RP-HPLC法同时测定穿山龙中薯蓣皂苷和纤细皂苷的含量

目的建立测定不同产地穿山龙供试品中薯蓣皂苷和纤细皂苷含量的方法。方法采用RP-HPLC法。色谱柱为Kromasil C8色谱柱(200 mm×4.6 mm,5μm),流动相为乙腈-水(体积比41∶59),检测波长为206 nm,柱温为室温。结果薯蓣皂苷质量浓度在0.10~1.00 g.L-1内、纤细皂苷质量浓度在0.02~0.20 g.L-1内与峰面积呈现良好的线性关系,相关系数均为0.9997(n=6)。其平均回收率分别为101.2%、100.7%,RSD分别为2.4%、2.0%(n=9)。供试品含量测定结果表明,饮片中薯蓣皂苷和纤细皂苷的含量显著高于新鲜药材烘干品。结论该方法简便、准确、重现性好,为穿山龙药材的质量控制提供了参考。     

HPLC法测定穿山龙中伪原薯蓣皂苷、薯蓣皂苷和延龄草苷的含量

目的:建立反相高效液相色谱法同时测定穿山龙中伪原薯蓣皂苷、薯蓣皂苷和延龄草苷的含量。方法:采用Zorbax SB-C18(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈(A)-水(B),梯度洗脱(0～18 min,25%A线性变为36%A;18～40min,36%A线性变为90%A;40～55 min,90%A保持不变),流速1.0 mL.min-1,检测波长210 nm,柱温30℃。结果:伪原薯蓣皂苷、薯蓣皂苷和延龄草苷的色谱峰面积与浓度呈良好的线性关系,线性范围分别为9.80～588.0μg.mL-1(r=0.9994),20.20～1212μg.mL-1(r=0.9995),5.330～319.8μg.mL-1(r=0.9999);平均回收率(n=9)分别为97.3%,99.4%,97.6%。结论:该方法简便、准确,为穿山龙药材的质量评价提供了方法。     

南重楼中纤细薯蓣皂苷的鉴别及含量测定

目的:建立南重楼药材中纤细薯蓣皂苷的定性鉴别及含量测定方法。方法:采用薄层色谱法定性鉴别南重楼药材中的纤细薯蓣皂苷,并采用高效液相色谱法测定纤细薯蓣皂苷的含量。色谱柱为Waters XTerra RP18,流动相为乙腈-水(梯度洗脱),流速为1.0 ml/min,检测波长为203 nm,柱温为30℃,进样量为10μl。结果:纤细薯蓣皂苷的TLC斑点清晰、分离度好。纤细薯蓣皂苷的进样量在0.22~11.10μg范围内与其峰面积积分值呈良好线性关系(r=0.999 9);精密度、稳定性、重复性试验的RSD<2%;平均加样回收率为100.81%,RSD为1.83%(n=3)。结论:本方法操作简便、结果准确、重复性好,可用于南重楼中纤细薯蓣皂苷的定性鉴别及含量测定。     

HPLC法测定不同产地穿山龙中薯蓣皂苷含量

目的采用高效液相色谱法测定不同产地穿山龙中薯蓣皂苷的含量,并对不同产地穿山龙进行质量评价。方法采用高效液相色谱法进行含量测定。色谱柱为Diamonsil C18(4.6 mm×250 mm,5μm);以乙腈-水(55∶45)为流动相;流速1.0 mL·min-1;检测波长为203 nm。结果通过高效液相色谱法测定不同产地穿山龙薯蓣皂苷的含量,吉林、内蒙古与山东泰山三地产穿山龙薯蓣皂苷含量均在3.0%以上,安徽、浙江两地含量1.36%¹.41%。结论北方产穿山龙薯蓣皂苷含量相对南方产要高,质量较好。     

HPLC法测定不同产地穿山龙中薯蓣皂苷的含量

目的对不同产地穿山龙药材中薯蓣皂苷的含量进行比较。方法采用HPLC法对不同产地穿山龙药材中的薯蓣皂苷进行了检测。结果不同地域穿山龙药材中薯蓣皂苷的含量有明显差异,伊春、兰州、陕南所产含量较高,其中伊春产穿山龙药材含量最高,为19.7mg·g-1。结论该方法简便、准确、重复性好,可以作为穿山龙药材的质量控制方法。     

穿龙薯蓣中甾体皂苷的分离与鉴定

目的 研究穿龙薯蓣Dioscorea ni pponica Makin根茎的化学成分.方法 采用硅胶柱色谱、ODS柱色谱以及制备HPLC等手段进行分离纯化,通过理化性质和波谱学技术进行结构鉴定.结果 从正丁醇层中分离得到5个甾体皂苷类化合物,分别鉴定为纤细皂苷(1)、薯蓣皂苷(2)、伪原薯蓣皂苷(3)、甲基原纤细皂苷(4)、原薯蓣皂苷(5).结论 其中化合物4为首次从穿龙薯蓣中分离得到.     

