Clinical characteristics of α-pyrrolidinovalerophenone (α-PVP) poisoning

Context: α-Pyrrolidinovalerophenone (α-PVP) is a synthetic cathinone that has been abused in recent years. The clinical presentation of acute α-PVP poisoning has not been well characterized.

Objective: To elucidate the clinical features of acute α-PVP poisoning.

Materials and methods: This retrospective case series included eight subjects that visited our hospital emergency department (ED) between March 2012 and November 2014 and had analytically confirmed blood α-PVP levels. Data related to subject demographics, clinical history, laboratory findings, blood drug levels, and outcome were collected.

Results: The median age of the eight study subjects was 27 (range; 21–63) years, and six were male. Drug preparations had been administered by rectal insertion (three subjects) or inhalation (five subjects). The time between drug exposure and presentation at the ED was 8.5 (1–24) h and blood α-PVP concentrations ranged from 1.0 to 52.5?ng/ml. Although psychiatric and neurological findings were reported before arrival at the ED in 5/8 and 7/8 subjects, respectively, these were only observed in 1/8 and 2/8 subjects, respectively, at the ED. Symptoms of high body temperature (3/8), tachycardia (5/8), hypertension (3/8), acid-base balance disorder (5/8), coagulopathy (4/6), blood creatinine phosphokinase?>190?U/l (6/8), and a blood lactate level?>?1.7?mmol/l (5/7) were observed. All subjects survived and were discharged.

Conclusions: This retrospective case series showed that after acute exposure to α-PVP, transient neuropsychiatric findings were accompanied by more persistent sympathomimetic physical findings, disorders of acid-base balance and blood coagulation, high blood creatinine phosphokinase, and hyperlactacidemia.     

盐酸文拉法辛缓释片Beagle犬体内药动学及生物等效性研究

目的 研究盐酸文拉法辛缓释片在Beagle犬体内的药动学和生物等效性。方法 8条健康Beagle犬随机分成2组,采用双周期、双交叉、单剂量分别ig盐酸文拉法辛缓释片受试制剂或参比制剂75 mg,清洗期为1周;建立血浆中盐酸文拉法辛液相色谱-质谱联用（LC-MS/MS）检测方法,进行方法精密度、准确度、提取回收率、基质效应、稳定性方法学验证;测定给药前（0 h）及给药后2、3、4、5、6、8、10、12、14、16、24、36、48、72 h血浆中盐酸文拉法辛血药浓度,运用DAS 2.1.1软件计算其药动学参数,并评价其生物等效性。结果 LC-MS/MS方法学经验证符合检测要求,受试制剂和参比制剂主要药动学参数分别如下：T_1/2分别为（7.16±2.34）和（6.95±1.57） h、Cmax分别为（522.89±201.12）和（515.22±159.29）ng/mL、T_max分别为（10.38±1.69）和（10.50±2.07）h、AUC_0-t分别为（8 398.64±3332.86）和（8 050.71±2103.15）ng·h/mL、AUC_0-∞分别为（8 701.60±3303.29）和（8 450.01±2273.45）ng·h/mL;以参比制剂为参考,受试制剂AUC_0-∞相对生物利用度为（101.0±13.1）%。结论 盐酸文拉法辛缓释片受试制剂与参比制剂在Beagle犬体内具有生物等效性。     

Regulated LC-MS/MS bioanalysis technology for therapeutic antibodies and Fc-fusion proteins using structure-indicated approach

In recent studies, the development of bioanalysis technologies using liquid chromatography-tandem mass spectrometry (LC-MS/MS) has attracted attention. Our developed nano-surface and molecular-orientation limited (nSMOL) proteolysis enables Fab-specific proteolysis and is optimal for LC-MS/MS analysis of antibody drugs and Fc-fusion proteins in biological samples. In this nSMOL method, antibodies and Fc-fusion proteins are held in pores of the particle and the subsequent proteolysis is carried out with protease-immobilized nanoparticles. The Fab of antibodies or fused region of Fc-fusion protein can be held to orient toward the reaction solution. The access of the immobilized protease is limited to a part in the structure of protein substrate on the particle surface. Thus, nSMOL proteolysis reacts selectively at the Fab complementarity-determining region of antibodies or N-terminal specific domain of Fc-fusion proteins and can be applied to both types of drugs. We have already evaluated drug concentrations in biological samples pretreated with nSMOL proteolysis using LC-MS/MS for more than twenty drugs, of which ten drugs have been fully validated and published. In this review, we discuss the development and application of LC-MS/MS bioanalysis, which enables the bioanalysis of therapeutic antibodies and Fc-fusion proteins by focusing on a structure-based approach.     

