Reversed－Phase High－Performance Liquid Chromatographic Determination of Drugs of Abuse

A high-performance liquid chromatographic method using uv detection for thedetermination of 10 drugs of abuse is described. The method is sample and sensitive. The linear rangesare 20￣100ng/ml with relative standard deviations of 0.7 ￣ 2.6% (n=7) and detection limits of 5￣ 10ng/ml.

Determination of the Antiretroviral Drug Zidovudine in Diluted Alkaline Electrolyte by Adsorptive Stripping Voltammetry at the Mercury Film Electrode

This paper describes a stripping method for the determination of zidovudine at the submicromolar concentration levels. This method is based on the controlled adsorptive accumulation of zidovudine at the thin-film mercury electrode, followed by a linear-sweep stripping voltammetry measurement of the surface species. Optimal experimental conditions include a NaOH solution of 2.0 × 10–3 mol●L–1 (sup-porting electrolyte), an accumulation potential of –0.30 V and a scan rate of 100 mV?s–1. The response of zidovudine is linear over the concentration range 0.01 - 0.08 ppm. After an accumulation time of 5 minutes, the detection limit was found to be 0.67 ppb (2.5 × 10–9 mol●L–1). More convenient methods to measure zidovudine concentration in the presence of the didanosine, acyclovir, nevirapine, lamivudine, and efavirenz, were also investigated. The presence of zidovudine together with ATP or ssDNA demonstrates the utility of this method.

Determination of Impurity Silicon in Anticancer Drug (Ge-132) by Electrothermally Atomised Atomic Absorption Spectrometry with Optical Temperature Control Accessory

The present paper describes the ashing and atomization processes in silicon analysis by electrothermally atomised atomic absorption spectrometry(EAAAS) with an uncoat-ed graphite tube, a pyrolytically coated graphite tube and a tungsten-coated graphitetube. The sensitivity and linear range of three graphite tubes were compared. By using optical temperature control accessory, the signals are enhanced by a factor of 2 and the germanium interferences in the determination of silicon are eliminated. The effects of time constant and carrier gas flow-rate on the determination of silicon were also tested. The sample can be directly analyzed in its aqueous solution without any pretreatment. The measurements of samples containing 0. 2 μg/mL and 0. 4 μg/mL silicon were run ten times and the variation coefficient is 4. 9% and 2.6%, respectively. The recovery tests for carboxyethyl germanium sesquioxide(Ge-132) synthesized and imported were performed, and the recoveries are 97. 0% and 110%, respectively. Keywords Carboxyethyl germanium sesquioxide, Electrothermally atomised atomic absorption spectrometry, Silicon

Construction of universal quantitative models for the determination of cefoperazone sodium/sulbactam sodium for injection from different manufacturers using near-infrared reflectance spectroscopy

To develop near-infrared （NIR） reflectance spectroscopic methods for the quantitative analysis of cefoperazone sodium/ sulbactam sodium from different manufacturers for injection powder medicaments. Various powders of cefoperazone sodium/ sulbactam sodium were directly analyzed by non-destructive NIR reflectance spectroscopy using the spectrometer EQUINOX55. Two quantitative methods via integrating sphere （IS） and fiberoptic probe （FOP） models were explored from 6 batches of commercial samples and 42 batches of laboratory samples at a content ranging from 30% to 70% for cefoperazone and 60% to 20% for sulbactam. The root mean square errors of cross validation （RMSECV） and the root mean square errors of prediction （RMSEP） of IS were 1.79% and 2.85%, respectively, for cefoperazone sodium, and were 1.86% and 3.08%, respectively, for sulbactam sodium; and those of FOP were 2.93% and 2.92%, respectively, for cefoperazone sodium, and were 2.23% and 3.01%, respectively, for sulbactam sodium. Based on the ICH guidelines and Ref. 12, the quantitative models were then evaluated in terms of specificity, linearity, accuracy, precision, robustness and model transferability. The non-destructive quantitative NIR methods used in this study are applicable for rapid analysis of injectable powdered drugs from different manufacturers.

