Crystallographic and NMR studies on sterols from green alga Chaetomorpha basiretorsa Setchell

Chemical investigation of the ethanol extract of the marine green alga Chaetomorpha basiretorsa Setchell led to the isolation of a new sterol stigmast-4,28-dien-3 alpha 6 beta-diol 1 in addition to the five known sterols of beta-lawsaritol 2, saringosterol 3, 24-hydroperoxy-24-vinyl - cholesterol 4, beta-stigmasterol 5, 29-hydroxystigmasta-5, 24(28) -dien-3 beta-ol 6. Compounds were isolated by normal phase silica gel and Sephadex LH - 20 gel colum chromatography, reverse phase HPLC and recrystalization. Their structures were elucidated by spectroscopic methods including MS, IR 1D/2D NMR and X-ray analysis. Cytotoxicity of compounds was screened by using the standard WIT method. All these compounds were isolated from the green alga Chaetomorpha basiretorsa Setchell for the first time and they were inactive (50% inhibitory concentration was greater than 10 mu g /cm(3)) against KB, Bel -7402, PC - 3M, Ketr 3 and MCF - 7 cell lines.

The chemical constituents from red alga Gymnogongrus flabelliformis Harv.

Eight compounds were isolated from red alga Gymnogongrus flabelliformis Harv. In normal phase silica gel, Sephadex LH-20 gel column chromatography, reverse phase HPLC, and recrystallization. Based on MS and 1D NMR spectroscopic data, their structures were determined as: stigmast-4-en-3-one (I), cholest-4-en-3-one (II), cholesterol (III), uracil (IV), uridine (V), adenosine (VI), succinic acid (VII), and 5-hydroxy-4-methyl-5-pentyl-2,5-dihydro-furan-2-on (VIII). All of them were obtained from this species for the first time. Cytotoxicity of these compounds was screened using standard MTT method, but all the compounds were inactive (IC50 > 10 mu g/ml).

Simultaneous Determination of Multi-Organotin Compounds in Seawater by Liquid-Liquid Extraction-High Performance Liquid Chromatography-Inductively Coupled Plasma Mass Spectrometry

The hyphenated technique of high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry (HPLC-ICP-MS) was applied to the simultaneous determination of five organotin compounds (trimethyltin, dibutyltin, tributyltin, diphenyltin and triphenyltin) in seawater samples. Agilent TC-C18 column was used for the separation, the mobile phase of HPLC was CH3CN : H2O: CH3COOH=65 : 23 : 12 (phi), 0.05% TEA, and pH value was adjusted to 3.0 by diluent ammonia. The flow rate was 0.6 mL . min(-1). Five mixed organotin compounds in a mix standard solution from 100 to 0.5 mu g . L-1 were applied for the method assessment. The experimental results indicate that the correlation coefficient of calibration curves (R-2) for each organotin compound was over 0.998 and the detection limits of the five organotin compounds were lower than 3 ng . L-1. Different mixed organic solvents including dichloromethane or toluene were used for extraction of organotin and the extraction condition of organotin from seawater was optimized. The 100 mL seawater acidized by hydrochloric acid was extracted by 10 mL carbon dichloride (CH2Cl2) with 2% tropolone for 10 min twice. Extracted organic solvents were mixed And blown to one drop by nitrogen with the rate of 1.7 mL . min(-1), then 1 mL acetonitrile was added to the drop for redissolving the organotin compounds. Finally, the mixed redissolution was filtered by 0.22 mu m organic filter membrane before analysis. it was found that the only organotin compound in seawater was triphenyltin (TPHT) and the content was 53.2 ng . L-1. The recoveries test from the standard addition for diphenyltin (DPHT), dibutyltin (DBT), tributyltin (TBT) and triphenyltin (TPHT) were over 80%. However, the recovery for trimethyltin (TMT) was relatively low and the value was 50%. The reason might be attributed to the decomposition or adsorption of those compounds during the extraction procedure. Further study on this subject is in progress.

