GC法测定大蒜油中二烯丙基三硫和二烯丙基二硫的含量

目的:建立测定大蒜油中二烯丙基三硫和二烯丙基二硫含量的GC分析方法,同时将GC与HPLC分析相比较,考察两种方法测定结果是否有差异.方法:色谱条件:DM-WAX石英弹性毛细管色谱柱(30 m×0.25 mm,df0.25 μm,Dikma Technologies);载气:氮气;流速:50 ml/min;检测器FID:H2 50 mL/min,Air 500 mL/min;内标物为十七烷.结果:二烯丙基三硫和二烯丙基二硫分别在0.08～0.40 mg/mL和0.15～0.73 mg/mL范围内与峰面积呈良好线性关系;平均回收率分别为98.84%和99.28%,RSD分别为2.61%和1.95%.结论:本法操作准确,可用于大蒜油中二烯丙基三硫和二烯丙基二硫的含量测定;样品经GC与LPLC含量测定,结果无显著性差异.     

大蒜油中二烯丙基二硫与二烯丙基三硫的热稳定性实验研究

目的 :探讨大蒜油中二烯丙基三硫 (简称为DATS)、二烯丙基二硫 (简称为DADS)的热稳定性。方法 :采用气相色谱法、高效液相色谱法测定大蒜油在 100℃加热不同时间后DATS ,DADS的相对含量变化以及单一的DATS ,DADS在 100℃加热 0.5h后 ,DATS相对含量变化 ,比较其热稳定性。结果 :大蒜油 100℃加热 0.5h后 ,DATS相对含量下降 19.45% ,DADS相对含量升高 12.71% ;单一的DATS ,DADS(含量在 90%以上 )在 100℃加热 0.5h后 ,DATS相对含量下降 6.28% ,DADS相对含量下降 2.56%。结论 :大蒜油中DADS热稳定性较DATS为好。     

RP-HPLC法测定脂降宁片中葛根素、二苯乙烯苷的含量

目的:建立高效液相色谱法测定脂降宁片中葛根素、二苯乙烯苷的含量。方法:采用kromasil C18柱(4.6 mm×250 mm,5μm);流动相为甲醇(A)-水(B),梯度洗脱;流速1.0 mL/min;检测波长为250nm及320nm;柱温:30℃。结果:葛根素进样量在232～29μg范围内(r=0.9998),二苯乙烯苷进样量在220～19.5μg范围内(r=0.9997)线性关系良好,平均回收率葛根素为97.2%(RSD=0.8%),二苯乙烯苷为97.6%(RSD=0.7%)。结论:该方法操作简便,分离度高,重复性好,可同时测定脂降宁片中葛根素、二苯乙烯苷的含量。     

不同产地虎杖中各成分含量的比较

目的:比较不同产地虎杖中各成分的含量。方法:采用梯度洗脱的高效液相色谱法测定虎杖苷、白藜芦醇、白藜芦醇苷、大黄素及大黄素甲醚的含量,色谱柱为C18柱,检测波长为306nm,流速为1.0m L/min,柱温为30℃;采用显色法测定虎杖中总黄酮含量。结果虎杖苷进样量在0.1452~1.4520μg(r=0.9999)范围内、白藜芦醇在0.01525~0.1525μg(r=0.9998)、白藜芦醇苷在0.01253~0.6265μg(r=0.9998)、大黄素在0.02358~0.2358(r=0.9998)范围内、大黄素甲醚在0.004892~0.2446μg(r=0.9999)范围内线性关系良好,各成分平均回收率分别为98.2%(RSD=1.0%,n=3)、98.5%(RSD=2.0%,n=3)、99.0%(RSD=1.5%,n=3)、100.2%(RSD=1.9%,n=3)、99.7%(RSD=1.0%,n=3);总黄酮的测定以芦丁为对照,测定波长500nm,在7.24~43.44g/L范围内线性关系良好(r=0.9999),平均回收率为98.5%(RSD=1.5%,n=3)。结论:不同产地虎杖中各成分的含量是有差别的。     

不同产地硫熏和烘干金银花中绿原酸含量的高效液相色谱测定

目的 比较不同产地硫熏和烘干金银花中绿原酸含量变化,考察硫熏对金银花内在化学成分的影响.方法 采用高效液相色谱(HPLC)法测定绿原酸的含量,C18柱,流动相:乙腈-0.4％磷酸溶液=13:87,流速1.0 ml·min-1;柱温30℃;检测波长327 nm.结果 绿原酸在0.0836～0.418μg范围内呈良好线性关系,r=0.999 9,平均回收率(RSD=0.49％).结论 硫熏后金银花绿原酸含量明显增加,不同产地之间的含量差异也较大.     

