以离子液体为流动相添加剂的HPLC法测定30批不同产地吴茱萸中8种生物碱的含量

目的 建立吴茱萸药材中去氢吴茱萸碱、二氢吴茱萸次碱、吴茱萸碱、吴茱萸次碱、1-甲基-2-[(Z)-5-十一碳烯]-4-(1H)-喹诺酮、吴茱萸卡品碱、1-甲基-2-[(6Z,9Z)-6,9-十五碳二烯]-4-(1H)-喹诺酮、二氢吴茱萸卡品碱8种生物碱的定量分析方法.方法 采用色谱柱为Waters symmetry C...     

反相高效液相色谱法测定吴茱萸中吴茱萸碱和吴茱萸次碱含量

目的建立测定吴茱萸中吴茱萸碱及吴茱萸次碱含量的反相高效液相色谱法。方法采用Boston Symmetrix ODS-R C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.04%辛烷磺酸钠溶液(50∶50),流速为1 mL/min,检测波长为225 nm。结果该法准确可靠,重现性好。结论反相高效液相色谱法可以同时测定吴茱萸中吴茱萸碱及吴茱萸次碱的含量。     

吴茱萸化学成分的分离与鉴定

目的对吴茱萸(Euodia rutaecarpa)的化学成分进行研究。方法采用硅胶柱色谱、ODS柱色谱、Sephadex LH-20柱色谱和制备高效液相色谱等方法对吴茱萸果实体积分数75%乙醇提取物进行分离纯化,通过波谱数据分析对化合物进行结构鉴定。结果从体积分数75%乙醇提取物中分离得到16个化合物,分别鉴定为苜蓿素-7-O-β-D-吡喃葡萄糖苷(tricin-7-O-β-D-glucopyranoside,1)、淫羊藿苷C(epimedoside C,2)、phellodensin F(3)、异鼠李素-3-O-β-D-半乳糖苷(isorhamnetin-3-O-β-D-galactoside,4)、槲皮素(quercetin,5)、儿茶素(catechin,6)、cinchonain(7)、猪毛菜碱A(salsoline A,8)、1β,4β-dihydroxyeudesman-11-ene(9)、柠檬苦素(limonin,10)、6β-乙酰氧基-5-表柠檬苦素(6β-acetoxy-epilimonin,11)、吴茱萸苦素(rutaevine,12)、吴茱萸苦素乙酸酯(rutaevine acetate,13)、吴茱萸内酯醇(evodol,14)、加洁茉里苦素(jangomolide,15)、isolimonexic acid(16)。结论化合物1、3、6~9为首次从吴茱萸属植物中分离得到。     

土茯苓中2个新化学成分

采用大孔树脂柱色谱、硅胶柱色谱、凝胶色谱及半制备液相色谱分离技术,对土茯苓Smilax glabra Roxb.乙酸乙酯部位化学成分进行了系统的分离纯化。结合分离单体化合物的理化性质,通过MS、NMR和CD等波谱数据鉴定其结构。从土茯苓乙酸乙酯部位中分离得到20个化合物,包括10个黄酮(1、3～11)、8个酚类(2、12～18)和2个呋喃衍生物(19、20),分别鉴定为土茯苓苷A (1)、土茯苓酮A (2)、5-羟基-3′,4′,7-三甲氧基二氢黄酮(3)、柚皮素(4)、槲皮素7-O-α-L-鼠李糖苷(5)、新落新妇苷(6)、新异落新妇苷(7)、异落新妇苷(8)、圣草素-7-O-β-D-吡喃葡萄糖苷(9)、柚皮素-7-O-α-L-鼠李糖苷(10)、山柰酚3-O-[4′′′′′-O-乙酰基-α-L-鼠李糖基-(1→6)]-[β-D-吡喃葡萄糖基-(1→2)]-β-D-吡喃葡萄糖苷(11)、5-羟基麦芽酚(12)、3,4,5-三甲氧基苯基-1-O-β-D-吡喃葡萄糖苷(13)、2-(3′,4′-二羟基苯基)-1,3-苯骈二氧戊环-5-甲醛(14)、kompasinol A (15)、ica...     