Determination of Resistance to Seven Insecticides in Plutella xylostella L. in Fields of Northern Hunan

The resistance of field populations of Plutella xylostella, from the three vegetable producing areas （Nianyuxu Town of Yueyang City, Canggang Town of Changde City and Shatou Town of Yiyang City） in northern Hunan, to seven insecticides was determined using leaf dipping method in door. The results showed that Plutella xylostella showed an extremely high-level resistance to beta-cypermethrin （resistance ratio, RR=257.13）, a high-level resistance to abamectin （RR=135.15） and indoxacarb （RR=103.08） and a moderate-level resistance to chlorfenapyr and emamectin benzoate. But Plutella xylostella is relatively sensitive to chlorantraniliprole and Bacillus thuringiensis （Bt）. Therefore, the prevention of Plutella xylostella in northern Hunan should focus on the alternative use of chlorfenapyr, emamectin benzoate, chlorantraniliprole and Bacillus thuringiensis and avoid the use of beta-cypermethrin so as to delay the generation and development of resistance to insecticides in Plutella xylostella.

Fast Determination of Vitamin B₂Based on Molecularly Imprinted Electrochemical Sensor

Under the condition of weak acidity of pH 5.2, a sensitive vitamin B2 electrochemical sensor based on molecularly imprinted nonconducting polymer of o-aminophenol by potentiostatic polymerization in the presence of template(vitamin B2) on a glassy carbon electrode was prepared, and its performance was studied. The sensor exhibited good sensitivity and selectivity to VB2. The detection limit went down to 2.3851nM, and a linear relationship between the current incremental and the concentration was found in the range of 10~120nM. And the sensor could use in detection of VB2 real sample for a long time and show good reproducibility. The average recovery rate to VB2 was 98.41%.

Determination of Polydatin in Rat Serum and Its Changes by UPLC Method

[Objectives]To establish a UPLC-UV method for the determination of polydatin in the serum of Wistar rats.[Methods]Acquity UPLC BEH shield RP18 column(1.7μm,2.1 mm×100 mm,Waters Corporation,USA)was used as the analytical column,acetonitrile-water(55∶45)was used as the mobile phase,the flow rate was 0.5 mL/min,and the column temperature was 30℃and the detection wavelength was 306 nm.[Results]The linear range of the established serum sample analysis method was 1.0-20.0μg/mL,and the correlation coefficient was r=0.9994;the intraday and interday RSD of Wistar rat serum was less than 3.0%,and the accuracy was higher than 90%.[Conclusions]This method is sensitive,accurate,and rapid.It is suitable for monitoring the concentration of polydatin in serum after intragastric administration,and can also be used for pharmacokinetics and bioavailability studies.

Molecular characterization of antimicrobial multi-drug resistance in non-typhoidal Salmonellae from chicken and clam in Mangalore, India

Salmonella enterica has been documented as one of the leading causes of salmonellosis throughout the world and is most commonly associated with the consumption of contaminated food products. Thus, this research was aimed at studying the antimicrobial susceptibility pattern and detection of quinolone resistance in Salmonella spp isolated from food of animal origin. Thirty-six Salmonella isolates comprising 8 from poultry and 28 from seafood（clams） were identified, serotyped and characterized for their antimicrobial susceptibility against 10 different antibiotics. Plasmid DNA was isolated from all the isolates by alkaline lysis, quinolone resistant non-typhoidal S. Weltevreden were examined for mutation in the DNA gyrase coding gene. Among the 36 Salmonella isolates, 20 were S. weltevreden（8 from poultry and 12 from seafood） and 16 were S. Typhimurium（from seafood）. All the isolates showed multiple resistance to nalidixic acid, tetracycline, co-trimoxazole and nitrofurantoin, but, interestingly, the isolates were 100% susceptible to ampicillin, chloramphenicol and gentamicin. Resistant isolates from the study carried the genes responsible for resistance to respective antibiotics. The strain S130 isolated in the study showed single point mutation,Asp87Gly, at position 87 in quinolone resistance determining region. It revealed mutation in quinolone resistance determining region as a cause for quinolone resistance in non-typhoidal Salmonellae. The occurrence of genes accountable for plasmid mediated resistance to quinolones（viz., qnrA, qnrB and qnrS） in plasmid of non-typhoidal Salmonellae isolates provides evidence for plasmid mediated quinolone resistance.

