Determination of Risperidone and 9-hydroxyrisperidone in Human Plasma by LC-MS/MS / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of risperidone (RIS) and 9-hydroxyrisperi-done (9-OH-RIS) in human plasma. METHODS:After liquid-liquid extraction of plasma sample,using AF2672 as internal stan-dard(IS),LC-MS/MS was adopted. The determination was performed on XtimateTM C18 column with mobile phase consisted of ace-tonitrile-10 mmol/L ammonium acetate solution(containing 0.1％ formic acid)(37:63,V/V,pH=3.2)at the flow rate of 0.25 mL/min. The column temperature was 40 ℃,and the sample size was 6 μL. The ESI was equipped and quantitative analysis was operated in positive ion and MRM mode. The mass transition ion-pairs were followed as m/z 411.26→191.02 for RIS,m/z 427.21→206.91 for 9-OH-RIS and m/z 418.00→175.95 for IS. RESULTS:The linear range of RIS was 0.2-50.0 ng/mL(r=0.9997)and 9-OH-RIS was 1.0-200.0 ng/mL (r=0.9987). RSDs of inter-day and intra-day were all lower than 15％,and the method recoveries were 92.42％-104.81％ and 94.51％-100.57％;matrix effects were 98.33％-107.09％ and 91.05％-105.80％;extraction recoveries were 78.11％-92.62％ and 76.32％-85.09％,respectively. The plasma concentrations of RIS and 9-OH-RIS in 78 schizophrenic patients were(13.58±8.31)and(25.62±15.52)ng/mL. CONCLUSIONS:The developed method is simple,sensitive and specific,which is suitable for routine drug monitoring and acute toxic analysis in patients receiving risperidone.

The Correlation between Multidrug Resistance Gene Polymorphism and Plasma Concentration of Risperi-done and Its Metabolite / 中国药房

OBJECTIVE:To discuss the correlation between multidrug resistance gene (MDR1) polymorphisms and plasma concentration of risperidone,9-hydroxyrisperidone and total active substance in Han patients with schizophrenia. METHODS:78 Han inpatients with schizophrenia in Mental Health Institute of Second Xiangya Hospital of Central South University from Dec. 2011 to Jan. 2013 were selected,LC-MS/MS was conducted to determine the plasma concentration of risperidone,9-hydroxyrisperi-done and total active substance,PCR-LDR was adopted to determine the genotyping of C1236T,G2677T and C3435T of patients, and one-way ANOVA was used to detect the correlation between C1236T,G2677T and C3435T polymorphisms and plasma concen-tration/dose calibration ratio (C/D) value of risperidone,9-hydroxyrisperidone and total active substance. RESULTS:The average plasma concentrations of risperidone,9-hydroxyrisperidone and total active substance for 78 patients were(13.58±8.31),(25.62± 15.52)and(39.24±25.76)ng/ml,respectively;the distribution frequencies of C1236T,G2677T and C3435T met the Hardy-Wein-berg equilibrium(P>0.05);the risperidone C/D value of patients with C12367T CT and TT genotype were higher than those with CC genotype,risperidone and total active substance C/D values of patients with G2677T TT genotype were higher than those with GG genotype,the differences were statistically significant (P0.05). CONCLUSIONS:The MDR1 C1236T and G2677T polymorphisms are associated with plasma concentration of risperidone and total active substance in Han patients with schizophrenia.

Effects of lipopolysaccharide on the tryptophan-kynurenine metabolic pathway / 中华行为医学与脑科学杂志

Objective To investigate the effects of lipopolysaccharide(LPS) on the tryptophan-kynurenine(TRP) metabolic pathway in rat brains and provide new evidence for the relationship between inflammation and depression.Methods Rats in LPS group were given a single dose of 3.5 mg/kg LPS.while the rats in control group were given the same dosage of saline.The dialysis in ventro-hippocampus were collected by microdialysis within 8 hours and then the TRP,KYN and KA were detected by LC-MS/MS.And the expression of indoleamine 2,3-dioxygenase was detected by Western-blot.Results The level of TRP((550.15± 107.96) pmol/L) and KYN ((337.95±62.73) pmol/L) showed a time-dependent increase after administration LPS 4 h compared with the control group(TRP (368.38±59.31) pmol/L,KYN (172.80±43.96) pmol/L),while KA level in the circulation exhibited a trend to decrease,especially at 7 h ((3.47±0.62) pmol/L,P＜0.05).The ratio of KYN/TRP significantly increased at about 5 h (0.69±0.11,P＜ 0.05),and an ratio of KA/KYN (0.02±0.00) was dramatically decreased after administering LPS 4 h compared with the control group (0.05±0.01)(P＜0.05).Most of the analytes showed more dramatic changes around 4 h to 8 h.LPS group(1.48±0.37) increased the protein expression of indoleamine 2,3-dioxygenase compared with the control group(1.00±0.24) (P＜0.01).Conclusions LPS may cause tryptophan metabolic abnormalities and accelerate the way of kynurenine metabolism,leading to decreased the kynurenic acid status.

