胶束电动毛细管色谱法测定穿心莲中穿心莲内酯和脱水穿心莲内酯

采用胶束电动毛细管色谱法分离测定了中药穿心莲中穿心莲内酯和脱水穿心莲内酯。电泳条件 :以2 0mmol L磷酸缓冲溶液 1 0mmol LSDS(含 5 %甲醇 ,V/V ,pH 6.8)为电泳介质 ,未涂层石英毛细管 (75 μmi.d.× 5 0cm ,有效分离长度 4 2 .2cm)为分离通道 ,压力进样 (2 5 0kPa·s) ,2 0kV恒压电泳 (2 5℃ )分离 ,检测波长 2 4 0nm。在 1 0 .4～93 .5mg/L ,和 1 0 3～ 92 .7mg L范围内 ,对两种内酯分别进行了定量分析。加样回收率穿心莲内酯为 1 0 2 % ,脱水穿心莲内酯为 1 0 2 %。     

胶束毛细管电泳在线扫集技术同时分离测定穿心莲内酯和脱水穿心莲内酯的研究

采用胶束毛细管电泳在线扫集技术分离测定了中药穿心莲中脱水穿心莲内酯和穿心莲内酯.电泳条件:以20 mmol/L H3BO310 mmol/L NaH2PO4-50mmol/L SDS(含体积分数20%甲醇,pH 2.4)为电泳运行缓冲溶液,未涂层石英毛细管(58 cm×50 μm i.d.,有效长度为41.2 cm)为分离通道,重力进样,进样高度为11 cm,-20 kV恒压,检测波长为246 nm.富集倍数可以达到200倍以上.在5.70～91.20 mg/L,和3.96～31.68 mg/L范围内呈良好的线性关系,对两种内酯分别进行了定量分析.加标平均回收率脱水穿心莲内酯为100.80%,穿心莲内酯为98.06%.     

微乳液毛细管电动色谱法快速测定消炎利胆片中的穿心莲内酯和脱水穿心莲内酯

建立了微乳液毛细管电动色谱同时分析消炎利胆片中穿心莲内酯和脱水穿心莲内酯的方法。考察了缓冲溶液的浓度、pH值、十二烷基硫酸钠(SDS)以及助表面活性剂的含量对分离测定的影响。在由乙酸乙酯-SDS-正丁醇-30 mmol/L硼砂缓冲液(pH 9.5)（质量比为0.5∶0.6∶6.0∶92.9）组成的微乳液体系中,两种内酯在6 min内完成分离。该法简便、快速、选择性好,用于实际样品中穿心莲内酯和脱水穿心莲内酯的分析,获得了满意的结果。     

穿心莲内酯的溶剂浮选

应用溶剂浮选法对穿心莲乙醇提取液中的穿心莲内酯进行分离富集,优化了穿心莲内酯的浮选条件.实验表明:浮选溶剂为乙酸乙酯,水相与有机相体积比为2: 1,加入水相体积4%的乙醇,溶液pH 7,氮气流速200 mL/min,浮选时间40 min为最佳浮选条件.在最佳实验条件下,穿心莲内酯平均收率为94.1%; RSD为1.1%,平均标准加入回收率95.1%; 富集倍数为9.7倍.溶剂浮选法分离穿心莲内酯不仅能得到满意的回收率和富集倍数,并能有效地除去水溶性杂质,提高目标物纯度.与溶剂萃取法相比, 溶剂浮选法能节约有机溶剂和操作时间.     

新型脱水穿心莲内酯环磷酸酯类衍生物的合成及其抗肿瘤活性

以穿心莲内酯为先导化合物,在其3-位和19-位进行结构修饰,设计并合成了3个新型的脱水穿心莲内酯环磷酸酯类衍生物(1a～1c),其结构经1HNMR,IR和MS表征。体外抗肿瘤活性测试结果表明,1a～1c对舌癌Tca-8113细胞具有较好的体外抑制活性。     

穿心莲内酯衍生物的合成研究进展

穿心莲内酯是中药穿心莲的主要二萜类活性成分之一. 近年来药物及有机化学工作者发现穿心莲内酯及其衍生物具有抗肿瘤, 抗HIV, α-葡萄糖苷酶抑制等活性, 由于其广泛的药理活性而倍受人们的关注. 作者对穿心莲内酯衍生物的合成进行了综述.     