Differential Pulse Voltammetric Studies on Lamivudine:An Antiretroviral Drug

Lamivudine (also known as 3TC) is a dideoxynucleoside analogue, which undergoes intracellular phosphorylation in the putative active metabolite, lamivudine triphosphate. Lamivudine triphosphate prevents HIV replication by competitively inhibiting viral reverse transcriptase. Lamivudine has been extensively used in the treatment of HIV patients owing to its antiretroviral activity. For the determination of lamivudine in pharmaceuticals, an analytical methodology using voltammetry was developed. Lamivudine was reduced at a hanging mercury drop electrode (HMDE) at –1.16 V vs Ag/AgCl at pH 2.0. The influence of electroanalytical parameters such as scan rate (20 mV.s–1), amplitude (50 mV), nature of the support electrolyte (Clark-Lubs), and pH (2.0) on the voltammetric signal was optimized. Under these optimized conditions, the method had been validated using pharmaceutical formulations. The lamivudine peak current varied linearly with its concentration from 1.15 to 10.40 mg.L–1, detection and determination limits of 0.46 and 1.0 mg.L–1, respectively, and recovery of 95.15% with a relative standard deviation of 1.10%.

HIV-1整合酶基因序列分析方法验证

目的 验证实验室自建人类免疫缺陷病毒1型(HIV-1)整合酶基因序列分析方法。该方法可用于评估HIV-1整合酶区段基因型耐药水平。方法 根据世界卫生组织自建基因序列分析方法验证的建议,从20份HIV-1阳性样本中提取RNA,扩增HIV-1整合酶区基因片段,并测序。通过与病毒质量保证(VQA)共识进行比对,评估实验室自建的HIV-1整合酶基因序列分析方案的准确性,通过扩增成功率评估其灵敏度,通过同一样本的重复检测结果评估其精密度和重现性。结果 20份样本与VQA共识的核苷酸一致率均>98%;10个高病毒载量(>10 000拷贝·mL^(-1))样本和5个低病毒载量(1 000~5 000拷贝·mL^(-1))样本的扩增成功率均为100%;4个样本的同批次5复孔和5个样本5次检测的结果均符合90%的样本配对比较核苷酸一致率>98%的要求。结论 该HIV-1整合酶基因序列分析方法的准确性、灵敏度、精密度和重现性均符合要求,适用于HIV-1整合酶基因序列分析。

论药品犯罪中“假药”的认定

在《中华人民共和国刑法修正案(十一)》删除了假药的概念之后,如何认定药品犯罪中的假药成为理论界与实务界共同关注的问题。该问题是一个典型的行刑衔接问题,法秩序统一性原理对其解决具有指导意义。违法相对论是对法秩序统一性原理最贴切的理解,因此,既不能赞成药品犯罪中的假药与《中华人民共和国药品管理法》规定的假药完全一致的观点,也不能肯定可以完全脱离《中华人民共和国药品管理法》的规定独立认定药品犯罪中假药的观点,而应当以《中华人民共和国药品管理法》规定的假药为前提,在此基础上进行实质危害性判断。一方面,只要某种药品不是《中华人民共和国药品管理法》规定的假药,即使其对人体健康有实质危害性,也不属于药品犯罪中的假药;另一方面,若某种药品属于《中华人民共和国药品管理法》规定的假药,但不会给人体健康带来实质危害,那么,也不能将其认定为药品犯罪中的假药。