闽产白花鬼针草乙酸乙酯部位化学成分研究

摘要：目的:研究闽产白花鬼针草乙酸乙酯部位化学成分。方法:以硅胶色谱、Sephadex LH-20色谱及C18反相硅胶色谱分离纯化乙酸乙酯部位的化学成分,以1H-NMR、13C-NMR以及质谱鉴定化学结构。结果:从白花鬼针草乙酸乙酯部位分离、纯化、鉴定了6个黄酮类化合物,分别为:3,3’-二甲氧基槲皮素(1)、2’-羟基-4,4’-二甲氧基查耳酮(2)、山奈酚(3)、槲皮素-3-甲氧基-7-O-β-D-吡喃葡萄糖苷(4)、槲皮素-3,3’-二甲氧基-7-O-β-D-吡喃葡萄糖苷(5)、奥卡宁4’-O-β-D-(2″,4″,6″-三乙酰基)-吡喃葡萄糖苷(6)。结论:6个化合物均首次从白花鬼针草中分离得到,其中,3,3’-二甲氧基槲皮素(1)、槲皮素-3-甲氧基-7-O-β-D-吡喃葡萄糖苷(4)、槲皮素-3,3’-二甲氧基-7-O-β-D-吡喃葡萄糖苷(5) 3个化合物属于首次从鬼针草属药用植物中分离鉴定。     

酸浆宿萼的黄酮苷类化学成分

目的研究茄科植物酸浆(Physalis alkekengiL.var.francheti(Mast.)Makino)宿萼的化学成分。方法用体积分数为80%的乙醇溶液加热回流提取,回收乙醇,浓缩后用水混悬,依次用环己烷、乙酸乙酯、正丁醇萃取;将正丁醇层通过聚酰胺柱色谱,Sephadex LH-20柱色谱、反相ODS开放柱色谱及反相制备HPLC等手段,共分离得到5个黄酮苷类化合物;利用其理化性质和波谱学分析数据,鉴定化合物结构。结果分离得到的5个黄酮苷类化合物,分别鉴定为木犀草素-7-O-β-D-葡萄糖苷(1),木犀草素-4′-O-β-D-葡葡萄糖苷(2),槲皮素-3-O-β-D-葡葡萄糖苷(3),槲皮素-7,3-二-O-β-D-葡葡萄糖苷(4),木犀草素7,3′-二-O-β-D-葡葡萄糖苷(5)。结论木犀草素-4′-O-β-D-葡萄糖苷(2),槲皮素-3,7-二-O-β-D-葡萄糖苷(4),木犀草素7,3′-二-O-β-D-葡萄糖苷(5)为首次从该属植物中分离得到,槲皮素-3-O-β-D-葡萄糖苷(3)为首次从该植物中分离得到。     

沙蓬黄酮类和香豆素类化学成分的分离与鉴定

目的对蒙药沙蓬的化学成分进行研究。方法采用大孔树脂柱色谱、反复硅胶柱色谱、Seph-adex LH-20柱色谱、ODS柱色谱和半微量制备高效液相色谱分离纯化,根据ESI-MS、1H-NMR和13C-NMR等谱学数据进行结构鉴定。结果从沙蓬全草的乙醇提取物的正丁醇萃取物和乙酸乙酯萃取物中分离得到7个黄酮类和2个香豆素类化合物,分别鉴定为芦丁(quercetin-3-O-rutinoside,1)、异鼠李糖-3-O-β-D-芸香糖苷(isorhamnetin 3-O-β-D-rutinoside,2)、tamarixetin 3-O-rutinoside(3)、槲皮素-3-O-β-D-吡喃葡萄糖苷(quercetin 3-O-β-D-glucopyranoside,4)、异鼠李素-3-葡萄糖苷(isorh-amnetin-3-glucoside,5)、二氢木蝴蝶素A(dihydrooroxylin A,6)、5,2'-二羟基-6,7-亚甲二氧基二氢黄酮(5,2'-dihydroxy-6,7-methylenedioxyflavanone,7)、异莨菪亭(isoscopoletin,8)和东莨菪亭(scopoletin,9)。结论化合物3、6、7为首次从藜科植物中分离得到,化合物1、2、4、5、8、9为首次从沙蓬属植物中分离得到。     

