Tulin Lu

Analgesic and anti-inflammatory activity and pharmacokinetics of alkaloids from seeds of Strychnos nux-vomica after transdermal administration: effect of changes in alkaloid composition.

ETHNOPHARMACOLOGICAL RELEVANCE Strychnos nux-vomica L. (Loganiaceae) is grown extensively in southern Asian countries. The dried seed of this plant, nux vomica, has been clinically used in Chinese folk medicine for improving blood circulation, relieving rheumatic pain, reducing swelling and treating cancer. AIM OF THE STUDY This study was carried out to investigate the effect of removing most strychnine from the total alkaloid fraction (TAF) extracted from nux vomica on analgesic and anti-inflammatory activity and pharmacokinetics after transdermal administration. MATERIALS AND METHODS Most strychnine was removed from TAF and the resulted modified total alkaloid fraction (MTAF) was obtained. The contents of strychnine and brucine in TAF and MTAF were determined. Then the analgesic and anti-inflammatory activity of TAF, MTAF, brucine and strychnine dissolved in hydrogel was compared after transdermal administration. Furthermore, in vitro and in vivo transdermal absorption profiles of brucine after administration of TAF, MTAF and brucine dissolved in hydrogel were also compared. RESULTS In contrast to TAF, most strychnine was removed from MTAF and the ratio of brucine to strychnine was adjusted from 1:1.8 to 2.7:1. MTAF showed significant analgesic activity in all the chemical-, thermal- and physical- induced nociception models, which indicated the presence of both centrally and peripherally mediated activities. MTAF also showed significant anti-inflammatory activity against xylene-induced ear edema. But TAF and strychnine demonstrated little activity in all those pharmacological tests. Brucine showed to be effective in acetic acid-induced writhing and xylene-induced ear edema test. Brucine in MTAF was absorbed more completely than it alone at the same dosage of brucine after transdermal administration. CONCLUSIONS The results from the present study appeared to support the viewpoint that most strychnine should be removed from TAF to improve analgesic and anti-inflammatory activity. The relatively higher pharmacological activity of MTAF compared to brucine alone is partly due to the enhanced transdermal absorption of brucine.

Effects of unprocessed versus vinegar-processed Schisandra chinensis on the activity and mRNA expression of CYP1A2, CYP2E1 and CYP3A4 enzymes in rats.

ETHNOPHARMACOLOGICAL RELEVANCE Schisandra chinensis (SC) is a well-known traditional Chinese herbal medicine that has been used in clinical practices for thousands of years. However, the differences between the effects of unprocessed and vinegar-processed Schisandra chinensis (VSC) on cytochrome P450 (CYP450) activities are poorly understood. AIM OF THE STUDY To evaluate the differences between processed and unprocessed SC on the metabolism of CYP1A2, CYP2E1 and CYP3A4 substrates in rats using a cocktail method based on a developed and validated HPLC method. We also investigate the influence of processing on the levels of CYP mRNA. MATERIALS AND METHODS Three probe substrates (theophylline, dapsone and chlorzoxazone) were delivered simultaneously into rats treated with single or multiple doses of processed or unprocessed SC extract. The plasma concentrations of the three probes were profiled by HPLC, and their corresponding pharmacokinetic parameters were calculated. Real-time RT-PCR was performed to determine the effects of processed and unprocessed SC on the mRNA expression of CYP1A2, CYP2E1 and CYP3A4 in the liver. RESULTS Treatment with single or multiple doses of either extract of SC induced CYP3A4 enzyme activity and inhibited CYP1A2 enzyme activity in rats. Furthermore, the inhibitory effect of SC was more potent after vinegar processing than without vinegar processing. CYP2E1 enzyme activity was induced after treatment with a single dose but was inhibited after multiple doses. The mRNA expression results were in accordance with the pharmacokinetic results. CONCLUSIONS These results provide useful scientific data for the safe clinical application of either extract of SC in combination with other drugs, which should lack the side effects induced by other herb-drug interactions.

Quality control and producing areas differentiation of Gardeniae Fructus for eight bioactive constituents by HPLC-DAD-ESI/MS.

Gardeniae Fructus (G.Fructus), the fruit of Gardenia jasminoides Ellis (Rubiaceae), is a commonly used traditional Chinese medicine (TCM) that has been used for the treatment of hepatitis, jaundice, hypersonic, diabetes and hematuria. Numerous researches have demonstrated that the major active constituents in G.Fructus were responsible for the majority of medical effects of this fruit and their quantification were important for the quality control of G.Fructus. However, in the current quality control standard, only geniposide was used as characteristic marker of G.Fructus, which could not reflect the overall quality of this fruit. In order to identify more chemical makers for improving the quality control standard and evaluate producing areas differentiation of G.Fructus, in the present study, a novel and sensitive high-performance liquid chromatography-diode array detector coupled to an electrospray tandem mass spectrometer (HPLC-DAD-ESI/MS) was developed for the simultaneous determination of 8 major constituents, including geniposidic acid (1), chlorogenic acid (2), genipin-1-β-gentiobioside (3), geniposide (4), genipin (5), rutin (6), crocin-1 (7), crocin-2 (8) in G.Fructus. Moreover, chemometric analysis techniques with principal component constituent analysis (PCA) and cluster analysis (CA) involved were introduced in statistical analysis of 8 investigated constituents in the 34 batches samples to discriminate the samples from different producing areas. The results indicated that the contents of the 8 major bioactive constituents in G.Fructus varied significantly among different producing areas. From results of the loading plot from PCA analysis, genipin-1-β-gentiobioside may have more influence in discriminating the sample from different producing areas, and which was found to be the most abundant bioactive component besides geniposide in all the 34 batches samples, suggesting that it should be added as chemical marker for further investigation on the pharmacological actions and the quality control of G.Fructus.

