皮肤外用制剂要求在皮肤局部发挥药效,口服给药药物的生物利用度研究方法对于皮肤外用制剂并不适用.目前血管收缩测定法已被FDA认可,但该法只适用于糖皮质激素类药物,其他外用制剂仍需通过临床对比试验来评价其生物等效性.FDA仍在努力寻找其他的研究方法,如胶带粘贴法、皮肤微透析法、开流微量灌注及体外研究方法.本文对以上方法的研究进展进行综述.

目的:考察醋酸曲安奈德微透析探针体外回收率的影响因素,探索提高微透析探针回收率的方法.方法:采用高效液相色谱法考察温度、灌流液流速、探针类型以及灌流液成分(羟丙基-β-环糊精、脂肪乳、牛血清白蛋白)对醋酸曲安奈德体外回收率的影响.结果:醋酸曲安耐德探针回收率随温度增高而增加;随灌流液流速增加而降低;不同类型的探针对醋酸曲安耐德回收率不同;灌流液成分会对醋酸曲安奈德回收率产生较大影响.结论:采用PBS(pH 7.4)溶液添加1％或2％牛血清白蛋白为灌流液,醋酸曲安奈德体外探针回收率得到较大提高,且回收率和传递率相等.

Objective To determine the intracutaneous concentration of Ftibamzone given percutaneously to test animals and investigate its percutaneous penetrability and the intracutaneous drug level.Methods An HPLC method combined with homogeneization was used to determine the intracutaneous concentration of Ftibamzone after topical administration.Results The percutaneous absorption and penetrability of Ftibamzone was relatively low.Conclusion The results of the test can give reference to the study of dermatopharmacokinetics of Ftibamzone in human.

Objective To evaluated the influence of multi-drug resistance gene 1(MDR1) C3435T,G2677T/ A and C1236T polymorphism on the pharmacokinetics of isotretinoin in 21 heathy male subjects.Methods Blood samples were collected from 21 healthy male subjects who received a single oral dose of 40 mg isotretinoin.The genotypes of MDR1 C3435T,G2677T/A and C1236T of the samples were detected by allele-specific amplification(ASA-PCR).The pharmacokinetic parameters of isotretonoin were compared between the different genotypes of MDR1 by statistical comparisons method.High performance liquid chromatography-electrospray ionization mass spectrometry(HPLC-MS) was used for the quantification of isotretinoin in human plasma which was standardized by dosage and body weight.Pharmacokinetic parameters were determined by non-compartmental analysis.Results For C1236T,the Cmax,Tmax,AUC 0-60 of carriers with CT and TT was remarkably higher than those with CC(P=0.057,P=0.252,P =0.049,respectively),the T1/2 and MRT of carriers with CT and TT was remarkably lower than those with CC(P=0.011 and P=0.035).For G2677T/A,there was no significant pharmacokinetic parameters difference among the different genetype subgroups(P0.05).For C3435T,there was no significant pharmacokinetic parameters differences among different genetype subgroups(P0.05).In addition,the Tmax of carriers with CC and CT was higher than those with TT(P=0.03).Conclusion It suggests that the MDR1 C1236T polymorphism influenced the human absorption of oral isotretinoin.Although there is genetic linage diequilibrium between C3435T and G2677T/A which can influences the isotretinoin absorption,the genetic polymorphisms of MDR1 C3435T and G2677T/ A had no effect to isotretinoin pharmacokinetic parameters in this research.

AIM To investigate the bioequivalence of two compound chlorzoxazone tablets in healthy volunteers.METHODS Single oral doses of two compound chlorzoxazone tablets(chlorzoxazone 250 mg and paracetamol 300 mg) were given to 20 healthy male volunteers in an open,randomized and crossover study. The plasma concentrations of chlorzoxazone and paracetamol were determined by HPLC-MS method with glipizide and acetophenetidin employed as the internal standard,and the pharmacokinetic parameters were calculated with BAPP2.2.RESULTS The major pharmacokinetic parameters of test and reference tablets in the singledose study were as follows;for paracetamol,ρ_(max) were(5.1±1.5) vs.(5.2±1.7) mg·L~(-1);t_(max) were(0.6±0.4) vs.(0.6±0.4) h;t_(1/2)were(3.0±0.4) vs.(3.0±0.5) h;AUC_(0-12)were(16.4±4.7) vs.(17.0±5.5) mg·h·L~(-1);AUC_(0-∞) were(17.6±5.1) vs.(18.2±6.0) mg·h·L~(-1),respectively.In the case of chlorzoxazone,ρ_(max) were(7.0±1.3) vs.(7.0±1.3) mgΛL~(-1);t_(max)were(0.9±0.6) vs.(1.0±0.5) h;t_(1/2) were(0.9±0.1) vs. (0.9±0.1) h,AUC_(0-∞)were(16.6±3.5) vs.(17.0±4.1) mg·h·L~(-1),AUC_90-∞) were(16.7±3.5) vs.(17.0±4.1) mg ? h ? L~(-1),respectively.There were no significant differences in pharmacokinetic parameters between two compound chlorzoxazone tablets(P0.05).The relative bioavalibility of paracetamol and chlorzoxazone in compound chlorzoxazone tablets were(99.3±14.3)%and(99.5±14.8)%,respectively.CONCLUSION The test tablet is bioequivalent to the reference tablet for chlorzoxazone and paracetamol in healthy volunteers.

