OBJECTIVE To study the anti-inflammatory and anti-tussive effects of Qingfei Dayuan granules (, QFDY), and to evaluate the acute and sub-chronic toxicity of QFDY. METHODS Anti-inflammatory effects were evaluated by murine model of xylene induced ear edema in mice. Ear swelling degree was calculated and tumor necrosis factor-α, interleukin-1β and interleukin-6 were determined. Anti-tussive evaluations were carried out in the mouse cough model induced by ammonia liquor. Latent period cough and number of cough within 3 min were counted. In acute toxicity study, the rats were randomly divided into test group and solvent control group. Body weighs, food intakes and general clinical signs were monitored. In the sub-chronic toxicity study, QFDY was administered to rats at 0, 4, 8 and 16 g/kg per day for 28 and 30 d of post treatment was conducted. Mortalities, clinical signs, body weight changes, food intakes, ophthalmological examinations, hematological parameters, biochemical indicators, electrolyte indicators, urinalyses and histopathological examinations were monitored. RESULTS QFDY significantly inhibited the development of ear edema in anti-inflammatory assay and decreased cough frequency caused by ammonia liquor. The results presented a dose-effect relationship. In acute toxicity study, no abnormality exhibited at dose of 24.0 g/kg per day during the 14-d observation period. In the sub-chronic toxicity study, higher reticulocyte count, lymphocyte and lower Cl-, blood urea nitrogen were analyzed compared with the solvent control group. But the differences were considered to be incidental and not clinically toxic. Obvious dose-effect relationship of urine color was observed, and the three test groups at the end of the experiments resulted in significant increase in urobilinogen, bilirubin, ketone body and urine leukocyte. However, all the positive indicators returned to normal in the recovery period. Therefore, no toxicological changes were found during the study period. CONCLUSION QFDY showed significant anti-inflammatory and anti-tussive effects in mice. The lethal dose (LD50) of per oral QFDY in rats was estimated to be more than 24.0 g/kg per day and the no observed adverse effect level was over 16 g/kg per day, which suggested that QFDY is relatively safe for oral medication at the present dose on rats. Our experimental results provide a reference for the further development and research of QFDY.

This study aimed to explore the physiological activity of phosphorylated soybean polypeptide chelated calcium (PSPCC) and improve its calcium chelation capacity. To be specific，the preparation process of PSPCC was optimized by single factor test and response surface methodology. And the effect of phosphorylated soybean peptide chelated calcium on osteoblast activity was evaluated by in vitro experiments. The results showed that, the optimum process of PSPCC was as follows: Mass ratio of sodium tripolyphosphate to soybean polypeptide 1:2, phosphorylation reaction temperature 52 ℃, phosphorylation reaction pH7.0, phosphorylation reaction time 9.7 h, mass ratio of phosphorylated soybean polypeptide to calcium chloride 2:1, chelation reaction pH8.0, chelation reaction temperature 50 °C, and chelation reaction time 1.5 h. The maximum calcium chelation amount of PSPCC was 107.25±0.10 mg/g under the above optimal conditions. The results of MTT assay showed that the relative proliferation rate of osteoblasts in the PSPCC group (group D) was 1.6 times that of the soybean polypeptide group (group A) (the third day). In particular, the ALP staining experiment showed that the positive rate of staining in group D was 33.6 times that of the blank group. In addition, the ALP activity of each sample was detected by the ALP kit on the 7th day, and the results showed that the ALP activity of the group D was 6.2 times that of the blank group. And it was found that the number of calcium nodules in the group D was 94 times higher than that of the blank group through the alizarin red staining experiment. It was reasonable that the preparation process of PSPCC was optimized by response surface methodology, which was preliminarily proved that PSPCC had a significant effect on promoting proliferation and differentiation of osteoblasts (P<0.05), and the effect of PSPCC on promoting proliferation and differentiation was higher than that of other samples. This result could provide a theoretical basis for the further exploitation and utilization of PSPCC.

