Modulatory Effect of Distillate of Ocimum sanctum Leaf Extract（Tulsi）on Human Lymphocytes Against Genotoxicants

Objective To study the modulatory effect of distillate of Ocimum sanctum (traditionally known as Tulsi) leaf extract (DTLE) on genotoxicants. Methods In the present investigation, we studied the antigenotoxic and anticlastogenic effect of distillate of Tulsi leaf extract on (i) human polymorphonuclear leukocytes by evaluating the DNA strand break without metabolic activation against mitomycin C (MMC) and hexavalent chromium (Cr 6) and (ii) human peripheral lymphocytes (in vitro) with or without metabolic activation against mitomycin C (MMC), hexavalent chromium (Cr 6) and B[a]P by evaluating chromosomal aberration (CA) and micronucleus assay (MN). Three different doses of DTLE, 50 μL/mL, 100 μL/mL, and 200 μL/mL were selected on the basis of cytotoxicity assay and used for studying DNA strand break, chromosomal aberration and micronucleus emergence. The following positive controls were used for inducing genotoxicity and clastogenicity: MMC (0.29 μmol/L) for DNA strand break, chromosomal aberration and 0.51 μmol/L for micronucleus assay; Potassium dichromate (Cr 6) 600 μmol/L for DNA strand break and 5 μmol/L for chromosomal aberration and micronucleus assay; Benzo[a]pyrene (30 μmol/L) for chromosomal aberration and 40 μmol/L for micronucleus assay. The active ingredients present in the distillate of Tulsi leaf extract were identified by HPLC and LC-MS. Results Mitomycin C (MMC) and hexavalent chromium (Cr 6) induced statistically significant DNA strand break of respectively 69% and 71% (P<0.001) as revealed by fluorometric analysis of DNA unwinding. Furthermore, the damage could be protected with DTLE (50 μL/mL, 100 μL/mL, and 200 μL/mL) on simultaneous treatment. Chromosomal aberration and micronucleus formation induced by MMC, Cr 6 and B[a]P were significantly protected (P<0.001) by DTLE with and without metabolic activation. Conclusion Distillate of Tulsi leaf extract possesses antioxidants contributed mainly by eugenol, luteolin and apigenin as identified by LC-MS. These active ingredients may have the protective effect against genotoxicants.     

高效液相色谱法测定食品中大豆异黄酮

目的：建立高效液相色谱法测定食品中大豆异黄酮（包括染料木黄酮和大豆苷元）的含量，探讨样品的提取方法及测定影响因素。方法：样品经0.50mol/L盐酸回液和乙醇提取大豆异黄酮后，采用甲醇－0.01mol/L乙酸铵溶液（pH4.5)(60 40)作为流动相分离待测物质，等度洗脱，采用紫外检测器于254nm波长下测定。结果：染料木黄酮的检出限为0．04μg/ml，大豆苷元为0．05μg/ml，样品的加标回收率为91．4％－113．1％，日内和日间相对标准偏差分别为1．1％－2．8％和2．9％－5．1％。结论：本法简便、快速，易于推广普及，为大豆及其制品和保健食品中大豆异黄酮的检测和膳食摄取植物性雌激素提供了分析方法。     

HPLC测定人尿液三苯双脒次级代谢物对苯二甲酸浓度

目的建立测定人尿液三苯双脒次级代谢物对苯二甲酸（TPA）浓度的高效液相色谱法（HPLC）。方法以阿魏酸为内标（internal standard, IS）,志愿者尿样稀释后经Ultimate XB0C18柱（4.6mm×200mm,5μm）、流动相甲醇－1%醋酸（35∶65,V/V）分离,在检测波长为240nm、流速为0.8mL/min、柱温为30℃条件下采用HPLC分析。结果在1～400μg/mL内,TPA与IS的峰面积比值与浓度线性关系良好（r=0.9954）,定量下限为1μg/mL,低、中、高浓度的相对回收率分别为（106.0±4.4）%、（107.4±4.2）%和（108.5±2.4）%,批内和批间相对标准偏差（RSD）小于8%。结论方法符合生物样品分析要求,可用于三苯双脒次级代谢物TPA尿液浓度的测定。     