Reduced variability in tacrolimus pharmacokinetics following intramuscular injection compared to oral administration in cynomolgus monkeys: Investigating optimal dosing regimens

Tacrolimus is one of the most commonly used immunosuppressive agents in animal models of transplantation. However, in these models, oral administration is often problematic due to the lowered compliance associated with highly invasive surgery and due to malabsorption in the intestinal tract. Therefore, we carried out a study to determine the pharmacokinetics of tacrolimus after intramuscular (IM) injection and to determine the optimal IM dosing regimens in primate models. Six male cynomolgus monkeys (Macaca fascicularis) were used in the study. Doses of 0.1 mg/kg and 5 mg were administered via IM injection and oral administration, respectively, once to determine single-dose pharmacokinetics and once daily for 5 days to determine multiple-dose pharmacokinetics. According to pharmacokinetic model estimates, the inter- and intra-individual variabilities in bioavailability following IM injection were remarkably reduced compared with those following oral administration. Monte Carlo simulations revealed that C_peak, C_trough and AUC would also have less variability following IM injection compared with oral administration. In this study, we found that the pharmacokinetic characteristics of tacrolimus were more constant following IM injection compared with oral administration. These results suggest that IM injection can be an alternative route of administration fin non-human primate model studies.     

高效液相色谱-串联质谱法测定人血清中文拉法辛和O-去甲基文拉法辛浓度的不确定度评价

目的：评价高效液相串联质谱（Liquid chromatography tandem mass spectrometry,LC-MS/MS）法测定人血清中文拉法辛和O-去甲基文拉法辛浓度的不确定度。方法：对可能会引入不确定度的步骤进行分析,包括测量重复性、样品称量、溶液配制、样品处理、仪器允差、基质效应、提取回收、标准曲线拟合等,评估扩展不确定度。结果：文拉法辛低（12 ng·mL^-1）、中（120 ng·mL^-1）、高（300 ng·mL^-1）浓度质控样品的拓展不确定度分别为：U_L=0.847 ng·mL^-1,U_M=7.518 ng·mL^-1,U_H=20.776 ng·mL^-1;O-去甲基文拉法辛低（60 ng·mL^-1）、中（600 ng·mL^-1）、高（1 500 ng·mL^-1）浓度质控样品的拓展不确定度分别为：U_L=11.666 ng·mL^-1,U_M=91.479 ng·mL^-1,U_H=254.523 ng·mL^-1（P=95%,k=2）。结论：LC-MS/MS法测定人血清中文拉法辛和O-去甲基文拉法辛浓度的不确定度主要来源于标准曲线拟合、基质效应和提取回收过程。选择合适的同位素氘代内标浓度能够有效降低标准曲线拟合过程引入的不确定度。     

基于LC-MS/MS分析鹿角方的入血移行成分

目的：研究鹿角方的入血移行成分及规律,初步阐明其药效物质基础。方法：通过LC-MS/MS法分析大鼠含药血清,明确鹿角方的原型入血成分;及不同给药剂量下各移行成分相对峰面积与剂量的关系,解析移行规律。结果：通过与标准品保留时间、离子碎片信息的比对,鉴定鹿角方中13种成分（马钱苷、柚皮苷、橙皮苷、朝霍定A、朝霍定B、朝霍定C、淫羊藿苷、补骨脂素、异补骨脂素、宝霍苷Ⅰ、红景天苷、莫诺苷、特女贞苷）均能以原型入血;随给药剂量上升,柚皮苷、橙皮苷、补骨脂素、异补骨脂素、特女贞苷的相对峰面积与剂量呈线性关系,而马钱苷、莫诺苷、红景天苷、宝霍苷Ⅰ呈非线性关系,其中马钱苷、莫诺苷、补骨脂素、异补骨脂素、宝霍苷Ⅰ、红景天苷易入血。结论：马钱苷、莫诺苷、补骨脂素、异补骨脂素、宝霍苷Ⅰ、红景天苷可能是鹿角方体内直接作用的主要物质,实验结果为鹿角方药效物质基础的研究提供借鉴。     