Speciation Analysis of Organotin Compounds in Shellfish by Hyphenated Technique of High Performance Liquid Chromatography-Inductively Coupled Plasma Mass Spectrometry

The hyphenated technique of high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry(HPLC-ICP-MS) was applied to the simultaneous determination of five organotin compounds in the shellfish samples. Agilent TC-C-18 column was selected, mobile phase of the HPLC was CH3CN:H2O: CH3COOH = 65:23:12 (V/V), 0. 05% TEA, pH = 3.0 at flow rate 0.4 mL/min. Five mixed organotin standards from 100 mu g/L to 0. 5 mu g/L was used for the method evaluation. The experimental results indicate that the linearity (R-2) for each compound was over 0.998. The shellfish samples were treated by supersonic extraction with mobile phase for 30min. Four organotin compounds including dibutyltin (DBT), tributyltin (TBT), diphenyltin (DphT) and triphenyltin (TPhT) in shellfish samples were detected with method mentioned above. It was found that the domain compounds in the samples were tributyltin (TBT) and triphenyltin (TPhT). The recoveries test from the standard addition for trimethyltin (TMT tributyltin (TBT), and triphenyltin (TPhT) were, over 80%. However, the recoveries for diphenyltin (DPhT) and dibutyltin (DBT) were relatively low, 37.3% and 75.2% respectively. The reason might be attributed to the decomposition of those compounds during the extraction procedure. The further study on this subject is under the progress.

GC-MS法测定藏木香栽培品种挥发油的化学成分

[目的]基于气相色谱-质谱（GC-MS）法测定藏木香栽培品种挥发油的化学成分。[方法]采用水蒸气蒸馏法从藏木香栽培品种中提取挥发油,并用GC-MS联用仪对其挥发油的化学成分进行研究。[结果]分离并确认了37种成分,其主要成分是桉叶油二烯5,11（13）-内酯-8,12,异-榄香烯,异-喇叭烯,桉叶油二烯4,11（13）-内酯-8,12。[结论]分析获得的主要化学成分及其功效为藏木香这一天然药用植物资源的人工规范化栽培和进一步综合开发利用提供了科学依据。

RP-HPLC测定几种花类药材黄酮含量

目的：为充分利用花类药材，建立马蔺花、桃花、月季等15种花类药材中黄酮含量测定的RP—HPLC方法，并比较它们的含量。方法：花类药材用甲醇提取，反相高效液相色谱法分析药材中槲皮素、山柰酚、异鼠李素3种黄酮苷元。结果：采用KromasilC_18柱（4．6mm×250／mm，5μm），以甲醇-0．1％磷酸（50：50）为流动相，可使3种黄酮苷元达到基线分离，其中马蔺花含槲皮素1．536％，桃花含山柰酚0．572％，茶花含异鼠李素0．029％，皆为听测样品中最高。结论：本实验首次采用RP—HPLC法测定这15种花类药材的总黄酮含量，本方法操作简便，快速，准确，可作为黄酮的定量分析方法。

青海栽培和野生唐古特大黄蒽醌类成分的HPLC对比分析

采用HPLC法测定青海栽培唐古特大黄中的5种蒽醌含量,并和野生唐古特大黄药材进行了比较.结果表明,二、三、四年龄栽培唐古特大黄中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚5种蒽醌总量分别为1.21%,2.01%,1.62%,其中三、四年龄栽培唐古特大黄已达到<药典>规定的药用标准;野生大黄的总蒽醌含量远高于栽培大黄为3.64%.

藏药提宗龙胆和线叶龙胆花中四种苦苷类成分的HPLC测定

应用HPLC分析方法同时测定藏药提宗龙胆和线叶龙胆两种植物花中落干酸、獐牙菜苦苷、龙胆苦苷、獐牙菜苷4种苦苷类成分的含量.采用Econosphere C_(18)色谱柱(250×4.6 mm,5 μm),以甲醇-0.5%乙酸为流动相进行梯度洗脱,流速为1.0 mL/min;检测波长为245 nm.结果表明,除提宗龙胆中未检出獐牙菜苦苷外,其它成分均在两种植物中存在,但含量存在一定的差异.

藏药短管兔耳草中松果菊苷和麦角甾苷的RP-HPLC分析

建立了藏药短管兔耳草中松果菊苷和麦角甾苷含量的高效液相色谱分析法.采用Waters XTerra RP18色谱柱(150mm×4.6 mm,5μm),以甲醇-1%冰醋酸溶液(28:72,V:V)为流动相,流速1.0 mL/min,检测波长330 nm,柱温30℃,在20 min内分离检测了该两种化合物.松果菊苷和麦角甾苷进样量分别在0.077～4.950μg(r=0.999 9)和0.085～5.450μg(r=0.999 9)内呈良好线性,平均加样回收率分别为98.35%和92.50%,RSD分别为2.35%和2.86%.所建立的方法简便、快捷、结果准确可靠,重现性好,可用于藏药短管兔耳草的质量控制,并为兔耳草属植物中苯丙素苷类化合物的分离分析提供一定的参考.

