Monoterpene Glycosides Research Articles

Involvement of the Hydroperoxy Group in the Irreversible Inhibition of Leukocyte-Type 12-Lipoxygenase by Monoterpene Glycosides Contained in the Qing Shan Lu Shui Tea.

We have previously found two novel monoterpene glycosides, liguroside A and liguroside B, with an inhibitory effect on the catalytic activity of the enzyme leukocyte-type 12-lipoxygenase in the Qing Shan Lu Shui tea. Here, two new monoterpene glycosides, liguroside C and liguroside D which inhibit this enzyme, were isolated from the same tea. The spectral and chemical evidence characterized the structures of these compounds as (5E)-7-hydroperoxy-3,7-dimethyl-1,5-octadienyl-3-O-(α-l-rhamnopyranosyl)-(1′′→3′)-(4′′′-O-trans-p-coumaroyl)-β-d-glucopyranoside and (2E)-6-hydroxy-3,7-dimethyl-2,7-octadienyl-3-O-(α-l-rhamnopyranosyl)-(1′′→3′)-(4′′′-O-trans-p-coumaroyl)-β-d-glucopyranoside, respectively. These ligurosides, which irreversibly inhibited leukocyte-type 12-lipoxygenase, have a hydroperoxy group in the monoterpene moiety. Additionally, monoterpene glycosides had the same backbone structure but did not have a hydroperoxy group, such as kudingoside A and lipedoside B-III, contained in the tea did not inhibit the enzyme. When a hydroperoxy group in liguroside A was reduced by using triphenylphosphine, the resultant compound, kudingoside B, showed a lower inhibitory effect on the enzyme. These results strongly suggest the involvement of the hydroperoxy group in the irreversible inhibition of the catalytic activity of leukocyte-type 12-lipoxygenase by the monoterpene glycosides contained in the Qing Shan Lu Shui tea.

Chemical constituents isolated from Clematis akebioides (Maximowicz) Veitch

A phytochemical investigation of the aerial parts of Clematis akebioides (Maximowicz) Veitch led to the isolation of eighteen compounds, including two benzenoids (1, 2), one linear monoterpene glycoside (3), and fifteen triterpenoid saponins (4–18). The structural elucidation of these compounds was determined by using spectroscopic methods and comparison with the literature. All of these compounds were obtained from C. akebioides for the first time, and compounds 1, 2 and 18 were isolated from the Ranunculaceae family for the first time. Furthermore, the chemotaxonomic significance of these compounds was discussed.

Characterization of the chemical constituents in Hongjingtian injection by liquid chromatography quadrupole time-of-flight mass spectrometry.

Hongjingtian injection is made from Rhodiola wallichiana and used in the treatment of stable angina pectoris associated with coronary heart disease. In this study, the chemical constituents in Hongjingtian injection were comprehensively studied using liquid chromatography quadrupole time-of-flight mass spectrometry. A total of 49 compounds were identified or assumed, including 10 organic acids, nine phenylethanoids, 10 phenylpropanoids, two flavonoid glycosides, seven monoterpene glycosides, seven octylglycosides and four other types of compounds. The structures of seven compounds were confirmed by comparing their retention times, MS and UV spectra with the corresponding authentic standards. Amongst the 49 compounds, 35 were firstly found in R. wallichiana, while they have been reported in other species of the genus Rhodiola, including Rhodiola crenulata, Rhodiola sacra, Rhodiola rosea and Rhodiola kirilowii. The possible fragmentation pathways in the mass spectrometry of the major types of compounds are proposed and summarized. Our study demonstrates a rapid method for characterizing the chemical constituents present in the Hongjingtian injection, which could also be applied to the identification of chemical constituents in other TCM formulae containing R. wallichiana.

Optimization of Ultrasound-Assisted Extraction of Monoterpene Glycoside from Oil Peony Seed Cake.

