高效液相色谱-串联质谱法检测绿茶中硝苯地平

风险监测显示市面上陆续出现了添加硝苯地平等二氢吡啶类降血压药物的绿茶,而我国目前尚未有绿茶中硝苯地平的权威检测方法。本试验根据实际需要建立了液相色谱串联质谱法检测绿茶中硝苯地平的方法。首先对绿茶样品进行前处理,采用甲醇超声提取,经过CARB-NH2固相萃取柱净化;然后进行色谱分析,以Inertsil ODS-3(150 mm×2.1 mm,5.0μm)为液相色谱柱,乙腈、0.2%乙酸水溶液为流动相,流速为0.3 mL/min;质谱采用正离子扫描,多反应监测方式检测绿茶中的硝苯地平含量。试验考察了方法学中的线性方程、检出限和定量限、加标回收率、重复性等指标。结果表明:在1~20 ng/mL范围内线性良好,相关系数为0.9996;检出限为13.8μg/kg,定量限为46.0μg/kg;加标回收率在2、4、8 ng/mL三个水平分别为83.5%、86.3%、89.6%,相应水平重复性依次为3.98%、3.43%、1.16%;所考察的指标均满足方法学要求。该方法操作简单、准确、可行,适用于绿茶中硝苯地平的检测。     

MS-222对鹦鹉鱼麻醉效果的研究Anesthetic Effect of MS-222 on Amphilophus 高仁法 刘 毅 陈舒畅 周鑫蔚 曲乐天 夏 腾 徐 晔 许建和*

本文研究了MS-222在不同浓度下对鹦鹉鱼的麻醉效果,观测了其不同麻醉和复苏阶段的表现,探讨了急性麻醉和模拟长途运输中适宜的麻醉浓度。结果表明MS-222对于鹦鹉鱼有很好的麻醉效果,起到了有效的降低应激的作用。急性麻醉中,MS-222的理想麻醉浓度为135 mg/L,长途运输中MS-222的适宜浓度为40 mg/L。     

Pharmacokinetics of ceftriaxone in freshwater crocodiles (Crocodylus siamensis) after intramuscular administration at two dosages

One of the major obstacles to the successful treatment of infectious disease in freshwater crocodile species is incorrect dosing of antibiotics. There are few reports on pharmacokinetics and dosage regimens of antimicrobial drugs in crocodiles. The purpose of the present study was to clarify the pharmacokinetic characteristics of ceftriaxone (CEF) in Siamese freshwater crocodiles (Crocodylus siamensis). Freshwater crocodiles, Crocodylus siamensis, in breeding farms were treated with a single intramuscular administration of CEF at two dosages, 12.5 and 25 mg/kg body weight (b.w.). Blood samples were collected at preassigned times up to 168 hr. The plasma concentrations of CEF were measured by a validated method through liquid chromatography tandem-mass spectrometry. CEF plasma concentrations were quantified up to 72 and 96 hr after low- and high-dose administration, respectively. The C_max values of CEF were 24.61 ± 5.15 µg/ml and 26.39 ± 2.81 µg/ml at dosages of 12.5 and 25 mg/kg b.w., respectively. The AUC_last values increased in a dose-dependent fashion. The half-life values were not statistically different between the groups (around 20 hr). The average binding percentage of CEF to plasma protein was 53.78 ± 2.11%. Based on the pharmacokinetic data, susceptibility break-point and the surrogate PK-PD index (T > MIC, 0.2 μg/ml), i.m. administration of CEF at a dose of 12.5 mg/kg b.w. might be appropriate for initiating treatment of susceptible bacterial infections in freshwater crocodiles.     