RP-HPLC法同时测定辣椒碱透皮贴剂中4种生物碱类成分的含量

目的建立RP-HPLC法同时测定辣椒碱透皮贴剂中辣椒碱、二氢辣椒碱、降二氢辣椒碱和高二氢辣椒碱4种生物碱类成分含量。方法采用Kromasil C_(18)(250 mm×4.60 mm,5μm)柱,以乙腈-水(体积比为42∶58)为流动相,流速为1.0 mL·min~(-1),柱温为40℃,检测波长为280 nm。结果 4种生物碱类成分的色谱峰与邻近色谱峰均能良好的分离,辣椒碱和二氢辣椒碱的质量浓度分别在2.65～79.50 mg·L~(-1)(r=0.999 7)、1.08～64.80 mg·L~(-1)(r=0.999 9)内与峰面积呈良好的线性关系,平均回收率分别为100.2%(RSD=1.5%)、99.5%(RSD=0.7%)。结论该方法可用于辣椒碱透皮贴剂的质量控制。     

谷精草中1个新的黄酮苷类化合物

采用大孔吸附树脂、反相硅胶、葡聚糖凝胶及微孔树脂等多种色谱技术,从谷精草水提取中分离得到10个化合物。通过NMR、ESI-MS、HR-ESI-MS、UV、IR等波谱技术并结合化学方法对其进行了结构鉴定,分别为6-methoxyquercetin-3-O-(2′′′-vanilloyl)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (1)、丁香脂素-4′-O-β-D-葡萄糖苷(2)、芦丁(3)、1-O-feruloylglycerol (4)、邻苯二酚(5)、吐叶醇(6)、β-D-(6-O-trans-feruloyl) fructofuranosyl-α-D-O-glucopyranosied (7)、二氢阿魏酸(8)、鸟嘌呤核苷(9)和槲皮素-3-O-β-D-龙胆二糖苷(10),化合物1为新化合物,化合物2、4～10为首次从谷精草属植物中分离得到,化合物3为首次从该植物中分离得到。分子对接结果显示化合物1具有潜在的肿瘤坏死因子-α (TNF-α)转换酶抑制活性。体外抗炎活性实验证实,化合物1在1、10和100μmol·L^-1     

毛樱桃叶化学成分的分离与鉴定

目的对毛樱桃(Cerasus tomentosa(Thunb.)Wall.)叶的化学成分进行研究。方法采用减压柱色谱、Sephadex LH-20、ODS柱色谱、硅胶柱色谱以及制备高效液相色谱等方法对毛樱桃叶的体积分数为70%的乙醇提取物进行提取分离,根据理化性质及波谱数据鉴定其结构。结果从毛樱桃叶的体积分数为70%的乙醇提取物中分离得到10个化合物,分别鉴定为(+)-丁香树脂醇[(+)-syringaresinol,1]、(+)-丁香树脂醇-4-O-β-D--吡喃葡萄糖苷[(+)-syringaresinol-4-O-β-Dglucopyranoside,2]、7S,8R-二氢去氢双松柏醇(7S,8R-dihydrodehydrodiconiferyl alcohol,3),7S,8R-5-甲氧基二氢去氢双松柏醇(7S,8R-5-methoxydihydrodehydrodiconiferyl alcohol,4)、山奈酚(kaempferol,5)、槲皮素(quercetin,6)、(-)-表儿茶素[(-)-epicatechin,7)、白蜡树精-6-β-D--半乳糖苷(fraxinol-6-β-D-galactopyranoside,8)、1-(4-羟基-3-甲氧基苯基)-2-[4-(3-羟丙基)-2-甲氧基苯氧基]丙烷-1,3-二醇{1-(4-hydroxy-3-methoxyphenyl)-2-[4-(3-hydroxypropyl)-2-methoxyphenoxy]propane-1,3-diol,9}、5,5'-二甲氧基-7-氧化落叶松脂醇(5,5'-dimethoxy-7-oxolariciresinol,10)。结论化合物2、4、10为从李属植物中首次分离得到,化合物1、3、8、9为从该植物中首次分离得到。     