两个穿心莲内酯苷的波谱学研究

利用柱色谱对穿心莲地上部分的提取物进行分离,得到3,14-二去氧穿心莲内酯苷和14-去氧穿心莲内酯苷。利用核磁共振氢谱(1H-NMR)、碳谱(13C-NMR)以及双量子滤波相关谱(DQFCOSY)、总相关谱(TOCSY)、异核多量子相干谱(HMQC)、异核多键相关谱(HMBC),解析这两个化合物的结构。确定了1H-NMR的氢信号和13C-NMR的碳信号的归属,并且用NOESY对3,14-二去氧穿心莲内酯苷的立体结构进行了研究。     

鲁米诺化学发光测定穿心莲中穿心莲内酯的含量

本文将过氧化氢氧化穿心莲内酯中羟基的有机反应与Ｌｕｍｉｎｏｌ－Ｈ２Ｏ２Ｃｏ２＋化学发光反应偶合为一体，快速灵敏地对穿心蓬内酯进行了测定，其最低检测限为１．８９×１０－８ｇ／ｍＬ，线性范围为２×１０－８ｇ／ｍＬ～１０×１０－７ｇ／ｍＬ，回收率为９９．４５％，发光值稳定，效果良好。     

15-二氘脱水穿心莲内酯的合成和氢-氘交换速率考察

以脱水穿心莲内酯为原料,氧化氘作为氘源,四氢呋喃为助溶剂,三乙胺催化反应,分离纯化得到反应产物,其结构经¹H NMR、 ¹³C NMR和HR-MS(ESI-TOF)确证,并考察了产物在不同pH值的氢氘交换速率。结果显示,产物表征鉴定为15-二氘脱水穿心莲内酯,15位H-D交换速率在pH 分别为6, 7和8时较慢,随pH值升高加快。     

大孔吸附树脂提取穿心莲总内酯的研究

研究了大孔吸附树脂提取和纯化穿心莲总内酯的方法，采用HPLC法测定了穿心莲内酯的含量；考查了乙醇浓度对浸提效果的影响，并经动态吸附筛选了4种树脂；最后确定以ADS－7作为提取分离穿心莲总内酯的树脂，此树脂吸附量较高，脱附容量且能与杂质分离，有利于得到质量较好的穿心莲总内酯产品。     

反相高效液相色谱法测定野生与栽培花锚药材及其制剂中的花锚甙和去甲氧基花锚甙

 建立了花锚药材及其制剂中两种抗肝炎有效成分花锚甙和去甲氧基花锚甙的反相高效液相色谱测定方法。采用甲醇回流提取进行样品处理,在乙腈 磷酸水溶液为流动相作梯度洗脱、ODS柱、检测波长为254 nm条件下,花锚甙和去甲氧基花锚甙均可达到基线分离。两种成分在0.68～3.40 g/L ,0.36～1.8 g/L 时,其峰面积与浓度成良好的线性关系,加标回收率为95%～105%。该法适用于花锚药材及其制剂的质量分析检验。     

On-line coupling of dynamic microwave-assisted extraction with high-performance liquid chromatography for determination of andrographolide and dehydroandrographolide in Andrographis paniculata Nees

A novel technique based on dynamic microwave-assisted extraction (DMAE) coupled on-line with high-performance liquid chromatography (HPLC) through a flow injection interface has been developed for determination of andrographolide and dehydroandrographolide in Andrographis paniculata Nees. A TM(010) microwave resonance cavity built in the laboratory was applied to concentrating the microwave energy. An extraction vessel was placed in microwave irradiation zone. The extraction was performed in a recirculating system. When a number of extraction cycles were completed, the fractional extract (20muL) was driven to the analytical column by 65% aqueous methanol and was measured by diode array detector (DAD) at 225nm. The optimized extraction conditions are follows: extraction solvent 60% aqueous methanol; microwave forward power 80W; extraction time 6min; extraction solvent flow-rate 1.0mLmin(-1). The detection and quantification limits obtained are 0.5 and 1.7microgmL(-1) for andrographolide and 0.6 and 1.9microgmL(-1) for dehydroandrographolide, respectively. The within-day and between-day precision (RSD) are 2.1% and 3.7% for andrographolide and 1.7% and 4.1% for dehydroandrographolide, respectively. Mean recoveries for andrographolide and dehydroandrographolide are 97.7% and 98.7%, respectively. Compared with ultrasonic extraction used in the Chinese pharmacopoeia, the proposed method was demonstrated to obtain higher extraction yield in a shorter time. In addition, only small quantities of solvent (5mL) and sample (10mg) were required.     

