveratrol were quantitatively determined by HPLC/DAD method in rat tissues including heart, liver, spleen, lung and kidney after oral administration of S. china extract. The separation was performed on a Zorbax SB-C18 column (4.6 mm x 250 mm, 5 microm) with acetonitrile-water using linear gradient elution and UV detection at wavelength of 320 nm. The flow rate was 1.0 mL x min(-1) and the column was kept at 40 degrees C.</p><p><b>RESULT</b>The HPLC/DAD method for the simultaneous determination of oxyresveratrol and resveratrol in rat tissues was developed.</p><p><b>CONCLUSION</b>The method has been successfully applied for the distribution study of two active constituents in rat through oral administration of S. china extract.</p>

Metabolic study of ginsenoside Re in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To systemically study the metabolism of ginsenoside Re in rats.</p><p><b>METHOD</b>Six SD rats were used divided into 3 groups. After oral administration of ginsenoside Re at a dosage of 100 mg x kg(-1), feces were collected at 12, 24, 36, 48, and 60 h. The microbial transformation metabolites of ginsenoside Re were used as standard references. HPLC-ESI-MS/MS analysis was used in the metabolism studies.</p><p><b>RESULT</b>Six metabolites of ginsenoside Re were detected in rat feces by HPLC-ESI-MS/MS analysis. Their structures were identified as 20(S)-ginsenoside Rg2, 20(S)-ginsenoside Rh1, 20(R)-ginsenoside Rh1, ginsenoside F1, 3-oxo-ginsenoside Rh1 and protopanaxatriol.</p><p><b>CONCLUSION</b>Ginsenoside Re has the same metabolic pathway with microorganisms, to some extent, justified the use of microbial models for mammalian metabolism studies.</p>

Progress in biotransformation of triptolides and bufadienolides / 北京大学学报(医学版)

The latest results from our research group in the biotransformation of triptolides and bufadienolides were reviewed. The trends in the development of biotransformation in the future were also briefly discussed.

Chemical constituents of Buddlejae officinalis and their bioactivities / 中草药

Object To investigate the chemical constituents of Buddlejae officinalis Maxim. , the relationship between their bioactivities, and the medicinal application of the herb. Methods Various chro-matographic techniques were employed for the isolation and purification of the constituents, including silica gel, Sephadex LH-20, and pre-HPLC. The structures of 11 isolated chemical compounds were elucidated by chemical and spectral analysis (NMR, IR, UV, and MS). The aldose reductase inhibiting test and test on DNA topoisomerase Ⅳ inhibitory effect were applied to evaluate the bioactivity of both the purified compounds and the crude fraction. Results Eleven compounds were isolated from the extracts of B. offic-inalis. They are four flavones and their glycosides; apigenin (Ⅰ), linarin (Ⅱ), apigenin-7-O-?-L-rhamnopyranosyl (1-6)-?-D-glucopyranoside (Ⅲ), and luteolin-7-O-?-D-glucoside (Ⅶ); four phenylen-thanoid glycosides: verbascoside (Ⅴ), isoacteoside (Ⅵ), cistanoside F, a and b (Ⅶ-1 and 2), campneo-side Ⅱ , a and b (Ⅷ-1 and 2); three triferpenoid saponins: mimengoside A (Ⅸ), mimengoside B (Ⅹ), songaroside A (Ⅺ). Some of the them (such as Ⅱ ,Ⅲ , and Ⅵ) showed the aldose reductase inhibitory activities with a higher inhibitory rate than that of quercetin (the positive control), others (such as Ⅰ,Ⅱ , Ⅴ, Ⅵ, and Ⅺ) displayed the inhibititory activities against the DNA topoisomerase Ⅳ . Conclusion Among all the isolated compounds, Ⅲ , Ⅶ-1 and 2, Ⅷ-1 and 2, and Ⅺ are separated for the first time both from the plant and the genus. Their bioactivities of bacteriostatic and against aldose reductase with DNA topoisomerase Ⅳ as target are tested for the first time and are related to the medicinal application of B. officinalis.

Effect of rare-earth elements La3+ on growth of hairy roots and normal roots of Rheum palmatum and their yield of anthraquinone / 中草药

Object To study the effect of rare-earth elements La3+ on growth of hairy roots and normal roots of Rheum palmatum L and their yield of anthraquinones. Methods The biomass and an- thraquinone yield of two hairy clones DH5c, DH7a, and normal root NOR cultures of R. palmatum were evaluated statistically after various concentration of La3+ were administrated into culture medium. Results Significant differences of biomass yield and anthraquinone yield were shown among six La3+ concentrations, in which 10 mg/L showed obvious inhibitory effect for root growth and 1. 0 mg/L was the best for anthraquinone yield. In general, aloe-emodin and rhein were predominant in five anthraquinones after treated with the rare-earth element. Conclusion The growth of hairy root cultures of R. palmatum and their anthraquinone yield are greatly promoted by addition of La3+ to the medium.