HPLC Determination of Captopril in Human Plasma with Pre-column Derivation and Solid-phase Extraction and Studies on Its Pharmacokinetic and Relative Bioavailability

A new pre-column derivation HPLC method with solid-phase extraction to determine captopril in human plasma was established. Derivation products were extracted by a solid-phase extraction method after the reagent, p-a-dibromoacetophenone(p-BPB), was added in the plasma samples. The samples were analyzed in a VP-ODS column with UV-detector. The calibration curve of captopril was linear within the range of 5~1000 ngmL-1 with r=0.9987, the recovery of this method was 98.652.04%, within day and between day RSD were no more than 3.4% and 8.4% respectively. To study the pharmacokinetics and the relative bioavailability of captopril tablets, two formulations of captopril tablets were given to 18 healthy male volunteers according to a randomized 2-way cross-over design with a 1-week washout period. The respective AUC0~6 , Cmax and Tmax values of the two formulations were 424.5125.7 and 439.4113.3 mghL-1; 505.9244.6 and 504.8172.2 mgL-1; 0.6620.181 and 0.5280.176 h. Results from statistics analysis showed that there were no significant difference between the AUC0~6 , Cmax and Tmax values of the two formulations, The relative bioavailability of tablets I with respect to II was 96.114.6% from AUC0~6 measurement. Bioequivalance was observed between the two tablets.

Optimal ultrasonic extraction condition and determination of polysaccharides in Tilia amurensis flowers

Seven extracting temperatures （25, 35, 45, 55, 65, 75, 85℃） and extracting time （30, 40, 50, 60, 70, 80, 90 min） were designed for selecting the optimal extracting time and extracting temperature for ultrasonic extraction of T. amurensis flowers. Polysaccharides from T. amurensis flowers were isolated and determined by spectrophotometry. Results show that the optimal ultrasonic temperature was 75℃ and ex tracting time 52 min. The content of polysaccharides in T. amurensis flowers measured by anthrone--H2SO4 colorimetry under 580 nm, was 9.74% with 0.47% of relative standard deviation （RSD, n=3）. This study demonstrated that ultrasonic extraction method was simple, and the color of the treated samples was stable in 4 h. The average recovery value for the polysaccharides measured was 99.48%±1.01%, with 0.112% of RSD （n=3）.

Extraction and RP-HPLC determination of taxol in rat plasma, cell culture and quality control samples

A rapid, sensitive, selective and validated reverse phase high-performance liquid chromatography （RP-HPLC） method for the estimation of paclitaxel in micro-sample of rat plasma and in culture of cancer cells was per- formed in this study. The mobile phase consisted of an （80： 20： 0.1, v/v/v）. Column elution at a flow rate of 1 optimized mixture of methanol：water： trifluroacetic acid mL/minute with UV detection at 225 nm at room tern- perature was used. The RP-HPLC method was successfully applied for the determination of paclitaxel in plasma samples and in culture of cancer cells with nano-quantity of estimation. The validation studies were performed in accordance with the International Conference on Harmonization （ICH） guidelines. The intra- and inter-day pre- cision showed that the coefficients of variation ranged from 1.07% to 4.27% at different levels of concentrations. To the best of our knowledge, this study also reported for the first time the optimization of different solvents for effective extraction of paclitaxel wherein tert.-butyl methyl ether （TBME）： diethyl ether （DEE） in 50：50 v/ v composition was found most efficient with extraction efficiency ranging between 77.99% and 91.74% and be- tween 76.14 and 93.66% in the plasma and cell culture, respectively. This proposed method was successfully ap- plied to study the pharmacokinetics of paclitaxel and the influence of verapamil and all-trans retinoic acid （atRA） on paclitaxel pharmacokinetics in rat models. This proposed method might emerge as a valuable aid in the labo- ratory monitoring of paclitaxel in a variety of in vitro as well as in vivo scenarios.