Approach based on high-performance liquid chromatography fingerprint coupled with multivariate statistical analysis for the quality evaluation of Gastrodia Rhizoma

Gastrodia Rhizoma is a Traditional Chinese Medicine applied in the treatment of stroke, numbness of limb, headache and dizziness. However, its clinical effect is threatened by sulfur-fumigation used in the process of storage. This article employs content determination coupled with high-performance liquid chromatography fingerprint to investigate the effect of sulfur-fumigation on Gastrodia Rhizoma so as to evaluate the quality of Gastrodia Rhizoma. The result was that most active ingredient in Gastrodia Rhizoma decreased after sulfur-fumigation and the fingerprints analyzed by mathematical statistics between sulfur-fumigated Gastrodia Rhizoma and unfumigated Gastrodia Rhizoma have substantial differences, which reveals that sulfur-fumigation has a significant influence on the quality of Gastrodia Rhizoma. The conclusion of hierarchical clustering analysis, principal component analysis and partial least squares could validate each other, which implies that the method of mathematical statistics applied for assessing the quality of Gastrodia Rhizoma is effective and stable. The method not only affords a viable strategy for distinguishing Gastrodia Rhizoma whether sulfur-fumigated or not and assessment of the quality of Gastrodia Rhizoma, but also provides a reference for other herbal medicine that suffers from sulfur-fumigation.

Pharmacokinetics and liver distribution study of unbound curdione and curcumol in rats by microdialysis coupled with rapid resolution liquid chromatography (RRLC) and tandem mass spectrometry

A sensitive, specific, convenient and endogenous interference-free microdialysis sampling method coupled with RRLC-MS was successfully developed and applied to the determination of protein-unbound curdione and curcumol in biological samples. Microdialysis probes were simultaneously inserted into the jugular vein toward heart and the median lobe near the center of liver of rats under anesthesia. The separation was accomplished on a Zorbax SB-C18 column (2.1 mm × 50 mm, 1.8 μm) with a gradient elution and chromatography was conducted with RRLC system. Analytes were detected by positive ion electrospray tandem mass spectrometry and quantified on the basis of extracted ion chromatography (EIC) peak area signal. The calibration curves were linear over the range of 3.3-213.2 ng/mL for curdione and 8.1-519.2 ng/mL for curcumol. All the validation data, such as accuracy, precision, stability and matrix effect were satisfactory and within the required limits. The validated method was successfully applied to the pharmacokinetic study of curdione and curcumol in rat blood and liver after oral administration of Rhizoma Curcumae extracts. The results could provide a meaningful basis for better understanding of the intracorporal process of Rhizoma Curcumae, which would be helpful for further study both in clinic and laboratory.

Chemical analysis of raw and processed Fructus arctii by high-performance liquid chromatography/diode array detection-electrospray ionization-mass spectrometry

Background: In traditional Chinese medicine (TCM), raw and processed herbs are used to treat the different diseases. Fructus Arctii, the dried fruits of Arctium lappa l. (Compositae), is widely used in the TCM. Stir-frying is the most common processing method, which might modify the chemical compositions in Fructus Arctii. Materials and Methods: To test this hypothesis, we focused on analysis and identification of the main chemical constituents in raw and processed Fructus Arctii (PFA) by high-performance liquid chromatography/diode array detection-electrospray ionization-mass spectrometry. Results: The results indicated that there was less arctiin in stir-fried materials than in raw materials. however, there were higher levels of arctigenin in stir-fried materials than in raw materials. Conclusion: We suggest that arctiin reduced significantly following the thermal conversion of arctiin to arctigenin. In conclusion, this finding may shed some light on understanding the differences in the therapeutic values of raw versus PFA in TCM.