HPLC法测定不同产地北沙参药材中香豆素的含量

目的:建立HPLC法同时测定不同产地北沙参中3种香豆素的含量.方法:色谱柱为Phenomenex-C18(250mm×4.6 mm,5μm),流动相为甲醇-水(65:35),流速为1.0 ml/min,检测波长248 nm.结果:补骨脂素在0.043μg～1.72μg(r=0.9999),欧前胡素在0.033μg～1.32 μg(r=0.9999),异欧前胡素在0.030μg～1.20 μg(r=0.9998)内线性关系良好;补骨脂素、欧前胡素和异欧前胡素平均回收率分别为97.59%、98.23%、96.94%,RSD分别为1.16%、1.36%、0.98%.结论:不同产地北沙参中3种香豆素含量存在明显差异,可为评价不同产地北沙参质量提供依据.     

RP-HPLC法测定大蒜油中二烯丙基三硫和二烯丙基二硫

大蒜油是百合科葱属植物大蒜Allium sativumL.的地下鳞茎外皮干燥后经水蒸气蒸馏提取而得到的挥发油。大蒜油具有抗菌、调血脂、降血糖、降压利尿、抗血小板凝集、增加纤维蛋白溶解性、保肝、抗衰老、抗肿瘤等多种功能[1,2],特别是对心血管、肿瘤有很好的生理活性,使其在新药开发、保健品生产上存在着很大潜力。我国每年约有20t大蒜油出口,也给大蒜油的生产厂家带来巨大的商机。但我国目前没有完善的大蒜油生产质量标准,仅依据挥发油的物理常数,使生产和出口受到一定的限制。为此本实验选择大蒜油中主要活性成分二烯丙基三硫和二烯丙基二硫…     

HPLC法测定八角莲中3种活性成分含量的研究

目的建立用高效液相色谱法测定八角莲中槲皮素、鬼臼毒素、山柰素含量的方法,比较不同产地八角莲药材3种化合物的含量。方法色谱柱:Waters Sunfire C_(18)柱(5μm,4.6×150 mm);流动相:甲醇-0.4%磷酸溶液(45:55);流速:1.0 mL/min;检测波长:290 nm;进样量:10μL;柱温30℃。结果槲皮素、鬼臼毒素、山柰素的线性范围分别为0.00973μg-9.73μg(r=0.9999)、0.01001μg~(-1)0.01μg(r=0.9999)、0.00998μg-9.98μg(r=0.9999),平均回收率分别为98.35%(RSD=2.6%)、100.25%(RSD=2.3%)、100.80%(RSD=2.7%)。结论该方法简便可行,重复性好,可用于同时测定八角莲药材中槲皮素、鬼臼毒素、山柰素3种成分的含量,为八角莲药材的质量控制提供依据。     

RP-HPLC同时测定西洋参总皂苷转化产物中人参皂苷Rg1、Rh1、Rd、Rg3和C-K的含量

目的:建立RP-HPLC法同时测定西洋参总皂苷转化产物中人参皂苷Rg1、Rd、Rh1、Rg3和C-K的含量。方法:采用WelchromC18色谱柱(4.6mm×250mm,5μm);流动相为乙腈(A)-水(B),梯度洗脱;流速为1.0mL/min;柱温为30℃;检测波长为203nm。结果:人参皂苷Rg1,Rh1,Rd,Rg3和C-K的线性范围分别为0.740~7.400μg(r=0.9999);0.536~5.360μg(r=0.9999);1.128~11.280μg(r=0.9999);0.3080~3.080μg(r=0.9999);0.376~3.7600μg(r=0.9999)。平均加样回收率分别为97.12%(RSD=1.22%),98.50%(RSD=0.87%),98.00%(RSD=1.06%),98.12%(RSD=1.66%)和96.65%(RSD=1.48%)。结论:该法操作简便,结果可靠,适用于西洋参转化产物的含量测定。     