Ultrasound-assisted extraction (UAE) of total monoterpene glycosides extract (TMGE) from oil peony seed cakes was investigated. The extraction yield was optimized using response surface methodology (RSM). The chemical constituents of the monoterpene glycosides extract were isolated by repeated column chromatography, and the contents of the main isolated monoterpene glycosides in the oil peony seed cakes were determined by HPLC. The optimum conditions were as follows: a liquid-to-solid ratio of 27 mL/g, ultrasonic extraction time of 16 min, ultrasonic extraction temperature of 26 °C, and ethanol concentration of 67%. Under these conditions, the extraction yield of TMGE was 10.24%. Twenty monoterpene glycosides were isolated from the oil peony seed cakes, and compounds 11-12, 16 and 20 showed strong inhibitory activities on NO production. TMGE from oil peony seed cakes can also to be used as promising immunosuppressive drug due to its high content of monoterpene glycosides and immune-inhibitory activity. PRACTICAL AAPPLICATION: The peony seed oil was authorized as a new food by the Ministry of Health of the People's Republic of China. Peony seed cake is one of the most important by-products in the preparation of peony seed oil, and accounts for approximately 40% of the total mass of the peony seed. Total monoterpene glycosides are the main active ingredient of oil peony seed cake. This research has optimized the extraction conditions of total monoterpene glycoside from seeds cake of Paeonia ostii, which will provide useful reference information for further studies, and offer related industries with helpful guidance in practice.

Chemical Constituents from Scindapsus officinalis (Roxb.) Schott. and Their Anti⁻Inflammatory Activities.

One new monoterpene glycoside (1), one new phenyl glycoside (2), one new caffeoyl derivative (3), were isolated from Scindapsus officinalis (Roxb.) Schott., along with four known compounds (4–7). Structures of the isolated compounds were elucidated by extensive analysis of spectroscopic data, especially 2D NMR data and comparison with literatures. All isolates were evaluated for anti-inflammatory activity against nitric oxide (NO) production in vitro. Compounds 3 and 7 exhibited moderate inhibitory effects on NO production with IC50 values of 12.2 ± 0.8 and 18.9 ± 0.3 μM, respectively.

Profiling and identification of chemical components of Shenshao Tablet and its absorbed components in rats by comprehensive HPLC/DAD/ESI-MSn analysis

AbstractShenshao Tablet (SST), prepared from Paeoniae Radix Alba (PRA) and total ginsenoside of Ginseng Stems and Leaves (GSL), is a traditional Chinese medicine (TCM) preparation prescribed to treat coronary heart disease. However, its chemical composition and the components that can migrate into blood potentially exerting the therapeutic effects have rarely been elucidated. We developed an HPLC/DAD/ESI-MSn approach aiming to comprehensively profile and identify both the chemical components of SST and its absorbed ingredients (and metabolites) in rat plasma and urine. Chromatographic separation was performed on an Agilent Eclipse XDB C18 column using acetonitrile/0.1% formic acid as the mobile phase. MS detection was conducted in both negative and positive ESI modes to yield more structure information. Comparison with reference compounds (tR, MSn), interpretation of the fragmentation pathways, and searching of in-house database, were utilized for more reliable structure elucidation. A total of 82 components, including 21 monoterpene glycosides, four galloyl glucoses, two phenols from PRA, and 55 ginsenosides from GSL, were identified or tentatively characterized from the 70% ethanolic extract of SST. Amongst them, seven and 24 prototype compounds could be detectable in the plasma and urine samples, respectively, after oral administration of an SST extract (4 g·kg−1) in rats. No metabolites were observed in the rat samples. The findings of this work first unveiled the chemical complexity of SST and its absorbed components, which would be beneficial to understanding the therapeutic basis and quality control of SST.