A rapid liquid chromatograph-electro-spray ionization(ESI) tandem mass spectrometric method was developed for the quantification of lafutidine in human plasma.Separation was performed using a Kromasil CN column(4.6 mm×150 mm,5 μm)maintained at 30℃ and the mobile phase consisting of a mixture of methyl alcohol and water(containing 20 mmol·L-1 ammonium acetate and 0.05% formic acid)(97∶3) was delivered at a flow rate of 1.0 mL·min-1;The analytes were analyzed by ESI in a multiple reaction monitoring(MRM) mode.The precursor to product ion transition of m/z 432.20→351.15 and m/z 275.10→230.05 were used to measure lafutidine and IS,respectively.The linearity ranged from 1.98 to 396 ng·mL-1(r=0.9991) and the limit of detection of lafutidine in human plasma was 1.98 ng·mL-1.The intra-and inter-assay precisions,accuracy and absolute recovery met the requirements of bioanalytical method,significant matrix effect was absent.The method is selective and suitable for the quantification of lafutidine in human plasma clinically.

Objective: To develope a rapid liquid chromatography tandem mass spectrometry method for the quantification of terbinafine in human plasma.Methods: Separation was performed using a C18 column(4.6 mm×150 mm,5 μm)maintained at 30℃ and the mobile phase consisting of a mixture of methyl alcohol and water(containing 20 mmol·L-1 ammonium acetate)(95∶5) was delivered at a flow rate of 1.0 mL·min-1.The analytes were analyzed by electro-spray ionization(ESI) in a multiple reaction monitoring(MRM) mode.The precursor to product ion transition of m/z 292.00→141.10 and m/z 530.95→82.10 were used to measure terbinafine and the internal standard(ketoconazole),respectively.Results: The linearity ranged from 10.3 to 2054 ng·mL-1(r=0.9978);the limit of quantification of terbinafine in human plasma was 10.3 ng·mL-1.The intra-and inter-assay precisions,accuracy and absolute recovery met the requirement of a bioanalytical method,the method was absent of matrix effects.Conclusion: The method is selective and suitable for the clinical quantification of terbinafine in human plasma.

Objective To investigate the hair changes in post-menopausal women,and analyse its relationship with estrogen receptor β gene polymorphism.Methods Healthy post-menopausal women were recruited,images of the areas in vertical and temporal region were obtained using hair detector;hair growth rate,hair density,white hair rate were counted by image analyzing software.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was employed to determine estrogen receptor β RsaⅠand AluⅠgene polymorphism.Results The distribution of genotypes was in Hardy-Weinberg equilibrium(P0.05).The differences of white hair rate were statistically significant in RsaⅠpolymorphism.Hair growth rate,hair density had no significance among different subgenotypes.Conclusion The RsaⅠpolymorphism of estrogen receptor β is associated with white hair growth in post-menopausal women.

Microneedles are a novel technique for transdermal drug delivery.They can overcome the barrier of stratum comeum and promote transdermal drug delivery,also have the advantages of painlessness,convenience and capability of cooperating with other techniques.Many safe and effective types of microneedles have been developed for the transdermal delivery of Various drugs,e.g.sugar micro needle and membrane-coated microneedle,which has extended the application range ofmicroneedles.

OBJECTIVE To study the pharmacokinetics and bioequivalence of terbinafine tablet in healthy volunteers.METHODS A single oral dose(250 mg of test and reference tablet) were given to 19 healthy volunteers in a randomized crossover study.The concentrations of terbinafine in plasma were determined by HPLC.The pharmacokinetics parameters were calculated and the bioavailability and bioequivalence of two tablets were evaluated.RESULTS After a single dose,the pharmacokinetics parameters for test and reference terbinafine tablets were as follows: Cmax were(1.66±0.62) μg·mL-1 and(1.55±0.66) μg·mL-1;Tmax were(1.5±0.7) h and(1.4±0.6) h;t1/2(Kel) were(12.65±3.07) h and(14.24±3.65) h,MRT were(10.21±3.13) h and(11.56±3.62) h,AUC0-48 were(5.98±2.45) μg·h·mL-1 and(6.76±3.14) μg·h·mL-1,AUC0-∞ were(6.32±2.58) μg·h·mL-1 and(7.20±3.27) μg·h·mL-1,The relative bioavailability of test terbinafine tablet was(95.1±22.5)%(AUC0-48) and(93.9±21.6)%(AUC0-∞).CONCLUSION The pharmacokinetics parameters of test and reference terbinafine tablets were of no statistical difference,the two tablets are bioequivalence.