Characterization of Chemical Composition of Pericarpium Citri Reticulatae Volatile Oil by Comprehensive Two-Dimensional Gas Chromatography with High-Resolution Time-of-Flight Mass Spectrometry

Pericarpium Citri Reticulatae (Chenpi in Chinese) has been widely used as an herbal medicine in Korea, China, and Japan. Chenpi extracts are used to treat indigestion and inflammatory syndromes of the respiratory tract such as bronchitis and asthma. This thesis will analyze chemical compositions of Chenpi volatile oil, which was performed by comprehensive two-dimensional gas chromatography with high-resolution time-of-flight mass spectrometry (GC × GC-HR-TOFMS). One hundred and sixty-seven components were tentatively identified, and terpene compounds are the main components of Chenpi volatile oil, a significant larger number than in previous studies. The majority of the eluted compounds, which were identified, were well separated as a result of high-resolution capability of the GC × GC method, which significantly reduces, the coelution. β-Elemene is tentatively qualified by means of GC × GC in tandem with high-resolution TOFMS detection, which plays an important role in enhancing the effects of many anticancer drugs and in reducing the side effects of chemotherapy. This study suggests that GC × GC-HR-TOFMS is suitable for routine characterization of chemical composition of volatile oil in herbal medicines.

Comparative pharmacokinetics and tissue distribution of schisandrin, deoxyschisandrin and schisandrin B in rats after combining acupuncture and herb medicine (schisandra chinensis)

Recently, combination therapy with acupuncture and medicine as a practical strategy to treat diseases has gained increasing attention. The present study aimed to investigate whether acupuncture stimulation at ST.36 had a potential impact on the pharmacokinetics and tissue distribution of lignans. An HPLC-ESI/MS analytical method was established and successfully applied to a comparative study of drug concentration in plasma and tissues of three lignans. The parameters area under the plasma concentration-time curve from time zero to the final measurable point and from time zero to infinity, and peak concentration were significantly increased, with a prolonged mean residence time and a corresponding decrease in clearance in comparision with the Schisandra-alone group. Additionally, tissue concentrations of three lignans were improved in the group with acupuncture, especially in liver. The results indicated that acupuncture has a synergistic effect on the pharmacokinetics and tissue distribution of the three lignans, which could postpone their elimination, resulting in a longer blood circulating time in rat plasma and prolonged residence time in target tissues, leading to higher tissue concentration. The findings provide some scientific evidence for the mechanism of the combined use of acupuncture and herbal medicine. Furthermore, we suggest that acupuncture and its combination with herbal medicine should be investigated further as a possible adjuvant therapy in clinical treatment for liver injury.

Simultaneous determination of eleven characteristic lignans in Schisandra chinensis by high-performance liquid chromatography.

Background: Schisandra chinensis, one of the well-known traditional Chinese herbal medicines, is derived from the dry ripe fruits of Schisandra chinensis (Turcz.) Baill. according to the 9th China Pharmacopeia. Lignans are the main components isolated from extracts of S. chinensis and their content varies depending on where S. chinensis was collected. We have established a qualitative and quantitative method based on the bioactive lignans for control of the quality of S. chinensis from different sources. Materials and Methods: To develop a high-performance liquid chromatography method, an Elite ODS C18 column (250 mm Χ 4.6 mm, 5μm) at a column temperature of 30°C and flow rate of 1.0ml/min using acetonitrile (A) and water (B) as the mobile phase with a linear gradient and the peaks were monitored at 217 nm. Results: All calibration curves showed good linearity (r ≥ 0.9995) within test ranges. This method showed good repeatability for the quantification of these eleven components in S. chinensis with intra- and inter-day relative standard deviations less than 0.43% and 1.21%, respectively. In the recovery test, results of accuracy ranged from 99.51% to 101.31% with RSD values less than 2. Conclusion: The validated method can be successfully applied to quantify the eleven investigated components in 22 samples of S. chinensis from different sources.

Effect of acupuncture on target tissue distribution of Schisandra lignans

Background Recently, the combination of acupuncture and Chinese medicine as a practical strategy to treat diseases is receiving considerable attention worldwide as they are usually found to exhibit intriguing therapeutic effectiveness. The current study aimed to study the adjunct effect of acupuncture on target tissue distribution of schisandra lignans when acupuncture is combined with Schisandra chinensis. Methods A simple and reliable high performance liquid chromatography-electrospray tandem-mass spectrometry (HPLC-ESI-MS) method for simultaneous analysis of three bioactive lignans (schisandrin, deoxyschisandrin and schisandrin B) in rat tissues was established. Using this analytical method we evaluated whether acupuncture had a synergistic effect on the tissue distribution of schisandra lignans. Results Tissue concentrations of the three lignans in the group receiving acupuncture were significantly higher than those in the schisandra only group, suggesting that acupuncture may potently increase tissue concentrations of schisandra lignans. The highest concentrations of the three lignans occurred in the liver compared with other tissues, and tissue concentrations in the heart, spleen, lungs and kidneys were increased by 315%, 203%, 250% and 224%, respectively. In addition, retention times of the lignans in tissues were prolonged for a relative long time. Conclusions Our date indicate that the combined use of acupuncture and Schisandra chinensis could produce a synergistic effect which could play a beneficial role on promoting the tissue distribution of lignans. This has supported our initial hypothesis. The HPLC-MS method showed good sensitivity in quantifying the three schisandra lignans in different tissues.