我国生产、销售假药犯罪特点及侦查对策研究——以2017—2021年裁判文书为分析样本

当前,生产、销售假药犯罪形势日益严峻。应用统计学方法,运用Spss27.0、Excel等软件基于2017—2021年788份涉网药品安全犯罪裁判文书以及1510名被告人样本的实证研究发现,我国生产、销售假药犯罪呈现犯罪地域性特征突出,数量显著减少,犯罪人以青壮年为主,文化水平特征突出,犯罪团伙地缘性、家族式特征突出,药物来源涉及种类复杂多样,危害后果两极化,量刑特征较为突出等特征。分析得出生产、销售假药犯罪侦查存在以下问题:犯罪主体团伙化,亲密性高致使案件突破难;假药成分多样、鉴定复杂,犯罪认定难,取证难度大;量刑轻刑化趋势致使犯罪屡禁不止。对此提出如下几点侦查对策:深挖犯罪线索,以“三维”信息打开假药案件突破口;加强综合侦查研判,以犯罪人社会关系摸排犯罪网络;组建专业侦查人才队伍,科学设置“预检分流”环节,健全药品保管制度;全面收集主客观证据,厘清各犯罪主体责任,提出合理量刑建议;提高资源保障力度,搭建合作交流平台,构建多元共治体系。

一测多评法同时测定不同配伍比例枳实-厚朴药对中11种成分的含量

目的建立一测多评法同时测定不同配伍比例枳实-厚朴药对中辛弗林、紫丁香苷、木兰花碱、芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、柠檬苦素、柚皮素、和厚朴酚、厚朴酚的含量。方法枳实、厚朴分别按1∶1、1∶2、2∶1比例配伍。分析采用Agilent 5TC-C_(18)色谱柱(4.6 mm×250 mm,5μm);流动相甲醇-0.1%磷酸,梯度洗脱;体积流量1.0 mL/min;柱温30℃;检测波长280 nm。以橙皮苷峰为内标,计算其他10种成分相对校正因子,测定其含量。结果11种成分在各自范围内线性关系良好(r≥0.9990),平均加样回收率97.04%~99.45%,RSD 1.22%~2.48%,一测多评法所得结果与外标法接近,配伍比例为1∶1各成分含量最高。结论该方法简便准确,重复性好,可用于枳实-厚朴药对质量控制。2种药材配伍比例为1∶1时,更有利于各成分提取。

耐药肺结核患者血清miR-129-3p、miR-144-3p检测的临床价值

目的探讨血清miR-129-3p和miR-144-3p检测在耐药肺结核中的临床价值。方法收集2021年1月—2022年6月川北医学院附属三台医院行新辅助放化疗的140例耐药肺结核患者(耐药肺结核组),根据治疗6个月的疗效,分为有效组(92例)和无效组(48例);另选取同期与耐药肺结核患者一般资料相匹配的140名体检健康者作为对照组。比较各组血清miR-129-3p、miR-144-3p表达的差异。采用多因素Logistic回归分析评估耐药肺结核患者疗效的影响因素;采用受试者工作特征(ROC)曲线分析治疗2个月后血清miR-129-3p、miR-144-3p水平和白蛋白、白细胞计数预测耐药肺结核患者疗效的效能。结果与对照组比较,耐药肺结核组血清miR-129-3p、miR-144-3p水平显著降低(P<0.05)。治疗2个月,无效组血清miR-129-3p、miR-144-3p水平均显著低于有效组(P<0.05)。血清miR-129-3p、miR-144-3p、白蛋白和白细胞计数是耐药肺结核患者疗效的影响因素(P<0.05)。治疗2个月后血清miR-129-3p、miR-144-3p、白蛋白、白细胞计数联合预测耐药肺结核患者疗效的曲线下面积为0.956,敏感性为87.50%,特异性为92.39%,优于4项指标单独检测(P<0.05)。结论耐药肺结核患者血清miR-129-3p、miR-144-3p水平显著降低。miR-129-3p、miR-144-3p联合白蛋白、白细胞计数对预测耐药肺结核患者疗效有较高的参考价值。