Paeonia veitchii seeds as a promising high potential by-product: Proximate composition, phytochemical components, bioactivity evaluation and potential applications

To assess the application value of the seeds of the traditional medicine Chishao (Radix Paeonia Rubra, the root of Paeonia veitchii Lynch), the thousand seed weight, oil and kernel contents of the seeds, composition and contents of the main fatty acids, the physical and chemical characteristics of the seeds oil, and proximate composition and phytochemical components of the seeds shell and seeds oil residue were evaluated in this study. In addition, bioactivities such as antitumour and antimicrobial activity of the oligostilbene compound from P. veitchii seeds shell, and the inhibitory activities of the monoterpene glycoside compounds in P. veitchii seeds oil residue were also evaluated using different in vitro biochemical assays. The results showed that the seeds of P. veitchii had high oil content, and the seeds oil was characterized by very high levels of unsaturated fatty acids. The seeds shell contained large amounts of cellulose, lignin, and oligostilbene compounds, and high levels of monoterpene glycosides and crude protein were found in the P. veitchii seeds oil residue. The results of the biological tests showed that total oligostilbene compounds from P. veitchii seeds shell had antibacterial activity and moderate anticancer effects in vitro, and the total monoterpene glycoside compounds in P. veitchii showed strong inhibitory activities against NO production. As a medical plant and industrial crop with broad prospects, P. veitchii can be planted in most regions of China. In the past, it was planted solely because of the medical use of the roots. The above proximate and phytochemical profile, and bioactivity results show that the seeds of P. veitchii have high utilization value with potential in the development of value-added products, representing another valuable part of P. veitchii in the near future.

Paeoniflorin regulates GALNT2-ANGPTL3-LPL pathway to attenuate dyslipidemia in mice

N-acetylgalactosaminyltransferase 2-Angiopoietin-like protein 3-lipoprotein lipase (GALNT2-ANGPTL3-LPL) pathway may be a useful pharmacologic objective for dyslipidemia. The present study was conducted to test the effect of paeoniflorin, a monoterpene Glycoside, on dyslipidemia in mice. Fifty mice were randomly divided into five groups (n = 10): three groups of apolipoprotein E-null (ApoE-/-) mice treated with paeoniflorin (10 or 20 or 30 mg/kg/day), untreated ApoE-/- mice group, and C57BL/6J control group. Six weeks after treatment, expression of hepatic ANGPTL3, hepatic GALNT2 and adipose tissue LPL, lipid levels in the liver and blood were quantified. Treatment with paeoniflorin (10 or 20 or 30 mg/kg) obviously down-regulated expression of ANGPTL3 and up-regulated expressions of GALNT2 and LPL concomitantly with elevated plasma high-density lipoprotein cholesterol level, reduced plasma concentrations of low-density lipoprotein cholesterol, total cholesterol, triglyceride, malonaldehyde, and 8-isoprostane. The present results suggest that paeoniflorin regulates GALNT2-ANGPTL3-LPL pathway to attenuate dyslipidemia in mice.

Combining liquid chromatography and tandem mass spectrometry approaches to the study of monoterpene glycosides (aroma precursors) in wine grape.

Monoterpene-glycosides are important aroma precursors that, undergo hydrolysis, confer intense floral notes to the wines. Therefore, the knowledge of the nature of the sugar residues and the structure of these molecules is of great interest. In present study, liquid chromatography (LC) separation coupled with different mass spectrometry (MS) experiments for the characterization of these compounds were explored. The LC parameters were tuned to optimize the resolution between the analytes present in grape sample extracts. Twenty principal peaks with a relative abundance >1% were selected and divided in 4 classes characterized by different molecular weight. In general, positive ionization of the studied compounds displayed the [M+NH4 ]+ ion as base peak. On the contrary, a distribution between [M+Cl]- and [M+HCOO]- species was observed in negative ion mode. However, a clear differentiation between the studied compounds was only possible by combining both LC and tandem MS (MS/MS). Indeed, by applying a series of energy resolved MS/MS experiments and monitoring both positive and negative ions, a structural characterization of the analytes was achieved. The proposed LC-MS/MS approach provided the profile of monoterpenol-diglycosides and allowed the identification of a series of isobaric terpene-diglycosides in grape. The study of their MS/MS spectra indicated the structure of geranic and/or nerolic acid aglycones. To verify the interest of studied compounds, a preliminary evaluation of the intensity of signals of these glycosides were carried out. The obtained results showed a significant difference between the grape samples collected in two different vintages.

