Simultaneous determination of atropine, scopolamine, and anisodamine from Hyoscyamus niger L. in rat plasma by high-performance liquid chromatography with tandem mass spectrometry and its application to a pharmacokinetics study.

In order to investigate the pharmacokinetics of tropane alkaloids in Hyoscyamus niger L., a sensitive and specific high-performance liquid chromatography with tandem mass spectrometry method for the simultaneous determination of atropine, scopolamine, and anisodamine in rat plasma is developed and fully validated, using homatropine as an internal standard. The separation of the four compounds was carried out on a BDS Hypersil C18 column using a mobile phase consisting of acetonitrile and water (containing 10 mmol ammonium acetate). Calibration curves were linear from 0.2 to 40 ng/mL for atropine, scopolamine, and from 0.08 to 20 ng/mL for anisodamine. The precision of three analytes was <5.89% and the accuracy was between -1.04 to 2.94%. This method is successfully applied to rat pharmacokinetics analysis of the three tropane alkaloids after oral administration of H. niger extract. The maximum concentration of these three tropane alkaloids was reached within 15 min, and the maximum concentrations were 31.36 ± 7.35 ng/mL for atropine, 49.94 ± 2.67 ng/mL for scopolamine, and 2.83 ± 1.49 ng/mL for anisodamine. The pharmacokinetic parameters revealed areas under the curve of 22.76 ± 5.80, 16.80 ± 3.08, and 4.31 ± 1.21 ng/h mL and mean residence times of 2.08 ± 0.55, 1.19 ± 0.45, and 3.28 ± 0.78 h for atropine, scopolamine, and anisodamine, respectively.

Integrated pharmacokinetics of five protoberberine-type alkaloids in normal and insomnic rats after single and multiple oral administration of Jiao-Tai-Wan.

ETHNOPHARMACOLOGICAL RELEVANCE: Jiao-Tai-Wan (JTW), an important herbal formula consists of Rhizoma coptidis and Cortex cinnamomi powder, is a famous prescription which has been used for centuries to treat insomnia in Traditional Chinese Medicine. The purpose of this study is to compare the pharmacokinetic properties of five protoberberine-type alkaloids (i.e. berberine, palmatine, coptisine, epiberberine and jatrorrhizine), the main bioactive constituents in JTW, between normal and insomnic rats. We also investigate the differences between single-dose and multiple-dose pharmacokinetics of five protoberberine-type alkaloids. MATERIALS AND METHODS: The insomnic rat models were induced by intraperitoneal injection of one-dose para-chlorophenylalanine acid (PCPA). Quantification of five protoberberine-type alkaloids in rat plasma was achieved by using a rapid LC-MS/MS method. Plasma samples were collected at different time points to construct pharmacokinetic profiles by plotting drug concentration versus time and estimate pharmacokinetic parameters. An unpaired Student׳s t test was used for comparisons with SPSS 17.0. RESULTS: The five protoberberine-type alkaloids of single-dose normal groups had slow absorption and low bioavailability, as well as a delay of peak time. In the single-dose oral administration, the Cmax and Tmax of five ingredients in insomnic rats had significant differences compared with those of normal rats. In the multiple-dose oral administration, the pharmacokinetic parameters of five protoberberine-type alkaloids varied greatly in insomnic rats. In the normal rats, there were significant differences (P<0.05) in the principal pharmacokinetic parameters such as Cmax and Tmax between single-dose and multiple-dose oral administration. In the insomnic rats, the five ingredients of multiple-dose groups showed better absorption than the single-dose groups. Particularly, three peaks were observed in multiple-dose model group of plasma-concentration curves. CONCLUSIONS: The pharmacokinetic behavior of five protoberberine-type alkaloids was described in this paper. In both normal groups and model groups, the pharmacokinetic behavior of multiple-dose had significant differences comparing with the single-dose; either single-dose or multiple-dose, the pharmacokinetic behavior of insomnic rats had significant differences comparing the normal rats. Multiple dosing may improve the absorption of JTW in insomnic rats, which will increase the bioavailability and bring into active role in therapeutical effect.

Determination of tacrine-6-ferulic acid in rat plasma by LC-MS/MS and its application to pharmacokinetics study.