高效液相色谱法测定藿香正气胶囊中橙皮苷的含量

目的建立高效液相色谱法测定藿香正气胶囊中橙皮苷的含量。方法采用高效液相色谱方法。色谱柱为十八烷基硅烷键合硅胶为填充剂〔DiamosilTM(钻石),5.0μm,200 mm×4.6 mm〕,流动相为甲醇-0.5%醋酸溶液(30∶70),检测波长为284 nm,流速为1.0 mL/min。结果橙皮苷在5.592～74.56μg/mL范围内呈良好的线性关系,回归方程为A=21 168.38C-8 066.67,R=0.999 99。回收率为99.32%。结论该方法简单、快速、准确,能消除干扰,可用于测定藿香正气胶囊中橙皮苷的含量。     

HPLC法测定大鼠肝微粒体中阿司匹林酯酶活性及体外动力学研究

目的 探索大鼠肝微粒体中阿司匹林酯酶的活性及体外动力学.方法 以阿司匹林为底物,大鼠肝微粒体匀浆在37℃温育40min,乙腈终止反应.反应液离心取上清液过滤后进行RP-HPLC分析,测定代谢系统中阿司匹林（ASA）和水杨酸（SA）的浓度.通过Lineweaver-Burk的双倒数法进行曲线回归求算Vmax、Km.结果 不同浓度阿司匹林在蛋白浓度0.33mg/mL下孵育40min,测得动力学参数为Km=4.42mmol·L-1和Vmax=0.736 mmol·min-1·mL-1.结论 HPLC法稳定可靠,灵敏度高,可用于阿司匹林酯酶的体外动力学研究.     

高效液相色谱法测定苍辛滴鼻剂中绿原酸及木兰脂素的含量

目的 运用高效液相色谱法对苍耳子中的绿原酸及辛夷中的木兰脂素进行含量测定.方法 绿原酸采用高效液相色谱法,色谱柱为Shim-Pack CLC ODS C18柱(250×4.6 mm,5μm);乙腈-0.4％磷酸溶液(10∶ 90)为流动相;检测波长为327nm;流速:1.0 mL/min.木兰脂素的色谱条件为:用辛基键合硅胶为填充剂Hypersil C8柱(250×4.6 mm,5μm)为色谱柱;乙腈-四氢呋喃-水(35∶ 1∶64)为流动相;检测波长为278 nm;流速:1.0 mL/min.结果 绿原酸在12.99 ～ 129.90 μg/mL(r =0.999 9)范围内及木兰脂素在19.67 ～ 196.70 μg/rmL(r =0.999 9)范围内线性关系良好,绿原酸回收率为99.28％,RSD为1.15％(n=9);木兰脂素回收率为99.42％,RSD为0.68％(n=9),3批样品中绿原酸的平均含量为0.818 4、0.818 5、0.818 9 mg/mL;木兰脂素的平均含量为0.163 7、0.163 4、0.164 1 mg/mL.结论 高效液相色谱法简便、灵敏、准确,可作为苍辛滴鼻剂的定量质量控制.     

反相高效液相色谱法同时测定血清中5种水溶性维生素

目的 建立血清中水溶性维生素B_1(VB_1)、B_6(VB_6)、烟酸、烟酰胺和叶酸的高效液相色谱同时测定方法.方法 取全血40 μL,用200 μL甲醇沉淀蛋白,冰箱冷冻(-18℃)30 min后离心(8000 r/min,8 min),上清液通过固相萃取柱进一步净化,收集洗脱液,50℃水浴中氮气吹至0.20 mL,取50 μL进样色谱柱分析.对样品的固相萃取条件和色谱条件进行优化后,用于测定人血清样品中5种水溶性维生素的含量.结果 优化后的固相萃取条件;固相萃取柱AGT cleanert~(TM) ODSC_(18)柱;洗脱液:甲醇(2.4 mL)-水(0.6 mL)混合液;洗脱流速:0.5 mL/min.本法的色谱分离条件为:分离柱Phenomenex C18 Luna色谱柱(250×4.6 mm,5 μm);柱温:室温(20℃);流动相:0.05 mol/L KH_2PO_4溶液(pH6.0)-甲醇梯度洗脱;流速:0.9 mL/min;紫外检测波长:266 nm.5种水溶性维生素标准曲线的线性范围为0～100 ng,相关系数均大于0.999,相对标准偏差(RSD)小于5%,方法 检出限(信噪比S/N=2)为0.076～0.170 μg/mL,加标回收率为80.2%～115.0%.人血清样品中除烟酰胺未检出外,其余4种均检出.结论 所建立的方法 快速、简便、灵敏、准确,可用于人血清中水溶性维生素VB_1、VB_6、烟酸和叶酸的同时测定,但烟酰胺的检测灵敏度有待提高.     

