Application of molecular imprinting polymers in separation of active compounds from plants.

Molecular imprinting technique is becoming an appealing and prominent strategy to synthesize materials for target recognition and rapid separation. In recent years, it has been applied in separation of active compounds from various plants and has achieved satisfying results. This review aims to make a brief introduction of molecular imprinting polymers and their efficient application in the separation of various active components from plants, including flavonoids, organic acids, alkaloids, phenylpropanoids, anthraquinones, phenolics, terpenes, steroids, and diketones, which will provide some clues to help stimulating research into this fascinating and useful area.

Spectrum-effect relationship between UHPLC-Q-TOF/MS fingerprint and promoting gastrointestinal motility activity of Fructus aurantii based on multivariate statistical analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Nowadays, gastrointestinal motility disorders (GMD) have reduced the quality of people's daily life worldwide, but there is still a lack of effective western medicine treatment. Fructus aurantii (FA), a representative regulating-qi herbal medicine, has been widely used to treat GMD in China for thousands of years, but it is not clear that which specific components contribute to the efficacy. AIM OF THE STUDY: The efficacy differences of various fractions of FA on normal mice and GMD rats were compared, so as to find out the main effective fraction of FA, and to screen the main regulating-qi components based on spectrum-effect relationship and multivariate statistical analysis. MATERIALS AND METHODS: The fingerprints of different fractions of FA were established and main compounds were identified with UHPLC-Q-TOF/MS technique. The promoting gastrointestinal motility activities of FA were evaluated by defecation test, gastric emptying and intestinal propulsion test in mice, and further investigated according to the biochemical analysis of 5-HT, SP, MLT, GAS and VIP in GMD rats' plasma. One-way ANOVA was used to find out the difference of efficacy. The active components were screened through spectrum-effect relationship with PCA-X, Pearson bivariate correlation analysis and OPLS analysis. CONCLUSIONS: Ethyl acetate fraction is the main active fraction, and nine compounds are the major regulating-qi components. The developed spectrum-effect analysis can be used for the screening of bioactive components in natural products with high accuracy and reliability.

Comparison of the chemical constituents of raw Fructus Aurantii and Fructus Aurantii stir-baked with bran, and the biological effects of auraptene.

ETHNOPHARMACOLOGICAL RELEVANCE: Fructus Aurantii (FA) is a Chinese herbal medicine commonly used in clinical practice to improve gastrointestinal motility, treat dyspepsia, and relieve constipation. More than 20 processing methods of FA have been recorded, among which FA stir-baked with bran is the earliest, most time consuming, and the most popular one. Raw FA has a strong ability to promote qi-moving and has middle-energizer-soothing effects; therefore, it is often used to relieve hypochondrium distension and pain, and to relax the stagnation of the liver Qi. FA stir-baked with bran is more effective in nourishing the stomach and curing indigestion. AIM OF THE STUDY: In this study, the chemical composition and differences between raw FA and FA stir-baked with bran were systematically compared. The chemical components that increased after stir-baking FA and bran were separated and their pharmacodynamic characteristics were determined. Lastly, the processing mechanism of FA was further explained. MATERIALS AND METHODS: Twelve main chemicals in raw FA and FA stir-baked with bran were compared using high-performance liquid chromatography (HPLC). The main differential components were identified, separated, purified, and then analyzed using pharmacodynamic tests. The intestine-pushing test, in vitro smooth muscle test, and in vitro acetylcholinesterase (AchE) activity test in mice were performed to explain the mechanism of auraptene in improving gastrointestinal motility. RESULTS: Using HPLC, the primary chemical that differed between raw FA and FA stir-baked with bran was identified as auraptene. The processed FA was extracted, separated, and purified to obtain pure auraptene. The intestine-pushing test in mice showed that low (0.6 mg·kg-1) and medium doses (1.2 mg·kg-1) of auraptene could promote peristalsis of the small intestine, whereas a high dose (2.4 mg·kg-1) inhibited peristalsis. In vitro studies on the smooth muscle of mice showed that a low dose of auraptene (0.2 mmol·L-1, 10-800 µL) could promote contraction, whereas a high dose (0.2 mmol·L-1, >1000 µL) had the opposite effect. Auraptene has a mechanism of action similar to that of the acetylcholinesterase inhibitor, neostigmine. Additionally, auraptene could inhibit AchE activity in vitro. CONCLUSIONS: Auraptene is the main chemical constituent that differs between raw FA and FA stir-baked with bran. Pharmacodynamic tests showed that auraptene has a cholinergic effect, by virtue of its role as an acetylcholinesterase inhibitor. Moreover, auraptene could dually regulate the gastrointestinal smooth muscle. Auraptene was present in low levels and its content varied in FA stir-baked with bran, depending on the origin and source of FA, and the treatment procedures it was subjected to. In the Chinese Pharmacopoeia, the recommended dose of FA stir-baked with bran is a low dose of 3-10 g, which effectively promotes small-intestinal peristalsis. The mechanism of action is attributed to an increase in the relative content of acetylcholine by the inhibition of AchE activity to promote gastrointestinal motility. The increased levels of auraptene in FA stir-baked with bran are the main reason and the primary purpose for the change in its medicinal properties. This technique, therefore, has potential to be used as one of the main processing mechanisms of raw FA.