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS) for Identification of Bacterial Isolates From Horses

Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) is used for bacterial identification by analyzing the spectra of isolates and comparing them against a database of reference spectra; it is known for its rapidity and accuracy. Although MALDI-TOF MS is used for identification of bacterial isolates from animals, not all animal pathogens are identified correctly. In this study, we used a commercial MALDI-TOF MS identification system to examine 3724 bacterial isolates from horses and their environments. Isolates that could not be identified with MALDI-TOF MS were identified by 16S rRNA gene sequence taxonomic analysis. MALDI-TOF MS could identify 86.2% of the isolates from horses to the species level, showing that this method could be successfully applied for bacterial identification in horses. However, some species known to be equine pathogenic agents including Taylorella equigenitalis and Rhodococcus equi were difficult to identify with MALDI-TOF MS, which might be the result of an inadequate reference database. Some Prevotella, Staphylococcus, and Streptococcus isolates, which could not be identified with either MALDI-TOF MS or 16S rRNA gene sequencing analysis, formed clusters in the 16S rRNA phylogenic tree, and might be unknown species isolated from horses. Adding the spectra of isolates identified in this study to an in-house database might make MALDI-TOF MS a more useful tool for identifying equine isolates.     

气质联用法(GC-MS)检测禾谷镰刀菌代谢指纹分析

代谢指纹分析(Metabolic fingerprint analysis)是代谢组学研究的重要分析模式,广泛应用于各类动植物体的非靶标的代谢产物的探索研究中,其在真菌微生物方面的研究却很少。采用气质联用法(GC-MS)对衍生化后的禾谷镰刀菌样品进行指纹图谱分析,比较不同培养时间、培养环境、不同衍生化方法对代谢产物的影响,选择最佳条件,鉴定出63种代谢产物。这为真菌类物质的代谢组学研究提供了参考依据。     

环酰菌胺在土壤中的吸附及降解特性

为评价环酰菌胺在土壤中的生态风险,采用超高效液相色谱-串联质谱(UPLC-MS/MS)方法测定了土壤和水中环酰菌胺的残留量,研究了该农药在红壤和水稻土中的吸附及降解特性,并对其淋溶特性进行了分析,评估了该农药对地下水的污染风险。结果表明:环酰菌胺在红壤和水稻土中的吸附符合Freundlich吸附等温线方程,KOC值分别为373.69和726.86 mL/g,水稻土对环酰菌胺的吸附能力强于红壤。好氧条件下,环酰菌胺在红壤和水稻土中的降解半衰期分别为0.63和5.06 d,积水厌氧条件下的降解半衰期分别为6.80和9.24 d,表明环酰菌胺在好氧条件下降解较快。环酰菌胺在红壤和水稻土中的地下水污染指数(groundwater ubiquity score)分别为1.19和1.10,表明其对地下水的污染风险较低。结果可为环酰菌胺的生态风险评估提供参考。     

In vitro organogenesis of Abutilon indicum (L.) Sweet from leaf derived callus and assessment of genetic fidelity using ISSR markers

This study reports on in vitro regeneration of Abutilon indicum plantlets through callus mediated organogenesis. The leaf explants implanted on Murashige and Skoogs (MS) medium supplemented with 4.52 µM 2, 4-Dicholorophenoxy acetic acid (2,4-D) and 8.88 µM 6 Benzyladenine (BA) showed highest response (70.3%) for callus proliferation, but these callus did not showed any morphogenetic differentiation on the same medium even after 12 weeks. Whereas, subsequent sub-culture of this green proliferated callus on MS medium added with 2.68µM α-Napthalene acetic acid (NAA), 8.88µM BA and 543 µM Adenine sulphate showed the highest frequency (62.2%) of multiple shoot-buds production and also elongation of shoots. Well developed shoots were efficiently rooted in vitro on half strength MS medium supplemented with 7.38 µM Indole-3-butyric acid (IBA). Seventy per cent of in vitro regenerated plantlets were successfully established in garden and were morphologically alike to the donor plants. The genetic homogeneity of these in vitro regenerated plantlets was also affirmed by inter simple sequence repeat (ISSR) analysis using eight ISSR primers. This standardised in vitro organogenesis protocol supplements a good platform for the conservation of A. indicum germplasms and also caters for the needs of the herbal industry.     

