Establishment of Detection Methods for Dexamethasone Sodium Phosphate in Injections by UPLC-PDA

A detection method of dexamethasone sodium phosphate in houttuynia injection,bupleurum injection,vitamin C injection,lincomycin hydrochloride injection,and asragulus polysacharin injection was established by UPLC-PDA.The Waters ACQUITY UPLC^® BEH C18 column(2.1 mm×50 mm,1.7 μm) was used,the mobile phase consisted of 0.75% triethylamine phosphatic liquor/solution (pH 3.0)-methyl alcohol-acetonitrile(50:45:5),the flow rate was 0.3 mL/min,the column temperature had remained 25℃,photodiode array detector wavelength range was 190 to 400 nm,the recorded wavelength was 242 nm,the injection volume was 10 μL.The linear range of dexamethasone sodium phosphatel was 10.0 to 200.0 mg/L (R²=0.9999),the mean recovery and RSD for dexamethasone sodium phosphate in houttuynia injection,bupleurum injection,vitamin C injection,lincomycin hydrochloride injection and asragulus polysacharin injection were 97.54% and 1.14%,101.59% and 0.19%,100.92% and 0.92%,99.82% and 0.73%,100.08% and 0.62%,respectively.This method was accurate,easy and fast,it was suitable for detection of dexamethasone sodium phosphatein houttuynia injection and other four kinds of injections.     

改良背主动脉注射法对鸡胚发育与转基因表达效率的影响

背主动脉注射是生产转基因鸡的经典方法,该方法不需换壳培养,但壳内注射技术难度大,并且无法实时观察后期胚胎发育情况。本实验对背主动脉注射法进行了改进,将增强型绿色荧光蛋白(EGFP)腺相关病毒(Adeno-associated Virus,AAV)壳外注射至150枚HH 14~16期(Hamburger-Hamilton Stage 14~16)鸡胚背主动脉中,再进行换壳培养,继续孵化至出壳,观察和分析改良背主动脉注射法与传统背主动脉注射法和胚盘下腔注射法对发育8、14、18、21 d鸡胚存活率、孵化率以及EGFP阳性检出率的影响。结果表明:改良背主动脉注射组鸡胚存活率均高于传统背主动脉注射组与胚盘下腔注射组(P<0.05或P<0.01);改良背主动脉注射组的鸡胚孵化率(37%)高于传统背主动脉注射组(16%)与胚盘下腔注射组(28%)(P<0.01);荧光蛋白手电筒检测显示,改良背主动脉注射组鸡胚EGFP阳性率(17%)明显高于传统背主动脉注射组(13%)与胚盘下腔注射组(12%)(P<0.01)。综上,背主动脉壳外注射结合换壳培养提高了转基因鸡胚胎孵化率和EGFP阳性检出率,对提高转基因鸡效率具有重要价值。     

林可霉素注射液非法添加盐酸左旋咪唑的HPLC检测方法研究

建立了兽用林可霉素注射液中非法添加盐酸左旋咪唑的高效液相色谱检测方法。采用Waters Atlantis? T3 C18(4.6×250mm, 5μm)色谱柱分离,以磷酸二氢钠溶液-甲醇(70:30)作为流动相进行等度洗脱（流速1.00mL/min）,用二极管阵列检测器在230nm波长处进行测定。考察了此色谱条件下方法的专属性、线性、回收率、精密度、检测限和耐用性。结果表明,盐酸左旋咪唑在进样浓度为0.02mg/mL～0.4mg/mL时呈良好的线性关系,R2=0.9999;重复性试验RSD为0.6%;平均回收率为99.7%;检测限为 0.0002mg/mL,定量限为0.0016mg/mL。该法准确、简便、可行,适用于林可霉素注射液中盐酸左旋咪唑的检测。     