A rapid and simple high performance liquid chromatographic method coupled with tandem mass spectrometry (LC–MS–MS) via electrospray ionization (ESI) has been developed and validated to separate and simultaneously quantify sodium ferulate (SF), salicylic acid (SA), cinnarizine (CIN) and vitamin B1 (VB1) in human plasma. Gemfibrozil (GEM) was used as the internal standard (IS) for SF and SA, whereas lomerizine (LOM) was used as the IS for CIN and VB1. The plasma samples were prepared by one-step protein precipitation followed by an isocratic elution with 10 mM ammonium acetate buffer (pH = 5.0): acetonitrile (35:65, v/v,) on an Agilent Zorbax SB-CN column (150 mm × 2.0 mm ID, 5 μm). The precursor and product ions of these drugs were monitored on a triple quadrupole mass spectrometer, operating in the selected reaction monitoring mode (SRM) with polarity switch, in the negative-ion mode for SF, SA and GEM, in the positive-ion mode for CIN, VB1 and LOM. The method was validated over the concentration range of 1.5–1,000 ng mL−1 for SF, 20–5,000 ng mL−1 for SA, 2–500 ng mL−1 for CIN, 1–30 ng mL−1 for VB1. The intra- and inter-batch precisions were less than 15% of the relative standard deviation. The recoveries for analytes and IS achieved from spiked plasma samples were consistent and reproducible. The validated LC–MS–MS method has been successfully applied to the pharmacokinetic study of sodium ferulate and aspirin capsule in healthy volunteers.

Objective: To assess the feasibility of retrodialysis method by calculating the recovery of microdialysis probe in percutaneous absorption in vivo.Methods:Zero-net-flux method was used to assess the recovery and delivery of probe in vitro.Retrodialysis method was used to calculate the recovery of probe in vivo.The in vivo percutaneous absorption process of triptolide was performed using pigs.Results:In the in vitro experiment,the recovery of probe was(64.29±2.34)%.The result from recovery was consistent with that from delivery.In the in vivo experiment,the recoveries of probe before and after experiment were(51.47±3.51)% and(51.96±4.97)%,respectively.The concentration of triptolide in dermis quickly reached to a steady state after administration of triptolide ointment but rapidly dropped following removal of the ointment.Conclusion:The recovery of probe calculated by retrodialysis method was stable throughout all experiments,indicating that the retrodilaysis method could be applied to assessing the recovery of probe in vivo.

依美斯汀(Emedastine,EME)由日本Kanabo-Kowa公司联合开发,1993年8月首先在日本上市,商品名DAREN,是新的第二代H1受体拮抗剂.依美斯汀可选择性地作用于H1受体,临床用于治疗荨麻疹、湿疹或皮炎、瘙痒症及痒疹、过敏性鼻炎等.本文就其药物作用机制、药代动力学、不良反应及应用前景作一简要介绍.

AIM: To develop an analytical method based on LC-MS/MS for the quantification of lisinopril in human plasma and the assessment of bioequivalence for two kinds of lisinopril tablets in Chinese healthy male volunteers. METHODS: Plasma samples of 20 healthy male volunteers, who were administrated with a single oral dose of lisinopril test tablets (10 mg) and reference tablets (10 mg) according to a random 2-way crossover design, were prepared by a simple protein precipitation and then were determined by LC-MS/MS method. RESULTS: The main pharmacokinetic parameters of lisinopril test tablets and reference tablets were as follows: cmax(54±s 21) and (53±23)μg·L-1, tmax(6.4±1.4) and (6.7±1.1) h, AUC0-36(569±244) and (566±269)μg·h·L-1, AUC0-∞(589±250) and (587±268)μg·h·L-1, respectively. Calculating by AUC0-36, the relative bioavailability of lisinopril test tablets was (102±8) %. CONCLUSION: The analytical method based on LC-MS/MS for quantification and assessment of bioequivalence is simple, rapid and sensitive, proving the bioequivalence of lisinopril test tablets to the reference tablets.