PHYTOCHEMICAL CONSTITUENTS AND ANTIOXIDANT EFFECT OF SOLANUM ROSTRATUM SPECIES FROM ALGERIA T

Objective: The aim of this study was to isolate the constituents and evaluate the antioxidant activity of Solanum rostratum Dunal (Solanaceae) from Algeria. Methods: In this study, phytochemical analyses of the chloroform, ethyl acetate, and n-butanol extracts obtained from the aerial parts of S. rostratum were performed by column chromatography, thin-layer chromatography, and high-performance liquid chromatography techniques. The antioxidant activity was performed by 2,2-diphenyl-1-picrylhydrazyl scavenging assay method.Results: The identification and structure elucidation of the isolated compounds were compared with their nuclear magnetic resonance spectra and the literature led to identify one monoterpene glycoside (linalyl-β-glucopyranoside) (1) and three flavonoid glycosides: Apigenin-7-O-glucoside (2), astragalin (3) and (isorhamnetin-3-O-glucoside (4).Conclusion: This is the first report on the isolation and structure elucidation of compound (1) (linalyl-β-glucopyranoside) and compound (2) (apigenin-7-O-glucoside) from this species. In addition, antioxidant effect of the chloroform extract from S. rostratum was evaluated. The chloroform extract exhibited the remarkable radical scavenging ability (IC50=0066±0.001 mg/mL).

HPLC-DAD analysis of 15 monoterpene glycosides in oil peony seed cakes sourced from different cultivation areas in China

The oil peony seed cake, one of the most important by-products during the seed oil preparation, contains numerous monoterpene glycosides. The development and optimization of a simple and reproducible method for simultaneous qualitative and quantitative determination of monoterpene glycosides from oil peony seed cakes would be a valuable work for its further utilization. In this study, an HPLC method using DAD detector was developed to analyse fifteen monoterpene glycosides in oil peony seed cakes. The optimum resolution of the fifteen monoterpene glycosides was achieved on an ODS-AQ column. Acetonitrile (solvent A) and 0.5% potassium dihydrogen phosphate solution (pH 2.8) (solvent B) were used as the mobile phase by a gradient elution. Paeoniflorin showed the highest content amongst 15 monoterpene glycosides. It accounted for more than half of the total content of the fifteen monoterpene glycosides (TCFMG) in oil peony seed cake, followed by oxypaeoniflorin and albiflorin. The samples of different cultivation areas contain the different contents of monoterpene glycosides, mainly determined by their geographical variation. The higher level contents of monoterpene glycoside are believed to have more potential in medicinal application. The method thus could be used to control the quality of products from oil peony seed cakes.

Neuroprotective effect of paeoniflorin on okadaic acid-induced tau hyperphosphorylation via calpain/Akt/GSK-3β pathway in SH-SY5Y cells

Abnormal phosphorylation of tau, one of the most common symptoms of dementia, has become increasingly important in the study of the etiology and development of Alzheimer’s disease. Paeoniflorin, the main bioactive component of herbaceous peony, is a monoterpene glycoside, which has been reported to exert beneficial effects on neurodegenerative disease. However, the effect of paeoniflorin on tauopathies remains ambiguous. SH-SY5Y cells were treated with okadaic acid (OA) for 8 h to induce tau phosphorylation and no cell death was observed. Optical microscopy results showed that paeoniflorin ameliorated okadaic acid induced morphological changes, including cell swelling and synapsis shortening. Western blotting data illustrated that paeoniflorin reversed okadaic acid induced tau hyperphosphorylation, which was enhanced by inhibiting the activities of calpain, Akt and GSK-3β. Transmission electron microscopy results showed that paeoniflorin alone can reduce the number of autophagosomes and stabilize the microtubule structure. In addition, calpastain and paeoniflorin enhance the effect of paeoniflorin on stabilizing microtubules. In addition, calpastain markedly enhanced the effect of paeoniflorin on reversing okadaic acid-lowered fluorescence intensity of both MAP-2 and β III-tubulin, two microtubule-associated proteins. This study shows that paeoniflorin protected SH-SY5Y cells against okadaic acid assault by interfering with the calpain/Akt/GSK-3β-related pathways, in which autophagy might be involved. Besides, paeoniflorin is found to relieve the stress response of the microtubule structure system caused by okadaic acid treatment. The results presented in this study suggest that paeoniflorin potentially plays an important role in tauopathies.