将苦荞麦在不同条件下进行萌发,研究分析萌发过程中黄酮类成分含量变化情况,并对萌发后的苦荞麦与未萌发的苦荞麦进行提取对比试验研究.结果表明,萌发后苦荞麦的黄酮类成分含量明显增加,最佳萌发条件为25℃浸泡16h,25℃萌发6d;苦荞麦萌发后进行提取,与未萌发苦荞麦相比有利于苦荞黄酮的提取溶出,提取液中黄酮类成分浓度和提取转移率均高于未萌发苦荞麦,制备的苦荞提取物黄酮类成分含量和收率也高于未萌发苦荞麦.

为优化大蒜活性成分大蒜素和大蒜多糖的提取工艺,并对直肠功能影响进行初步探究.通过单因素实验结果进行正交组合设计,确定溶剂、料液比、温度和时间对提取大蒜活性成分的影响,通过小鼠直肠切片观察大蒜活性成分对小鼠直肠组织形态结构的影响.结果表明:大蒜素最佳提取条件为料液比 1∶8(g∶mL)、80%乙醇 85℃下提取 2 次,2.0 h/次,大蒜多糖最佳提取条件为料液比 1∶8(g∶mL)、60%乙醇65℃下提取 2 次,1.0 h/次,该条件下提取的大蒜素和大蒜多糖含量理论最大值分别为 50.36 和 479.47 mg/g,与实际值 50.23 和 480.90 mg/g基本一致;药物组有明显的直肠功能改善.大蒜活性成分的最佳提取工艺可靠,大蒜素和大蒜多糖可使直肠损伤小鼠的绒毛长度、隐窝深度得以显著性提高,改善小鼠的消化吸收能力.

为研究桑叶提取物的主要活性成分与其体外抗氧化与α-葡萄糖苷酶抑制活性之间的关系,采用热循环提取与有机膜、树脂纯化相结合的方式制备了4个规格的桑叶提取物,分析了4种桑叶提取物的主要有效成分含量差异,并比较了它们的体外抗氧化与α-葡萄糖苷酶抑制活性.研究发现,桑叶提取物(总黄酮≥20％)中DNJ、黄酮、多酚、多糖分别为0、22.47％、16.85％、30.63％,桑叶提取物(DNJ≥1％)中DNJ、黄酮、多酚、多糖分别为1.37％、5.34％、5.00％、10.46％,桑叶提取物(DNJ≥5％)中DNJ、黄酮、多酚、多糖分别为5.43％、18.63％、13.67％、25.25％,桑叶提取物(DNJ≥10％)中DNJ、黄酮、多酚、多糖分别为13.28％、14.29％、11.66％、16.89％.除桑叶提取物(DNJ≥1％)外,另外3种成分均具有较好的DPPH自由基和ABTS自由基清除能力及ORAC自由基吸收能力,且结果具有一致性.提取物对α-葡萄糖苷酶的抑制活性与DNJ的含量正相关,但非线性相关,且与其黄酮、多酚、多糖含量无显著相关性.桑叶提取物中还存在其他非黄酮类、酚类、多糖类的未知成分对α-葡萄糖苷酶具有抑制活性.

以苦荞麦为原料,研究苦荞黄酮的最佳提取工艺,并对苦荞黄酮进行分离纯化工艺研究.结果表明,苦荞最佳提取工艺为乙醇体积分数60％,提取温度80℃,乙醇体积为苦荞麦质量的4倍,提取时间2 h,提取次数2次,该提取条件下苦荞黄酮提取率最高,对苦荞提取液采用50 nm微滤膜初步除杂,以及800 D或1 000 D的纳滤膜进一步去除分子量较大的杂质,浓缩除去乙醇并进行水沉分离得苦荞黄酮粗提物,再经过反溶剂重结晶,最终得到95％含量以上高纯度苦荞黄酮提取物.

