高效液相色谱法测定珠芪复肝颗粒中没食子酸的含量

<正>珠芪复肝颗粒是由珠子草、黄芪、甘草等组成的中药复方制剂,有清热解毒、舒肝利胆、扶正祛邪等作用,临床用于乙型病毒性肝炎的治疗。该制剂中主药珠子草主要含有多酚类、黄酮类,其中多酚类成分没食子酸为珠子草的主要特征性有效成分。为了有效控制该制剂的质量,本实验建立了以高效液相色谱法(HPLC)测定制剂中没食子酸含量的方法。     

槟榔多酚提取物中总多酚和儿茶素类的含量测定

目的 建立槟榔多酚提取物中总多酚和儿茶素类含量的测定方法,为槟榔多酚提取物的质量控制提供参考。方法 采用紫外分光光度法对槟榔提取物中总多酚的含量进行测定;高效液相色谱法对儿茶素类物质进行含量测定。结果 紫外分光光度法测定槟榔提取物中总多酚含量在9.8～58.8μg/ml范围内线性关系良好,平均加样回收率为98.44%,RSD为1.4%;在高效液相色谱测定中,儿茶素、表儿茶素和原儿茶酸3种主要成分完全分离,在各自范围内线性关系良好,平均加样回收率为99.17%～101.67%,RSD为1.2%～2.5%。结论 建立的含量测定方法简单、稳定可靠,可用于槟榔多酚提取物的定量分析,为槟榔多酚提取物的质量控制提供实验依据。     

贵州产黄珠子草总黄酮毒性及抗CCl4、AAP中毒小鼠肝损伤实验

黄珠子草为大戟科叶下珠属(Phyllanthus)植物,贵州产叶下珠属植物黄珠子草(Phyllanthus Virgatus Forst．f．)广泛分布于兴义、兴仁、织金、贞丰等地。我们采用贵州产黄珠子草已进行其总黄酮的提取分离方法与含量测定,但资料查新未见有关贵州产黄珠子草总黄酮毒性与抗肝损伤等方面的研究报道。为此我们特进行了贵州产黄珠子草总黄酮毒性及抗CCl4、AAP中毒小鼠肝损伤实验,现将其研究结果报告如下。     

HPLC同时测定珠子草中没食子酸、短叶苏木酚和鞣花酸的含量

目的 建立同时测定珠子草中没食子酸、短叶苏木酚和鞣花酸的HPLC法,并对珠子草该3种成分进行含量测定。方法 色谱柱：Shimazu C₁₈(150 mm×4.6 mm,5 μm);流动相：0.1%磷酸(A)-乙腈(B)梯度洗脱,流速为0.8 mL·min^-1;柱温：室温;检测波长：270 nm;进样量10 μL。结果 没食子酸、短叶苏木酚和鞣花酸含量分别在0.6~9.6 μg、0.525~8.4 μg、0.475~7.6 μg内与峰面积呈良好的线性关系。没食子酸的平均含量为0.196 2%,短叶苏木酚为0.518 2%,鞣花酸为0.411 0%。结论 本方法操作简便,准确度高,可用于珠子草中没食子酸、短叶苏木酚和鞣花酸3种成分含量的测定。     

不同干燥方法对三叶青活性成分含量的影响

目的 探讨不同干燥方法对三叶青活性成分含量的影响。方法 选取三叶青中总黄酮、总多酚和多糖3种主要活性成分作为评价指标,分析测定三叶青鲜品及冷冻干燥、真空干燥、热风干燥3种不同干燥方法的三叶青总黄酮、总多酚和多糖含量。结果 三叶青鲜品和3种干燥方法的总黄酮含量分别为7.68,7.73,6.65,6.36 mg/g,总多酚含量分别为8.69,8.88,8.32,6.51 mg/g,多糖含量分别为6.39%,6.52%,4.87%,5.11%;传统的热风干燥方法对总黄酮、总多酚和多糖造成的含量损失分别为17.18%,25.09%,20.03%,真空干燥对总黄酮、总多酚和多糖造成的含量损失分别为13.41%,4.25%,23.79%,冷冻干燥的三叶青活性成分含量与三叶青鲜品相当,几乎没有损耗。结论 不同的干燥方法对三叶青活性成分含量影响很大,冷冻干燥能最大程度地保留三叶青活性成分,适合作为三叶青干燥加工方法。     