HPLC法同时测定美洲大蠊标准汤剂中6种成分含量

目的:建立HPLC法同时测定美洲大蠊标准汤剂中尿嘧啶、次黄嘌呤、尿苷、肌苷、鸟苷、原儿茶酸的含有量。方法:采用WATERS Symmetry Shield C₁₈色谱柱(250 mm×4.6 mm, 5μm);甲醇-0.1%磷酸水作为流动相,梯度洗脱;体积流量0.6 mL/min;检测波长254 nm;柱温20℃。结果:测定显示尿嘧啶、次黄嘌呤、尿苷、肌苷、鸟苷、原儿茶酸分别在0.041～0.341μg(r=0.9994)、0.097～0.810μg(r=0.9994)、0.020～0.168μg(r=0.9999)、0.277～2.308μg(r=0.9999)、0.016～0.135μg(r=0.9997)、0.131～1.096μg(r=0.9998)范围内线性关系良好,平均加样回收率分别为99.93%(RSD=1.18%)、98.03%(RSD=1.05%)、97.17%(RSD=1.29%)、100.19%(RSD=1.91%)、98.94%(RSD=1.11%)、100.70%(RSD=1.05%)。十批美洲大蠊标准汤剂各成分含量范围分别为：尿嘧啶0....     

Diallyl disulfide induces MUC5B expression via ERK2 in human airway epithelial cells

Garlic has been shown to have antimicrobial, hypolipidemic, antithrombotic, antitumor and immunostimulatory properties. The medicinal effects of garlic are derived from the flavonoid and organosulfur components. Diallyl disulfide (DADS), an organosulfur, is the main component responsible for the diverse biological effects of garlic. However, the effects of DADS on mucin gene expression in airway epithelial cells have not been reported to date. Therefore, this study was performed to investigate the effects and brief signaling pathway of DADS associated with MUC5B expression in NCI-H292 epithelial cells using RT-PCR, ELISA, western blot, immunocytochemistry and cell transfection with siRNA. DADS induced MUC5B expression and activated the phosphorylation of ERK1/2 MAPK. In addition, U0126 inhibited DADS-induced MUC5B expression and DADS-activated phosphorylation of ERK1/2 MAPK. Moreover, the immunopositive cells for MUC5B protein did not appear after treatment of DADS with U0126, and the knockdown of ERK2 MAPK by ERK2 MAPK siRNA significantly blocked DADS-induced MUC5B mRNA expression. However, DADS did not activate the phosphorylation of p38 MAPK, and SB203580 did not inhibit DADS-induced MUC5B expression. This is the first study to show that DADS-induced MUC5B expression appears to be regulated by activation of the ERK2 MAPK signaling pathway in human NCI-H292 airway epithelial cells.     

Effects of different garlic-derived allyl sulfides on peroxidative processes and anaerobic sulfur metabolism in mouse liver

Biological activity of garlic has been attributed to organosulfur compounds, most of all to oil-soluble allyl sulfides, such as diallyl sulfide (DAS), diallyl disulfide (DADS) and diallyl trisulfide (DATS). This study evaluated the effectiveness of garlic-derived allyl sulfides in influencing peroxidative processes, levels of thiols and sulfane sulfur and its metabolic enzymes in normal mouse liver cells. Various allyl sulfides (DAS, DADS and DATS) dissolved in corn oil were given intraperitoneally to mice for 10 days. After sacrificing the mice, biochemical assays were performed in liver homogenates and in plasma in order to establish liver function. All allyl sulfides under study had a beneficial effect in the mouse liver since they decreased reactive oxygen species and malondialdehyde levels and increased glutathione S-transferase activity and non-protein sulfhydryl group level. Moreover, DADS and DATS elevated total sulfane sulfur pool and activity of sulfane sulfur biosynthetic enzymes. The increase in sulfane sulfur level entailed augmentation of its antioxidant and regulatory capacities. Garlic-derived allyl sulfides exhibited antioxidant action in the liver and elevated anaerobic cysteine metabolism leading to the formation of sulfane sulfur-containing compounds. Thus, DADS and DATS showed beneficial action in the liver, which can be used for protection of normal liver cells during chemotherapy or for alleviation of liver damage.     