RP-HPLC-DAD法同时测定毛冬青根中4种三萜皂苷和2种木脂素的含量

建立了反相高效液相色谱-二极管阵列检测法同时测定毛冬青根中的6种活性成分（tortoside A,（＋）-丁香脂素,毛冬青皂苷B3,毛冬青皂苷A1,毛冬青皂苷B1,毛冬青素A）的含量。RP-HPLC分离用反相C18分析柱,流动相以乙腈-0.1%磷酸水体系梯度洗脱。流速1.0 mL/min,检测波长为210 nm。6个指标成分达到基线分离,标准曲线具有良好的线性（r2〉0.9996）,精密度、稳定性和重复性符合分析方法学要求。加样回收率范围为99.00%–104.52%。最后对15个批次的毛冬青药材含量测定结果进行了聚类分析。结果表明该方法不仅可以对毛冬青药材进行质量控制,而且也为毛冬青药材的道地性分析提供了科学依据。     

Simultaneous determination and pharmacokinetic analysis of seven alkaloids and two flavonoids from rat plasma by HPLC-DAD after oral administration of Wuzhuyu decoction

A simple and sensitive high-performance liquid chromatographic method was developed for the simultaneous determination and pharmacokinetic analysis of seven alkaloids dehydroevodiamine (DHED), 10-hydroxyrutaecarpine (HDR), evodiamine (EDM), rutaecarpine (RCP), 1-methyl-2-n-nonyl-4(1H)quinolone (MNQ), evocarpine (ECP), and dihydroevocarpine (DHE), and two flavonoids isorhamnetin-7-O-rutinoside (RIM) and diosmetin-7-O-β-d-glucopyranoside (GRD) in rat plasma after oral administration of Wuzhuyu decoction. The flow rate was kept at 1.0?ml/min and the detection wavelength was set at 300?nm. The calibration curves were linear in the range of 0.5013-30.076?μg/ml for DHED, 0.2161-21.608?μg/ml for RIM, 0.161-12.876?μg/ml for HDR, 0.2146-21.457?μg/ml for GRD, 2.0464-40.928?μg/ml for EDM, 1.0398-31.194?μg/ml for RCP, 0.5970-35.818?μg/ml for MNQ, 0.8371-20.928?μg/ml for ECP, and 0.5167-31.003?μg/ml for DHE. The precision (relative standard deviation (RSD), %) for all was less than 10% and the accuracy (relative error (RE), %) was within ±?10%. The results demonstrated that the assay had remarkable reproducibility with acceptable accuracy and precision. The lower limit of quantifications for the compounds in plasma ranged from 0.12 to 0.23?μg/ml and the lower limit of detections ranged from 0.024 to 0.076?μg/ml. This validated method has been successfully applied in the pharmacokinetics study of seven alkaloids and two flavonoids after orally administrating the Wuzhuyu decoction to rats.     

Simultaneous determination and pharmacokinetic analysis of seven alkaloids and two flavonoids from rat plasma by HPLC–DAD after oral administration of Wuzhuyu decoction

A simple and sensitive high-performance liquid chromatographic method was developed for the simultaneous determination and pharmacokinetic analysis of seven alkaloids dehydroevodiamine (DHED), 10-hydroxyrutaecarpine (HDR), evodiamine (EDM), rutaecarpine (RCP), 1-methyl-2-n-nonyl-4(1H)quinolone (MNQ), evocarpine (ECP), and dihydroevocarpine (DHE), and two flavonoids isorhamnetin-7-O-rutinoside (RIM) and diosmetin-7-O-β-d-glucopyranoside (GRD) in rat plasma after oral administration of Wuzhuyu decoction. The flow rate was kept at 1.0 ml/min and the detection wavelength was set at 300 nm. The calibration curves were linear in the range of 0.5013–30.076 μg/ml for DHED, 0.2161–21.608 μg/ml for RIM, 0.161–12.876 μg/ml for HDR, 0.2146–21.457 μg/ml for GRD, 2.0464–40.928 μg/ml for EDM, 1.0398–31.194 μg/ml for RCP, 0.5970–35.818 μg/ml for MNQ, 0.8371–20.928 μg/ml for ECP, and 0.5167–31.003 μg/ml for DHE. The precision (relative standard deviation (RSD), %) for all was less than 10% and the accuracy (relative error (RE), %) was within ± 10%. The results demonstrated that the assay had remarkable reproducibility with acceptable accuracy and precision. The lower limit of quantifications for the compounds in plasma ranged from 0.12 to 0.23 μg/ml and the lower limit of detections ranged from 0.024 to 0.076 μg/ml. This validated method has been successfully applied in the pharmacokinetics study of seven alkaloids and two flavonoids after orally administrating the Wuzhuyu decoction to rats.     