HPLC Determination of the Major Non-protein Amino Acids and Common Biogenic Amines in Lathyrus sativus Using a Novel Extraction Method

An assay is presented for simultaneously determining 5 biogenic amines and the major non-protein amino acids: the toxin β-N-oxalyl-L-α,β-diaminopropanoic acid (β-ODAP), its isomer α-ODAP and homoarginine in Lathyrus sativus extracts using the HPLC system after derivatization with para-nitrobenzyloxycarbonyl chloride (PNZ-Cl). However, it is more worthy of noting that this paper also describes a new extraction method using 0.2 mol/L HClO4. The new method has some advantages: shorter extraction-time, simultaneous extraction of free amino acids and polyamines, better inhibiting the isomerization of β-ODAP to α-ODAP, and so on.

Optimum of Mobile Phase Condition for Resolving Isoflavones in RP-HPLC

In reversed-phase high performance liquid chromatography （RP-HPLC）, the mobile phase condition for separating eight isoflavones （daidzin, glycitin, genistin, 6＇-o-acetyl daidzin, 6＇-o-malonyl genistin, daidzein, glycitein and genistein） was optimized using the HCI （High-Purity Separation Laboratory, Department of Chemical Engineering, Inha University） program software. The optimum composition of mobile phase for the separation of the eight isoflavones was obtained. The elution profiles were calculated by the plate theory based on the equations of retention factor, In k=A＋BF＋CF2, where F was the volume percentage of acetonitrile with 0.1% acetic acid （AA）. The first mobile phase composition was water with 0.1% AA/acetonitrile with 0.1% AA （88%/12%, by volume）, followed at 9min later by the second composition of mobile phase which was step-changed to 85%/15%, at 19rain by the third composition which was step-changed to 73%/27%, at 30min when it was changed to 65%/35% and finally it was maintained in isocratic mode to the end of the run time at 50rain. Although, using step gradient mode to separate the isoflavones, the calculated and experimented data were not achieved very good agreement, we could estimate the closed retention time before experiment. And the agreement between the experimental data and the calculated values was relatively good using isocratic separation for eight isoflavones, but the retention time is very long.

Studies on Optimal Extraction Conditions of Flavonoid in Leaves of Black Currnt

Employing the orthogonal design,the optimal extraction conditions for flavonoid in leaves of black currant were determined by colorimetric estimation,which were extraction agent of 75% EtOH-H 2O,the material ratio(ratio of weight of fresh leaves to volume of the solvent)of 1:16,the temperature of 35℃,the time of 1 h and times of three.A total extraction rate of over 97% and the flavonoid contents of 179 33 mg/100 g were obtained under the optimal extraction conditions.

Cloud point extraction coupled with HPLC-UV for the determination of phthalate esters in environmental water samples

A method based on cloud point extraction was developed to determine phthalate esters including di-ethyl-phthalate （DEP）, di- （2-ethylhexyl）-phthalate （DEHP） and di-cyclohexyl-phthalate （DCP） in environmental water samples using high-performance liquid chromatography separation and ultraviolet detection （HPLC-UV）. The non-ionic surfactant Triton X-114 was chosen as extraction solvent. The parameters affecting extraction efficiency, such as concentrations of Triton X-114 and Na2SO4, equilibration temperature, equilibration time and centrifugation time were evaluated and optimized. Under the optimum conditions, the method can achieve preconcentration factors of 35, 88, 111 and detection of limits of 2.0, 3.8, 1.0 ng/ml for DEP, DEHP and DCP in 10-ml water sample, respectively. The proposed method was successfully applied to the determination of trace amount of phathalate esters in effluent water of the wastewater treatment plant and the lixivium of plastic fragments.

Response surface optimization of microwave-assisted extraction for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii

Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved by soaking the sample with 70% methanol（1∶15,v/v）for 30 min,and then microwave irradiation for 11 min at a power of 600 W.Coupling the extraction process with HPLC-fluorescence presented good recovery,satisfactory precision,and good linear relation.Compared with a method from the Chinese Pharmacopoeia,the proposed method enables higher extraction efficiency and more accurate analytical results.It can be of potential value in quality assessment of Radix Puerariae thomsonii medicinal materials.