Effects of paeoniflorin on neurobehavior, oxidative stress, brain insulin signaling, and synaptic alterations in intracerebroventricular streptozotocin-induced cognitive impairment in mice

Paeoniflorin (PF) is a natural monoterpene glycoside in Paeonia lactiflora pall with anti-diabetic, antioxidant, anti-inflammatory, and neuro-protective properties. This study was designed to investigate the neuroprotective effects of PF against cognitive deficits induced by intracerebroventricular (ICV) injection of streptozotocin (STZ) in mice. STZ was injected twice intracerebroventrically (3 mg/kg ICV) on alternate days (day 1 and day 3) in mice. Daily treatment with PF (10 mg/kg per day, intraperitoneally) starting from the first dose of STZ for 21 days showed an improvement in ICV-STZ induced cognitive deficits as assessed by novel object recognition and Morris water maze (MWM) test. PF significantly attenuated STZ induced mitochondrial dysfunction manifested by dramatically elevated cytochrome c oxidase activity and ATP synthesis, and restoration of the mitochondrial membrane potential (MMP), and oxidative stress in hippocampus and in the cortex compared to control. Moreover, PF treatment also markedly increased synaptic density in the CA1 region of the hippocampus compared to control. Furthermore, PF ameliorated defective insulin signaling by up-regulating p-PI3K and p-Akt protein expression while downregulating p-IRS-1 protein expression. Taken together, the outcomes of the current study suggest the therapeutic potential of PF in the cognitive deficits induced by ICV-STZ.

In Vitro and In Vivo Antioxidant Activities of the Flowers and Leaves from Paeonia rockii and Identification of Their Antioxidant Constituents by UHPLC-ESI-HRMSn via Pre-Column DPPH Reaction.

The genus Paeonia, also known as the “King of Flowers” in China, is an important source of traditional Chinese medicine (TCM). Plants of this genus have been used to treat a range of cardiovascular and gynecological diseases. However, the potential pharmacological activity of one particular species, Paeonia rockii, has not been fully investigated. In the first part of the present study, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic) acid (ABTS), reducing power assays, and metal ion chelating assays were used to investigate the in vitro antioxidant activities of Paeonia rockii. In the second portion of the study, a mouse model of d-galactose-induced aging was used to validate the antioxidant effects of the flowers from Paeonia rockii in vivo. Lastly, potential antioxidant constituents were screened and identified by ultra-high pressure liquid chromatography and electrospray ionization coupled with high-resolution mass spectrometry (UHPLC-ESI-HRMSn) combined with the DPPH assay. Results indicated that the flowers and leaves exhibited stronger antioxidant activity than ascorbic acid in vitro. The therapeutic effect of Paeonia rockii was determined in relation to the levels of biochemical indicators, such as 8-iso-prostaglandin F2α (8-iso PGF2α) in the serum, superoxide dismutase (SOD), protein carbonyl, malondialdehyde (MDA), and glutathione (GSH) in the liver and brain, after daily intra-gastric administration of different concentrations of extracts (100, 200 and 400 mg/kg) for three weeks. The levels of 8-iso PGF2α (p < 0.01) and protein carbonyl groups (p < 0.01) were significantly reduced, whereas those of SOD (p < 0.05) had significantly increased, indicating that components of the flowers of Paeonia rockii had favorable antioxidant activities in vivo. Furthermore, UHPLC-ESI-HRMSn, combined with pre-column DPPH reaction, detected 25 potential antioxidant compounds. Of these, 18 compounds were tentatively identified, including 11 flavonoids, four phenolic acids, two tannins, and one monoterpene glycoside. This study concluded that the leaves and flowers from Paeonia rockii possess excellent antioxidant properties, highlighting their candidacy as “new” antioxidants, which can be utilized therapeutically to protect the body from diseases caused by oxidative stress.