Research progress on Nrf2-ARE signaling pathway involved in liver disease pathological mechanism / 中国药理学通报

Nuclear factor erythroid-2 related factor 2 ( Nrf2 ) is an important nuclear transcription factor which protects cells a-gainst oxidative stress injury. Upon exposure to reactive oxygen species ( ROS) or electrophilic stress, Nrf2 can translocate into the nucleus, and then bind to the antioxidant response element ( ARE) , regulating the expression of several antioxidant enzymes and phase Ⅱ detoxifying enzymes which aimed at the detoxifica-tion and elimination of harmful exogenous chemicals, resulting in the facilitation of hepatoprotection. Oxidative stress is the com-mon pathogenesis of many liver diseases, while the Nrf2-ARE signaling pathway is extremely important in the prevention and progression of many liver diseases. Nrf2 has more recently been implicated as a new therapeutic target in treating liver diseases. Here, we focus on the most common liver diseases and the devel-opment of these conditions where activation of Nrf2 may alleviate disease progression, so as to provide reference for related re-search in the future.

Endothelial progenitor cells: the new target of anti-atherosclerosis drugs / 中南大学学报(医学版)

Endothelial progenitor cells (EPCs) are a kind of progenitor cells with high potential of proliferation, which exist in the bone marrow, umbilical cord blood, and peripheral blood. Under certain conditions, EPCs can differentiate into mature vascular endothelial cells. Many studies have shown that EPCs could delay the onset and development of atherosclerosis by promoting the repair of the endothelium and neovascularization. EPCs have also been considered to be a biological marker for cardiovascular diseases. Recent investigations demonstrate that EPCs can mediate the effect of some anti-atherosclerosis drugs. This paper reviews the role of EPCs in atherosclerosis and the influence of drugs on EPC function. The feasibility and the problem of using EPCs as a treatment strategy are also discussed.

Effect of 18alpha-glycyrrhizic acid and 18beta-glycyrrhizic acid on P-gp function and expression in Caco-2 cells / 中国中药杂志

<p><b>OBJECTIVE</b>The aim of the present study was to evaluate the modulating effect of glycyrrhizic acid C-18 epimers, 18alpha-glycyrrhizic acid (alpha-GL) and 18beta-glycyrrhizic acid (beta-GL) on both P-glycoprotein (P-gp) activity and expression in Caco-2 cell.</p><p><b>METHOD</b>The effects of P-gp activity were analyzed by rhodamine (Rhd 123) accumulation test, and those of P-gp expression were analyzed by flow cytometry and real-time PCR.</p><p><b>RESULT</b>At middle and high concentrations (10, 60 micromol x L(-1)), alpha-GL inhibited the function of P-gp and with on dose dependent while beta-GL induced the function of P-gp at three test concentrations with no dose dependent too. At middle and high concentrations (10, 60 micromol x L(-1)), alpha-GL down-regulated the expression of MDR1 mRNA. At high concentrations (60 micromol x L(-1)), beta-GL up-regulated the expression of MDR1 mRNA; At high concentrations (60 micromol x L(-1)), beta-GL induced the expression of P-gp protein while alpha-GL has no effect on the expression of P-gp protein at three test concentrations.</p><p><b>CONCLUSION</b>The effects of alpha-GL and beta-GL on the expression of MDR1 mRNA and CYP3A mRNA showed the same trend. The character that epimers of GL act on CYP3A and P-gp show similar stereo selectivity whether relate to PXR need further study.</p>

RP-HPLC study of resveratrol derivative (BTM-0512) in rat plasma and tissue distribution / 药学学报

This paper described a rapid and se nsitive HPLC method to analyze (E)-3, 5,4'-trimethoxystilbene (BTM-0512) in rat plasma and tissues. The analysis used a BDS Hypersil C18 analytical column (250 mm×4.6 mm ID, 5 μm) and acetonitrile / water as the mobile phase. The UV detection wavelength was 319 nm. Proteins were precipitated with acetonitrile and diethylstilbestrol as internal standard. The method was validated according to State Food and Drug Administration of China and ICH of Technical Requirements for Registration of Pharmaceuticals for Human Use Guidelines. The limit of interday precision values (%RSD) were in the range of 2.6% -5.1% and 2.4% -4.8%, respectively.Mean accuracy and absolute recoveries of BTM-0512 ranged from 95.3% - 100. 1% and 95.9% -100.9% for plasma and tissues, respectively. This method can be quite useful for BTM-0512 pharmacokinetic and tissue distribution studies, for purpose which multiple plasma and tissue samples can be analyzed quickly with high reproducibility.

Study on Relative Bioavailability of Cefaclor Capsules in Human Body / 中国药房

OBJECTIVE:To evaluate the relative bioavailability of2kinds of preparations of cefaclor capsules.METH?ODS:The subjects'blood concentrations were determined by HPLC at different time after administered randomly crossover with single oral dose of cefaclor testing preparation or reference preparation.The actual values of C max and T max were adopted;AUC was calculated by trapezoid planimetry;the bioequiavailability of the2preparations were evaluated by two-sides and one-side t-test.RESULTS:The AUC 0～4 of the testing preparation and reference preparation were respectively(13.44?3.06)(?g?h)/ml and(14.19?3.28)(?g?h)/ml;their respective AUC 0～∞ were(13.80?3.08)(?g?h)/ml and(14.62?3.33)(?g?h)/ml;their respective C max were(11.65?2.39)?g/ml and(12.37?2.41)?g/ml;their respective t max were(0.57?0.24)h and(0.66?0.19)h.The relative bioavailability of testing preparation was(95.62?13.51)%.CON?CLUSION:The2preparations are bioequivalent.