高效液相色谱法快速测定脑梗塞模型大鼠脑组织中多胺的含量

目的:建立一种简便、灵敏和快速分离测定大鼠脑组织中腐胺、精脒和精胺等多胺含量的高效液相色谱法。方法:采用苯甲酰氯作为衍生化试剂,以1,6-己二胺为内标,采用polygocyl(250×4.6 mm 5μm)C18色谱柱,流动相为甲醇-水(62:38 V/V),流速1.0 mL/min,检测波长229 nm。结果:本法线性范围是1～2 000/mL,回收率大于90%。日内和日间变异系数为1.5%～4.2%。最低定量浓度为1 nmol/mL。结论:该方法符合生物样品分析要求,为脑组织中多胺的含量测定提供了一种简便、准确、灵敏度高的分析方法。     

Anticlastogenic Effect of Redistilled Cow＇s Urine Distillate in Human Peripheral Lymphocytes Challenged With Manganese Dioxide and Hexavalent Chromium

Objective To study the anticlastogenic effect of redistilled cow＇s urine distillate （RCUD） in human peripheral lymphocytes （HLC） challenged with manganese dioxide and hexavalent chromium. Methods The anticlastogenic activity of redistilled cow＇s urine distillate was studied in human polymorphonuclear leukocytes （HPNLs） and human peripheral lymphocytes in vitro challenged with manganese dioxide and hexavalent chromium as established genotoxicants and clastogens which could cause induction of DNA strand break, chromosomal aberration and micronucleus. Three different levels of RCUD： 1 μL/mL, 50 μL/mL and 100 μL/mL, were used in the study. Results Manganese dioxide and hexavalent chromium caused statistically significant DNA strand break, chromosomal aberration and micronucleus formation, which could be protected by redisfilled cow＇s urine distillate. Conclusion The redistilled cow＇s urine distillate posseses strong anfigenotoxic and antielastogenic properties against HPNLs and HLC treated with Cr^＋6 and MnO2. This property is mainly due to the antioxidants present in RCUD.     

Enhancement of urinary elimination of 3-bromobenzanthrone metabolites by oral supplementation of ascorbic acid in guinea pigs

Objective 3-Bromobenzanthrone (3-BBA), an anthraquinone intermediate dye, is extensively used in textile industry. Since, our prior studies have shown that 3-BBA caused significant depletion of ascorbic acid (AsA) levels, the effect of exogenous supplementation of AsA on the urinary elimination of 3-BBA metabolitcs was investigated. Method Guinea pigs were treated with singleoral dose of 3-BBA (50 mg/kg b. wt,) in groundnut oil while another group was treated with single oral dose of 3-BBA (50 mg/kg b. wt.) along with 3 day prior and post oral supplementation of AsA. Control groups were either treated with groundnut oil or AsA alone. Urine from individual animals was collected, extracted and analysed on HPTLC. Results The highest elimination of 3-BBA (75ug) was found to be in 0-24 h urine fraction which decreased to 18ug and 5 ug in the two subsequent 24 hourly fractions of urine. Exogenous supplementation of AsA increased the total urinary elimination of 3-BBA by almost 77%. A total of 10 fluorescent metabolites excluding the parent compound were eliminated in the urine of guinea pigs treated with 3-BBA. Densitometric scanning of chromatogram showed different peaks at Rf 0.18, 0.22, 0.27, 0.34, 0.40, 0.48, 0.56, 0.66, 0.72, 0.80, and 0.95 which were eliminated and marked as urinary metabolite 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and 11 respectively. AsA not only significantly enhanced the elimination of 3-BBA metabolites but also modified the pattern of metabolites drastically in 0-6 h, 6-24 h and 24-48 h urine fractions. Conclusion These results indicate that AsA may be useful in protecting the toxicity of 3-BBA by fascilitating the urinary metabolite(s) excretion of 3-BBA.     