Constituents and Anti-Hyperuricemia Mechanism of Traditional Chinese Herbal Formulae Erding Granule.

Erding granule (EDG) is a traditional Chinese medicine that has recently been identified as having anti-hypouricemic effects. However, the active components and underlying mechanism for this new indication have not been elucidated. Therefore, we compared the effects of different EDG extracts (water, 50% ethanol and 95% ethanol) on serum uric acid concentrations in the hyperuricemia model mouse. We also analyzed the constituents of different extracts by ultra-high performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS) to observe the variation between the active and inactive products. Extract activity and target site were evaluated by assessing uric acid- and inflammation-suppressing effects along with evaluating ability to regulate the uric acid transporter. The results showed that the 50% ethanol extract (EDG-50) had an obvious serum uric acid concentration lowering effect compared with water (EDG-S) and the 95% ethanol extract (EDG-95). UHPLC-Q-TOF-MS/MS analysis showed that EDG-50 was compositionally different to EDG-S and EDG-95. EDG-50 showed dose-dependent effects on reducing uric acid, suppressing inflammation and regulating uric acid transporters. Moreover, western blot analysis showed that EDG-50 down-regulated GLUT9 and URAT1 expression, and up-regulated OAT1 expression. Therefore, our findings enable the preliminarily conclusion that EDG-50 lowers serum uric acid concentrations, mainly by down-regulating the expression of GLUT9 and URAT1 proteins and up-regulating the expression of OAT1 proteins. This provides a research basis for clinical use of EDG as an anti-hyperuricemic agent.

Tissue Distribution and Anti-Lung Cancer Effect of 10-Hydroxycamptothecin Combined with Platycodonis Radix and Glycyrrhizae Radix ET Rhizoma.

10-Hydroxycamptothecin (HCPT) is a broad-spectrum chemotherapeutic drug, although its side effects and multidrug resistance (MDR) limit its clinical application. A range of drug delivery systems have been utilized to overcome its shortcomings and maintain its therapeutic efficacy, however the use of the transport effect of traditional Chinese medicines (TCMs) to improve the distribution of chemotherapeutic drugs has not been widely reported. Platycodonis Radix (JG) and Glycyrrhizae Radix ET Rhizoma (GC) are common TCMs in clinics and are often combined as drug pairs to act as "transport agents". In the present study, the effect of JG and GC (JGGC) on the distribution of HCPT in tissues and its antitumor efficacy after being combined as a therapy were investigated, for which ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was used. Furthermore, the effect on the protein expression of multidrug resistance proteins (P-gp and LRP), and the immunomodulatory and synergistic antiapoptotic effect on Lewis lung cancer-bearing C57BL/6J mice were also evaluated. The results demonstrate that JGGC significantly increased the area under the concentration time curve (AUC) and mean residence time (MRT) and reduced the clearance rate (CL) of HCPT. In addition, the combined use of JGGC decreased the levels of LRP, P-gp and Bcl-2/Bax when treated with HCPT. JGGC also significantly elevated the levels of RBCs, PLTs, HGB, IL-2, and IFN-γ, and decreased IL-10 levels. In summary, an increased concentration of HCPT in tissues was observed when it was combined with JGGC through inhibition of efflux protein, with a synergistic enhancement of the anticancer effect observed through promotion of apoptosis and immunity due to a reversion of the Th1/Th2 shift. Our findings provide a reference for the feasibility of combining JGGC with chemotherapy drugs in clinical applications.

Study on anti-hyperuricemia effects and active ingredients of traditional Tibetan medicine TongFengTangSan (TFTS) by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.

