Pharmacokinetics and penetration into synovial fluid of systemical and electroporation administered sinomenine to rabbits

Sinomenine is an anti‐rheumatoid arthritis (RA) drug derived from the Sinomenium acutum. The major site of RA treatment is within the synovial compartment. However, the pharmacokinetic and penetration into synovial fluid (SF) of sinomenine have not been reported. In our study, the pharmacokinetics and penetration into SF of systemic and electroporation administered sinomenine were investigated by microdialysis incorporated with HPLC‐MS/MS. Sinomenine went into plasma and SF more rapidly with higher peak concentration (C_max) by intramuscular injection compared with oral administration. The area under the concentration–time graph (AUC_0–∞) of intramuscularly injected sinomenine was 1,403,294.75 ± 125,534.567 ng min/mL in plasma and 456,116.37 ± 62,648.36 ng min/mL in SF, which were equivalent with those for an oral dose. These results indicated that equal amounts of sinomenine could penetrate into SF by the two administration routes, and the permeation ratios were approximately 1:3. The AUC_0–∞ and C_max were lower with electroporation compared with systemic administration, but the C_SF/C_Plasma (concentration of sinomenine in SF vs that of plasma) at 90, 120, 150, 180, 240 and 480 min by electroporation was 3‐ to 10‐fold higher relative to systemic administration. This illustrated that sinomenine can be targeted into joints by electroporation, and electroporation is a potential technique for sinomenine's transdermal delivery. Copyright © 2014 John Wiley & Sons, Ltd.     

Comparative pharmacokinetic study of the main components of cortex fraxini after oral administration in normal and hyperuricemic rats

Cortex Fraxini is an important traditional Chinese herbal medicine used for the treatment of gout and hyperuricemia. An efficient and rapid ultra‐performance liquid chromatography mass spectrometry method was developed and validated for simultaneous quantitation of six coumarins (aesculin, fraxin, aesculetin, fraxetin, sopoletin and 7‐hydroxycoumarin) in normal and hyperuricemic rats plasma after oral administration of Cortex Fraxini. The method could successfully be applied for pharmacokinetics studies. The pharmacokinetic behavior of six coumarins in normal and hyperuricemia rats plasma was determined. Results showed that, for some of analytes, the pharmacokinetic parameters (AUC_0–t , AUC_0–∞, C _max, T _max and CL ) were significantly different between normal and hyperuricemic rats. The different pharmacokinetic parameters might result from renal impairment or a change of metabolic enzymes in the pathological state. The pharmacokinetic study in pathological state could provide more useful information to guide the clinical use of traditional Chinese herbal medicine.     

In vitro and in vivo identification of metabolites of magnoflorine by LC LTQ‐Orbitrap MS and its potential pharmacokinetic interaction in Coptidis Rhizoma decoction in rat

Magnoflorine, an important aporphine alkaloid in Coptidis Rhizoma, is increasingly attracting research attention because of its pharmacological activities. The in vivo and in vitro metabolism of magnoflorine was investigated by LC LTQ‐Orbitrap MS. In vivo samples including rat urine, feces, plasma and bile were collected separately after both oral (50 mg kg^?1) and intravenous administration (10 mg kg^?1) of magnoflorine, along with in vitro samples prepared by incubating magnoflorine with rat intestinal flora and liver microsome. As a result, 12 metabolites were found in biological samples. Phase I metabolites were identified in all biological samples, while phase II metabolites were mainly detected in urine, plasma and bile. In a pharmacokinetic study, rats were not only dosed with magnoflorine via oral (15, 30 and 60 mg kg^?1) and intravenous administration (10 mg kg^?1) but also dosed with Coptidis Rhizoma decoction (equivalent to 30 mg kg^?1 of magnoflorine) by intragastric administration to investigate the interaction of magnoflorine with the rest of compounds in Coptidis Rhizoma. Studies showed that magnoflorine possessed lower bioavailability and faster absorption and elimination. However, pharmacokinetic parameters altered significantly (p < 0.05) when magnoflorine was administered in Coptidis Rhizoma decoction. Oral gavage of Coptidis Rhizoma decoction decreased the absorption and elimination rates of magnoflorine, which revealed that there existed pharmacokinetic interactions between magnoflorine and the rest of ingredients in Coptidis Rhizoma. Copyright © 2015 John Wiley & Sons, Ltd.     