超高效液相色谱-串联质谱法测定水稻中噁唑酰草胺残留量

建立了检测水稻植株、稻壳、糙米、田水基质中噁唑酰草胺残留量的超高效液相色谱-串联质谱(UPLC-MS/MS)的分析方法。待测样品经乙腈匀浆提取、经PSA净化后,经ACQUITY UPLC BEH C₁₈色谱柱(50 mm×2.1 mm,1.7 μm)分离,以甲醇-水为流动相梯度洗脱,采用电喷雾电离串联质谱正离子多反应检测(MRM)模式测定,外标法定量。结果表明：噁唑酰草胺在0.001~0.05 μg/mL浓度范围内,在水稻植株、糙米、稻壳、田水基质中,供试农药的质量浓度与其相应的峰面积间呈良好线性相关,相关系数(r)分别为0.998、0.996、0.994和0.996;在0.005、0.05、0.5 mg/kg 3个水平下进行添加回收试验,噁唑酰草胺在水稻植株、稻壳、糙米、田水基质中的回收率分别在85.8%~114.6%、80.6%~106.4%、84.2%~95.0%和92.8%~109.3%之间,相对标准偏差(RSD)分别在2.7%~6.7%、3.8%~9.2%、2.9%~4.6%和5.0%~12.3%之间,定量限(LOQs)为0.005 mg/kg。该方法简单、快速、准确、灵敏,所得结果满足农药残留检测的要求,适用于水稻植株、糙米、稻壳、田水中噁唑酰草胺的残留检测。     

Pharmacokinetics of tolfenamic acid in green sea turtles (Chelonia mydas) after intravenous and intramuscular administration

The present study aimed to evaluate the pharmacokinetic features of tolfenamic acid (TA) in green sea turtles, Chelonia mydas. Green sea turtles were administered single either intravenous (i.v.) or intramuscular (i.m.) injection of TA, at a dose of 4 mg/kg body weight (b.w.). Blood samples were collected at preassigned times up to 168 hr. The plasma concentrations of TA were measured using a validated liquid chromatography tandem mass spectrometry method. Tolfenamic acid plasma concentrations were quantifiable for up to 168 hr after i.v. and i.m. administration. The concentration of TA in the experimental green sea turtles with respect to time was pharmacokinetically analyzed using a noncompartment model. The C_max values of TA were 55.01 ± 8.34 µg/ml following i.m. administration. The elimination half-life values were 32.76 ± 4.68 hr and 53.69 ± 3.38 hr after i.v. and i.m. administration, respectively. The absolute i.m. bioavailability was 72.02 ± 10.23%, and the average binding percentage of TA to plasma protein was 19.43 ± 6.75%. Based on the pharmacokinetic data, the i.m. administration of TA at a dosage of 4 mg/kg b.w. might be sufficient to produce a long-lasting anti-inflammatory effect (7 days) for green sea turtles. However, further studies are needed to determine the clinical efficacy of TA for treatment of inflammatory disease after single and multiple dosages.     

SPE-UPLC-MS/MS测定葡萄酒中的高氯酸盐

为建立前处理简单、定量准确、灵敏度高的葡萄酒中高氯酸盐的快速测定和分析方法,本研究使用酸化甲醇提取葡萄酒样品,最终选择WAX固相萃取柱前处理净化,亲水性聚丙烯滤膜(GHP)过滤,经Phenyl-Hexyl色谱柱分离,以电喷雾离子源(ESI)在多反应监测(MRM)负离子模式下测定葡萄酒中高氯酸盐。结果表明,该方法的检测时间为12 min,检出限和定量限分别为0.15 μg·L^-1和0.5 μg·L^-1,在1、2、5、10和20 μg·L^-1水平的加标回收率介于94.5%~105%之间,相对标准偏差(RSD)小于3.17%。该方法的建立可为葡萄酒生产企业和相关监管部门提供一定的技术支撑。