Determination of combined p-hydroxy benzoic acid preservatives in a liquid pharmaceutical formulation by HPLC

This paper describes a reversed-phase high performance liquid chromatographic (RP-HPLC) assay method for the determination of combined p-hydroxy benzoic acid (ethylparaben (EP), methylparaben (MP) and propylparaben (PP)) preservatives in a liquid pharmaceutical formulation. The chromatographic separation was achieved with potassium phosphate buffer (pH 7.05)-methanol (47.5:52.5, v/v) as mobile phase, a Spherisorb C(18) column (250 mm x 4.6mm) and UV detection at 254 nm. The analysis time was <8 min. The method was validated with respect to linearity, precision, accuracy, selectivity, specificity and ruggedness. The calibration curves showed good linearity over the concentration range of 2-140 microg/ml. The correlation coefficient were >0.9999 in each case. The relative standard deviation (R.S.D.) values for intra- and inter-day precision studies were <1%. The procedure describe here is simple, selective and is suitable for routine quality control analysis and stability tests.     

HPLC analysis of 5H-benzo[a]carbazole with antifungal activity

A sensitive and simple high-performance liquid chromatographic (HPLC) method for the assay of 6,11-dihydro-2-methoxy-5H-benzo[a]carbazole (1) and 6,11-dihydro-2-methoxy-11-[2-(1-piperidinyl)]ethyl-5H-benzo[a]carbazole (2) was developed. The procedure is based on the use of the reversed-phase high-performance liquid chromatographic (RP-HPLC) method with UV detector. Each analysis required no longer than 11 min. A linear relationship between the concentration of both the drugs and the UV absorbance at 254 nm was obtained. This linearity was maintained over the concentration ranged from 5 to 80 microg/ml. The detection limits were found to be 1.6 and 0.7 ng for compounds 1 and 2. The quantitation limits were found to be 5.3 and 2.5 ng for compounds 1 and 2, respectively. For recovery studies, several determinations were carried out. Recovery values ranged from 98 to 102.1% for compound 1 and from 98.4 to 101.6% for compound 2. Method precision was also evaluated and RSD% found was less than 2%. This method was applied without any interference from degradation products.     

Determination and pharmacokinetic study of p-hydroxyphenethyl anisate following intravenous and oral administration in rats by RP-HPLC method

In the present study, we developed a rapid and specific reversed-phase high-performance liquid chromatographic (RP-HPLC) method for the quantification of p-hydroxyphenethyl anisate (HPA), one of the main bioactive constituents of the roots and rhizomes ofNotopterygium incisumand N. franchetii, in rat plasma after an intravenous (20 mg/kg, i.v.) and an intragastrical (200 mg/kg,i.g.) administration to rats, respectively. The method involved a plasma clear-up step using liquid-liquid extraction by EtOAc, followed by RP-HPLC separation and detection. Separation of HPAwas performed on an analytical Diamonsil^TM ODS C₁₈ column with the mobile phase of MeOH–H₂O at ratios of 75:25 (v/v) for i.v. and 70:30 (v/v) for i.g. administration. The flow-rate was 1.0 mL/min, and UV detection was performed at 256 nm. The calibration curves were linear over the ranges of 0.05–5.0 μg/mL (r² = 0.9984) for i.v. and 0.5–10.0 μg/mL (r² = 0.9995) for i.g. administrationin rat plasma. The extraction recoveries were in the range of 82.01%–87.97%. The intra- and inter-day precisions were between 1.71% and 3.99%, with accuracies ranging from 91.22% to 110.5%. The absolute bioavailability of an orally administered HPA in rats was about 48.17%. The developed method was suitable for the determination and pharmacokinetic studyof HPA in rat plasma.     

