Pharmacokinetics of articaine hydrochloride and its metabolite articainic acid after subcutaneous administration in red deer (Cervus elaphus)

AIM: To develop and validate a simple and sensitive method using liquid chromatography-mass spectrometry (LC-MS) for quantification of articaine, and its major metabolite articainic acid, in plasma of red deer (Cervus elaphus), and to investigate the pharmacokinetics of articaine hydrochloride and articainic acid in red deer following S/C administration of articaine hydrochloride as a complete ring block around the antler pedicle.

METHODS: The LC-MS method was validated by determining linearity, sensitivity, recovery, carry-over and repeatability. Articaine hydrochloride (40?mg/mL) was administered S/C to six healthy male red deer, at a dose of 1?mL/cm of pedicle circumference, as a complete ring block around the base of each antler. Blood samples were collected at various times over the following 12 hours. Concentrations in plasma of articaine and articainic acid were quantified using the validated LC-MS method. Pharmacokinetic parameters of articaine and articainic acid were estimated using non-compartmental analysis.

RESULTS: Calibration curves were linear for both articaine and articainic acid. The limits of quantifications for articaine and articainic acid were 5 and 10?ng/mL, respectively. Extraction recoveries were >72% for articaine and >68% for articainic acid. After S/C administration as a ring block around the base of each antler, mean maximum concentrations in plasma (C_max) of articaine were 1,013.9 (SD 510.1) ng/mL, detected at 0.17 (SD 0.00) hours, and the C_max for articainic acid was 762.6 (SD 95.4) ng/mL at 0.50 (SD 0.00) hours. The elimination half-lives of articaine hydrochloride and articainic acid were 1.12 (SD 0.17) and 0.90 (SD 0.07) hours, respectively.

CONCLUSIONS AND CLINICAL RELEVANCE: The LC-MS method used for the quantification of articaine and its metabolite articainic acid in the plasma of red deer was simple, accurate and sensitive. Articaine hydrochloride was rapidly absorbed, hydrolysed to its inactive metabolite articainic acid, and eliminated following S/C administration as a ring block in red deer. These favourable pharmacokinetic properties suggest that articaine hydrochloride should be tested for efficacy as a local anaesthetic in red deer for removal of velvet antlers. Further studies to evaluate the safety and residues of articaine hydrochloride and articainic acid are required before articaine can be recommended for use as a local anaesthetic for this purpose.     

黄连素口服纳米乳的研制、质量及安全性评价

研究选择适宜的油相、表面活性剂和助表面活性剂,利用伪三元相图,以新型纳米乳为载体,对黄连素的传统剂型进行改进,研制出适合口服的纳米乳制剂;以离心(13000r·min-1,30min)稳定性和乳滴粒径作为主要的评价标准,对纳米乳的黏度、电导率、折光率、粒径和初步稳定性等方面进行质量评价;用小鼠灌胃的急性毒性实验对纳米乳的安全性进行评价。结果表明研制出的黄连素口服纳米乳为澄清透明的液体,透射电镜下观察为球状液滴,平均粒径为56.8nm;高速离心稳定、高温和强光条件下考察10d,其含量和粒径均未发生明显变化;小鼠灌胃结果,纳米乳无毒性。由此证明研制出的黄连素口服纳米乳是一种质量稳定、安全性高的良好药物传递系统。     

小檗碱和川芎嗪对猪卵母细胞体外成熟中一氧化氮生成量的影响

试验以mTCM-199添加抗生素为对照组,以分别添加0.1μg/mL小檗碱(berberine,BR)和0.5μg/mL川芎嗪(ligustrazine,Lig)为试验组,比较各组卵母细胞体外成熟中一氧化氮(NO)生成量和卵母细胞的成熟效果,从而探讨中药单体成分对猪卵母细胞体外成熟过程中NO生成量的影响。结果表明:体外成熟前22 hNO生成量,对照组与Lig组差异极显著(P<0.01)、与BR组差异显著(P<0.05),Lig组与BR组差异显著(P<0.05);体外成熟后22 h NO生成量,对照、Lig及BR三组间均无显著差异(P>0.05),但Lig组的NO生成量少于BR组和对照组;体外成熟培养0～44 h不同组别NO生成量整体呈下降趋势,Lig组与BR组卵母细胞成熟率显著高于对照组(P<0.05)。结果表明卵母细胞体外成熟培养过程中,NO生成量随体外培养时间增加呈递减趋势,中药单体成分BR和Lig对卵母细胞成熟均起到一定的促进作用。低浓度NO代谢量可能对卵母细胞成熟有一定促进作用。     

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.     