藤茶中杨梅素、二氢杨梅素及其乙酰化衍生物在SD大鼠体内药动学研究

目的：建立LC-MS/MS法测定SD大鼠血浆中杨梅素（myricetin,MY）和二氢杨梅素（dihydromyricetin,DMY）的浓度,将其运用到药动学研究,并评价DMY乙酰化衍生物是否改善生物利用度。方法：SD大鼠随机分为4组,分别灌胃DMY （100 mg·kg^-1,相当于9 375.0 μmo·L^-1）的0.5%羧甲基纤维素钠生理盐水溶液,以及等摩尔剂量的DMY-1、DMY-2、MY。于给药前0.0 h和给药后0.08,0.17,0.25,0.33,0.50,0.75,1.0,2.0,4.0,6.0,8.0,12.0 h尾静脉断尾采血200 μL,血浆样品经0.1%甲酸-乙腈沉淀蛋白,内标法定量,DAS3.0和SPSS统计分析得到药动学参数和曲线。结果：低、中、高的质控样品日内、日间精密度RSD均小于10.2%;DMY和MY的基质效应分别为98.3%~105.5%、98.0%~108.1%;萃取回收率分别为99.4%~103.0%、95.0%~99.7%,均符合生物样品分析检测的要求;DMY-2和DMY-1组相比于DMY组,相对生物利用度分别为47.22%,10.70%,表明口服灌胃给药DMY相对生物利用度要高出乙酰化衍生物组。结论：本研究建立测定大鼠血浆中DMY和MY的LC-MS/MS法,具有灵敏度高、准确性高、重复性好等优点,能够满足生物样品分析检测的要求,可应用于MY、DMY及其衍生物的药动学研究,也可为DMY结构修饰和新药开发提供参考依据。     

Silicone Oil-Free Polymer Syringes for the Storage of Therapeutic Proteins

Prefilled syringes are a popular choice for the delivery of biopharmaceuticals. However, glass syringes might not be the optimal primary packaging material for all biopharmaceuticals. There is evidence that the necessary lubricant silicone oil in glass syringes can interact with proteins and can be shed from the surface into the product solution. In recent years, silicone oil-free polymer syringes were developed. Despite several advantages, however, a major shortcoming of these polymer systems is their relatively high gas permeability, which might be a limitation for the storage of oxygen sensitive biopharmaceuticals. So far, no long-term protein stability studies regarding such polymer systems have been published. In this study, 2 therapeutic proteins were stored in glass syringes and in silicone oil-free polymer syringes. In addition, polymer syringes stored in nitrogen-filled aluminum pouches or covered with oxygen-tight labels were included. Similar chemical protein stability was achieved at 4°C for all syringes. However, in contrast to the polymer syringes, high particle counts were observed in the glass syringes. Polymer syringes stored in nitrogen-filled aluminum pouches presented a promising alternative for the storage of biopharmaceuticals as they do not expose patients to silicone oil and silicone oil-protein aggregates.     

The Impact of Endogenous Breast Cancer Resistance Protein on Human P-Glycoprotein-Mediated Transport Assays Using LLC-PK1 Cells Transfected With Human P-Glycoprotein

Lilly Laboratories cell porcine kidney 1 (LLC-PK1) cells transfected with human P-glycoprotein (LLC-PK1-P-gp) are widely used in transport assays to identify drug candidates that function as substrates of this efflux transporter. Endogenous transporters expressed in LLC-PK1 cells may complicate the interpretation of findings from P-gp-mediated transport assays. We investigated the impact of porcine breast cancer resistance protein (Bcrp) in P-gp-mediated transport assays in LLC-PK1 cells. Porcine Bcrp mRNA was detected in both LLC-PK1 wildtype (WT) and LLC-PK1-P-gp cells by quantitative RT-PCR. To investigate the activity and impact of porcine Bcrp, we conducted transport assays using 6 typical BCRP substrates in LLC-PK1 cells. Efflux ratios (ER) of the 6 BCRP substrates in LLC-PK1 WT cells were >2, and were reduced in the presence of the BCRP inhibitor Ko143. The efflux activities of the 6 BCRP substrates were confirmed using MDCKII cells transfected with human BCRP. Net ERs of prazosin and fluvastatin, dual substrates of P-gp and BCRP, determined by dividing ERs in LLC-PK1-P-gp cells by those in LLC-PK1 WT cells, were <2, but increased to >2 in the presence of Ko143. These results indicated that endogenous Bcrp in LLC-PK1 cells was involved in the transport of BCRP substrates and may interfere with the identification of P-gp substrates.     

LC-MS/MS法同时测定消毒产品中6种抗生素残留

目的建立同时测定消毒产品中甲硝唑、盐酸金霉素、土霉素二水合物、盐酸米诺环素、红霉素及盐酸四环素的高效液相色谱-串联质谱(LC-MS/MS)检测方法。方法样品经甲醇超声提取,0.45μm聚四氟乙烯膜过滤后,选择WatersSymmetryC18(2.1mm×150mm,3.5μm)为分析柱,5mmol/L乙酸铵缓冲溶液、甲醇和乙腈为流动相,用LC-MS/MS正离子扫描多反应监测模式检测。结果上述6种抗生素在0～2000ng/ml范围内线性关系良好,相关系数均大于0.998,回收率为74.7%～114%,相对标准偏差为1.6%～20.2%。该法成功用于115件皮肤、黏膜使用的膏霜剂型抗(抑)菌制剂消毒产品中抗生素残留量的检测。结论该法操作简单,灵敏度高,选择性强,可用于膏霜剂型消毒产品中抗生素残留量的测定。