TongFengTangSan (TFTS), a traditional Tibetan medicine comprising of Tinospora sinensis (TS), Terminalia chebula Retz (TC) and Trogopterori faeces (TF), is used to treat joint diseases like gout, gout arthritis, swelling, pain etc. Despite the significant therapeutic effects of TFTS, its pharmacological components have not been analyzed so far. Therefore, the chemical composition of the effective part of TFTS was analyzed by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS). The results show that the ethanol extract (EE) of TFTS was more effective in reducing the serum uric acid (SUA) and XOD (Serum and Liver) levels in a hyperuricemic rats model compared to the TFTS raw powder (RP). UHPLC-Q-TOF-MS/MS identified a total of 106 compounds in the positive and negative ion mode, of which 87 were from TC, 13 from TS and 6 from TF. In addition, 106 compounds contained 57 tannins, 6 triterpenoids, 10 alkaloids, 7 flavonoids, 22 organic acids and 4 phenylpropanoids. The preliminary results indicate that the EE of TFTS includes the active anti hyperuricemic substances. The present study first investigated the efficacy and the active components of TFTS in hyperuricemic treatment, and further summarized the diagnostic ion and neutral loss patterns of MS/MS cracking of tannic compounds. These findings lay the foundation for the further study and clinical application of TFTS.

Acute and subacute toxicity of the extract of Aristolochiae fructus and honey-fried Aristolochiae fructus in rodents.

Aristolochiae Fructus (AF) and honey-fried Aristolochiae Fructus (HAF) have been used in China for thousands of years as an anti-tussive and expectorant drug. Few clinical cases were reported associated with the toxicity of AF and HAF, although relatively high contents of aristolochic acids (AAs) were found in them. This work was designed to compare the acute and subacute toxicity of AF and HAF in order to provide references for safe clinical use and to evaluate the possibility of reducing toxicity of AF by honey-processing. The extracts of the herb were fed to mice or rats via gastric tube. Various toxic signs and symptoms, body weights, serum biochemical assay, organ weights and histopathology were used to evaluate the toxic effects. The median lethal dose (LD50) of AF and HAF are 34.1±7.2 g/kg/d and 62.6±8.0 g/kg/d with a 95% average trustable probability (p=0.95), respectively. The subacute results showed a dose-dependant relationship of the toxicity of AF and HAF. Even in the high dose groups, only moderate toxicity was observed. Honey-frying and decoction with water can decrease the contents of AAs, and attenuate the toxic effects of AF. But sufficient attention should be still paid to the safety of AF and HAF due to the existence of AAs.

[Effect of honey-toasting on the constituents and contents of aristolochic acid analogues in aristolochiae fructus].

OBJECTIVE: To evaluate the degree of de-toxification of Aristolochiae Fructus by honey-toasting technology from chemical viewpoint. METHODS: The contents of aristolochic acid analogues (AAs) in Aristolochiae Fructus and its honey-toasted product were determined by HPLC, and the degree of de-toxification was evaluated comprehensively. RESULTS: After honey-toasted, the contents of AAs decreased to varying degrees, and some new compounds were found. CONCLUSION: The constituents and contents of Aristolochiae Fructus change after honey-toasted, which indicate honey-toasting can reduce the toxicity of Aristolochiae Fructus.

Simultaneous quantification of polymethoxylated flavones and coumarins in Fructus aurantii and Fructus aurantii immaturus using HPLC-ESI-MS/MS.

The major lipid-soluble constituents in Fructus aurantii (zhiqiao) and Fructus aurantii immaturus (zhishi) are polymethoxylated flavones (PMFs) and coumarins. In the present study, a high-performance liquid chromatography with electrospray ionization tandem mass spectrometry method was developed to quantify PMFs (nobiletin, tangeretin, 5-hydroxy-6,7,8,4'-tetramethoxyflavone, and natsudaidai) and coumarins (marmin, meranzin hydrate, and auraptene) simultaneously. PMFs and coumarins were detected by electrospray ionization tandem mass spectrometry in positive ion mode and quantified with multiple reaction monitor. Samples were separated on a Diamonsil C18 (150 mm × 4.6 mm, 5 µm) column using acetonitrile and formic acid-water solution as a mobile phase in gradient mode with a flow rate at 0.5 mL/min. All calibration curves showed good linearity (r² > 0.9977) within the test ranges. Variations of the intraday and interday precisions were less than 4.07%. The recoveries of the components were within the range of 95.79%-105.04% and the relative standard deviations were less than 3.82%. The method developed was validated with acceptable accuracy, precision, and extraction recoveries and can be applied for the identification and quantification of four PMFs and three coumarins in citrus herbs.