目的 分析念珠菌酵母相全细胞水解生成的D-葡萄糖与D-甘露糖含量比值。方法 采用双波长薄层扫描法，测定65株8种医学上重要的念珠菌标准株及酿酒酵母标准菌株8株的酵母相全细胞完全酸水解产物中D-葡萄糖与D-甘露糖含量比值。同时测定了46株念珠菌临床分离株，并与标准株进行了比较。结果 经Tukey法统计处理，可将9种菌分成5个同质性亚群。经t检验比较，酿酒酵母和8种念珠菌的结果差异均有显著性；而念珠菌临床分离株与标准株结果差异均无显著性。结论 该比值结果能在属水平区分念珠菌和酿酒酵母，并可将8种念珠菌分为4个同质性亚群；且经临床分离株验证，重复性较好。

Objective To analyse the ratio of D-glucose to D-mannose hydrolyzed from whole yeast phase cells of 8 medically important species of genus Candida.Methods The ratio of D-glucose to D-mannose of 65 standard strains and 46 clinical isolates of 8 medically important species of genus Candida,8 standard strains of Saccharomyces cerevisiae.were determined by dual-wavelength thin-layer scanning technique.Results The results analyzed by Tukey test showed that 9 species could be classified into 5 homogeneous subsets.Analyzed by t test,there were significant differences between S.cerevisiae and 8 species of Candida,but there was not significant differences between clinical isolates and standard strains of some species of genus Candida.Conclusion This index could identify Candia from S.cerevisiae on genus level and classify 8 medically important species of genus Candida into 4 homogeneous subsets.

AIM: To investigate the expression of chemokine receptor CXCR3 mRNA in peripheral blood lymphocytes and neutrophils in patients with progressive plaque psoriasis and its correlation with Psoriasis Area and Severity Index (PASI).METHODS: Reverse transcriptase-polymerase chain reaction(RT-PCR) was applied to semi-quantitatively analyze CXCR3 mRNA expression in peripheral blood lymphocytes and neutrophils from 33 psoriatic cases,16 cases with about 70% remission of the 33 patients after treatment and 30 healthy controls.The correlation between CXCR3 mRNA expression and PASI was evaluated.RESULTS: CXCR3 mRNA level in lymphocytes from psoriatic patients was 1.12±0.81,markedly higher than that in normal controls(0.28±0.28,P<0.01)and treated patients(0.57±(0.61,) P<0.05).Furthermore,a positive correlation was noticed between CXCR3 mRNA expression and PASI(r=0.516,P<(0.05).) However,there was no statistic significance in CXCR3(mRNA) level in neutrophils among pretreated psoriatic patients((1.72)±1.68),normal controls(1.27±1.17,P>0.05) and treated patients(1.27±1.01,P>0.05).CONCLUSION: The increased expression of CXCR3 in lymphocytes may be associated with migration and aggregation of lymphocytes in psoriatic lesion,suggesting that this chemokine receptor be involved in the pathogenesis of progressive plaque psoriasis.

Objective: To investigate the expression of chemokine receptors on neutrophils in plaque type psoriasis and in progressive stage its clinical significance. Methods: Reverse transcription-polymerase chain reaction(RT-PCR) technique was applied to semi-quantitatively analyze CXC type chemokine receptors CXCR1-3 and CC type chemokine receptors CCR1 and CCR5 mRNA expression on peripheral blood neutrophils in 33 cases of psoriatic patients, 16 of the 33 patients who were treated until 70% of the skin lesions resolved, and 30 healthy controls. The correlations between CXCR1, CXCR2 and CCR1 mRNA expression and psoriatic area and severity index (PASI) were evaluated, respectively. Results: The expression levels of CXCR1, CXCR2 and CCR1 mRNA on neutrophils from psoriatic patients were 1.36±1.19, 1.45±0.87 and 1.43±1.28, respectively. which were significantly higher than those in normal controls[0.55±0.37(P 0.01), 0.74±0.58 and 0.68±0.37( both P 0.001), respectively] and post-treatment patients[0.46±0.33(P 0.001), 0.63±0.58(P 0.01) and 0.73±0.86(P 0.05), respectively]. A positive correlation was observed between both CXCR1 and PASI, and CXCR2 and PASI. Conclusions: The increased expression of CXCR1/2 and CCR1 maybe associated with migration and aggregation of neutrophils in psoriatic lesional epidermis, suggesting that these three chemokine receptors be involved in the pathogenesis of plaque type psoriasis in progressive stage.

近20年来,微透析技术在皮肤方面的应用日趋广泛和深入.微透析技术是一种膜取样技术,并需要连接分析装置.这一技术最初用于神经生理学研究,用于药物经皮渗透及皮肤中药物浓度的研究中,称为经皮微透析.就近10年来微透析技术在经皮药代动力学方面研究中的应用作一综述.