Comparative pharmacokinetics of nine major bioactive components in normal and ulcerative colitis rats after oral administration of Lizhong decoction extracts by UPLC–TQ–MS/MS

Lizhong decoction (LZD), a classic formula, has been used to treat ulcerative colitis (UC) for thousands of years in clinical practice. However, the pharmacokinetic characteristics of its major bioactive components in rats under different physiological and pathological states are not clear. Thus, in this study, a rapid and sensitive analytical method, ultra‐performance liquid chromatography coupled with mass spectrometry (UPLC–MS/MS) method, was developed and applied to simultaneously determine glycyrrhizic acid, liquiritin, isoliquiritin, glycyrrhizin, isoliquiritigenin, 6‐gingerol, ginsenoside Rg1, ginsenoside Rb1 and ginsenoside Re in normal and UC rats after oral administration of LZD extract. A Waters BEH C₁₈ UPLC column was used for chromatographic separation, while acetonitrile and 0.1% formic acid were selected as mobile phase. The linearity of nine analytes was >0.9920. Inter‐ and intra‐day accuracy was ≤ 11.4% and precision was from 1.1 to 12.7%. Additionally, stable and suitable extraction recoveries were also obtained. The established method was validated and found to be specific, accurate and precise for nine analytes. Furthermore, it was successfully applied to the pharmacokinetic investigation of nine major components after oral administration of LZD extracts to normal and model rats, respectively. The results showed that the pharmacokinetic parameters (C_max, T_max, AUC_0–t, AUC_0–∞) in the plasma of UC rats were significantly different from those of normal rats, which could provide a reference for the clinical application of LZD.     

LC Determination and Pharmacokinetic Study of Sinomenine in Dog Plasma

A simple HPLC method has been developed for determination of sinomenine in dog plasma and has been used to evaluate the pharmacokinetics of sinomenine tablets in dogs. Chromatographic separation was performed on a reversed-phase column with 0.78% (w/v) NaH₂PO₄-acetonitrile, 88:12 (v/v), as mobile phase, delivered at a flow rate of 1.5 mL min^?1. Detection was performed at 265 nm. The limit of quantification was 5.0 ng mL^?1. The calibration range was from 5.0 to 1000 ng mL^?1. The developed method was applied to pharmacokinetic studies of sinomenine sustained-release tablets (test preparation) and sinomenine conventional tablets (reference preparation) in six dogs. Pharmacokinetic data t _max, C _max, AUC _0-t , AUC _0-∞, and t _1/2 for both preparations were determined from plasma concentration-time profiles. The method was sufficiently sensitive, simple, and repeatable for use in pharmacokinetic studies.     

Screening and identification of Caulis Sinomenii bioactive ingredients with dual‐target NF‐κB inhibition and β2‐AR agonizing activities

Caulis Sinomenii (CS) is a valuable traditional medicine in China. Its extract can act as an anti‐inflammatory agent and a vascular smooth muscle relaxant. However, the underlying mechanisms remain unknown. In this study, we developed a simple dual‐target method based on ultra‐performance liquid chromatography/quadrupole time‐of‐flight mass spectrometry combined with a dual‐target bioactive screening assay for anti‐inflammatory and antispasmodic activities to characterize the chemical structure of various bioactive compounds of CS rapidly. Seven potential NF‐κB inhibitors were identified, including laudanosoline‐1‐O‐xylopyranose, 6‐O‐methyl‐laudanosoline‐1‐O‐glucopyranoside, menisperine, sinomenine, laurifoline, magnoflorine and norsinoacutin. Furthermore, IL‐6 and IL‐8 assays confirmed the anti‐inflammatory effects of these potential NF‐κB inhibitors, in which laudanosoline‐1‐O‐d ‐xylopyranose and menisperine were revealed as novel NF‐κB inhibitors. Among the seven identified alkaloids, three potential β₂‐adrenergic receptor agonists, including sinomenine, magnoflorine and laurifoline, were characterized using a luciferase reporter system to measure for the activity of β₂‐adrenergic receptor agonists. Finally, sinomenine, magnoflorine and laurifoline were identified not only as potential NF‐κB inhibitors but also as potential β₂‐adrenegic receptor agonists, which is the first time this has been reported. Molecular dynamic simulation and docking results suggest that the three dual‐bioactive constituents could not only inhibit Pseudomonas aeruginosa PAK strain‐induced inflammatory responses via a negative regulation of the Braf protein that participates in MAPK signaling pathway but also activate the β₂‐adrenegic receptor. These results suggest that CS extract has dual signaling activities with potential clinical application as a novel drug for asthma.     