创新抗高血压药物中间体Boc-L-脯氨酸冰片酯质量分析

采用高效液相色谱法(HPLC)法对创新抗高血压药物中间体Boc-L-脯氨酸冰片酯的含量进行了分析。依据药典方法,对Boc-L-脯氨酸冰片酯进行性状及检验,采用柱色谱法,进行Boc-L-脯氨酸冰片酯供试品的纯化;采用HPLC法,色谱柱为UltimateTMXB-C18柱(4.6×250 mm,5μm),流动相为乙腈和纯水[V(乙腈)∶V(水)=90∶10],流速为0.8 mL/min,进样量为10μL,检测波长为210 nm,柱温为30℃,标定对照品,进行系统适应性试验、分析方法的验证和供试品的含量测定。供试品主峰的分离良好;Boc-L-脯氨酸冰片酯对照品的纯度为99.41%;系统适应性试验结果显示,该色谱系统满足实验要求;范围为800~1200μg/mL;线性回归方程:A=1171.8c-61(r=0.996);检测限为10μg/mL;回收率为94.2%~104.6%。按照《中国药典》(2020版)原料药分析方法指导原则,以HPLC建立了一种专属强、灵敏度高的分析方法,为Boc-L-脯氨酸冰片酯的含量测定提供了一种合理的思路。

胶体金免疫层析法高通量测定动物肌肉组织中多兽药残留

[目的]本文旨在建立一种胶体金免疫层析技术高通量测定猪肉、鸡肉组织中8种磺胺类、3种四环素类、2种喹诺酮类和甲氧苄啶药物残留的方法。[方法]动物肌肉组织中磺胺类、四环素类、喹诺酮类和甲氧苄啶经磷酸缓冲液提取,并经含Tween 20的磷酸缓冲液稀释后,与胶体金标记的单克隆抗体结合,抑制抗体与硝酸纤维素膜检测线(T线)上的抗原结合,从而导致T线颜色变化。通过比较T线和质控线(C线)颜色深浅,对试样中多兽药残留进行定性判定。[结果]本试验优化了pH值、抗体添加量、抗原包被量、Tween 20添加量和冻干保护剂种类等关键因素,磺胺类、四环素类、喹诺酮类和甲氧苄啶等药物的检出限分别为50、100、50、35μg·kg^(-1),灵敏度≥97%,假阳性率≤2%,假阴性率≤3%,且与现有标准方法检测结果一致。[结论]该方法操作简便,灵敏度和准确度高,检测时间短且稳定性好,可用于动物肌肉组织中多兽药残留的高通量快速检测。

抗血小板药物的质量研究进展

抗血小板药物近年来呈现蓬勃发展的态势,其质量控制标准也在不断发展和完善。本综述基于《中华人民共和国药典》2020年版(ChP 2020)、《美国药典》43版(USP 43)、《日本药典》17版(JP 17)等国内外及相关指导原则等参考资料,全面归纳了抗血小板药物在药典中的收录情况,对常见的抗血小板药物的质量控制方法进行总结,对含量测定和体内药物分析方法等项目进行全面分析,期望为抗血小板药物的研究与开发提供参考。