淡紫青霉ACCC31890对纤细薯蓣皂苷的生物转化

摘 要 目的：研究淡紫青霉菌对纤细薯蓣皂苷的生物转化,分离鉴定转化产物并进行药理活性探讨。方法: 使用硅胶柱色谱和半制备液相色谱分离转化产物,依据质谱(MS）、核磁共振(NMR)波谱分析进行结构鉴定,并对产物进行了体外抗炎活性的研究。结果: 从转化粗提物中分离得到三个依次脱糖基产物,分别为5R-spirost-5-ene-3-ol-O-β-D-Glucopyranoside (1→3)-β-D-Glucopyranosyl（1）,延龄草苷（2）和薯蓣皂苷元（3）,转化率分别为1%,1%,45%;体外抗炎活性研究表明三种转化产物对LPS刺激巨噬细胞分泌NO,IL-6和MCP-1具有不同程度的抑制作用,其抗炎活性随着糖链的逐步水解有所增加,以终产物薯蓣皂苷元的抗炎活性最强。结论:淡紫青霉菌转化纤细薯蓣皂苷得到薯蓣皂苷元的产率较高,糖基的数量对甾体皂苷类化合物抗炎活性有着一定的影响。     

HPLC法测定中药白英中薯蓣皂苷元的含量

目的用HPLC法测定不同来源白英中薯蓣皂苷元的含量。方法选用Apollo C18色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈水(体积比为90∶10),检测波长为210 nm,柱温为室温。结果薯蓣皂苷元的质量浓度在0.04~0.20 g.L-1内与峰面积呈良好线性关系,方法的平均回收率为104.5%(n=9);不同来源的白英药材中薯蓣皂苷元的含量有较大差异。结论方法操作简便、准确、灵敏度高,为白英药材质量控制提供了定性和定量依据。     

HPLC法测定薯蓣属3种植物中薯蓣皂苷的含量

目的:建立薯蓣属3种植物福州薯蓣、绵萆薢、穿龙薯蓣根茎中薯蓣皂苷的含量测定方法。方法:药材粉末经95%乙醇回流提取、水饱和正丁醇萃取后,采用HPLC法进行测定。色谱柱:Kromasil C_8(250mm×4.6mm,5μm),流动相:乙腈-水(41∶59),流速:1.0mL·min~(-1),柱温:室温,检测波长:203nm。结果:薯蓣皂苷与其他成分分离良好。线性范围为10.0-200.0μg·mL~(-1)(r=0.9997)。高、中、低浓度的平均回收率分别为97.9%,99.1%,102.5%。结论:方法简便、准确、重现性好,可用于薯蓣属药材的质量控制。     

内生真菌固体发酵提高穿山龙中薯蓣皂苷元的研究

目的 分离提取穿龙薯蓣内生真菌,筛选能够发酵提高穿山龙药材中薯蓣皂苷元的菌种.方法 采用PDA培养基分离纯化穿龙薯蓣内生真菌,将内生真菌与121℃湿热灭菌后的穿山龙药材粗粉在28℃条件下固体发酵培养15d,以HPLC法测定发酵物中薯蓣皂苷元,并与穿山龙原药材、灭菌空白药材以及未经灭菌药材的发酵物进行比较.结果 分离筛选的穿龙薯蓣内生真菌C39能够稳定提高灭菌后穿山龙药材中薯蓣皂苷元的水平,提高幅度达原药材的70％～75％.结论 采用固体发酵的方法可以使穿龙薯蓣内生真菌C39稳定提高穿山龙药材中薯蓣皂苷元.     

