叶下珠提取工艺探讨

用双波长薄 层扫描法测定了 叶下珠常温 水浸 法,煎 煮法,５５ ％ 乙 醇渗 漉提 取法 及５５ ％ 乙 醇回 流提取法４ 种提 取工艺及提取液 中有效成分的 含量。提示,用渗 漉法提取最佳     

银杏黄酮的酶法提取工艺研究

本文研究了银杏黄酮的酶法提取工艺，银杏叶原料经纤维素酶预处理后浸提，总黄酮得率显著提高，得率可达到2．01％，其酶解过程的最优参数为：料液中酶浓度为0．125g/L，酶与底物配比为1：1200，酶解温度45℃，自然pH值，酶解时间2h。     

叶下珠提取工艺的研究

目的优选叶下珠的提取工艺.方法采用正交试验法,以总黄酮含量为检测指标优选叶下珠提取工艺的最佳条件.结果影响提取的主次因素为D＞A＞B＞C(A为乙醇浓度,B为乙醇用量,C为提取时间,D为提取次数).最佳提取条件为7倍量70%乙醇,提取3次,每次2 h.结论优选得到的工艺稳定可行.     

超声波法碱水提取叶下珠黄酮的研究

目的 研究超声波法碱水提取叶下珠黄酮的工艺.方法 以pH、处理时间、料液比为主要考察因素,以浸取样品中总黄酮的含量为主要考察指标,确定最佳提取条件.结果 最佳提取工艺为:Ca(OH)2的pH值为11,超声波提取时间为40 min,料液比为1:50.结论 与传统的乙醇提取法进行对比显示,用超声波提取法具有快速、高效等特点.     

微波辅助提取麦冬黄酮的工艺研究

目的:研究微波辅助法提取麦冬黄酮的工艺条件。方法:采用单因素试验,探讨料液比、微波处理时间、微波强度以及乙醇体积分数对麦冬黄酮提取量的影响,并与传统回流法进行比较。结果:微波提取优化工艺条件为:乙醇体积分数为90%,料液比为1:14(g/ml),微波功率为中档,微波提取时间4min。微波处理后再进行回流提取较不经微波处理得到提取物的黄酮含量高出39.7%。结论:微波辅助法是一种提取麦冬黄酮的有效方法。     

鸡爪槭黄酮提取工艺研究

目的:以鸡爪槭叶片为原料,对鸡爪槭叶片黄酮提取工艺进行优化研究。方法:通过单因素试验和L16(45)正交试验法对影响鸡爪槭叶片黄酮提取的溶剂浓度、料液比、浸提温度、反应时间等因素进行研究。结果:鸡爪槭黄酮的最佳提取工艺条件为:80%的乙醇,料液比1:25,浸提温度55℃,反应时间2h,pH值为13,浸提2次。在此最佳工艺条件下,对其叶片粉末进行浸提,得到黄酮的得率为3.634%。结论:此鸡爪槭黄酮最佳提取工艺明显优于一般鸡爪槭黄酮提取工艺,值得推广使用。     

纤维素酶在三七提取工艺中的应用

为考察纤维素酶对中药材中皂甙类成分提取效果的影响 ,实验选用三七 ,用乙醇溶液进行提取 ,以提取液中固形物含量和三七总皂甙含量为考察指标 ,对加入纤维素酶与不加纤维素酶两种提取方法进行比较研究 ,结果比较满意。为纤维素酶的工业化应用提供了一定依据。1　实验仪器与材料　纤维素酶粗品 (活性单位≥ 150 0ｕ/ｇ) ,由华美生物工程公司提供 ;人参皂甙Ｒｇ1 、Ｒｂ1 、Ｒｅ和三七皂甙Ｒ1 对照品 ,由中国药品生物制品检定所提供 ;三七药材从四川省中药材公司购进 ,经鉴定为Ｐａｎａｘｎｏｔｏｇｉｎｓｅｎｇ(Ｂｕｒｋ .)Ｆ .Ｈ .Ｃｈｅｎ…     

叶下珠总黄酮的半仿生提取工艺

目的 研究半仿生法提取叶下珠总黄酮的工艺条件.方法 利用正交实验对叶下珠总黄酮的半仿生法提取工艺进行了优选,以黄酮提取得率为考察指标,考察提取温度、提取时间和料液比三因素对总黄酮提取得率的影响.结果 实验得到的最佳提取工艺条件为,料液比1∶20,在85℃下每次提取1.5h,分别以pH 2.0、7.5、8.3的水溶液为提取溶剂各提取一次.在此条件下,黄酮的提取得率较高,均值为1.505％.结论 研究结果可为进一步开发利用叶下珠植物资源提供科学依据.     