目的:研究板栗壳提取物对SD大鼠的致畸毒性.方法:将受孕SD大鼠随机分为低、中、高3个剂量板栗壳提取物组,阴性对照组和阳性对照组,每组18～20只.低、中、高剂量组分别按0.37、1.11和3.33 g/kg给予板栗壳提取物;阴性对照组给予蒸馏水;阳性对照组按13 mg/kg给予维生素A.于妊娠期的第6～15天灌胃给予受试物或对照物,第20天麻醉后处死孕鼠.观察母鼠和胎鼠的生长发育、受孕率、生殖能力、外观、内脏和骨骼畸形等指标.结果:与阴性对照组比较,阳性对照组第9天至第20天孕鼠体质量偏低,体质量增量降低,胎鼠体质量减轻、身长明显降低,其胎鼠外观畸形率、外观畸形窝数率、内脏畸形率、内脏畸形窝数率、骨骼畸形率、畸形窝数率明显升高,差异有统计学意义(P<0.05),表明阳性对照维生素A对大鼠致畸作用明显,模型成功建立.与阴性对照组比较,板栗壳提取物各剂量未观察到对母鼠及胎鼠的毒性和生殖力的影响,未观察到外观、内脏和骨骼畸形,差异均无统计学差异(P>0.05).结论:在本试验条件下,板栗壳提取物对大鼠母体毒性和胚胎发育毒性未观察到有害作用剂量(NOAEL)大于3.33 g/kg,对大鼠无致畸作用.

目的:确定从艾叶中分离、纯化出的白色晶体为高纯度白坚木皮醇,并检测其纯度.对比分析9个产地艾叶中白坚木皮醇含量差异.方法:采用提取、结晶、重结晶的方法进行分离、纯化,通过核磁共振和高分辨-电喷雾离子源-质谱(HR-ESI-MS)数据鉴定其化学结构和相对分子质量,使用高效液相色谱-蒸发光散射检测器法检测其纯度.结果:从艾叶中分离、纯化出一种白色晶体,根据其HR-ESI-MS数据鉴定其化学结构为白坚木皮醇,纯度>99％.9个产地艾叶中的白坚木皮醇含量差异有统计学意义(P<0.05),这可能与产地和采收时间有关.结论:提供了一种工业化获取高纯度白坚木皮醇的方法,增加了艾叶的附加值.

The yield of total alkaloids from Artemisia argyi leaves was used as the index, and the optimal range conditions of solid-liquid ratio, compound enzyme addition amount, enzymolysis time, enzymolysis pH, ultrasonic time, ultrasonic power, ethanol concentration and ultrasonic temperature were obtained through single factor experiments. The Plackett-Burman method was employed to screen out the factors that had significant effects on the yield of total alkaloids from Artemisia argyi leaves. The Box-Behnken method was used to optimize the extraction process to obtain the optimun extraction conditions. Finally, the inhibitory effect and the minimum inhibitory concentration of the total alkaloid extract of Artemisia argyi leaves against Escherichia coli and Staphylococcus aureus were investigated. The results showed that the important factors affecting the yield of total alkaloids from Artemisia argyi leaves were ultrasonic time, compound enzyme addition amount and enzymolysis time. The optimum extraction conditions were as follows: Ultrasonic time 40 min, compound enzyme addition 1.60%, enzymolysis time 1.5 h, solid-liquid ratio 1:25 g/mL, enzymolysis pH6.0, ultrasonic power 160 W, ethanol concentration 80%, ultrasonic temperature 60 ℃. Under the optimized conditions, the highest yield of total alkaloids was 0.720±0.05 mg/g. The total alkaloids from Artemisia argyi leaves had antibacterial activity against Escherichia coli and Staphylococcus aureus, and their minimum inhibitory concentrations were 3.2 and 1.6 mg/mL, respectively. The actual value of the extraction process and the predicted value had a high degree of fit, which was reliable for prediction of the extraction of the total alkaloids from Artemisia argyi leaves, and the obtained total alkaloids from Artemisia argyi leaves had a certain antibacterial activity.

优选丹参中丹酚酸B和丹参酮类的最佳提取工艺.以丹酚酸B和丹参酮类包括丹参酮ⅡA、丹参酮Ⅰ和隐丹参酮为综合评价指标,采用单因素试验和正交试验考查溶剂体积分数、溶剂倍数、提取温度、提取时间对提取工艺的影响.最佳提取条件为6倍量70％乙醇在80℃下提取3次,每次2 h.稳定性研究和中试验证结果表明所得工艺合理可行,可作为丹参中丹酚酸B和丹参酮类的提取工艺.

