Simultaneous determination of cinnamaldehyde and its metabolite in rat tissues by gas chromatography-mass spectrometry.

Cinnamaldehyde (CA), an active ingredient isolated from the traditional Chinese medicine Cortex Cinnamomi, has a wide range of bioactivities. To clarify the distribution characteristics of CA, a selective and sensitive method utilizing gas chromatography-mass spetrometry was initially developed for simultaneously determining the concentration of CA and its metabolite cinnamyl alcohol in rat tissues. Selected ion masses of m/z 131, 105 and 92 were chosen, and separation of the analytes was performed on a DB-5 ms (30 m × 0.25 mm, 0.25 µm, thickness) capillary column by gas chromatography-mass spectrometry. The calibration curves demonstrated good linearity and reproducibility over the range of 20-2000 and 20-4000 ng/mL for various tissue samples. Recoveries ranged from 86.8 to 107.5%, while intra- and interday relative standard deviations were all <11.3%. The analysis method was successfully applied in tissue distribution studies for CA and cinnamyl alcohol. As CA and cinnamyl alcohol may inter-convert to one another, simultaneous determination of both analytes provides a comparative and accurate data for tissue study. The concentrations of CA and cinnamyl alcohol remaining in spleen were the highest among the main organs, including heart, liver, spleen, lung, kidney and brain. In addition, there was no long-term accumulation of CA in rat tissues.

[Protective effect of Sankang Capsule on immunity-deficiency mice induced by cyclophosphamide].

OBJECTIVE: To study the immunomodulatory effect of Sankang Capsule( SKC) on the immunity-deficiency mice. METHODS: The immunity-deficiency model was induced by intraperitoneal injection (ip) of cyclophosphamide (CTX) at the dose of 100 mg/ kg in mice which were randomly divided into normal control group, immunity-deficiency model group, SKC treated group (250 mg/kg, 500 mg/kg) and positive control group (500 mg/kg). The number of WBC in peripheral blood, the weight of immune organs, the phagocytosis activity of macrophage, the content of serum hemolysin and the proliferation of T lymphocyte in the spleen were measured. RESULTS: The number of WBC, the weight of immune organs, the phagocytosis activity of macrophage, the content of serum hemolysin and the proliferation of T lymphocyte in the model group were all decreased compared with those of normal control group. After the administration of SKC, the index mentioned above was increased compared with those of model group. CONCLUSION: SKC has a protective effect on the immunity-deficiency mice, which may be related to the increasing of cellular immunity, humoral immunity and nonspecific immunity.

Effect of sinomenine on morphine dependence in isolated guinea pig ileum.

OBJECTIVE: To investigate the effects of sinomenine on morphine withdrawal response and acetylchline (Ach)-induced contracture in isolated guinea pig ileum. METHODS: The withdrawal contracture was elicited by subjecting isolated ileum incubated with morphine (3 micromol/L) at 37.5 degrees Celsius for 4 h to naloxone (1 micromol/L) treatment. Sinomenine (10, 50, 250 micromol/L) and nimodipine (Nim, 0.1 micromol/L) were administered 1 min before and after naloxone in morphine-dependent ilea bathed in Krebs solution containing morphine, to observe the changes in the withdrawal contracture of the ileum. The effect of sinomenine (10, 50, 250 micromol/L) on the contracture of untreated ileum in Krebs solution elicited by acetylcholine was also observed. RESULTS AND CONCLUSION: Naloxone-induced withdrawal contracture or acetylcholine-induced contracture of the ileum was significantly decreased in a dose-dependent manner, indicating that sinomenine can inhibit morphine withdrawal symptoms in guinea pigs.

Pharmacokinetic study of cinnamaldehyde in rats by GC-MS after oral and intravenous administration.

A selective and sensitive method utilizing gas chromatography-mass spectrometry was developed for simultaneous determination of cinnamaldehyde, cinnamyl alcohol, and methyl cinnamate in rat plasma. Cinnamaldehyde and cinnamyl alcohol can inter-convert to one another in rats, thus simultaneous quantifying both analytes provided a reliable and accurate method of assessment. Three qualifying ions (131 m/z, 105 m/z and 92 m/z) were chosen for simultaneous quantification of cinnamaldehyde and its metabolites. In this study, the calibration curves demonstrated a good linearity and reproducibility over the range of 20-2000ng/ml (r(2)≥0.999) for all analytes. Furthermore, the sensitivity of gas chromatography-mass spectrometry revealed sufficient lower limit of quantitation and detection of 20ng/ml and 5ng/ml, respectively, in the pharmacokinetic analysis. The intra- and inter-day precision variations were less than 10.4% and 12.2%, respectively, whilst accuracy values ranged from -8.6% to 14.8%. All analytes were stable in plasma and in processed samples at room temperature for 24h with no significant degradation after three freeze/thaw cycles. A small amount of the administered cinnamaldehyde had long half-life of 6.7±1.5h. In this study, gas chromatography-mass spectrometry was demonstrated to be a powerful tool for the pharmacokinetic studies of rats after intravenous and oral administration of cinnamaldehyde.

Immunoliposome co-delivery of bufalin and anti-CD40 antibody adjuvant induces synergetic therapeutic efficacy against melanoma.

Liposomes constitute one of the most popular nanocarriers for improving the delivery and efficacy of agents in cancer patients. The purpose of this study was to design and evaluate immunoliposome co-delivery of bufalin and anti-CD40 to induce synergetic therapeutic efficacy while eliminating systemic side effects. Bufalin liposomes (BFL) conjugated with anti-CD40 antibody (anti-CD40-BFL) showed enhanced cytotoxicity compared with bufalin alone. In a mouse B16 melanoma model, intravenous injection of anti-CD40-BFL achieved smaller tumor volume than did treatment with BFL (average: 117 mm(3) versus 270 mm(3), respectively); the enhanced therapeutic efficacy through a caspase-dependent pathway induced apoptosis, which was confirmed using terminal deoxynucleotidyl transferase-mediated dUTP-Fluorescein nick end labeling and Western blot assay. Meanwhile, anti-CD40-BFL elicited unapparent body-weight changes and a significant reduction in serum levels of tumor necrosis factor-α, interleukin-1ß, interleukin-6, interferon-γ, and hepatic enzyme alanine transaminase, suggesting minimized systemic side effects. This may be attributed to the mechanism by which liposomes are retained within the tumor site for an extended period of time, which is supported by the following biodistribution and flow cytometric analyses. Taken together, the results demonstrated a highly promising strategy for liposomal vehicle transport of anti-CD40 plus bufalin that can be used to enhance antitumor effects via synergetic systemic immunity while blocking systemic toxicity.