RP-HPLC determination of recombinant human interferon omega in the Pichia pastoris fermentation broth

A rapid and valid reversed-phase high performance liquid chromatography (RP-HPLC) method for determination of recombinant human interferon omega (rhIFNomega) in the yeast Pichia pastoris fermentation broth was developed. The method is based on the hydrophobicity of rhIFNomega followed by RP-HPLC separation with UV detection. The chromatography analysis was performed on EC 250/4 NUCLEOSIL 300-5 C18 (250 mm x 4 mm i.d., 300 A, with a particle size of 5 microm) column. The compositions of the mobile phase A and B were 999:1 (v/v) water/TFA and 999:1 (v/v) acetonitrile/TFA at a flow rate of 1.0 ml min(-1). Detection was done by spectrophotometry at 280 nm and the column temperature was 30+/-1 degrees C. Calibration curve was linear (r=0.9986, n=7) in the range of 0.074-0.555 mg ml(-1) for rhIFNomega and the regression equation was y=2.02 x 10(6)x-1.27 x 10(5). Limit of detection for rhIFNomega was 0.053 mg ml(-1). The values of R.S.D. (%) of intra-day and inter-day precision were <5.65 and <5.68 (n=6), respectively. The R.S.D. (%) values and the average recovery rate of recovery experiment were <1.23 (n=3) and 97.97%.     

Quantification of the active ingredient quercetin of “Guangdong Wang-bu-liu-xing” by high-performance liquid chromatography with UV detection

High-performance liquid chromatography with UV detection was used for the quantification of the flavonoid quercetin, the active compound found in “Guangdong Wang-bu-liu-xing”. The method was developed and demonstrated to provide superior performance over other commonly documented methods. This HPLC assay achieved high specificity through the use of a reversed-phase C18 column eluted with a mobile phase of methanol-0.4% (v/v) phosphoric acid (50:50, v/v) over the course of 30 min. UV detection at 360 nm was used. This analytical method provided excellent precision and a mean recovery of 99%, demonstrated by repeated analysis of 11 sample groups. Because of its high performance and simplicity, this HPLC assay can be readily utilized as a practical method for the quality control of active compounds extracted from Guangdong Wang-bu-liu-xing.     

Analysis of new serotonergic anxiolytics by liquid chromatography

A simple isocratic procedure was developed for the analysis of new serotonergic anxiolytics and the related compounds in bulk materials, pharmaceutical formulations and in biological samples. The system may be applied for the assay of other serotonergic anxiolytics of related structure such as buspirone. The liquid chromatographic assay utilizes a reversed-phase C18 column, a mobile phase consisting of a mixture (55:45, v/v) of (A) buffer potassium dihydrogen phosphate (0.05 M) containing sodium lauryl sulphate (0.005 M) and (B) acetonitrile. A fluorescence detection is used with lambda ex 237 nm; lambda cm 374 nm. The accuracy, precision and sensitivity of the proposed method are established. Standard curves are linear with respect to concentration in the range 0.05-7.5 micrograms ml-1. The method also allows the separation and identification of related compounds at concentrations below 0.01%.     

膜荚黄芪毛状根中异黄酮成分的反相高效液相色谱分析

对膜荚黄芪毛状根中6种异黄酮成分进行了反相高效液相色谱法测定。色谱柱为NucleosilC₁₈柱,流动相为甲醇—水(3∶2和3∶1,v/v),检测波长为254nm和280nm。6种异黄酮成分为:10-羟基-3,9-二甲氧基紫檀烷,(3R)8,2’-二羟基-7,4’-二甲氧基异黄烷,芒柄花素(7-羟基-4’-甲氧基异黄酮),8,3’-二羟基-7,4’-二甲氧基异黄酮,2’-羟基-3’,4’-二甲氧基异黄烷-7-O-葡萄吡喃糖甙,毛蕊异黄酮(7,3’-二羟基-4’-甲氧基异黄酮)。异黄酮浓度在2.5～12.5μg范围内与峰面积有良好的线性关系;加样回收率为96.47%～103.33%;精密度试验相对标准偏差为2.57%～6.52%;测得黄芪毛状根中6种异黄酮成分的含量在0.0005%～0.0065%之间。