Inhibition by Diallyl Trisulfide,a Garlic Component, of Intracellular Ca~(2 ) Moilization without Affecting Inositol-1,4,5-trisphosphate(IP_3) Formation in Activated Platelets

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In vitro anti-Helicobacter pylori activity of diallyl sulphides and protocatechuic acid

The in vitro inhibitory effects of diallyl disulphide (DADS), diallyl trisulphide (DAT), roselle calyx extract and protocatechuic acid (PA) on the growth of Helicobacter pylori (15 susceptible, 11 clarithromycin-resistant and 9 metronidazole-resistant strains) were studied. The inhibition zone was determined after each agent had been heated at 25, 60, 100 degrees C for 60 min. The minimal inhibitory concentration (MIC) of each agent was determined by the tube dilution assay. The results showed that heat treatment did not affect the anti-H. pylori activity of DADS, DAT, roselle calyx extract and PA, and the MIC values of these agents against test H. pylori strains were in the range 8-64 mg/L. The time-kill study assay for DAT and PA at 1x MIC was monitored in Muller Hinton broth supplemented with 10% horse blood or mice stomach homogenate. Both DAT and PA inhibited the growth of all test H. pylori in broth and mice stomach homogenate (p < 0.05); however, the inhibitory effects of these two agents were less in mice stomach homogenate than in broth (p < 0.05). DAT at 4, 6, 8, 10, 12, 14 mg/L and PA at 8, 16, 24, 32, 40, 48 mg/L were used for urease activity assay. These two agents significantly reduced urease activity of test H. pylori strains (p < 0.05), in which DAT and PA at 1x MIC reduced the urease activity of H. pylori to 70% and 40%, respectively. These agents, based on their lower MIC values and heat tolerance, might be useful in the prevention or therapy of H. pylori.     

The Role of Diallyl Sulfides and Dipropyl Sulfides in the In Vitro Antimicrobial Activity of the Essential Oil of Garlic,Allium sativum L., and Leek,Allium porrum L.

The in vitro antibacterial activity of essential oils (EOs) obtained from fresh bulbs of garlic, Allium sativum L., and leek, Allium porrum L. ( Alliaceae), was studied. A. sativum (garlic) EO showed a good antimicrobial activity against Staphylococcus aureus (inhibition zone 14.8 mm), Pseudomonas aeruginosa (inhibition zone 21.1 mm), and Escherichia coli (inhibition zone 11.0 mm), whereas the EO of A. porrum (leek) had no antimicrobial activity. The main constituents of the garlic EO were diallyl monosulfide, diallyl disulfide (DADS), diallyl trisulfide, and diallyl tetrasulfide. The EO of A. porrum was characterized by the presence of dipropyl disulfide (DPDS), dipropyl trisulfide, and dipropyl tetrasulfide. The antimicrobial activities of the DADS and DPDS were also studied. The results obtained suggest that the presence of the allyl group is fundamental for the antimicrobial activity of these sulfide derivatives when they are present in Allium or in other species (DADS inhibition zone on S. aureus 15.9 mm, P. aeruginosa 21.9 mm, E. coli 11.4 mm). Copyright © 2012 John Wiley & Sons, Ltd.     