甲基原薯蓣皂苷对人肝微粒体中7种CYP450酶活性的影响

目的研究甲基原薯蓣皂苷对CYP450酶的7种亚型酶活性的影响。方法将MPD和CYP450酶7种亚型的特异性探针底物咖啡因(CYP1A2)、右美沙芬(CYP2D6)、甲苯磺丁脲(CYP2C9)、s-美芬妥因(CYP2C19)、氯唑沙宗(CYP2E1)、香豆素(CYP2A6)及咪达唑仑(CYP3A4)与人肝微粒体进行孵化反应,采用HPLC和LC-MS/MS法测定对应的7种代谢产物(1,7-二甲基黄嘌呤、去甲右美沙芬、4-羟基甲苯磺丁脲、4-羟基美芬妥因、6-羟基氯唑沙宗、7-羟基香豆素和1-羟基咪达唑仑)的浓度,通过与对照组比较,确定MPD对以上7种酶活性的影响。结果MPD在1~10μmol.L-1时对7种酶均无明显抑制作用,在100μmol.L-1时对CYP2D6有抑制趋势,但对其他6种酶无抑制作用,均无统计学意义(P>0.05)。结论MPD在与以上6种酶(CYP1A2、CYP2E1、CYP2C19、CYP3A4、CYP2C9和CYP2A6)代谢的药物联合用药时,发生药物相互作用的可能性较小。     

基于体外药效学结合分子对接技术研究穿山龙抗心肌缺血的作用机制

目的 通过液质联用技术、体外药效学及分子对接技术探究穿山龙抗心肌缺血的物质基础和作用机制。方法 利用UPLC-Q TOF-MS技术确定穿山龙醇提物中的主要成分,比较不同质量浓度（40、10、4、0.4、0.004 mg/mL）的穿山龙醇提物对H9c2心肌细胞缺氧/复氧（H/R）损伤后细胞活力的影响。应用AutoDock软件对潜在药效物质和关键靶点进行分子对接,相关结果采用Cytoscape 3.6.1进行构图和网络拓扑结构分析。结果 从穿山龙的醇提物中鉴定出7个化学成分,并证明穿山龙醇提物对H/R损伤H9c2心肌细胞具有一定的保护作用。筛选出5种（薯蓣皂苷元、薯蓣皂苷、对羟基苄基酒石酸、原薯蓣皂苷、薯蓣次皂苷A）与抗心肌缺血相关的活性成分,5种活性成分的分子对接结果显示潜在的药效物质与关键靶点内皮一氧化氮合酶（NOS）2、NOS3、环氧化酶/细胞色素C氧化酶多肽II（COX2）、热休克蛋白（HSP70）对接结果能量低于-8 kcal/mol。结论 穿山龙中的甾体皂苷类成分可能是其治疗心肌缺血的物质基础,穿山龙治疗心肌缺血是多成分与多靶点相互作用的结果。     

Potential nephrotoxic effects produced by steroidal saponins from hydro alcoholic extract of Tribulus terrestris in STZ-induced diabetic rats

Abstract

Chronic hyperglycemia leads to the development of microvascular complications like diabetic nephropathy. The present study investigated the potential effects of the hydroalcoholic extract of Tribulus terrestris, a plant of Zygophyllaceae family, on the renal complications in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced by administering STZ (90?mg/kg) to the 2-days old neonates. After 6 weeks of induction, diabetic rats were treated with 50?mg/kg hydroalcoholic extract of T. terrestris for 8 weeks. The anti-hyperglycaemic nature was confirmed by reduction in blood glucose and improvement in insulin levels. Diabetic renal injury associated with decrease in total proteins and albumin levels was observed to be improved by T. terrestris extract. Glomerular filtration rate along with inflammatory and growth factors, adiponectin and erythropoietin were also improved by the treatment, though the findings were not significant. However, the beneficial antidiabetic effects of T. terrestris extract in plasma were not observed in kidney histopathology. This was confirmed by the quantitative estimation of unhydrolyzed fraction of saponins (major component: protodioscin) in plasma and kidney samples of normal and diabetic rats. Hence, it can be concluded that 8 weeks treatment with T. terrestris extract produces potential toxic effects in kidney, which are independent of its anti-diabetic action.     