正交试验优选酸水法提取黄连中小檗碱的工艺及其抑菌活性的研究

采用正交试验设计,采用酸水法提取黄连中小檗碱,并测定提取的小檗碱对八株大肠埃希氏杆菌的体外最小抑菌浓度（MIC）。结果表明,最佳正交提取工艺为C3B3A1,即硫酸浓度0.2%、冷浸72h、用15%NaCl盐析,提取物对八株致病性大肠杆菌的MIC为0.50mg/mL-1.5mg/mL。提取的小檗碱对致病性大肠杆菌具有较好的抑菌作用。     

盐酸小檗碱对草鱼补体系统及补体c3作用的研究

为了探究盐酸小檗碱(berberine hydrochloride)对草鱼(Ctenopharyngodon idellus)补体c3的作用机理,用含0、0.06%、0.12%、0.24%盐酸小檗碱饲料投喂草鱼,在第7 d、14 d检测血清中补体c3含量,在第14 d检测肝脏中补体c3 mRNA表达量及急性感染嗜水气单胞菌(Aeromonas hydrophila)后存活情况。并进一步对盐酸小檗碱体内药代动力学和补体消耗试验进行了研究。结果显示;摄食盐酸小檗碱的草鱼血清中补体c3含量和肝脏补体c3 mRNA表达量均显著性增加(P0.05/P0.05),草鱼的存活率与对照组相比也有显著性提高(P0.01)。补体消耗试验结果显示,当盐酸小檗碱在血清中浓度小于5 mg/L时与对照组相比补体消耗不显著,当盐酸小檗碱浓度大于5 mg/L时,补体消耗达到显著性差异(P0.05),表明此时盐酸小檗碱与补体c3直接结合,激活补体。药代动力学实验发现,灌服盐酸小檗碱后,出现双峰现象,两峰值分别是0.243 mg/L和0.117 mg/L,此浓度远远低于与补体c3直接结合量最低量(5 mg/L)。结果表明,盐酸小檗碱对草鱼补体c3的作用机理可能是通过上调补体c3mRNA的表达增加补体c3数量,而非通过直接结合补体c3来改变其活性。     

复方盐酸土霉素注射液的制备

【目的】制备复方盐酸土霉素注射液,并对其质量浓度进行测定。【方法】通过预试验,确定复方盐酸土霉素注射液的配方组分及其大致用量。以焦亚硫酸钠、MgCl2、乙醇胺和复合有机溶媒为影响因素,以灭菌前后盐酸土霉素和甲硝唑质量浓度变化为考察指标,利用L9(34)正交试验确定复方注射液的最优配方。利用紫外分光光度法对注射液中甲硝唑和盐酸土霉素的质量浓度进行考察,并计算其回收率和精密度。【结果】复方盐酸土霉素注射液的最优配方为:每100mL注射液中含有2.5g甲硝唑、5.0g盐酸土霉素、3g氯化镁、0.2g焦亚硫酸钠、40mL复合有机溶媒(由N,N-二甲基甲酰胺和聚乙二醇400组成,体积比为2∶1)、4mL乙醇胺,其余为注射用水。甲硝唑和盐酸土霉素在1～30μg/mL质量浓度时与吸光度呈现良好的线性关系。制备的复方盐酸土霉素注射液中甲硝唑的平均回收率为(99.22±1.48)%,变异系数为1.49%;盐酸土霉素的平均回收率为(98.22±1.24)%,变异系数为1.26%。注射液中甲硝唑和盐酸土霉素的日内、日间精密度测定结果为:平均质量浓度较高,相对标准偏差较小,且不大于2.00%。【结论】确定的复方盐酸土霉素注射液配方合理,甲硝唑和盐酸土霉素稳定性好,且回收率高。     

超声提取永思小檗盐酸小檗碱工艺的优化

[目的]优选出永思小檗盐酸小檗碱的最佳超声提取工艺条件,为盐酸小檗碱在医药工业上的研发应用奠定基础.[方法]以乙醇为溶媒,采用超声提取法提取永思小檗盐酸小檗碱,并通过单因素试验和正交设计L9(34)考察乙醇浓度、提取时间、提取温度、溶媒倍数对永思小檗盐酸小檗碱提取效果的影响.[结果]综合考虑成本及提取效率,永思小檗盐酸小檗碱最佳超声提取工艺为:以150倍45%乙醇作溶媒,60℃超声提取一次,时间为40 min.[结论]在最佳工艺条件下,超声提取永思小檗盐酸小檗碱的平均含量达11.9633 mg/g,进一步证实以乙醇为溶媒、采用超声提取法提取小檗碱的可行性.     

仔猪肠黏膜上皮细胞体外培养及其对小檗碱适应性的研究

为了体外分离培养和鉴定仔猪肠粘膜上皮细胞,探讨小檗碱对体外培养的仔猪肠粘膜上皮细胞的安全剂量。本试验取刚出生未吃初乳的仔猪小肠的回肠段,用胰蛋白酶和胶原酶Ⅳ进行消化处理,获得原代仔猪肠粘膜上皮细胞,用免疫组化法对培养的仔猪肠粘膜细胞进行鉴定,用MTT检测法测定硫酸小檗碱对仔猪肠粘膜上皮细胞的安全浓度。结果表明:免疫组化法分离培养的细胞膜呈棕黄色,为粘膜上皮细胞。硫酸小檗碱对体外培养的仔猪肠粘膜上皮细胞的安全浓度为20μmol/L。     

不同产地益母草饮片中盐酸水苏碱含量差异分析

目的比较全国不同产地益母草饮片中盐酸水苏碱含量差异,为益母草的质量研究提供参考依据。方法按照2010版《中国药典》规定,采用HPLC-ELSD法测定各样品中盐酸水苏碱的含量。结果不同产地益母草饮片中盐酸水苏碱含量差异很大,在0.102%～2.142%之间。结论为保证益母草饮片的质量和疗效,建议选择规范化人工栽培的益母草。