Simultaneous determination of paeoniflorin from total glucosides of paeony in Sprague–Dawley rats and spontaneously hypertensive rats by high‐performance liquid chromatography–tandem mass spectrometry: in vivo and in vitro studies

Paeoniflorin is a well‐known monoterpene glucoside in the herbal drug that exhibits a number of biological activities. The pharmacokinetic characteristics of paeoniflorin from total glucosides of paeony in spontaneously hypertensive rats (SHR) are still unclear. It is essential to investigate the in vivo and in vitro pharmacokinetic differences of paeoniflorin from total glucosides of paeony in Sprague–Dawley (SD) and SHR. The in vivo pharmacokinetic data were analyzed using DAS 2.0 software and the in vitro metabolic characteristics were measured using rat hepatic microsomes. The concentration of paeoniflorin in biological samples was determined using high‐performance liquid chromatography–electrospray ionization tandem mass spectrometry method, which showed good precision and stability. The plasma concentration–time profiles of paeoniflorin following oral administration of total glucosides of paeony showed a single peak and there were significant differences in the mean values of AUC_(0–t), AUC_(0–∞), CL_z/F and T_max between SD and SHR (p < 0.05). The metabolic rate of paeoniflorin from total glucosides of paeony was slower in SHR than in SD rats (p < 0.05). The results might be useful in further applications of paeoniflorin and total glucosides of paeony. Copyright © 2016 John Wiley & Sons, Ltd.     

The effect of tripterygium glucoside tablet on pharmacokinetics of losartan and its metabolite EXP3174 in rats

Losartan and tripterygium glucoside tablet (TGT) are often simultaneously used for reducing urine protein excretion in clinic. However, it is unknown whether there is potential herb–drug interaction between losartan and TGT. The aim of this study was to investigate their potential herb–drug interaction, and clarify the mechanism of the effect of TGT on the pharmacokinetics of losartan and its metabolite EXP3174 in rats. The plasma concentrations of losartan and EXP3174 were determined by LC–MS, and the main pharmacokinetic parameters were calculated. The C _max, t _1/2 and AUC_{(0–t )} of losartan became larger after co‐administration, while the C _max and AUC_{(0–t )} of EXP3174 became smaller, suggesting that TGT could influence the pharmacokinetics of losartan and EXP3174. The effects of TGT and its main components on the metabolic rate of losartan were further investigated in rat liver microsomes. Results indicated that TGT and its two main ingredients could decrease the metabolic rate of losartan. Therefore, it was speculated that TGT might increase the plasma concentration of losartan and decrease the concentration of EXP3174 by inhibiting the metabolism of losartan. The results could provide references for clinical medication guidance of losartan and TGT to avoid the occurrence of adverse reactions.     

Metabolic profile of Kudiezi injection in rats by UHPLC coupled with Fourier transform ion cyclotron resonance mass spectrometry

In this study, a reliable and sensitive ultra‐high performance liquid chromatography coupled with fourier transform ion cyclotron resonance mass spectrometry method was developed for the systematic study of the metabolic profile of Kudiezi injection in rat plasma, bile, urine, and feces after intravenous administration of a single dose. The chromatographic separation was performed on an Agilent Eclipse Plus C₁₈ column (4.6 mm × 50 mm, 1.8 μm) and the identification of prototype components and metabolites was achieved on a Bruker Solarix 7.0 T ultra‐high resolution spectrometer in negative ion mode. Results indicated that a total of 76 constituents including 29 prototype compounds and 47 metabolites (10 phase I metabolites and 37 phase II metabolites) were tentatively identified. And the metabolic pathways of these prototype compounds including hydroxylation, dehydrogenation, glucuronidation, and sulfate conjugation. In conclusion, the developed method with high resolution and sensitivity was effective for screening and identification of prototypes and metabolites of Kudiezi injection in vivo. Moreover, these results would provide significant information for further pharmacokinetic and pharmacological research of Kudiezi injection in vivo.     