以白坚木皮醇的提取率为指标,通过单因素实验和响应面分析法对艾叶药渣中白坚木皮醇的乙醇提取工艺条件进行优化研究,考查了提取时间、料液比、提取温度、提取次数对白坚木皮醇提取率的影响,使用Box-Behnken试验设计方法对影响白坚木皮醇提取率的各因素进行显著性及交互作用分析,最终确定从艾叶药渣中提取白坚木皮醇的最佳工艺条件.结果表明,提取艾叶药渣中白坚木皮醇的最佳工艺条件为:每次提取时间2.5h,提取次数3次,料液比1:25,提取温度80℃,在这个最佳提取条件下白坚木皮醇提取率可达1.765±0.003％.实际值与预测值之间被证明具有很好的拟合性,因此该提取工艺条件可提高白坚木皮醇的提取率.研究结果指出了一条获取白坚木皮醇的新途径,提高了艾叶的附加值.

目的 考察从麦冬中提取纯化特定大小多糖的工艺.方法 考察提取次数对粗多糖提取的影响,确定最佳提取次数,再考察溶剂倍数、提取时间、提取温度的单因素提取率曲线,确定三个因素的最佳正交试验梯度,并进行正交实验,通过分析得出最佳提取工艺.通过不同浓度乙醇醇沉研究探索特定分子量麦冬多糖的纯化工艺.结果 试验显示最佳提取条件为12倍纯水、95℃提取3 h(两次),验证试验粗多糖提取率平均值为:83.33％,(RSD=1.14％,n=3),先通过80％浓度乙醇醇沉去大分子沉淀,再90％浓度乙醇醇沉去小分子上清,制备的麦冬多糖提取物,分子量为4000～9000 g·mol-1的占比高达74％,相比醇沉前提取液中4000～9000 g·mol-1的麦冬多糖占比18％,纯度大幅提高.结论 通过单因素实验确定了各因素合适的正交试验梯度,通过正交试验优化得到最佳提取条件,验证试验结果与正交试验基本吻合,通过两次不同浓度的乙醇醇沉,大幅提高了具有降血糖功能的多糖纯度,工艺简单可行.

目的 比较不同厂家茯苓配方颗粒的质量差异.方法 采用红外光谱结合共有峰率和变异峰率双指标序列法评价不同厂家茯苓配方颗粒与药材之间的相似性和差异性.结果 茯苓配方颗粒与茯苓药材共有特征峰范围为:3381～3394,2921～2934,1637～1646,1369～1395,1077～1080,558～595cm-1;不同厂家茯苓配方颗粒的共有峰率和变异峰率存在差异,共有峰率在50.0％～66.7％,变异峰率在0～71.7％.结论 茯苓配方颗粒与药材红外光谱的相似性和差异性不同,表明不同厂家茯苓配方颗粒质量存在明显差异,可为茯苓配方颗粒质量评价提供参考依据.

保元汤是《古代经典名方目录(第一批)》收载的经典名方之一,已具有500多年应用历史,现代广泛用于治疗元气不足、虚损劳怯等.通过分析明、清两代具有代表性的中医古籍结合现代研究,分析、总结了保元汤来源、组成、药效等,综述保元汤的古代和现代文献研究概况,为建立保元汤物质基准质量标准提供了参考.通过研究发现,保元汤早期用以治疗小儿痘疹类疾病,随着社会发展,历代医家通过在临床实践中不断摸索,赋予了保元汤新的内涵,使其临床治疗疾病的范围得到扩充.现代医家将保元汤及其加减方临床用于冠心病、再生障碍性贫血、慢性肾功能衰竭、慢性肾炎、白细胞减少等多种疾病,充分说明保元汤临床效果显著,具有潜在开发价值.