柱前紫外衍生-高效液相色谱法同时测定功能食品中八种糖醇和糖

目的 建立功能食品中赤藓糖醇、木糖醇、半乳糖醇、山梨醇、甘露醇、麦芽糖醇,以及葡萄糖和三氯蔗糖的柱前紫外衍生-高效液相色谱同时测定方法。 方法 食品中的糖醇等经水提取后,与苯甲酰氯反应生成紫外衍生物,衍生物经C18柱分离,柱温为30 ℃,甲醇-水梯度洗脱,流速1.00 mL/min,检测波长为232 nm。 结果 所测8种物质的标准曲线线性良好（r>0.999）,检出限低于2.2 μg/mL,加标回收率为89.6%~117.0%,日内相对标准偏差小于5%。 结论 该方法衍生反应简便快速、所需试剂经济易得,本法不需特殊检测器,易于推广,适用于功能食品中多种糖醇的同时测定。     

蛇床子等中药提取物对几种病菌生物活性的初步研究

以蛇床子、野菊花和白芷丙酮提取物为材料,测定了其对8种植物病原真菌的生物活性。结果表明:在供试质量浓度为0.05gm/L时,蛇床子提取物对8种供试菌中的6种病原真菌菌丝生长的抑制作用效果最好,抑制率均在65%以上,野菊花提取物抑菌活性次之,对其中3种供试菌抑制率均在65%以上;在供试质量浓度为0.1gm/L时,野菊花提取物对棉花枯萎病菌孢子萌发抑制作用效果较好,抑制率为92.5%。蛇床子和野菊花作为植物源杀菌剂值得进一步研究。     

The effect of the haematocrit value on the determination of glucose levels by reagent-strip methods

Twenty-one patients' blood samples, each with modified haematocrit concentrations of about 20%, 30%, 40%, 50% and 60%, were assayed for the presence of glucose by two reagent-strip methods--the Ames Glucometer with BG reagent strips and the Reflolux reflectance meter with BM-test strips. The reference method was the YSI blood glucose analyser. Both Ames and BM systems were found to have good precision. Ames results were falsely high (mean, 15%) at a haematocrit value of 20% and falsely low (mean, 21%) at a haematocrit value of 60%. BM results also overestimated (mean, 4%) and underestimated (mean, 5%) at the corresponding haematocrit levels. The results showed good agreement between Ames and BM methods and the YSI method for normal haematocrit concentrations of 40% to 50%. Clinicians should be aware of inaccuracies in glucose results by test-strip methods in patients with abnormal haematocrit concentrations.     

高效液相色谱法测定青皮药材中橙皮苷的含量

目的 建立高效液相色谱法(HPLC)测定青皮中橙皮苷含量的方法.方法 采用色谱柱为kromasilC 18(4.6 mm×250 mm,5μm);流动相为乙腈-0.5％磷酸(75∶25);检测波长285nm;流速1.0 mL/min,测定青皮中橙皮苷.结果 橙皮苷的进样量在11.2～89.6 μg/mL范围内线性关系良好(r=0.9997).橙皮苷平均回收率为99.07％(n=6),RSD为1.08％.结论 建立的HPLC法简便、快速、准确,重现性、稳定性好,回收率高,可为青皮药材的质量控制提供依据.     

高效液相色谱法测定复方阿司达莫人体血药浓度

目的：建立高效液相色谱法测定复方阿司达莫人体血药浓度。方法双嘧达莫色谱条件采用ODS C18（4．6mm×215;250mm ,5μm ）色谱柱;流动相甲醇-1％醋酸（25：75）;流速1．0 mL·min^-1;柱温30℃;检测波长285nm。水杨酸色谱条件ODS C18（3．9mm×150mm ,5μm）色谱柱;流动相为甲醇-1％醋酸（33：67）;柱温30℃;流速为1．0mL·min^-1;检测波长234nm。结果在0．05～8μg·mL^-1范围内,双嘧达莫最低定量限为0．05μg·mL^-1,线性关系良好（r＝0．9998,n＝6）,方法回收率80．14％～104．78％,低、中、高三种浓度质控样品的日内、日间变异小于2．5％;在0．1～8μg＆#183;mL -1范围内,水杨酸最低定量限为0．1μg·mL^-1,线性关系良好（r＝0．9999,n＝6）,方法回收率95．19％～100．06％,低、中、高三种浓度质控样品的日内、日间变异小于2．5％。结论该方法定量准确,灵敏度高,适用于复方阿司达莫人体药代动力学研究。     