Comparative study of thermal stability of healthy and focal segmental glomerulosclerosis plasma albumin

The thermodynamic parameters calculated from measurements obtained by differential scanning calorimetry from healthy and focal segmental glomerulosclerosis albumin are reported. The same values were determined by fluorescence spectra and by the second derivative absorption spectra and they resulted in agreement with values obtained from the calorimetry technique. Nevertheless the unfolding mechanism seems to be completely altered when pathological albumin is compared with healthy albumin. The C _p values measured by calorimetry show an increase with mild slope with healthy protein; on the contrary the slope intensely increase with pathological protein. Furthermore the λ_max of this molecule is lower and drastically decrease with the increase of temperature when compared with healthy one. Therefore the modification of cys 34 on pathological albumin is supposed to cause an alteration of the structure, the swelling and the unfolding mechanism.     

The pharmacokinetic study of sinomenine,paeoniflorin and paeonol in rats after oral administration of a herbal product Qingfu Guanjiesu capsule by HPLC

An accurate and reliable high‐performance liquid chromatography–diode array detector (HPLC‐DAD) method was developed and validated for determination of sinomenine (SI), paeoniflorin (PF) and paeonol (PA), which was further applied to assess the pharmacokinetics of SI, PF and PA in an anti‐arthritic herbal product, Qingfu Guanjieshu (QFGJS) capsule, in rats. Successful separation was achieved with a C₁₈ column and a mobile phase composed of acetonitrile and aqueous phase (containing 0.1% formic acid, adjusted with triethylamine to pH 3.5 ± 0.2). The method was validated with excellent precision, accuracy, recovery and stability in calibration ranges from 0.06 to 11.62 µg/mL for SI, from 0.09 to 35.70 µg/mL for PF, and from 0.15 to 4.53 µg/mL for PA (with r² > 0.999 for all three compounds). Our results showed that absorption of PF after administration of QFGJS was similar to that after oral administration of PF alone; the absorption of SI was decreased while the absorption of PA was increased after giving QFGJS orally compared with pure compounds. We may conclude that pharmacokinetic studies of complex herbal products are not only necessary but also feasible by using representative bioactive chemicals as indicators of establishing quality control standards and of determining pharmacokinetic behavior of herbal medicines. Copyright © 2014 John Wiley & Sons, Ltd.     

Study on the PK profiles of magnoflorine and its potential interaction in Cortex phellodendri decoction by LC-MS/MS

Magnoflorine, an aporphine alkaloid in Cortex phellodendri, is increasingly attracting research attention because of its antidiabetic effects. However, at present, little information on its pharmacokinetics (PK) in vivo is available. In this study, a sensitive, rapid, and selective method was developed to determine the magnoflorine content in rat plasma using liquid chromatography–tandem mass spectrometry. Following liquid–liquid extraction, the calibration curve showed good linearity within the concentration range of 2.93 to 1,500 ng ml^?1. The intra- and inter-day precisions were all below 7.8 %, and the accuracy ranged from 94.9 to 103.4 %. The method was successfully applied in investigating the PK of magnoflorine in rats. The compound had low bioavailability, a high absorption rate, and a high elimination rate. However, area under the curve, T _1/2, and MRT increased approximately twofold when the same dosage of the compound was administered in a C. phellodendri decoction (20.8 g kg^?1). Moreover, T _max was prolonged from 0.3 to 3.33 h. Furthermore, a comparison of coadministration of the mixture group, magnoflorine (40 mg kg^?1) and berberine (696.4 mg kg^?1), with the C. phellodendri decoction group, revealed that no statistical difference (P?>?0.05) was found in the parameter AUC, and certain similar changes in the PK trend to the herbal medicine group were also observed. These results suggested that oral administration of the herbal medicine decreased the absorption and elimination rates of magnoflorine and increased its bioavailability. Berberine played a significant role in interacting with magnoflorine and in affecting the PK profiles of magnoflorine in the C. phellodendri decoction group.