黄珠子草的化学成分研究

黄珠子草 (ＰｈｙｌｌａｎｔｈｕｓＶｉｒｇａｔｕｓＦｏｒｓｔ .ｆ .)在贵州广泛分布于兴义、兴仁、织金、贞丰等地 ,生长在1350米以下山坡、草地。贵州民间用叶下珠 (Ｐｈｙｌ ｌａｎｔｈｕｓＵｒｉａｎｒｉａＬ .又名山皂角 )、黄珠子草治疗泌尿系感染、肠炎、结膜炎、黄疸型肝炎等。黄珠子草(ＰｈｙｌｌａｎｔｈｕｓＶｉｒｇａｔｕｓＦｏｒｓｔ .ｆ .)据报道在体外有较好的抗ＨＢＶ作用[1] ,但未曾见有对其化学成分的报道。我们通过对黔产黄珠子草总黄酮 (含量2 4 0 2 14ｍｇ/ｇ)的测定 ,结果表明 ,黔产黄珠子草总黄酮含量较高 ,因此黔…     

RP-HPLC测定黄珠子草中短叶苏木酚的含量

目的:建立高效液相色谱法测定黄珠子草中短叶苏木酚的含量。方法:以 Thermo Hypersil-Keystone ODS C_(18)为分析柱,甲醇-1%醋酸(36:64)为流动相,流速1.0mL·min~(-1),检测波长278nm,采用外标法定量测定。结果:短叶苏木酚在10～100μg·mL~(-1)范围内线性关系良好(r=0.999 7);其平均回收率为98.77%,RSD 为0.98%(n=6)。结论:该方法简便可靠,快速,重现性好,可用于黄珠子草中短叶苏木酚的含量测定。     

枸杞多酚提取物中总多酚的测定

目的建立紫外分光光度法测定枸杞多酚提取物中的总多酚。方法采用紫外分光光度法测定枸杞多酚提取物中总多酚,用水稀释样品,加入显色剂Filin-Ciocalteu试剂,在760 nm处测定吸光度,用同样方法测定并绘制标准曲线。结果紫外分光光度法测定枸杞多酚提取物中总多酚的量分别为24.03%、26.38%、25.45%、28.27%、29.08%,线性范围1～8μg/mL,R2=0.999 5。平均回收率为103.5%,RSD<5%。结论该方法操作简单,准确可靠,适于枸杞多酚提取物中总多酚的测定。     

紫外分光光度法测定吉祥草中的总皂苷

目的 建立测定吉祥草中总皂苷含量的方法,并测定不同来源的吉祥草药材中总皂苷的含量.方法 采用UV法,在453nm处测定吉祥草中总皂苷的含量.结果 凯提皂苷元26-136μg与吸光度的线性关系良好(r=0.9996),方法的平均回收率为97.3%(n=9),RSD=1.8%;不同来源的吉祥草药材中总皂苷含量有较大差异.结论 所用测定方法简便、准确、灵敏、可靠,可控制吉祥草的药材质量.     

中药多酚提取物对α-葡萄糖苷酶抑制作用的研究

目的 探讨青钱柳和养心草两种中药的多酚提取物对α-葡萄糖苷酶活性的抑制作用.方法 制备两种闽产中药多酚提取物,采用Folin-Ciocalteu法测定多酚含量并研究其对α-葡萄糖苷酶活性的抑制作用.结果 两种提取物总多酚含量达10%以上,并能抑制α-葡萄糖苷酶活性.结论 这两种中药多酚提取物可作为α-葡萄糖苷 酶抑制剂.     

Phytochemicals from Phyllanthus niruri Linn. and their pharmacological properties: a review

This review discusses the medicinal plant Phyllanthus niruri Linn. (Euphorbiaceae), its wide variety of phytochemicals and their pharmacological properties. The active phytochemicals, flavonoids, alkaloids, terpenoids, lignans, polyphenols, tannins, coumarins and saponins, have been identified from various parts of P. niruri. Extracts of this herb have been proven to have therapeutic effects in many clinical studies. Some of the most intriguing therapeutic properties include anti-hepatotoxic, anti-lithic, anti-hypertensive, anti-HIV and anti-hepatitis B. Therefore, studies relating to chemical characteristics and structural properties of the bioactive phytochemicals found in P. niruri are very useful for further research on this plant as many of the phytochemicals have shown preclinical therapeutic efficacies for a wide range of human diseases, including HIV/AIDS and hepatitis B.     