不同剂量伊曲康唑制剂对犬血常规及肝肾功能的影响

旨在评价不同剂量伊曲康唑-环糊精制剂通过皮下注射途径对健康比格犬肝肾功能及血常规的影响。选用体重±10 kg的健康纯种比格犬32只,按试验要求随机分为4组,每组8只,建立稳定的生活制度。分别按照0.25 mg/kg、0.75 mg/kg和1.25 mg/kg剂量,皮下注射伊曲康唑制剂,3日1次,连续7次,对照组注射生理盐水。于试验过程中第0、5、10、15、19天采集相关样品,对犬临床表现、血常规指标、血液生化指标、尿常规指标及肝肾功能进行全面的评价。结果表明,给药组与对照组相比,除白细胞相关指标外,其余指标都在正常范围内。给药组在试验过程中均出现白细胞升高现象,但在试验结束后5 d内均恢复正常。因此,伊曲康唑-环糊精制剂作为犬皮下注射剂型,具有良好的安全性和耐受性,操作方便,动物依从性好,适合兽医临床应用。     

Polymorphism in promoter region of growth hormone receptor is associated with potential production capacity of insulin‐like growth factor‐1 in pre‐pubertal Holstein heifers

Insulin‐like growth factor‐1 (IGF‐1) is one of the important factors for growth, milk production and reproductive functions and mainly released from the liver in response to growth hormone (GH) via GH receptor (GHR) in cattle. Recently, some single nucleotide polymorphisms (SNPs) were identified in the bovine GHR gene. Some GHR‐SNPs were shown to be related to plasma IGF‐1 concentration in cattle. Hence, the capacity to IGF‐1 production in the liver might be affected by GHR‐SNP and associated with performance in the future. This study examined whether GHR‐SNP is associated with IGF‐1 production in the liver of pre‐pubertal heifers. In 71 Holstein calves, blood samples for genomic DNA extraction were obtained immediately after birth. To genotype the GHR‐SNPs in the promoter region, polymerase chain reaction (PCR) products were digested with restriction enzyme NsiI (cutting sites: AA, AG and GG). All heifers at 4 months of age were intramuscularly injected with 0.4 mg oestradiol benzoate. Blood samples were obtained from the jugular vein just before (0 h) and 24 h after injection. The number of AA, AG and GG at the NsiI site was 0, 17 and 54 respectively. In AG and GG, plasma GH concentrations were higher pre‐injection than 24 h post‐injection (p < 0.01). Moreover, plasma GH concentrations in AG post‐injection were higher than in GG (p < 0.05). In contrast, the GG genotype exhibited higher plasma IGF‐1 concentrations in pre‐injection than post‐injection (p < 0.01), although oestradiol did not change IGF‐1 concentration in the AG genotype. We conclude that the GG polymorphism in the promoter region of GHR is associated with a higher potential capacity of IGF‐1 production in the liver of cattle.     

欧盟食品动物注射部位兽药残留指南概述

2022年3月25日,欧盟药品管理局(EMA)兽医药品委员会(CVMP)发布了食品动物注射部位兽药残留指南,并于2022年8月1日正式生效。该指南代替旧版的注射部位兽药残留指南(EMA/CVMP/542/03-FINAL)。本文就新发布指南的背景、适用范围、法律依据、评价方法、残留研究及相关定义等内容进行介绍。     

烷基-二醇基硅胶填料的合成及对莠去净类除草剂的在线分离

三步合成了烷基-二醇基硅胶(ADS)色谱材料，首先用γ-缩水甘油醚氧丙基三甲氧基硅烷把功能基团键合到硅胶基体上，再把该环氧化硅胶与硬脂酸反应以制备酯型十八烷基反相填料，最后在20MPa压力下装成色谱预柱(35×4.6mm I.D)。用该预柱通过六通阀后接ODS分析柱(250×4.6mm I.D)，构成中心切割二维HPLC。该系统能对具有一定疏水性的化合物进行在线富集，实现环境及生物复杂样品直接进样分离分析。使用该系统对尿中的莠去净和西玛津残留进行了分离分析，以含有0.1%乙酸的酸性水缓冲液对加标尿液中莠去净类化合物保留在预柱上，干扰分析的大分子被排阻而无保留通过，其它干扰分子保留小而通过，再切换六通阀，以乙腈和0.1%乙酸水溶液进行梯度洗脱，进而在下一级ODS分析柱上进行分离（紫外检测225nm），两种除草剂的分离分析取得了良好的定性定量结果