Rapid Screening and Quantitative Determination of Active Components in Qing-Hua-Yu-Re-Formula Using UHPLC-Q-TOF/MS and HPLC-UV.

Qing-Hua-Yu-Re-Formula (QHYRF), a new herbal preparation, has been extensively used for treating diabetic cardiomyopathy. However, the chemical constituents of QHYRF remain uninvestigated. In the present study, rapid ultrahigh-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) was used to qualitatively analyze the components of QHYRF. Qualitative detection was performed on a Kromasil C18 column through the gradient elution mode, using acetonitrile-water containing 0.1% formic acid. Twenty-seven compounds were identified or tentatively characterized, including 12 phenolic acids, nine monoterpene glycosides, two flavonoids, three iridoids, and one unknown compound. Among these, six compounds were confirmed by comparing with standards. A high-performance liquid chromatography (HPLC) method was developed to simultaneously determine the following six active components in QHYRF: danshensu, paeoniflorin, acteoside, lithospermic acid, salvianolic acid B, and salvianolic acid C. These HPLC chromatograms were monitored at 254, 280, and 320 nm. The method was well validated with respect to specificity, linearity, limit of detection, limit of quantification, precision, stability, and recovery. The HPLC-UV method was successfully applied to 10 batches of QHYRF.

Isocoumarin derivatives and monoterpene glycoside from the seeds of Orychophragmus violaceus

Five new compounds, including four new isocoumarin derivatives [orychophramarin A–D (1–4)] and a new monoterpene glycoside [orychovioside A (5)], together with fourteen known compounds were isolated from the seeds of Orychophragmus violaceus. Their structures were established on the basis of chemical evidence and spectroscopic analysis. Compound 1 showed significant cytotoxicity against HCT-116 and Hela cell lines compared with the positive control group (Cisplatin) with IC50 values of 5.10 and 8.91μM, respectively. Compound 1 exhibited evident cell cycle retardation that arrested HCT-116 cells at G2 phase and induced apoptosis in HCT-116 cells in the further mechanism study.

Phytochemical variation among the traditional Chinese medicine Mu Dan Pi from Paeonia suffruticosa (tree peony)

Mu Dan Pi is a traditional Chinese medicine used to treat inflammation, cancer, allergies, diabetes, angiocardiopathy, and neurodegenerative diseases. In this study, the metabolome variation within Mu Dan Pi collected from 372 tree peony cultivars was systematically investigated. In total, 42 metabolites were identified, comprising of 14 monoterpene glucosides, 11 tannins, 8 paeonols, 6 flavonoids, and 3 phenols. All cultivars revealed similar metabolite profiles, however, they were further classified into seven groups on the basis of their varying metabolite contents by hierarchical cluster analysis. Traditional cultivars for Mu Dan Pi were found to have very low metabolite contents, falling into clusters I and II. Cultivars with the highest amounts of metabolites were grouped in clusters VI and VII. Five potential cultivars, namely, ‘Bai Yuan Qi Guan’, ‘Cao Zhou Hong’, ‘Da Zong Zi’, ‘Sheng Dan Lu’, and ‘Cheng Xin’, with high contents of monoterpene glycosides, tannins, and paeonols, were further screened. Interestingly, the majority of investigated cultivars had relatively higher metabolite contents compared to the traditional medicinal tree peony cultivars.