Development of andrographolide molecularly imprinted polymer for solid-phase extraction

A method employing molecularly imprinted polymer (MIP) as selective sorbent for solid-phase extraction (SPE) to pretreat samples was developed. The polymers were prepared by precipitation polymerization with andrographolide as template molecule. The structure of MIP was characterized and its static adsorption capacity was measured by the Scatchard equation. In comparison with C(18)-SPE and non-imprinted polymer (NIP) SPE column, MIP-SPE column displays high selectivity and good affinity for andrographolide and dehydroandrographolide for extract of herb Andrographis paniculata (Burm.f.) Nees (APN). MIP-SPE column capacity was 11.9±0.6 μmol/g and 12.1±0.5 μmol/g for andrographolide and dehydroandrographolide, respectively and was 2-3 times higher than that of other two columns. The precision and accuracy of the method developed were satisfactory with recoveries between 96.4% and 103.8% (RSD 3.1-4.3%, n=5) and 96.0% and 104.2% (RSD 2.9-3.7%, n=5) for andrographolide and dehydroandrographolide, respectively. Various real samples were employed to confirm the feasibility of method. This developed method demonstrates the potential of molecularly imprinted solid phase extraction for rapid, selective, and effective sample pretreatment.     

Determination of andrographolide and dehydroandrographolide in rabbit plasma by on-line solid phase extraction of high-performance liquid chromatography

The high-performance liquid chromatography (HPLC) coupled with on-line solid phase extraction (SPE) and ultraviolet (UV) detection was developed for determining andrographolide and dehydroandrographolide in rabbit plasma. Plasma samples (100 μL) were injected directly into a C18 SPE column and the biological matrix was washed out for 6 min using 15% aqueous methanol. By rotation of the switching valve, andrographolide and dehydroandrographolide were eluted in the back-flush mode and transferred to the analytical column by the chromatographic mobile phase consisted of methanol:acetonitrile (ACN):water (50:10:40; v/v). The UV detection was performed at 225 nm. The calibration curves showed excellent linear relationship (R ≥ 0.9993) over the concentration range of 0.05-5.0 μg mL⁻¹. The within- and between-day precisions (R.S.D.) of two analytes were in the range of 1.2-6.5% and the accuracies were between 92.0% and 102.1%. Their recoveries were all greater than 94%. The limits of detection were 0.019 μg mL⁻¹ for andrographolide and 0.022 μg mL⁻¹ for dehydroandrographolide. This method was successfully applied to the plasma concentration-time curve study after oral administration of Andrographis paniculata Nees extract in rabbit.     

Simultaneous determination of four andrographolides in Andrographis paniculata Nees by silver ion reversed-phase high-performance liquid chromatography

A rapid and sensitive silver ion high-performance liquid chromatographic (Ag[I]-HPLC) method is developed for the simultaneous determination of the biologically active diterpenoids andrographolide, 14-deoxy-11,12-didehydroandrographolide, 14-deoxyandrographolide, and neoandrographolide in Andrographis paniculata Nees. HPLC is carried out for determining andrographolide and its derivatives with methanol-water (55:45, v/v) as the mobile phase on a C18 column (5 microm, 150 mm x 4.6 mm i.d.) with UV detection at 205 nm. Four andrographolides are baseline separated in a novel way: by adding silver ions (0.005 mol L(-1)) to the previously mentioned mobile phase. Validation of the method challenges specificity, linearity, limit of detection, limit of quantitation, accuracy, and repeatability, and the results met the acceptance criteria for all analytes. The molecular mechanism of retention is demonstrated by comparing partition coefficients (logP) of different andrographolides and andrographolide-Ag(I) complexes. Thus, the method is successfully applied to characterize and determine the four andrographolides in Andrographis paniculata Nees extract and its commercial product.     