Effects of acute lanthanum exposure on calcium absorption in rats

After an acute exposure to lanthanum chloride, the pharmacokinetics of calcium uptake in rats was studied by radioactive ⁴⁷Ca tracer. The accumulated doses of calcium in the left femurs during 24 hours were determined. The results showed that the area under the curves (AUC), specific activity of maximal blood ⁴⁷Ca concentration (C _max ), distribution rate constant (K _a ) and the accumulated dose of calcium in the left femur decreased while time to C _max (T _peak ) increased with the rising dosage of lanthanum exposure. It indicated that lanthanum expose had a negative effect on calcium absorption.     

Randomized two‐way cross‐over bioequivalence study of two amoxicillin formulations and inter‐ethnicity pharmacokinetic variation in healthy Malay volunteers

The objectives of this study were to develop a new deproteinization method to extract amoxicillin from human plasma and evaluate the inter‐ethnic variation of amoxicillin pharmacokinetics in healthy Malay volunteers. A single‐dose, randomized, fasting, two‐period, two‐treatment, two‐sequence crossover, open‐label bioequivalence study was conducted in 18 healthy Malay adult male volunteers, with one week washout period. The drug concentration in the sample was analyzed using high‐performance liquid chromatography (UV–vis HPLC). The mean (standard deviation) pharmacokinetic parameter results of Moxilen® were: peak concentration (C_max), 6.72 (1.56) µg/mL; area under the concentration–time graph (AUC_0–8), 17.79 (4.29) µg/mL h; AUC_0–∞, 18.84 (4.62) µg/mL h. Those of YSP Amoxicillin® capsule were: C_max, 6.69 (1.44) µg/mL; AUC_0–8, 18.69 (3.78) µg/mL h; AUC0_0–∞, 19.95 (3.81) µg/mL h. The 90% confidence intervals for the logarithmic transformed C_max, AUC_0–8 and AUC_0–∞ of Moxilen® vs YSP Amoxicillin® capsule was between 0.80 and 1.25. Both C_max and AUC met the predetermined criteria for assuming bioequivalence. Both formulations were well tolerated. The results showed significant inter‐ethnicity variation in pharmacokinetics of amoxicillin. The C_max and AUC of amoxicillin in Malay population were slightly lower compared with other populations. Copyright © 2014 John Wiley & Sons, Ltd.     

Pharmacokinetic study of five ginsenosides using a sensitive and rapid liquid chromatography–tandem mass spectrometry method following single and multiple oral administration of Shexiang Baoxin pills to rats

Shexiang Baoxin pills (SBP) are a traditional Chinese medicine that are used for treating coronary heart disease. Ginsenosides are the main effective components of SBP, but a comprehensive and deep pharmacokinetic study of ginsenosides in SBP, including multiple dosing and linear or nonlinear properties, is lacking. This study was designed to investigate and compare the pharmacokinetic characteristics of ginsenosides in SBP at a single dose and in multiple doses. A liquid chromatography–tandem mass spectrometry (LC‐MS/MS) method was developed for the simultaneous determination of the ginsenosides Rg1, Re, Rb3, Rc and Rb1 in rat plasma. Rats were randomly assigned to receive a single dose of 4, 8 or 12 g/kg and multiple doses (4 g/kg) of SBP for 8, 15 or 22 consecutive days. The results revealed that ginsenosides, following a single oral dose of 4 or 8 g/kg, were absorbed rapidly, with a T_max ranging from 0.250 to 1.08 h. The AUC_0–t and C_max of the ppd‐type ginsenosides Rb3, Rc and Rb1 were greater than those of the ppt‐type ginsenosides Rg1 and Re. Nondose‐dependent exposure was observed at doses of 4–12 g/kg for all of the ginsenosides. After multiple dosing, the plasma levels of the ppt‐type ginsenosides decreased, whereas those of the ppd‐type ginsenosides did not change significantly. In conclusion, the LC‐MS/MS method was successfully applied to investigate the pharmacokinetics of ginsenosides after single and multiple oral administrations of SBP. The ginsenosides did not accumulate after multiple dosing. The ppd‐type ginsenosides displayed more favorable pharmacokinetic properties compared with the ppt‐type ginsenosides. Copyright © 2014 John Wiley & Sons, Ltd.     

