杜仲叶中绿原酸含量测定方法的建立

用紫外分光光度法与高效液相色谱法相结合,建立一种新的、简单快捷的杜仲液提取液绿原酸检测方法-紫外分光光度法直接测定杜仲叶提取液绿原酸含量。采用方法:(1)绘制绿原酸标准曲线:分别绘制紫外分光光度法、高效液相色谱法的绿原酸标准曲线并考察每种检测方法的线性关系;(2)研究两种检测方法是否存在关系:探索杜仲叶提取液中绿原酸吸光度值与对应峰面积是否存在关系。结果:(1)紫外分光光度法标准曲线方程:y=22.6442x-0.2039,r=0.9999(n=6)。绿原酸浓度在5~40μg/m L范围内呈良好线性关系;高效液相色谱法的标准曲线方程:y=0.0653x+0.6412,r=0.9994(n=6)。绿原酸浓度在5~40μg/m L范围内呈良好线性关系;(2)杜仲叶提取液中绿原酸吸光度值(紫外分光光度法)和对应峰面积(高效液相色谱法)存在关系,关系方程为:y=228.66x-14.538,r=0.9995(n=6)。结论:本试验建立了一种新的绿原酸检测方法-紫外分光光度法直接测定杜仲叶提取液绿原酸含量。     

超高效液相色谱-串联质谱法检测双黄连口服液中3种有效成分

应用超高效液相色谱-串联质谱技术,建立了同时测定双黄连口服液中3种有效成分(黄芩苷、绿原酸、连翘苷)的检测方法。采用BEH C18色谱柱,以0.2%甲酸-乙腈作为流动相进行梯度洗脱。在ESI负离子模式下,通过多反应监测模式进行测定。黄芩苷、绿原酸、连翘苷的线性范围分别为0.02~10μg/m L(r=0.9991)、0.02~10μg/m L(r=0.9998)、0.025~10μg/m L(r=0.9996)。定量限分别为0.02 mg/kg、0.02 mg/kg、0.025 mg/kg。3种成分的回收率为98%~101.7%,相对标准偏差均小于2.5%。该法快捷、准确、重复性好,可用于双黄连口服液中的3种有效成分含量的同时测定。     

沙葱异黄酮含量测定方法的研究

比较紫外分光光度法和高效液相色谱法测定沙葱异黄酮的含量,建立沙葱异黄酮含量测定的最佳试验方法。两种试验方法均以鸢尾苷为对照品,紫外分光光度法在其最大吸收峰295nm处进行测定。高效液相色谱法以Intertsil ODS-SP C18(250mm×4.6mm,5μm)为色谱柱,流动相为30%乙腈,柱温25℃,流速0.5ml/min,进样量10μl,检测波长295nm。试验结果表明,紫外分光光度法中,沙葱异黄酮含量比高效液相色谱法结果高出46%,相对标准偏差为5.7%,平均加样回收率为98.75%;高效液相色谱法中,沙葱异黄酮含量为4.320mg/kg,相对标准偏差为0.26%,平均加样回收率为99.99%。紫外分光光度法简便、快速,但测定误差大。高效液相色谱法虽然复杂,但比紫外分光光度法准确。     

紫外分光光度法和高效液相色谱法测定盐酸土霉素可溶性粉的含量

采用紫外分光光度法和高效液相色谱法测定盐酸土霉素可溶性粉中土霉素的含量。紫外分光光度法试验表明,用0.01 mol/L盐酸溶液作参比液,以268 nm为测定波长,土霉素在4~32μg/mL范围内吸光度与浓度呈良好的线性关系,相关系数r=0.9999,平均回收率(n=5)为101.04%,RSD=1.02%。高效液相色谱法试验表明,以0.05 mol/L草酸铵溶液-二甲基甲酰胺-0.2 mol/L磷酸氢二铵溶液(75∶20∶5)为流动相,以280 nm为检测波长,土霉素在0.02~0.14mg/mL范围内浓度与峰面积线性关系良好,相关系数r=0.9999,平均回收率(n=5)为100.52%,RSD=0.25%。紫外分光光度法、高效液相色谱法与抗生素微生物检定法比较,结果基本一致。     

HPLC法同时测定小叶丁香中4种护肝活性成分

采用高效液相色谱法同时测定小叶丁香中4种具有保肝护肝作用的活性成分,为小叶丁香作为畜禽饲料开发利用提供依据。将干燥的小叶丁香茎叶部位粉碎,用950 mL/L乙醇超声提取,浓缩,甲醇溶解,离心取上清液经微孔滤膜过滤,以乙腈-1 mL/L醋酸水溶液为流动相,进行梯度洗脱,在254 nm和330 nm双波长下测定,分析小叶丁香中橄榄苦苷、紫丁香苷、松果菊苷和连翘酯苷B的含量。数据显示,松果菊苷在5μg/mL～80μg/mL,橄榄苦苷在50μg/mL～800μg/mL,紫丁香苷和连翘酯苷B在50μg/mL～1 000μg/mL浓度范围内线性关系良好,平均加样回收率为95.2%～99.4%,相对标准偏差介于1.73%～2.83%。该方法前处理简单,灵敏,重复性良好。经测定,小叶丁香茎叶中橄榄苦苷、紫丁香苷、松果菊苷和连翘酯苷B的平均含量分别为0.709%、0.452%、0.059%和0.512%。     