Quality control of medicinal herbs Fructus gardeniae, Common Andrographis Herb and their preparations for their active constituents by high-performance liquid chromatography-photodiode array detection-electrospray mass spectrometry

Dehydroandrographolide, andrographolide and geniposide are the main active constituents of many herbal medicines, e.g., Fructus gardeniae, Common Andrographis Herb. They are used as the markers to control the quality of such herbal medicines and their herbal preparations. In this paper, a simple and sensitive high-performance liquid chromatographic (HPLC) method coupled with photodiode array detection (DAD) and electrospray mass spectrometry (ESI/MS) were developed to determine the three compounds simultaneously in extracts of medicinal herbs and herbal preparations produced by different companies. The extracts were separated on a C₁₈ reversed phase HPLC column, with a gradient solvent system, the time for the separation of the three target analytes was 10 min. The abundance ions were recorded using selected ion monitoring (SIM) mode with m/z 297.3, 297.3 and 411.1 for dehydroandrographolide, andrographolide and geniposide, respectively. The limit of detection for dehydroandrographolide, andrographolide and geniposide were 20, 30 and 150 ng mL⁻¹, respectively. The proposed method was successfully applied to the determination of the contents of the compounds in related to medicinal herbs and preparations.     

高效液相色谱-蒸发光散射检测法测定藜芦中的介藜芦碱和藜芦胺

建立了高效液相色谱-蒸发光散射检测(HPLC-ELSD)测定藜芦中介藜芦碱、藜芦胺含量的方法,并对4种藜芦属药材样品进行了测定。采用的色谱柱为Kromasil C18柱(250 mm×4.6 mm, 5 μm),以乙腈和0.1%三氟乙酸水溶液为流动相进行梯度洗脱,洗脱程序为:0～5 min, 20%乙腈; 5～30 min, 20%乙腈～40%乙腈, 30～40 min, 40%乙腈～20%乙腈; 40～45 min, 20%乙腈;流速为0.8 mL/min;柱温为35 ℃;采用ELSD检测,漂移管温度为98 ℃,载气流速2.2 L/min 。介藜芦碱和藜芦胺的线性范围分别为42.05～980 mg/L和43.77～1020 mg/L;平均回收率分别为99.2%和101.4%,相对标准偏差分别为1.7%和2.1% (n=6);信噪比为3时,测得介藜芦碱和藜芦胺最低检测限分别为18.37 mg/kg和21.50 mg/kg。该方法快速简便、灵敏度和分离度好,适用于藜芦药材中活性生物碱的测定。     

高效液相色谱-质谱法同时测定清热解毒口服液中的5种有效成分

建立了同时测定清热解毒口服液中的绿原酸、栀子苷、黄芩苷、连翘苷和靛玉红5种有效成分的高效液相色谱-电喷雾电离质谱（HPLC-ESI/MS）分析方法。采用Zorbax SB C18色谱柱,以含0.2%甲酸的0.4 mmol/L醋酸钠(A相)、乙腈(B相)为流动相进行梯度洗脱,在ESI正离子模式下,采用选择离子监测方法进行测定,用峰面积进行定量。结果表明,绿原酸、栀子苷、黄芩苷、连翘苷和靛玉红的线性范围分别为0.050～50 mg/L,0.020～20 mg/L,0.005～30 mg/L,0.010～15 mg/L和0.010～10 mg/L;检出限分别为0.010,0.005,0.001,0.002和0.003 mg/L。5种成分的加样回收率为97.0%～101.7%,相对标准偏差小于2.2%。该法快捷、准确、重复性好,可用于清热解毒口服液中的5种有效成分含量的同时测定。     

离子对色谱法同时分离测定吡啶及咪唑离子液体阳离子

建立了用紫外检测的反相离子对色谱梯度淋洗同时分离测定4种吡啶离子液体阳离子和5种咪唑离子液体阳离子的方法。实验采用ZORBAX Eclipse XDB-C18反相色谱柱,以离子对试剂水溶液(用柠檬酸调节pH值)+乙腈为流动相,考察了离子对试剂种类和浓度、乙腈浓度和色谱柱温度对保留的影响,探讨了相关保留规律,确定最佳色谱条件为:流速1.0 mL/min、柱温30℃,以1.0 mmol/L庚烷磺酸钠水溶液(pH 4.0)-乙腈为淋洗液进行梯度洗脱。在此条件下,4种吡啶阳离子和5种咪唑阳离子在15 min内达到基线分离。检出限(S/N=3)为0.31～0.54 mg/L,峰面积的相对标准偏差为0.10%～0.75%。将该方法用于实验室合成的离子液体样品分析,加标回收率为94%～98%。该方法准确、可靠,具有较好的实用性。