内部沸腾法提取叶下珠没食子酸

目的:优选内部沸腾法提取叶下珠没食子酸工艺条件.方法:用少量乙醇溶液为解吸剂润湿叶下珠粉末,使没食子酸充分解吸,加一定量热提取剂,使渗透到叶下珠组织内部的乙醇沸腾,强化提取过程.选取乙醇体积分数、解吸时间、乙醇用量、提取时间和提取温度等5个因素,4水平进行正交试验,采用高效液相色谱法测定没食子酸含量.结果:内部沸腾法提取叶下珠没食子酸的最佳工艺条件为1.6倍量60％乙醇解吸30 min,80℃提取15 min.结论:在最佳提取工艺条件下,叶下珠没食子酸提取得率可达0.970％,该法有较好的应用前景.     

正交试验设计优选栀子有效成分半仿生-纤维素酶提取工艺

目的优选半仿生-纤维素酶提取法提取栀子有效成分的最佳工艺条件。方法以栀子苷、总环烯醚萜得率和干浸膏收率为评价指标,以纤维素酶用量、酶解时间、酶解p H、料液比为考察因素,以正交试验设计优选最佳提取工艺。结果最佳提取工艺为水解酶用量2 mg·g-1,酶解时间2 h,酶解p H 6.5,料液比1:18。结论优选的工艺合理、可行,可为栀子后续研究提供参考。     

叶下珠高效液相色谱指纹图谱研究

目的:建立叶下珠药材的高效液相色谱指纹图谱.方法:采用反相高效液相色谱法,以柯里拉京为内参比峰.色谱条件为Kromasil C18柱(4.6 mm × 250 mm,5μm),流动相乙腈-体积分数为0.1％的磷酸水,梯度洗脱,流速1.0 mL·min-1,柱温30℃,检测波长280 nm.采用SPSS 16.0软件进行系统聚类分析;采用国家药典委员会颁布的中药色谱指纹图谱相似度评价系统(2009版)进行相似度评价.结果:在色谱指纹图谱中,确立了21个共有峰,指认了其中6个色谱峰,系统聚类分析将14批叶下珠药材分为两类,建立了叶下珠药材的共有模式,在14批叶下珠药材中有12批样品的指纹图谱相似度在0.90以上.结论:该方法简便、准确、重复性好,为评价叶下珠药材的质量提供了依据.     

Effects of Phyllanthus urinaria extract on HepG2 cell viability and oxidative phosphorylation by isolated rat liver mitochondria

Ethnopharmacological relevance

Phyllanthus urinaria is widely used as anti-inflammatory, anti-diarrheal and hepatoprotective medicines in Asian countries such as India, China and Thailand. In Thailand, Phyllanthus urinaria is traditionally used as an adjuvant or alternative medicine for cancer patients, including liver cancer. However, there is limited scientific evidence supporting its use in cancer particularly hepatocellular carcinoma.

Aim of the study

To investigate the cytotoxic effect of Phyllanthus urinaria extract on human hepatocellular carcinoma HepG2 cells and the effect on oxidative phosphorylation by isolated rat liver mitochondria.

Materials and methods

HepG2 cells and isolated rat liver mitochondria were treated with the 50% methanolic extract of Phyllanthus urinaria. Cytotoxicity of the extract was assessed by trypan blue exclusion and MTT assay. Rates of oxygen consumption of isolated mitochondria were determined with a Clark oxygen electrode.

Results

It was found that the hydromethanolic extract induced cell death of HepG2 cells in a dose-dependent fashion. The IC₅₀ of Phyllanthus urinaria extract measured by trypan blue exclusion and MTT assay were 431 ± 65 μg/ml and 445 ± 62 μg/ml, respectively. Morphological changes of the cells were also observed. With isolated rat liver mitochondria, the extract slightly stimulated mitochondrial state 4 respiration but profoundly depressed state 3 respiration and respiratory control ratio.

Conclusions

The extract impairs energy metabolism by acting as inhibitor of oxidative phosphorylation and weak mitochondrial uncoupler. These mitochondrial effects may play a role in the cytotoxic action of Phyllanthus urinaria extract on HepG2 cells. These results provide preliminary experimental evidence supporting the use of Phyllanthus urinaria against hepatocellular carcinoma and open the possibility of considering this plant an adjunctive medicine for the treatment of this deadly disease.     