Tacrine, as a drug for treating Alzheimer's disease (AD), has low efficacy owing to its single function and serious side effects. However, tacrine-6-ferulic acid (T6FA), the dimer which added ferulic acid to tacrine, has been found to be a promising multifunctional drug candidate for AD and much more potent and selective on acetylcholinesterase (AChE) than tacrine. The aim of the present work was to develop and validate an LC-MS/MS method with electrospray ionization for the quantification of T6FA in rat plasma using tacrine-3-ferulic acid (T3FA) as internal standard and to examine its application for pharmacokinetic study in rats. Following a single liquid-liquid extraction with ethyl acetate, chromatographic separation was achieved at 25 °C on a BDS Hypersil C18 column with a mobile phase composed of 1% formic acid and methonal (30:70, v/v) at a flow rate of 0.2 mL/min. Quantification was achieved by monitoring the selected ions at m/z 474.2 → 298.1 for T6FA and m/z 432.2 → 199.0 for T3FA. The method was validated to be rapid, specific, accurate and precise over the concentration range of 0.5-1000.0 ng/mL in rat samples. Furthermore, it was successfully applied for the pharmacokinetic measurement of T6FA with an oral administration at 40 mg/kg to rats.

Isolation, identification and activities of natural antioxidants from Callicarpa kwangtungensis Chun.

Reactive oxygen species leads to some diseases associated with oxidative stress. Callicarpa kwangtungensis Chun (CK) is a common remedy in traditional Chinese medicine and possesses diverse biological activities involving antioxidant properties; its main compounds phenylethanoid glycosides (PG) and flavonoids are always reported as antioxidants. In order to develop CK as a safe and activated antioxidant, our investigation was performed to validate antioxidant properties and assess which types of compounds (similar polarity or similar structure), even which compounds, played the role of antioxidants. The extracted compounds of CK were analyzed qualitatively and quantitatively by HPLC-DAD-ESI-Trap MS and UV for their contents and antioxidant activities. The correlations between antioxidant activities and known contents were respectively counted and a semi-quantitative experiment was designed to screen antioxidant compounds of CK with HPLC-UV. The n-butanol fraction (BF) showed the highest total phenolic and flavonoid contents (TPC, TFC), and three PG (forsythiaside B, poliumoside and acteoside) contents. BF showed the significantly best (P<0.05) activities in most assays. There were significant correlations (P<0.05) between DPPH•, ABTS(+)•, •O2(-) scavenging, Cu(2+)-chelating, anti-lipidperoxidation activities and TPC. BF also has significant antioxidant activities on CCl4-induced acute liver injury Mice and TBHP-reduced HepG2 cells. Nine PG (forsythiaside B, poliumoside, acteoside, alyssonoside, brandioside and their derivatives) and one flavone (rhamnazin) were screened out as antioxidants. BF in CK contained abundant polyphenolic, which reflected some definite antioxidant properties. The antioxidant compounds consisted at the least of nine PG and one flavone.

Simultaneous determination of borneol and its metabolite in rat plasma by GC-MS and its application to pharmacokinetic study.

A gas chromatography mass spectrometry (GC-MS) method has been developed and fully validated for the simultaneous determination of natural borneol (NB) and its metabolite, camphor, in rat plasma. Following a single liquid-liquid extraction, the analytes were separated using an HP-5MS capillary column (0.25 mm×30 m×0.25 µm) and analyzed by MS in the selected ion monitoring mode. Selected ion monitor (m/z) of borneol, camphor and internal standard was 95, 95 and 128, respectively. Linearity, accuracy, precision and extraction recovery of the analytes were all satisfactory. The method was successfully applied to pharmacokinetic studies of NB after oral administration to Wistar rats.

Simultaneous determination of three phenylethanoid glycosides from Callicarpae Caulis et Folium in rat plasma by LC-MS/MS and its application to PK study.

BACKGROUND: Callicarpae Caulis et Folium (CCF) is a traditional Chinese medicine usually used for hemostasis in clinics. In this study, a novel LC-MS/MS method was developed and validated for the simultaneous quantification of three phenylethanoid glycosides in rat plasma (verbascoside, forsythoside B and poliumoside), which are the major bioactive compounds of CCF; MS was operated in negative mode. RESULTS: This method was linear between 5.2 and 1010 ng/ml for poliumoside, 7.0 and 420 ng/ml for forsythoside B and 2.60 and 260.0 ng/ml for verbascoside. The MS/MS ion transitions monitored were m/z 769.4→160.5, m/z 755.3→593.3, m/z 623.1→160.5 and m/z 179.0→133.6 for poliumoside, forsythoside B, verbascoside and caffeic acid (IS), respectively. Linearity, accuracy, precision and extraction recovery of three analytes were all satisfactory. CONCLUSION: The method developed was sensitive, specific and rapid. It has been successfully applied in a PK study of three phenylethanoid glycosides after a single oral administration of CCF extract to rats.