金英黄归注射液制备检测及其对奶牛乳房炎的疗效

为了评价自制的金英黄归注射液的质量标准及临床疗效,本文将金银花等8味中药常规制备成奶牛乳区注射用药,通过急性毒性试验及热源检测评价其安全性,通过紫外分光光度法检测该注射液中多糖的含量,高效液相色谱法测定了绿原酸、大黄酸、大黄素的含量,并将其应用于兽医临床治疗奶牛乳房炎。结果显示,该注射液不引起家兔体温升高,LD50>5 g/kg,多糖浓度为0.48×10-1mg/mL,绿原酸、大黄酸、大黄素的含量分别是0.59 mg/mL、0.26×10-2mg/mL、0.17×10-1mg/mL,隐性乳房炎的奶牛通过3 d中药乳区注射用药治疗后治愈率达53.8%,有效率达92.3%;患隐性乳房炎奶牛乳区自愈率为0;健康奶牛乳区发病率为13.9%;治愈率、有效率经X2检验中药透皮剂治疗组与阳性对照组比较差异显著或极显著(P<0.05,P<0.01),临床型乳房炎的奶牛通过3 d中药乳区注射用药治疗后临床诊断其治愈率达88.9%,有效率达88.9%;CMT诊断治愈率达33.3%,有效率达88.9%。表明该注射剂无热源,几乎无毒性,有效成分检测指标可能是绿原酸,对奶牛乳房炎具有一定的临床疗效的注射剂。     

贞苓增免散的质量标准研究

通过对贞苓增免散进行定性鉴别、定量分析,建立该制剂的质量控制方法。本研究采用薄层色谱法对女贞子、茯苓进行定性鉴别;采用高效液相色谱法测定贞苓增免散中特女贞苷的含量,条件为:以十八烷基硅烷键合硅胶为填充剂、甲醇-水(40:60)为流动相、检测波长224 nm、流速1.0 mL/min、柱温室温、进样量10μL;同时采用硫酸-蒽酮法测定贞苓增免散中茯苓多糖的含量,以葡萄糖为对照品,硫酸-蒽酮溶液为显色剂,在620 nm处测定OD值。结果显示:薄层色谱斑点清晰、无阴性干扰、专属性强、分离度高,能有效鉴别女贞子、茯苓;加样回收试验测得特女贞苷与茯苓多糖的含量分别在0.03~0.30 g/L、0.052~0.100 g/L呈良好的线性关系,平均加样回收率分别为98.28%、99.20%,RSD分别为2.17%、2.22%;高效液相法测得贞苓增免散中特女贞苷含量为12.3 mg/g;硫酸-蒽酮法测得0.1 g/L中贞苓增免散中茯苓多糖含量为0.077 8 mg。本研究建立的方法简便易行、稳定性、重复性好,可有效控制贞苓增免散的质量。     

连翘子中连翘酯苷含量测定方法的研究

为了建立连翘子中连翘酯苷含量的测定方法,为连翘资源的综合开发利用提供理论依据,试验采用反向高效液相色谱法,色谱柱为Hypersil ODS2(250 mm×4.6 mm,5μm);流动相为甲醇-0.1%冰醋酸(35∶65);柱温为30℃;检测波长为290 nm;流速为1.0 mL/min。结果表明:连翘酯苷进样量在0.05~0.80μg范围内线性关系良好(r=0.999 8),平均回收率为98.81%,相对标准偏差(RSD)为1.60%(n=6)。说明该方法简便、灵敏、准确,可作为连翘子中连翘酯苷质量的控制手段。     

HPLC法同时测定茵栀解毒颗粒中3种有效成分的研究

为了建立同时测定中兽药茵栀解毒颗粒中绿原酸、栀子苷和黄芩苷的方法，研究采用高效液相色谱法，选用C₁₈色谱柱(250 mm×4.6 mm, 5μm),乙腈和0.05%磷酸溶液为流动相，流速1.0 mL/min,进样量10μL,柱温为30℃,检测波长为230 nm。结果表明，栀子苷在3.110～155.5μg/mL、黄芩苷在1.897～94.86μg/mL、绿原酸在2.478～123.9μg/mL浓度范围内线性关系良好，相关系数均大于0.999,平均加样回收率分别为97.1%、96.2%和95.3%。该试验操作简便、快捷、定性、定量准确，能满足中兽药茵栀解毒颗粒中3种有效成分同时检测的需要。     