余甘子对肿瘤细胞抑制作用及免疫调节的研究

目的 :运用中药血清药理学的研究方法,研究余甘子对S180荷瘤小鼠肿瘤存活率、主要免疫器官及红细胞免疫调节能力的影响。 方法 :余甘子采用鲜果榨汁经冷冻干燥保存。建立小鼠肿瘤模型,模型小鼠随机分组,小鼠末次给药后摘眼球取血,分离红细胞、淋巴细胞,进行肿瘤红细胞淋巴细胞混合花环实验;观察不同剂量给药组对小鼠肿瘤抑制率及脾、胸腺指数的影响。 结果 :与模型对照组比较,余甘子大剂量组红细胞促淋巴细胞黏附肿瘤细胞能力及抑制肿瘤存活率具极显著差异性;与阳性对照组比较,各剂量组对免疫器官的影响均具显著差异性。 结论 :余甘子在抑制肿瘤生长的同时,对免疫器官有极好的保护作用。     

Antidiabetic and antioxidant effect of various fractions of Phyllanthus simplex in alloxan diabetic rats

Aim of the study

To evaluate the antidiabetic and antioxidant effects of various fractions of Phyllanthus simplex on alloxan induced diabetes in rats.

Materials and methods

Hypoglycemic effect of Phyllanthus simplex fractions was evaluated in normal and diabetic rats. Diabetes was induced by intraperitoneal injection of alloxan monohydrate (120 mg/kg). Normal and diabetic rats were divided into different groups (six rats each group) and orally administered with petroleum ether (P.E.) (200 and 400 mg/kg), ethyl acetate (EtOAc) (100 and 200 mg/kg), methanol (125 and 250 mg/kg), water fraction (150 and 300 mg/kg) and glibenclamide (10 mg/kg) for 21 days. Blood samples were collected from overnight fasted normal rats on day 21, from overnight fasted diabetic rats at 7, 14 and 21 days of treatment and analyzed for blood glucose level. On day 22 blood samples were collected from diabetic rats to estimate biochemical parameters, rats were sacrificed by single stunning and tissues were excised to measure their antioxidant and glycogen status.

Results

In the normoglycemic rats, MeOH (125 and 250 mg/kg) and aqueous fractions (150 and 300 mg/kg) showed a significant (P < 0.05) hypoglycemic effect on day 21. In diabetic control rats, MeOH (125 and 250 mg/kg) and aqueous fractions (150 and 300 mg/kg) showed significant antihyperglycemic effect (P < 0.001). The active fractions (MeOH and aqueous) of Phyllanthus simplex also increased the body weight of diabetic rats significantly compared to the control group. The active fractions were able to normalize the marked alterations in antioxidant enzymes and antioxidant parameters levels in liver and kidney. Treatment with the active fractions also normalized the diabetic induced hyperlipidemia and liver glycogen.

Conclusions

These results demonstrate the antidiabetic and antioxidant potential of fractions of Phyllanthus simplex and suggests that the plant may have therapeutic value in diabetes and related complications.     

五指莲重楼的黄酮和甾体类化学成分研究

目的:研究五指莲重楼根茎中的化学成分。方法:采用溶剂法提取,利用正相硅胶色谱、葡聚糖凝胶SephadexLH-20、反相制备色谱等手段进行分离纯化,并通过现代波谱技术进行结构鉴定。结果:从乙酸乙酯层和正丁醇层中分离得到14个化合物,分别鉴定为胡萝卜苷(1),豆甾醇-3-O-β-D-吡喃葡萄糖苷(2),β-蜕皮激素(3),偏诺皂苷元-3-O-α-L-呋喃阿拉伯糖基(1→4)-[α-L-吡喃鼠李糖基(1→2)]-β-D-吡喃葡萄糖苷(4),薯蓣皂苷元-3-O-α-L-吡喃鼠李糖基(1→4)-α-L-吡喃鼠李糖基(1→4)-[α-L-吡喃鼠李糖基(1→2)]-β-D-吡喃葡萄糖苷(5),山柰酚(6),芦丁(7),杨梅素(8),4,2′,4′-trihydroxychalcone(9),异鼠李素-3-O-β-D-吡喃葡萄糖苷(10),异鼠李素-3-O-芸香糖苷(11),异鼠李素-3-O-龙胆双糖苷(12),5,5′,7-三羟基-4′,4,7″-三甲氧基-[3′-8″]双黄酮(13),5,5′,7,4′,4,7″-六羟基-[3′-8″]双黄酮(14)。结论:化合物1~3,6~14为首次从该种植物中分离得到,其中化合物7~9,13和14为首次从该属植物中分离得到。     