Solid phase extraction of methomyl and thiodicarb from environmental water with an activated carbon cartridge and their determination by HPLC

A solid phase extraction procedure of methomyl and thiodicarb from environmental water was presented. This method utilizes a 40—60 mesh activated carbon cartridge and high performance liquid chromatography (HPLC). The 40—60 mesh activated carbon works faster and yield higher adsorption efficiency. Detection limits of methomyl and thiodicarb in environmental water are 0 1 μg/L and 0 2 μg/L, respectively. Average recoveries of fortified methomyl and thiodicarb in water are in the range of 90 7%—98 8% and 88 9%—103 6%, respectively. The relative standard deviations are lower than 7%. This method is simple, rapid, accurate and precise.

Optimization of Extraction Conditions for Total Flavonoids from Fructus Aurantii Immaturus and Its Anti-UVB Radiation Activity

[Objectives]Optimum extraction conditions of total flavonoids from Fructus Aurantii Immaturus(TFFAI)and its resistance activity to ultraviolet radiation were investigated in present research.[Methods]The optimal extraction conditions of TFFAI were determined by single factor and orthogonal experiments,and the survival rate of TFFAI on HaCaT cells irradiated with UVB rays was investigated.It s antioxidant capacity was determined by ABTS.[Results]The results showed that the highest yield of TFFAI was obtained with 70%ethanol at a solid-to-liquid ratio of 1:50(w/v)and 40℃for 1.5 h by single-factor and orthogonal experiments.Total flavonoids(0.25-1.00 mg/ml)could significantly improve the survival rate of HaCaT cell line.Meanwhile,the maximum absorption peak of TFFAI was found at 283 nm,and in-vitro antioxidant experiment identified that TFFAI had a good clearance rate to ABTS.It suggestes that TFFAI could protect the cells from UVB damage by absorption of UVB rays and anti-oxidation.[Conclusions]TFFAI played a protective role on UVB irradiated cells through UVB ultraviolet absorption and antioxidant pathways.

Optimization of Extraction Conditions of Pomegranate Peel Polyphenols and Its Protective Effect on Acute Alcoholic Liver Injury in Mice

[Objectives]This study was conducted to explore the optimization of ultrasonic-assisted organic solvent extraction of pomegranate peel polyphenols(PPPs),and to study the protective effect of PPPs on acute alcoholic liver injury in mice.[Methods]The optimal extraction conditions of PPPs were determined by single factor and orthogonal experiments,and an acute alcoholic liver injury model in mice was established.Bifendate was used as the positive control group to investigate the protective effect of low,medium and high doses of PPPs on acute alcoholic liver injury.[Results]The optimum extraction process parameters were followed as 60%ethanol concentration,solid-liquid ratio of 1:40(w/v),extraction temperature of 50℃,and extraction time of 1.5 h,and the yield was 1.42%.The results of animal experiments showed that PPPs could effectively reduce the degree of alcoholic liver injury in mice,reduce the levels of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST),and reduce the inflammation and necrosis of liver tissue in mice.Meanwhile,the total polyphenols from pomegranate peel also significantly reduced the expression levels of malondialdehyde(MDA),tumor necrosis factor(TNF-α)and interleukin-6(IL-6)in mice,and increased the levels of superoxide dismutase(SOD)and reduced glutathione(GSH)in liver tissue of mice,indicating its antioxidant and anti-inflammatory effects,further illustrating its protective effect on alcoholic liver injury.[Conclusions]PPPs could reduce the expression levels of TNF-α,IL-6 and MDA in mice,and increase the expression levels of SOD and GSH to achieve the protective effect on acute alcoholic liver injury in mice.This study will provide new ideas for the development of new anti-alcoholic liver injury drug resources.