以人参为原料,用不同浓度乙醇提取,大孔树脂对提取液进行吸附,比较皂苷提取量及树脂对皂苷的吸附量,将解吸液干燥制得粉末,比较总皂苷含量及单位成本.另将提取液经过处理,考察在不同环境下的稳定性.结果表明:用55％乙醇时人参皂苷提取量最大,达74.3 mg/g;制得人参皂苷粉末中皂苷含量最高,为734 mg/g,综合单位生产成本也最低.各组提取液经过滤处理后,只有35％乙醇提取过滤液在冷、热、常温下稳定性均较好.

研究黑花生中有效成分的提取制备工艺,为黑花生农作物的高值化利用提供理论依据.采用单因素试验和正交设计试验方法,确定黑花生有效成分的最佳提取条件.以黑花生总黄酮为目标,确定了最佳提取条件为料液比1∶20(g∶mL),50％乙醇90℃下提取2次,1.0 h/次,含量为88.3 mg/g;以黑花生总多糖为目标,最佳提取条件为料液比1∶15 (g∶mL),20％乙醇80℃下提取2次,2.0 h/次,含量为667.2 mg/g.通过验证,确定了黑花生各有效成分的最佳提取条件,为黑花生的合理使用提供了依据.

目的:比较桑不同药用部位(桑叶、桑枝、桑白皮)总生物碱对α-葡萄糖苷酶活性的抑制作用.方法:采用采用蒸发光散射检测器(ELSD)-高效液相色谱法(HPLC)测定桑不同药用部位总生物碱中1-脱氧野尻霉素(DNJ)含量;以半数抑制浓度(IC50)为评价指标,以阿卡波糖为阳性对照,以对-硝基苯基-α-D-吡喃葡萄糖苷为底物,采用体外抑制模型评价桑不同部位总生物碱对α-葡萄糖苷酶活性的抑制作用.结果:桑不同药用部位总生物碱中DNJ质量分数为桑白皮生物碱(30.1％)>桑枝生物碱(25.8％)>桑叶生物碱(21.4％).桑不同药用部位总生物碱对α-葡萄糖苷酶抑制作用强度为桑枝生物碱>桑叶生物碱>桑白皮生物碱>阿卡波糖.结论:在桑枝、桑叶、桑白皮总生物碱中,以桑枝总生物碱对α-葡萄糖苷酶活性抑制作用最强,为桑资源开发辅助降血糖的药品和保健食品提供依据.

目的 建立黄连样品的HPLC特征图谱,阐述其量质传递规律.方法 采用相同色谱条件分别建立23批黄连药材、饮片、提取液、浓缩液、标准汤剂冻干粉、喷雾干膏粉和配方颗粒成品的HPLC特征图谱,比较其相似度;再对其进行定性、定量的量质传递分析,通过K值、损失率的大小和波动性来评价定量传递关系.结果 黄连特征图谱均含有8个色谱峰,其中,峰1为木兰花碱、峰3为非洲防己碱、峰4为药根碱、峰5为表小檗碱、峰6为黄连碱、峰7为巴马汀、峰8为小檗碱;23批样品的相似度均≥0.990,呈现良好的相关性;各过程中K值的波动稳定,实现了稳定的量质传递,损失率≤10％.结论 所用方法准确、稳定、可靠,通过HPLC特征图谱阐述了黄连配方颗粒的量质传递规律.

为研究从胡芦巴中提取总皂苷的最佳工艺,考查确定最佳提取次数后,以提取乙醇体积分数、溶剂倍数、提取时间、提取温度为4个因素进行单因素试验,确定各因素正交试验时适宜的因素水平,选用L9(34)正交表,以总皂苷提取率为指标,进行四因素三水平正交试验,筛选胡芦巴总皂苷最佳提取工艺.结果表明,胡芦巴总皂苷的最佳提取工艺(提取2次)为乙醇体积分数30％,溶剂倍数12倍,提取时间2.5 h,提取温度75 ℃,在此条件下,皂苷提取率可达81.02％(RSD=0.81％,n=3),得率1.12％,3组验证提取表明,正交试验得到的最佳工艺稳定可靠、提取率高,为胡芦巴在中药降血糖上的应用提供参考和依据.