靶向抗金葡多肽Ph-SA对β-内酰胺类抗生素的增效作用初探

目的探讨Ph-SA抑制金葡β-内酰胺酶产生,增强β-内酰胺类抗生素杀菌效果,对抗细菌耐药的可能性.方法实验分为1.43×10-4μmol/mL(10 μg/mL)Ph-SA、2.81×10-2 μmol/mL(10 μg/mL)青霉素、0.715×10-4μmol/mL(5μg/mL)Ph-SA+1.40×10-2μmol/mL(5μg/mL)青霉素和对照四个组.在耐青霉素金葡菌对数生长期加药,测定各组菌液的生长情况及β-内酰胺酶活性.同样方法测定Ph-SA与头孢唑啉合用的效果.结果1.43×10-4 μmol/mL Ph-SA对细菌β-内酰胺酶活性有显著的抑制作用而未表现出明显的杀菌效应;0.715×10-4μmol/mL Ph-SA+1.40×10-2μmol/mL青霉素组及0.715×10-4μmol/mL(5μg/mL)Ph-SA+1.05×10-2μmol/mL(5μg/mL)头孢唑啉组细菌生长情况分别与2.81×10-2μmol/mL青霉素组及2.10×10-2μmol/mL(10μg/mL)头孢唑啉组相当,而β-内酰胺酶活性均低于对照.结论Ph-SA可降低β-内酰胺酶活性,增强抗生素的杀菌效果,具有对抗细菌耐药的价值.     

不同厂家生产的独一味制剂中木犀草素含量的测定

目的 用高效液相色谱法测定不同厂家生产的独一味制剂中木犀草素的含量.方法 色谱柱为Dikma platisil C18(250mm×4.6 mm,5μm);流动相为甲醇-0.4％磷酸溶液(55∶45);流速为1.0 mL/min;检测波长为350 nm;柱温为35℃.结果 木犀草素含量的线性回归方程为Y=5.26+19...     

Analysis of chemical composition and bioactive property evaluation of Indian propolis

Objective

To analyze the chemical composition and to evaluate the bioactive potential of hydroalocoholic extract of propolis.

Methods

Ethanol extract of propolis was analyzed by GC-MS, HPTLC and HPLC methods and in vitro antioxidant, anticholinesterase and cytotoxicity assay were performed.

Results

GC-MS analysis revealed the presence of fatty acids, alcohols, and quercetin. Quercetin was identified and quantified by HPTLC and HPLC methods. Dose dependent DPPH and hydroxyl radical scavenging activity of hydroalcoholic extract of propolis was calculated as 16.20 and 34.33 µg/mL respectively. Inhibition of lipid peroxidation was significant and the IC₅₀ value was calculated as 55.56µg/mL. Anticholinesterase activity was less observed. The cytotoxic activity against both breast (MCF-7) and lung cancer (A543) cell lines were significant and the IC₅₀ value was calculated as 10 and 13 µg/mL respectively.

Conclusions

These findings showed that bioactive compounds present in propolis will alleviate many diseases and can be used for better human health.     

黄连解毒汤体外抗菌活性研究

目的探讨黄连解毒汤体外抗菌活性。方法采用连续倍比试管稀释法和琼脂扩散法测定黄连解毒汤的最低抑菌浓度（MIC）和最低杀菌浓度（MBC）。结果黄连解毒汤对副溶血弧菌、金黄色葡萄球菌的MIC分别为62．5mg/mL、125．0m咖L,MBC分别为125．0mg/mL、125．0mg／mL;黄连解毒汤对福氏志贺菌、铜绿假单胞菌、大肠埃希菌、肺炎链球菌、伤寒沙门菌MIC均〉1000mg／mL。结论黄连解毒汤在体外对副溶血弧菌及金黄色葡萄球菌有显著的抗菌活性。     

High-performance liquid chromatographic determination of urinary trans, trans-muconic acid excreted by workers occupationally exposed to benzene

INTRODUCTION Benzene exists ubiquitously as an environmental contaminant to which the general population are exposed, due to its occurrence in mineral oil and formation in many combustion processes[1]. It is mutagenic and carcinogenic in animal models and…