Validation of a LC method for the analysis of phenolic compounds from aqueous extract of Phyllanthus niruri aerial parts

A reversed-phase high-performance liquid chromatographic separation and quantitative method using a phosphoric acid-acetonitrile gradient was developed to analyze phenolic compounds present in aqueous extract from the aerial parts of Phyllanthus niruri. The chromatographic method was validated for linearity, precision and accuracy for both reference substance (gallic acid) and for three well resolved peaks from P. niruri aqueous extract. Both calibration curves were linear with correlation coefficients higher than 0.999. The reproducibility for the three peaks ranged from 2.3% to 4.6% and the accuracy for gallic acid in the aqueous extract was 103%. The method allowed the complete resolution of three peaks, one of them was identified by diode array detection as gallic acid. The analysis of the botanic morphological elements of the aerial parts from P. niruri showed that the leaves have a higher amount of phenolic compounds than the branches.     

Antihyperuricemic lignans from the leaves of Phyllanthus niruri

The methanol extract from the leaves of Phyllanthus niruri L. showed oral antihyperuricemic activity in potassium oxonate- and uric acid-induced hyperuricemic rats. Fractionation of the extract by resin chromatography led to the isolation of a less polar fraction which exhibited the highest reduction of plasma uric acid. Further antihyperuricemic-guided purification of the fraction afforded three lignans, phyllanthin (1), hypophyllanthin (2) and phyltetralin (3), of which 1 significantly reversed the plasma uric acid level of hyperuricemic animals to its normal level in a dose-dependent manner, comparable to that of allopurinol, benzbromarone and probenecid which are used clinically for the treatment of hyperuricemia and gout. Thus, the lignans of P. niruri are potential antihyperuricemic agents worthy of further investigation.     

Symptomatic treatment of acute tonsillo-pharyngitis patients with a combination of Nigella sativa and Phyllanthus niruri extract

Acute tonsillopharyngitis is characterized by tonsil or pharyngeal inflammation and mostly is a virus in origin; thus, treatment that covers both the inflammation and inadequate immune response against the pathogenic organism is needed. NSPN extract containing Nigella sativa and Phyllanthus niruri extracts has both antiinflammatory and immunomodulatory effects. A comparative, parallel, randomized, double-blind, placebo-controlled study with a treatment period of 7 days was conducted to examine clinical effectiveness of Nigella sativa and Phyllanthus niruri extract (NSPN extract). Of 200 enrolled patients, 186 patients completed the study, 12 patients withdrew and 2 patients were principally screened failure but inadvertently included. NSPN capsules, each containing 360 mg Nigella sativa and 50 mg Phyllanthus niruri extracts, were orally administered 3 times 1 capsule daily for 7 days. At Hour 5 or 6 of the first dosing of study medication, the sore throat assessed as swallowing pain and difficulty, was markedly alleviated in the NSPN group. In line with the significant alleviation of pain, from Days 0 to 2 of treatment, subjects in the NSPN group also needed significantly less escape aanalgesica therapy (paracetamol tablets) than those in the placebo group. At the end of treatment (Day 7), a significantly greater proportion of patients in the NSPN group than in the placebo group had their sore throat completely relieved. NSPN extract was also found to be safe and well tolerated in acute tonsillopharyngitis patients. This study proved significant benefits of NSPN extract in the treatment of acute tonsillopharyngitis as compared to placebo.     

大鼠血浆中儿茶素主要代谢产物没食子酸含量测定

目的 建立大鼠血浆中儿茶素主要代谢产物没食子酸含量检测方法.方法 采用改进的高效液相色谱法测定对大鼠口灌儿茶素后血浆中没食子酸含量,色谱柱为大连依利特BDS C18(250×4.6mm)色谱柱,流动相为乙腈:0.1%柠檬酸水溶液=5:95;柱温30℃,波长280nm,以茶碱作为内标.结果 改进的HPLC方法 测定儿茶素...     

Phylogeny of medicinal Phyllanthus species in China based on nuclear ITS and chloroplast atpB-rbcL sequences and multiplex PCR detection assay analysis

Plants of the genus Phyllanthus are commonly used in India, China and Korea for medicinal purposes. Although they are widely cultivated and marketed, there has been uncertainty about the efficacy of different species. A prerequisite of Good Agricultural Practice (GAP) is the authentication of relevant species, and this is now made unequivocal by applying DNA sequence tools. In this study the phylogenetic relationships among 18 Phyllanthus species found in China were investigated by DNA sequence analyses of the nuclear internal transcribed spacers (ITS1 and ITS2) along with the combined chloroplast ATPB and RBCL sequences. Cladistic analysis indicated that this genus is paraphyletic and strongly supports the notion that two problematic and confusing species, P. niruri and P. amarus, are two individual, albeit closely related, species. We have also developed an ITS rDNA-based multiplex PCR assay to differentiate three medicinally important species, P. amarus, P. niruri and P. urinaria.     