Aged garlic extract, garlic powder extract, S-allylcysteine, diallyl sulfide and diallyl disulfide do not interfere with the antibiotic activity of gentamicin

It was shown that aged garlic extract (AGE), garlic powder and the following garlic-derived compounds: S-allylcysteine (SAC), diallyl sulfide (DAS) and diallyl disulfide (DADS), ameliorate gentamicin (GM)-induced nephrotoxicity in rats. However, it was not established if the above mentioned extracts and compounds of garlic could interfere with the antibiotic action of GM. To address this point, AGE, garlic powder extract (GPE), SAC, DAS and DADS were assessed for their ability to interfere with the in vitro antibiotic activity of GM in Escherichia coli cultures. It was found that the above mentioned extracts and compounds of garlic were unable to decrease the antibiotic capacity of GM and even SAC, DAS and DADS alone inhibited the growth of Escherichia coli and enhanced the antibiotic effect of GM. Our data show that SAC, DAS and DADS are antibacterial compounds against E. coli and suggest that AGE, GPE, SAC, DAS and[sol ]or DADS may be administered along with GM-treatment to ameliorate GM-induced nephrotoxicity without interfering with its antibiotic activity.     

大蒜油固体脂质纳米粒大鼠体内药动学的研究

 目的研究大蒜油的血药浓度测定方法及其在大鼠体内的药动学。方法12只健康大鼠,单剂量(含大蒜油每只10mg)颈静脉注射给大蒜油固体脂质纳米粒和大蒜油注射液,在设计的时间点从颈静脉取血,采用气相电子捕获法同时测定大蒜油中指标性成分二烯丙基三硫化合物和二烯丙基二硫化合物在全血中药物浓度,药动学参数采用DAS2.0程序进行统计矩处理。结果大鼠颈静脉注射给药后,大蒜油的指标性成分DATS首先快速转化成DADS,然后继续降解;大蒜油固体脂质纳米粒中的DATS和DADS的MRT_0-t分别为7.3和16.9min,ρ_max分别为3574.3和21416.7μg·L^-1;大蒜油注射液的DATS和DADS的MRT_0-t分别为19.4和19.5,ρ_max分别为3692.8和9335.8μg·L^-1。t_max均为1min。结论本方法简单、快速,灵敏度和准确度较高,适合药动学研究。静注给药后大蒜油在大鼠体内表现为快速分布和消除,而大蒜油固体脂质纳米粒因被动靶向的关系消除比大蒜油注射液快。     

大蒜油固体脂质纳米粒中二硫化物和三硫化物在大鼠体内的组织分布

目的：考察大蒜油固体脂质纳米粒(GO-SLN)中二硫化物(DADS)和三硫化物(DATS)在大鼠体内的组织分布。方法：建立了测定大鼠体内大蒜油中二硫化物(DADS)和三硫化物(DATS)的气相-电子捕获法，色谱条件：恒温110 ℃，检测器300 ℃，汽化室180 ℃，载气N2(纯度>99.999％)，流速 1.0 mL·min-1，分流比1∶10；尾吹60 mL·min-1；进样量1 μL。并测定大鼠颈静脉注射GO-SLN和GO注射液后组织中的药物浓度。结果：此色谱条件下各组织的标准曲线、精密度等实验结果表明，该方法适于分析大鼠体内大蒜油中DADS和DATS含量。与GO注射液相比，GO-SLN在大鼠体内的分布特性有不同程度的改变，GO-SLN在各组织中分布均相对较高。结论：SLN能一定程度上提高药物的被动靶向性并延长药物在各组织中的作用时间。     

DADS对人结肠癌SW480细胞凋亡的影响

目的：探讨二烯丙基二硫（diallyl disulfide,DADS）诱导人结肠癌SW480细胞凋亡的作用。方法：采用MTT、细胞计数法,观察不同浓度和作用时间DADS对SW480细胞的生长抑制作用;施用流式细胞术检测不同浓度DADS对SW480细胞的凋亡率。结果：MTT法显示,DADS能明显抑制SW480细胞生长增殖,呈浓度和时间依赖性,细胞生长增殖速度明显减慢。细胞计数表明,当DADS浓度增加时,SW480细胞群体倍增时间延长,与对照组比较,差异有显著性意义（P〈0．05）。流式细胞仪分析显示,随着DADS处理浓度增加,使SW480细胞凋亡率逐渐增高,差异有显著性意义（P〈0．05）。结论：DADS具有诱导人结肠癌SW480细胞凋亡的作用。