A sensitive UPLC–MS/MS method for simultaneous determination of polyphenols and theaflavins in rat plasma: Application to a pharmacokinetic study of Da Hong Pao tea

A method based on ultra‐performance liquid chromatography–tandem mass spectrometry has been developed for the rapid and simultaneous determination of five catechins and four theaflavins in rat plasma using ethyl gallate as internal standard. The pharmacokinetic profiles of these compounds were compared after oral administration of five kinds of Da Hong Pao tea to rats. Biosamples processed with a mixture of β‐glucuronidase and sulfatase were extracted with ethyl acetate–isopropanol. Chromatographic separation was achieved by gradient elution using 10 mm HCOONH₄ solution and methanol as the mobile phase. Analytes were detected using negative ion electrospray ionization in multiple reaction monitoring mode. The lower limits of quantification were 1.0, 0.74 and 0.5 ng/mL for theaflavins, two catechins and three catechins, respectively. The validation parameters were well within acceptable limits. The average half‐lives (t_1/2) in blood of the reference solution group was much shorter than those of tea samples. The values of AUC_0–t and C_max of the polyphenols and theaflavins exhibited linear pharmacokinetic characteristics which were related to the dose concentration.     

Nine components pharmacokinetic study of rat plasma after oral administration raw and prepared Semen Cassiae in normal and acute liver injury rats

In China, Semen Cassiae has long been used to protect liver, brighten eyes, and relieve constipation. Prepared Semen Cassiae is produced from raw Semen Cassiae by processing, the two forms of Semen Cassiae have different clinical applications. Pathological state is an important factor affecting the efficacy of drugs, the pharmacokinetic behavior of drugs could be significantly changed when people or animal were under different pathological state. To clarify the effect of processing mechanism and pathological state for pharmacokinetic behavior, the pharmacokinetics of nine components of raw and prepared Semen Cassiae under normal and acute liver injury rats were examined. The results showed that the bimodal phenomenon appeared on the plasma concentration‐time profiles of obtusin, emodin, chrysophanol, aloe emodin and rhein. The T_max of aurantio‐obtusin, obtusin, chrysoobtusin, emodin, chrysophanol, aloe emodin, physcion in normal groups administrated prepared Semen Cassiae were shorter than those administrated raw Semen Cassiae. For the AUC_0–t, aurantio‐obtusin, obtusin, chrysoobtusin, chrysophanol, aloe emodin and physcione in model groups administrated prepared Semen Cassiae were significantly higher than other groups, unlike above components, rhein had poor absorption in model groups. The study would be useful for further studies on pharmacokinetics and clinical application of raw and prepared Semen Cassiae.     

Application of column‐switching HPLC method in evaluating pharmacokinetic parameters of zaltoprofen and its salt

The objective of this study was to compare the pharmacokinetic parameters of zaltoprofen and those of its sodium salt in rats. Zaltoprofen, a potent non‐steroidal anti‐inflammatory agent, was virtually insoluble in water, but its sodium salt had excellent water solubility. To investigate the effect of aqueous solubility differences upon their pharmacokinetic parameters, minicapsules containing the drug powders were administrated orally to rats, and blood samples were taken via the common carotid artery. A column‐switching high‐performance liquid chromatographic analytical procedure was developed and validated for the quantitation of zaltoprofen in rat plasma samples. Our study demonstrated that the time required to reach maximum plasma concentration (T_max) of zaltoprofen sodium was significantly reduced and its maximum plasma concentration (C_max) was increased 1.5‐fold, relative to the values for zaltoprofen. It is anticipated that the sodium salt of zaltoprofen will allow the rapid onset of the drug's action in the treatment of inflammatory diseases. Copyright © 2008 John Wiley & Sons, Ltd.