连翘酯苷A对内毒素致鸡脾淋巴细胞白介素-1β的影响

试验旨在研究内毒素和连翘酯苷A对鸡脾淋巴细胞中白介素-1β(IL-1β)水平的影响,将1日龄雏鸡常规饲养至50日龄时,心脏采血处死后,无菌剖检取脾,并用鸡脾淋巴细胞分离液将鸡脾淋巴细胞分为20个组,具体为5个时间点×4个剂量组=20个组。作用时间分别为0、6、12、24和48 h,作用剂量组分别为对照组、内毒素(LPS)组(5μg/m L)、连翘酯苷A预防低剂量组(5μg/m L LPS+200μg/m L连翘酯苷A)和连翘酯苷A预防高剂量组(5μg/m L LPS+400μg/m L连翘酯苷A)。按上述方法培养后,收集鸡脾淋巴细胞,采用酶联免疫吸附试验(ELISA)和实时荧光定量PCR(qRT-PCR)方法检测鸡脾淋巴细胞中IL-1β水平。ELISA结果显示:与对照组相比,LPS组鸡脾淋巴细胞中IL-1β蛋白含量升高;与LPS组相比,连翘酯苷A预防组鸡脾淋巴细胞中IL-1β蛋白含量下降,并与连翘酯苷A添加剂量呈正相关。qRT-PCR结果显示:与对照组相比,LPS组鸡脾淋巴细胞中IL-1βmRNA表达量升高;与LPS组相比,连翘酯苷A预防组鸡脾淋巴细胞中IL-1βmRNA表达量下降,并呈剂量时间效应。试验表明LPS可以诱导鸡脾淋巴细胞中IL-1β蛋白含量和mRNA表达量升高,而连翘酯苷A预防组鸡脾淋巴细胞中IL-1β蛋白含量和mRNA表达量下降,表明连翘酯苷A可以通过转录和翻译途径抑制LPS诱导的鸡脾淋巴细胞中IL-1β蛋白含量和mRNA表达量的升高,减轻炎症反应,发挥抗炎功能。     

商会自治的基石——商会自治规范研究

在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。     

癸氧喹酯干混悬剂含量测定的改进

采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。     

猪毛色遗传的研究进展

本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。     

Comparison of 2 methods of centesis of the bursa of the biceps brachii tendon of horses

REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.     

不同样本商品蜂蜜中硒含量的测定

用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10～0.82μg/g。表明:蜂蜜应视为天然富硒营养品。     

乳酸杆菌益生作用机制的研究进展

乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分（表面蛋白、脂磷壁酸和肽聚糖）与肠道受体（Ｃ型凝集素受体、Ｔｏｌｌ样受体和 Ｎｏｄ样受体）,阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。     

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.     

Mechanism of Action of Probiotic Function of Lactobacilli

乳酸杆菌作为益生菌广泛用于人和动物.本文综述了乳酸杆菌改善宿主健康的机制.乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道.文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制.     

Evaluation of serum concentrations of biomarkers of skeletal metabolism and results of radiography as indicators of severity of osteochondrosis in foals

OBJECTIVE: To determine whether serum concentrations of biomarkers of skeletal metabolism can, in conjunction with radiographic evaluation, indicate severity of osteochondrosis in developing horses. ANIMALS: 43 Dutch Warmblood foals with varying severity of osteochondrosis. PROCEDURE: 24 foals were monitored for 5 months and 19 foals were monitored for 11 months. Monthly radiographs of femoropatellar-femorotibial and tibio-tarsal joints were graded for osteochondral abnormalities. Serial blood samples were assayed for 8 cartilage and bone biomarkers. At the end of the monitoring period, foals were examined for macroscopic osteochondrosis lesions. RESULTS: Temporal relationships were evident between certain serum biomarkers and osteochondrosis severity in foals during their first year. Biomarkers of collagen degradation (collagenase-generated neoepitopes of type-II collagen fragments, type-I and -II collagen fragments [COL2-3/4C(short)], and cross-linked telopeptide fragments of type-I collagen) and bone mineralization (osteocalcin) were positive indicators of osteochondrosis severity at 5 months of age. In foals with lesions at 11 months of age, osteochondrosis severity correlated negatively with COL2-3/4C(short) and osteocalcin and positively with C-propeptide of type-II procollagen (CPII), a collagen synthesis marker. Radiographic grading of osteochondrosis lesions significantly correlated with macroscopic osteochondrosis severity score at both ages and was strongest when combined with osteocalcin at 5 months and CPII at 11 months. CONCLUSIONS AND CLINICAL RELEVANCE: The ability of serum biomarkers to indicate osteochondrosis severity appears to depend on stage of disease and is strengthened with radiography. In older foals with more permanent lesions, osteochondrosis severity is significantly related to biomarker concentrations of decreased bone formation and increased cartilage synthesis.