Metabolism and hepatotoxicity of N,N-dimethylformamide,N-hydroxymethyl-N-methylformamide,and N-methylformamide in the rat

The metabolism and hepatotoxicity ofN,N-dimethylformamide (DMF) and two of its metabolites,N-hydroxymethyl-N-methylformamide (HMMF) andN-methylformamide (NMF) were evaluated over a 4-day period in rats. DMF toxicity was dose dependent and delayed toxicity after the administration of a high DMF dose (13.7 mmol/kg) in comparison to a lower dose (4.1 mmol/kg) was observed. Treatment of rats with 13.7 mmol/kg DMF, HMMF, or NMF showed i) that DMF is more toxic than HMMF or NMR, and ii) that hepatotoxicity occurs later for DMF than for HMMF or NMF. Analysis of serum and urine samples demonstrated that DMF is first metabolized to HMMF, which is then partially converted to NMF. After HMMF administration, NMF was found both in serum and in urine. The time course of DMF and HMMF toxicity in relation to NMF formation fitted the hypothesis that the hepatotoxicity of DMF and HMMF is mediated via NMF. The degree of hepatotoxicity after HMMF and NMF treatment is similar. However, the degree of DMF hepatotoxicity is much higher than in the case of NMF or HMMF. The role of NMF as an obligatory intermediate in DMF and HMMF hepatotoxicity is discussed.     

Effect of bay K 8644, a calcium channel agonist,on dog cardiac muscarinic receptors

To investigate further whether the effects of the dihydropyridine (DHP) drugs on calcium channels are related to those of these drugs on muscarinic receptors, the binding characteristics of the DHP calcium channel agonist, Bay K 8644, on muscarinic receptors and calcium channels were compared to those of the DHP calcium channel antagonists, nicardipine and nimodipine in the dog cardiac sarcolemma. Bay K 8644, nicardipine and nimodipine inhibited the specific [³H]QNB binding with K _i values of 16.7μM, 3.5μM and 15.5μM respectively. Saturation data of [³H]QNB binding in the presence of these DHP drugs showed this inhibition to be competitive. Bay K 8644, like nicardipine and nimodipine, blocked the binding of [³H]nitrendipine to the high affinity DHP binding sites, but atropine did not, indicating that the muscarinic receptors and the DHP binding sites on calcium channels are distinct. The K _i value of Bay K 8644 for the DHP binding sites was 4 nM. Nicardipine and nimodipine (K _i :0.1–0.2 nM) were at least 20 times more potent than Bay K 8644 in inhibiting [³H]nitrendipine binding. Thus, the muscarinic receptors were about 4000 times less sensitive than these high affinity DHP binding sites to Bay K 8644. These results suggest that the DHP calcium agonist Bay K 8644 binds directly to the muscarinic receptors but its interaction with the muscarinic receptors is not related to its binding to the DHP binding sites on calcium channels.     

Anti-acne activities of pulsaquinone, hydropulsaquinone, and structurally related 1, 4-quinone derivatives

Quinone type compound, pulsaquinone 1, isolated from the aqueous ethanol extract of the roots of Pulsatilla koreana exhibited antimicrobial activities against an anaerobic non-spore-forming gram-positive bacillus, Propionibacterium acnes, which is related with the pathogenesis of the inflamed lesions in a common skin disease, acne vulgaris. Compound 1 was unstable on standing and thus converted to more stable compound 2, namely hydropulsaquinone by hydrogenation, whose activity was comparable to mother compound 1 (MIC for 1 and 2 against P. acnes: 2.0 and 4.0 μg/mL, respectively). Other structurally-related quinone derivatives (3–13) were also tested for structure-activity relationship against anaerobic and aerobic bacteria, and fungi. The antimicrobial activity was fairly good when the quinone moiety was fused with a nonpolar 6- or 7-membered ring on the right side whether or not conjugated (1,4-naphtoquinone derivatives 3–5), while simple quinone compounds 6–9 showed poor activity. It seems that the methoxy groups at the left side of the quinone function deliver no considerable antimicrobial effect.     

Homocysteinemia,hypertension, and family history of diabetes in a smoking male population in Saudi Arabia

Arabs have a lower incidence of atherosclerosis than other ethnicities, but few studies have examined homocysteine (HCYS) as a risk factor for cardiovascular disease in this population. Here, we investigated the association between serum HYCS levels and risk factors for cardiovascular disease (smoking, hypertension, and family history of diabetes) in Saudi males. A total of 50 smokers and 72 nonsmokers completed a general health questionnaire. In addition, their lipid profiles were measured using routine methods and HCYS levels by high-performance liquid chromatograph with electrochemical detection. Regression analysis showed negative associations between HCYS and glucose (r = −0.22; P < 0.05) as well as family history of diabetes (r = −0.21; P < 0.05). HCYS levels were similar between hypertensive and nonhypertensive smokers, but they were significantly elevated in hypertensive nonsmokers (P = 0.027) and lower in smokers with family history of diabetes (P = 0.01). Levels of HCYS among nonsmokers inversely correlated with history of diabetes and elevated glucose. Nonsmokers’ HCYS levels were significantly elevated in the presence of hypertension and correlated with diastolic blood pressure. Thus, HCYS may be a predictor of hypertension among nonsmokers. Until further trials are conducted, we recommend vitamin B6/folic acid supplementation for the Saudi hypertensive population as an adjuvant therapy.     