回心草提取工艺研究

目的:采用正交设计法,对回心草的提取工艺进行优化。方法:采用L9(34)正交试验,乙醇体积分数(A)、乙醇用量(B)、提取时间(C)作为3个因素,每个因素选择3个水平,以熊果酸转移率来确定最佳工艺条件。结果:B,A影响最大,C最小。结论:最佳提取条件是A1B3C2,即60%乙醇16倍量提取2次,每次2 h。     

In vitro and in vivo anti-hepatitis B virus activities of the lignan niranthin isolated from Phyllanthus niruri L.

Ethnopharmacological relevance

Niranthin is a lignan isolated from Phyllanthus niruri L. This plant has long been used in folk medicine for liver protection and antihepatitis B in many Asian countries. This study was designed to evaluate the anti-hepatitis B virus activity of niranthin using HepG2.2.15 cells and duck hepatitis B virus (DHBV) infected ducks as in vitro and in vivo models.

Materials and methods

Niranthin was isolated from Phyllanthus niruri L. (Euphorbiaceae) by extraction and chromatographic procedures and the anti-hepatitis B virus activity was evaluated both in vitro and in vivo. The human HBV-transfected liver cell line HepG2.2.15 was used in vitro assay. And the in vivo anti-hepatitis B virus activity was evaluated on the expression of HBV replication, HBsAg, HBeAg, ALT and AST on day 0, 7, 14, 17 after niranthin was dosed intragastricly (i.g.) once a day for 14 days at the dosages of 25, 50 and 100 mg/kg/day in the duck hepatitis B virus (DHBV) infected ducks.

Results

In the human HBV-transfected liver cell line HepG2.2.15, the secretion of HBsAg and HBeAg were significantly decreased after treatment with niranthin for 144 h, with IC₅₀ values for HBsAg of 15.6 µM, IC₅₀ values for HBeAg of 25.1 µM. In DHBV-infected ducklings, niranthin significantly reduced the serum DHBV DNA, HBsAg, HBeAg, ALT and AST. Furthermore, analysis of the liver pathological changes confirmed the hepatoprotective effect of niranthin.

Conclusion

The experimental data demonstrated that niranthin exhibits anti-hepatitis B virus activity both in vitro and in vivo.     

红花减压提取工艺优选

目的:优选红花减压提取工艺参数.方法:以羟基红花黄色素A的转移率和干膏率为考察指标,采用正交试验法对加水量、真空度、提取时间及提取次数等因素进行优选,并与常压煎煮法及文献方法进行比较.结果:优选的减压提取工艺为加12倍量水提取3次,每次20 min,系统真空为-0.045 ～-0.055 MPa.减压提取法优于常压煎煮法及文献方法.结论:该优选工艺稳定,可行.     

叶下珠化学成分研究

目的研究叶下珠Phyllanthus urinaria全草的化学成分。方法应用硅胶、Sephadex LH-20、RP_(18)、MCI-gel柱色谱和半制备HPLC等色谱方法对叶下珠的乙醇提取物进行分离纯化;根据理化性质和波谱学方法鉴定化合物的结构。结果从叶下珠95%乙醇提取物的石油醚和醋酸乙酯萃取部位中分离得到16个化合物,分别鉴定为没食子酸(1)、咖啡酸(2)、没食子酸乙酯(3)、没食子酸甲酯(4)、4-乙氧基苯甲酸(5)、邻苯二甲酸二异丁酯(6)、邻苯二甲酸二丁酯(7)、(4R,6R)-2,3-dihydromenisdaurilide(8)、(4R,6S)-2,3-dihydromenisdaurilide(9)、aquilegiolide(10)、menisdaurilide(11)、cassipourol(12)、(9Z,12Z)-nonadeca-9,12-dienoic acid(13)、亚油酸甲酯(14)、豆甾醇(15)、(8R,8'S,7S)-4'-(3"-methoxyrhamnopyranosyl)oxy-8'-hydroxy-3,3',4-trimethoxy-8-hydroxymethyl-lign-7,9'-lactone(16)。结论化合物5~7、10、11、13、14为首次从该植物中分离得到,化合物8、9、12、16为首次从该属植物中分离得到。