Determination of in Vitro Adhesion and Dissolution Rate of Total Puerariae Flavones(TPF)Bioadhesive Sustained-release Tablets / 中国药房

OBJECTIVE:To study the dissolution rate of total puerariae flavones(TPF)bioadhesive sustained-release tablet,and to determine its bioadhesive force to animal stomach and small intestine in vitro METHODS:Using rotating basket method,the dissolubility summation was determined with 0 1mol/L HCl as dissolution medium at speed of 100r/min,single-index model,Weibull distributing model,Higuchi equation and zero-class model were used to imitate the dissolution curve The biggest absolute error,the biggest relative error and AIC were used as comprehensive indices to select the best imitating model A new apparatus made by ourselves was used to compare the adhesive force between the sustained-release tablets and popular tablets in adhering to rabbits'stomach or small intestine RESULTS:The results showed that the dissolution rate in vitro imitated in single-index model was the best Bioadhesion study showed that there were obvious differences between bioadhesive sustained-release tablets and non-bioadhesive tablets in adhering with gastric mucosa and small intestines mucosa(P

Determination of Enalapril in Human Plasma With HPLC-MS / 中国药房

OBJECTIVE:To estabilish a HPLC-MS method for determining enalapril in human plasma METHODS:Alprozalam was added into plasma sample as an internal standard,then supernate of the sample was extracted through solid-phase extration column,washed with methanol and detected by HPLC-MS method:column,ODS C18;mobile phase,methanol-0 01% formic acid(45∶55);flow rate,0 8ml/min;capillary voltage,3 81kV;cone voltage,39 0V The selected ion was determined by EST RESULTS:The calibration curve was linear within the range of 2 5～400ng/ml r=0 9 996,the recovery was 102 2%,RSDs of intra -day and inter-day were 4 0% and 5 4%,respectively CONCLUSION:The method is accurate and sensitive with no endogenous interference It can be applied to studying the pharmacokinetics and bioavailability of enalapril tablets in humans

Experimental Study on the Factors Affecting in Vitro Dissolubility of Total Puerariae Flavones Bioadhesive Tablets / 中国药房

OBJECTIVE:To study the main factors affecting in vitro dissolubility of total puerariae flavones(TPF)bioad?hesive tablets.METHODS:Using HPMC,Carbopol(CP934NP)as bioadhesive and base materials,lactose as porogenic agent to prepare bioadhesive tablets;Basket-rotating method was adopted to determine the dissolubility while0.1mol/L HCl was used as dissolution medium,Rotational speed was100r/min.The accumulated dissolution was detected and the influence of the amount of HPMC,CP,kind of porogenic agent,amount of lactose,size of granules in pressed tablets and medium pH on dissolubility was observed.RESULTS&CONCLUSION:The amount of HPMC,CP and lactose,kind of porogenic agent,size of granules in tablets and medium pH can affect the dissolubility of bioadhesive tablets.

The Effect of Different Methods of Purification on the Quality of Total Flavones of Radix Puerariae / 中国药房

OBJECTIVE:To investigate the technique of purifying the total flavones of Radix Puerariae,so as to provide an experimental basis for extraction and purification of the flavones METHODS:Five purifying methods,including adsorption with macroreticular adsorption resin AB-8,D101,D201 and with polyamide,extration with n-butanol,were adopted,and the amount and purity of total flavones collected and preservation of the components of total flavones were compared RESULTS:By using the above-mentioned five methods,the percentages of the contents of total flavones were 89 4%,83 6%,91 0%,96 0% and 82 9% and the recoveries of products were 79 3%,64 2%,88 0%,55 7% and 77 2% respectively CONCLUSION:Among the five methods,adsorption with AB-8 macroreticular adsorption resin is the best,which may be used for purification of total flavones of Radix Puerariae

Study on Formulations and in Vitro Drug Release of Sustained-release Bioadhesive Danazol Suppository / 中国药房

OBJECTIVE:To screen the formulations of bioadhesive danazol suppository,and to study its drug release in vitro.METHODS:Hydroxypropyl methylcellulse(HPMC),polyethylene glycol(PEG)6000and PEG600were employed in the bioadhesive sustained-release formulation.Suppositories were prepared and the correlation between drug release rate from suppositories and the ratio of HPMC to PEGs used in the formulae was studied.RESULTS:HPMC retarded drug release,and drug gradually released in the12-hours period when the ratio of HPMC to PEGs was1∶6.5;the law of drug release was conformed to weibull models and mono-exponential models.CONCLUSION:HPMC,PEG6000and PEG600were suitable to the formulation of bioadhesive danazol suppositories,and HPMC could slow down the drug release from the this preparation;the best ratio of HPMC to PEGs was1∶6.5.