抱茎獐牙菜中当药黄素的HPLC测定研究

[目的]采用反相高效液相色谱法对抱茎獐牙菜中当药黄素的含量进行测定。[方法]色谱柱为Kromasil C18（250min×4．60mm，5μm），流动相为甲醇-浓度0．02％磷酸水溶液，梯度洗脱，流速为1ml／min，检测渡长为260nm。[结果]当药黄素在0．92-4．60μg范围内线性关系良好，r=0．99950平均加样回收率为98．73％，RSD为0.31％。[结论]抱茎獐牙菜中当药黄素在花的部位含量最高。该测定方法适应性广，可用于测定抱茎獐牙菜中的当药黄素。

HPLC测定藏药帕朱胶囊中胡椒碱

帕朱胶囊是藏医最常用的治疗胃寒性疾病药物之一。收载于《中华人民共和国卫生部药用标准•藏药》（第一册），主要由寒水石、河子、石榴子、胡椒和荜茇等药物组成；功能主治健胃散寒，除痰、破痞瘤，养荣强壮。现代临床试验表明，帕朱胶囊对中晚期肿瘤患者有改善细胞免疫及体液免疫功能的作用，同时还可以减轻患者疼痛症状。尽管帕朱胶囊长期以来被广泛使用，但是却没有很好的质量控制标准。我们采用超临界CO2萃取方法提取帕朱胶囊的可溶性成分，并用GC—MS法对提取部位进行化学成分分析，发现其主要成分为胡椒碱，含量占提取物的44，2％。因此，本实验用HPLC法对帕朱胶囊中所含胡椒碱进行测定，以期建立胡椒碱的科学检测方法，提高产品质量控制标准。

藏药帕朱胶囊超临界CO_2萃取成分的GC-MS分析

目的：分析治疗胃病常用的藏成药帕朱胶囊（寒水石,诃子,石榴子,胡椒,荜拔等）的主要化学成分。方法：采用超临界CO_2萃取方法提取帕朱胶囊的可溶性成分,并用气相色谱-质谱联用法对提取部位进行化学成分分析,峰面积归一化法计算各组分的相对含量。结果：共分离鉴定了22种化合物,主要成分含量为胡椒碱44.2%、亚油酸21.1%、甲基-甲撑基奥-异丙烯内酯10.5%、棕榈酸4.8%和γ-羟基-榄香烯酸内酯3.5%等。结论：藏药帕朱胶囊的主要化学成分为胡椒碱和亚油酸,两者含量共占全部提取物的65.3%

藏药独一味RP-HPLC色谱指纹图谱的建立及其质量评价

[目的]建立独一味药材HPLC指纹图谱，研究不同地区独一味药材的质量，为其质量控制提供有效的方法。[方法]采用RP-HPLC方法梯度洗脱，测定10批不同地区的独一味样品。[色谱备件]Kromasil C18柱（250mm×4．60mm，5μm），流动相为甲醇-0．4％的磷酸水溶液（55：45），梯度洗脱程序为0～70min，甲醇的体积分数由1％线性增加至15％；70～100min内，甲醇由15％线性增加至17％；100～130min内，甲醇由17％线性增加至20％；130～150min，甲醇由20％线性增加至100％；流速为1ml/min，检测波长为254n／n，柱温为30℃。[结果]确定了独一味药材中的7个共有峰。根据聚类分析结果，将独一味药材分为3类。[结论]该研究建立的方法重复性好，可用于不同地区独一味药材的质量评价，结合含量测定可用于独一味药材的质量的全面控制。

藏药材石榴籽超临界CO_2萃取成分的GC-MS分析

目的：用新方法提取藏药材石榴籽油,并对其进行化学成分分析,为石榴籽的药理研究和应用提供实验依据。方法：采用超临界CO_2萃取方法提取石榴籽的可溶性成分,并用气相色谱-质谱联用法对提取部位进行化学成分分析,峰面积归一化法计算各组分的相对含量。结果：共分离鉴定了10种化合物,主要成分含量为亚油酸40.0%、油酸22.7%、棕榈酸17.5%、硬脂酸9.0%及共轭亚油酸5.7%。结论：藏药材石榴籽的主要化学成分为亚油酸和油酸,两者含量共占全部提取物的62.7%。

藏药蕨麻油脂化合物的GC-MS分析

蕨麻,藏医谓之卓老沙曾,原植物为蔷薇科委陵菜属鹅绒委陵菜Potentilla anserine L.,为多年生草本.在高海拔地区蕨麻根的中下部形成块根,在温暖低平地区不形成膨大的块根[1].蕨麻全草入药,收敛止血,止咳利痰,治各种出血及下痢.块根入药,有健脾益胃、生津止渴、益气补血功效;治脾虚、腹泻、产后贫血、营养不良等症[2].