高效液相一测多评法测定鼻炎片中羧基苍术苷、苍术苷、连翘酯苷B、连翘酯苷A、连翘苷、升麻素苷、升麻素、5-O-甲基维斯阿米醇苷和亥茅酚苷

目的 建立高效液相一测多评（HPLC-QAMS）法测定鼻炎片中羧基苍术苷、苍术苷、连翘酯苷B、连翘酯苷A、连翘苷、升麻素苷、升麻素、5-O-甲基维斯阿米醇苷和亥茅酚苷。方法 以乙腈-0.2%磷酸为流动相,梯度洗脱,体积流量1.0 mL/min;Agilent HC-C₁₈（250 mm×4.6 mm,5 μm）色谱柱,柱温30℃;检测波长203 nm（测定羧基苍术苷和苍术苷）、275 nm（测定连翘酯苷B、连翘酯苷A和连翘苷）和254 nm（测定升麻素苷、升麻素、5-O-甲基维斯阿米醇苷、亥茅酚苷）。进行专属性试验、线性关系考察、精密度试验、稳定性试验、重复性试验、加样回收率试验;以连翘苷为内参物,建立该成分与羧基苍术苷、苍术苷、连翘酯苷B、连翘酯苷A、升麻素苷、升麻素、5-O-甲基维斯阿米醇苷、亥茅酚苷间的相对校正因子,并计算鼻炎片中9种成分的含量,同时与外标法（ESM）的测定结果进行比较,验证该方法的可行性。结果 建立了鼻炎片中9种成分的定量控制方法,方法学验证结果符合要求。鼻炎片中各成分的计算值与实测值无显著性差异。结论 HPLC-QAMS法适用于鼻炎片的多指标成分质量评价模式,可以为鼻炎片质量控制方法的建立提供参考。     

Chronic Effects of an Insecticide, Fenobucarb, on the Larvae of Two Mayflies, Epeorus latifolium and Baetis thermicus, in Model Streams

Effects of the insecticide fenobucarb on 2 mayfly species, Epeorus latifolium and Baetis thermicus, were examined in indoor model streams. Aqueous concentrations of fenobucarb residues in the model streams were 69–71% of its nominal level until 24 h, except at low concentrations (less than 2 g l^–1 in the water) in the water. Chronic effects of fenobucarb on the growth and emergence of Ephemeropteran larvae were examined over 2 months. For concentrations of 1 and 2 g l^–1, the numbers of individuals of the third larval growth stage (average head width 3.0 mm) gradually decreased up to 20 days after the applications; the number of emergent individuals also remained 20 to 25 by the end of the experiment. The number of individuals to emerge at 1 to 2 g l^–1 fenobucarb was restricted in the long-term experiment. These results suggest that fenobucarb affects the emergence of this species through disruption of the endocrine system. On the other hand, the number of emergent individuals of B. thermicus at low concentrations of fenobucarb was not significantly less than that in the control.     

自旋探针α-苯基-N-4-丁基硝酮纳米粒的制备及其对肝肿瘤细胞的亲和性研究

采用薄膜分散-超声法制备、优化自旋捕捉剂α-苯基-N-4-丁基硝酮(N-tert-butyl-α-phenylnitrone，PBN)脂质体，并研究其相关性质。以人肝肿瘤细胞SMMC-7721为实验对象，采用反相高效液相法(RP-HPLC)测定细胞内PBN的浓度，以此评价PBN脂质体对细胞的亲和性；考察了PBN脂质体对细胞生长的影响。结果显示，采用正交设计优化得到PBN脂质体的平均粒径137.5 nm，包封率71.52%，多分散系数0.286。PBN脂质体可稳定进入癌细胞内，同游离的PBN相比，PBN脂质体对肝肿瘤细胞更具亲和性，转染后癌细胞坏死率更高。表明自旋捕捉剂脂质体对肝肿瘤细胞具有较好的亲和性并能明显抑制其生长。为研究肿瘤自旋靶向性干预提供了实验依据。