复方苦味叶下珠胶囊的制备及质量控制

目的:制备复方苦味叶下珠胶囊并建立其质量控制方法。方法:方中药物成分的提取,苦味叶下珠采用醇提法,黄芪和甘草采用水煎煮法;用薄层色谱法鉴别苦味叶下珠,薄层扫描法测定甘草中甘草酸单铵盐的含量。结果:甘草酸单胺盐线性范围为2. 08μg～6 24μg ,加样回收率为99. 13 % ,RSD=0 .71 %。结论:制备工艺简单,质控方法可行。     

Protein isolate from the herb, Phyllanthus niruri L. (Euphorbiaceae), plays hepatoprotective role against carbon tetrachloride induced liver damage via its antioxidant properties.

Phyllanthus niruri L. (Euphorbiaceae) (P. niruri) is a well-known hepatoprotective herbal plant. In the present study, hepatoprotective potential of the protein isolate of P. niruri was investigated against carbon tetrachloride (CCl(4)) induced liver damage in vivo. Protein isolate of P. niruri was intraperitoneally injected in mice either prior to (preventive) or after the induction of toxicity (curative). Levels of different liver marker enzymes in serum and different anti-oxidant enzymes, as well as lipid peroxidation products and glutathione (GSH) in liver homogenates were measured in normal, control (toxicity induced) and protein isolate treated mice. Administration of CCl(4) increased the serum glutamate pyruvate transaminase (GPT) and alkaline phosphatase (ALP) levels of mice sera along with increased lipid peroxidation and reduced levels of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) in the liver. Treatment with the protein isolate of P. niruri significantly altered these changes to almost normal. The protein isolate also showed protective properties as was evidenced in histopathological studies. Results suggest that the protein isolate of P. niruri protects liver tissues against oxidative damage and somehow helps stimulating repair mechanism present in liver. It could be used as an effective hepatoprotector against CCl(4) induced liver damage.     

Inhibitory effects of methyl brevifolincarboxylate isolated from Phyllanthus niruri L. on platelet aggregation

A platelet-aggregatory inhibitor was isolated from the 50% MeOH extract of Phyllanthus niruri L. leaf. Its structure was determined to be methyl brevifolincarboxylate on the basis of the 1H-, 13C-NMR, and high-resolution mass spectral data. We compared the antiplatelet aggregatory effects of the constituent with adenosine, a well-known inhibitor of platelet aggregation. Platelet aggregation was induced by collagen or adenosine 5'-diphosphate as an activating agent; the extent of inhibition was monitored with a platelet aggregometer employing a laser-scattering method. The inhibitory effects of methyl brevifolincarboxylate were found to be as potent as adenosine that is known to act on an A2A subtype receptor.     

Hepatocurative and antioxidant profile of HP-1, a polyherbal phytomedicine

HP-1 a herbal formulation comprising of Phyllanthus niruri and extracts of Terminalia belerica, Terminalia chebula, Phyllanthus emblica and Tinospora cordifolia has been evaluated for hepatoprotective activity against carbon tetrachloride (CCl4) induced toxicity. Results show that HP-1 reversed the leakage of lactate dehydrogenase (LDH) and glutamate pyruvate transaminase (GPT) and prevented the depletion of glutathione (GSH) levels in a primary monolayer culture of rat hepatocytes (in vitro). HP-1 attenuated the serum toxicity as manifested in elevated levels of transaminases (glutamate oxaloacetate transaminase (GOT), and GPT) The antioxidative enzymes in liver (catalase and superoxide dismutase (SOD)) were restored to normal values after the oral administration of HP-1. HP-1 suppressed the formation of the superoxide anion radical and reduced CCl4 mediated lipid peroxidation (LPO). Silymarin and antioxidants (ascorbic acid, beta-carotene and alpha-tocopherol) were used for comparison. The present study showed that HP-1 is a potential hepatoprotective formulation with an additional attribute of being anti-peroxidative.