Three new monoterpene glycosides from oil peony seed cake

A phytochemical investigation of oil peony seed cake led to the isolation and identification of three new monoterpene glycosides called paeonirockosides A (1), B (2), and C (5), together with 21 known monoterpene glycosides. Their structures were elucidated through the extensive application of one- and two-dimensional nuclear magnetic resonance (1D and 2D NMR) spectroscopy and a comparison with authentic standards. The inhibitory activity of the isolated compounds against nitric oxide (NO) production was evaluated on lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. Compounds mudanpioside F (3), benzoylpaeoniflorin (11), benzoyloxypaeoniflorin (12), oxypaeonidanin (18) and pyrindylpaeoniflorin (22) and the 70% aqueous ethanol extract (AEE) exhibited potent inhibitory activity against NO production with IC50 values of 18.65, 23.32, 25.14, 40.8, 28.61 and 36.42μM, respectively. Monoterpene glycosides from peony seed cake can be developed into a promising immunosuppressive drug.

Screening, separation, and evaluation of xanthine oxidase inhibitors from Paeonia lactiflora using chromatography combined with a multi-mode microplate reader.

Natural products have become one of the most important resources for discovering novel xanthine oxidase inhibitors, which are commonly employed in the treatment of hyperuricemia and gout. However, to date, few reports exist regarding the use of monoterpene glycosides as xanthine oxidase inhibitors. Thus, we herein report the use of ultrafiltration coupled with liquid chromatography in the screening of monoterpene glycoside xanthine oxidase inhibitors from the extract of Paeonia lactiflora (P. lactiflora), and both high-performance counter-current chromatography and medium-pressure liquid chromatography were employed to separate the main constituents. Furthermore, the xanthine oxidase inhibitory activities and the mechanisms of inhibition of the isolated compounds were evaluated using a multi-mode microplate reader by Molecular Devices. As a result, three monoterpene glycosides were separated by combined high-performance counter-current chromatography and medium-pressure liquid chromatography in purities of 90.4, 98.0, and 86.3%, as determined by liquid chromatography. These three compounds were identified as albiflorin, paeoniflorin, and 1-O-β-ᴅ-glucopyranosyl-8-O-benzoylpaeonisuffrone by electrospray ionization tandem mass spectrometry, and albiflorin and paeoniflorin were screened as potential xanthine oxidase inhibitors by ultrafiltration with liquid chromatography. The evaluation results of xanthine oxidase inhibitory activity corresponded with the screening results, as only albiflorin and paeoniflorin exhibited xanthine oxidase inhibitory activity.

Simultaneous determination and pharmacokinetic study of twelve bioactive compounds in rat plasma after intravenous administration of Xuebijing injection by UHPLC-Q-Orbitrap HRMS

Xuebijing injection (XBJ) is a traditional Chinese herbal prescription widely used in the treatment of sepsis. Extensive studies revealed that the major bioactive constituents of XBJ injection, including phenolic acids, flavonoids, monoterpene glycosides, lactones and organic acids, play an important role in the treatment. In this study, a rapid, sensitive and accurate ultra high performance liquid chromatography – Q Exactive hybrid quadrupole – orbitrap high-resolution accurate mass spectrometry (UHPLC-Q-Orbitrap HRMS) method was developed for simultaneous determination of twelve bioactive compounds in rat plasma after intravenous administration of XBJ injection. A gradient elution for separation was achieved on a waters ACQUITY UHPLC® BEH C18 column (2.1mm×50mm, 1.7μm) column with acetonitrile-water (containing 0.1% formic acid) as mobile phase at a flow rate of 0.2mL/min. All compounds and IS were monitored by parallel reaction monitoring assay both in positive and negative ion mode. The developed method was validated for intra-day and inter-day accuracy and precision whose values fell in the acceptable limits. Extraction recoveries at three levels of QC concentrations were all more than 80% for all compounds and IS, and matrix effects were found in the range of 80.0–120.0%. Stability results showed that all analytes were stable at the investigated conditions. The validated method was successfully applied to a pharmacokinetic study of Xuebijing injection following intravenous administration of 2.5, 5.0, 10.0mL/kg to rats respectively. And the result indicated that the pharmacokinetic behavior of XBJ injection is positively related to dosage at the range of 2.5–10mg/kg. This study will provide a meaningful basis for evaluating the rationality of XBJ injection for clinical application.