两种不同取血部位制备桂枝汤含药血清对其抗炎作用影响差异的研究

目的基于两种不同取血部位制备桂枝汤含药血清,探讨其抗炎作用差异。方法取雄性SD大鼠按10倍临床剂量给药(桂枝汤汤剂生药1 g/mL),取门静脉血、外周血含药血清,采用快速液相-三重四级杆质谱联用仪(rapid resolution liquid chromatography-triple quadrupole mass spectrometry,RRLC-QQQ-MS/MS),建立桂枝汤汤剂、含药血清中8种化学成分含量测定方法,并对含量进行测定。取对数生长期小鼠脑微血管内皮细胞(brain-derived endothelial cells.3,bEnd.3)模型,用白细胞介素(interleukin,IL)-1β刺激,建立细胞炎症模型,使用bEnd.3细胞作为正常组。除正常组、模型组外,设置桂枝汤汤剂高、中、低剂量组(分别含5%、2.5%、1.25%桂枝汤汤剂),门静脉含药血清高、中、低剂量组(分别含20%、10%、5%门静脉含药血清)和外周血含药血清高、中、低剂量组(分别含20%、10%、5%外周血含药血清),ELISA法测定细胞外液中的前列腺素(prostaglandin,PG)E_(2)含量。结果含量测定结果显示汤剂中芍药苷、芍药内酯、甘草苷、甘草酸比例相对较高,血清中肉桂酸、6-姜酚含量相对较高,较汤剂中新出现甘草次酸等代谢物。门静脉血清中以上8种化合物含量均高于外周血血清。5%桂枝汤汤剂中甘草酸含量为798.13 ng/mL,20%门静脉含药血清中甘草酸含量为316.22 ng/mL,20%外周血中甘草酸含量为481.17 ng/mL,药物中成分的含量在同一数量级。与模型组比较,将高、中、低剂量桂枝汤汤剂添加到bEnd.3细胞培养体系中,高、中、低剂量桂枝汤组细胞外液中PGE_(2)的水平均明显高于模型组(P<0.05),而门静脉或外周血含药血清组细胞外液中PGE_(2)的水平均明显低于模型组(P<0.05),且门静脉血清的药效作用显著强于相同浓度的外周血血清(P<0.05)。结论桂枝汤汤剂经胃肠道给药后,门静脉含药血清相较外周血含药血清显示出更高的成分含量与抗炎作用。这一发现提示采用门静脉含药血清而非传统的外周血含药血清在中药方剂研究中的重要性。

脊髓性肌萎缩症患者利司扑兰治疗药物监测方法研究

目的建立脊髓性肌萎缩症患者利司扑兰治疗药物监测的方法。方法采用超高效液相色谱串联质谱法,色谱柱为Phe‐nomenex Kinetex XB C18柱(50 mm×3 mm,2.6µm),流动相为0.07%甲酸水溶液(pH 4.5)-0.07%甲酸乙腈溶液(pH 5.5),梯度洗脱,流速为0.3 mL/min,柱温为40℃,进样量为2µL;采用电喷雾离子源(正离子模式),多反应监测模式,利司扑兰的离子对m/z分别为402.2→319.2、402.2→374.4,RG7800的离子对m/z为417.3→360.2。结果利司扑兰质量浓度在1.95~125.00 ng/mL范围内与其峰面积与内标峰面积比值线性关系良好(R2=0.9991,n=8),定量限为1.95 ng/mL;基质效应试验结果的变异系数均小于12%;精密度、稳定性试验结果的RSD均小于20%,准确度分别为87%~108%、87%~113%。6例患者利司扑兰血药浓度为13.03~91.14 ng/mL。结论该方法操作简便、高效快速,可用于脊髓性肌萎缩症患者利司扑兰的治疗药物监测。

从单味中药的剂量选择探讨随证施量与临床用药安全

“随证施量”是“辨证论治”之后重要的一环,考虑患者体质、年龄、病程等个体化差异,是中医个体化治疗的体现。总结现代医家关于附子、大黄、水蛭、天花粉、淫羊藿五味常用中药在“随证施量”过程中超剂量用药情况,不同剂量针对疾病的轻重程度和相关指标的改善情况,希望能够拓宽药物临床剂量使用范围,为临床提供理论依据。