Determination of Triclocarban, Triclosan and Methyl-Triclosan in Environmental Water by Silicon Dioxide/Polystyrene Composite Microspheres Solid-Phase Extraction Combined with HPLC-ESI-MS

This paper developed a sensitive and efficient analytical method for triclocarban (TCC), triclosan (TCS) and Methyl-triclosan (MTCS) determination in environmental water, which involves enrichment by using silicon dioxide/polystyrene composite microspheres solid-phase extraction and detection with HPLC-ESI-MS. The influence of several operational parameters, including the eluant and its volume, the flow rate and acidity of water sample were investigated and optimized. Under the optimum conditions, the limits of detection were 1.0 ng/L, 2.5 and 4.5 ng/L for TCC, TCS, and MTCS, respectively. The linearity of the method was observed in the range of 5-2000 ng/L, with correlation coefficients (r2) >.99. The spiked recoveries of TCC, TCS and MTCS in water sampleswereachieved in the range of 89.5% -96.8% with RSD below 5.7%. The proposed method has been successfully applied to analyze real water samples and satisfactory results were achieved.

迷迭香提取物HPLC特征图谱研究

研究旨在运用高效液相色谱法(HPLC)建立饲用植物迷迭香提取物的特征图谱,为迷迭香提取物的辨别和质量控制提供新的方法。采用Sunfire C18色谱柱,以乙腈-0.1%甲酸水为流动相,以1.0 mL/min的流速、10µL的样品进样量、25℃的柱温、280 nm的紫外吸收波长作为液相色谱条件进行检测。结果显示:试验选取的15批迷迭香提取物色图谱中分离度、峰型较好的8个峰为特征峰,指认了迷迭香酸、鼠尾草酸、鼠尾草酚3个色谱峰,建立了迷迭香提取物的HPLC特征图谱。试验所建立的迷迭香提取物特征图谱实现了对其质量的稳定控制,可为迷迭香提取物质量评价提供参考。

Polysaccharides in Sipunculus nudus: Extraction Condition Optimization and Antioxidant Activities

Marine organisms constitute unlimited resource of bioactive substances due to their high biodiversity and represent a valuable source of new compounds. This study optimized the alkali-extraction conditions and antioxidant activities of soluble polysaccharides from the body wall of Sipunculus nudus. The effects of solid-liquid ratio, extraction duration, extraction temperature, and alkali concentration on the yield of S. nudus polysaccharides(SNP) were examined, according to which the optimal combination of extraction parameters was obtained by an orthogonal test. The relative influencing importance of different extraction parameters on the yield of SNP followed the order as solid-liquid ratio > extraction temperature > alkali concentration > extraction duration. The highest extraction yield, 1.98%, was achieved under an optimal extraction condition: temperature 60℃, solid-liquid ratio 1:6 g mL^(-1), duration 5 h, and alkali(Na OH) mass fraction 6%. The in vitro antioxidant activities examination showed that extracted SNP under this optimized condition had strong power in reducing certain hydroxyl and superoxide radical scavenging abilities. The promising results showed that extracted SNP could be a potent natural antioxidant.

Inquiry of Water-Soluble Polysaccharide Extraction Conditions from Grapefruit Skin

Water-soluble polysaccharide was isolated from grapefruit with the method of water-dissolving and ethanol-precipitating. By single factor test and orthogonal experimental method, the extraction conditions of the water-soluble polysaccharide were investigated, which contained liquid-solid ratio, temperature, time and pH. The results showed that pH was the most important factor of polysaccharide extraction, followed by temperature. The optimized extraction technology conditions were obtained. The liquid-solid ratio was 1:40, temperature was 90?C, time was 2.0 h, and pH was 2. The highest extraction rate of crude polysaccharide was 8.30%.

固相萃取-HPLC法研究格尔德霉素在Beagle犬药代动力学

目的:建立格尔德霉素在 Beagle 犬血浆中的同相萃取(SPE)-HPLC 测定方法,并研究其在犬体内的药代动力学。方法:18只 Beagle 犬分为0.25,0.5,1.0 mg·kg^(-1)低、中、高3个剂量组,单次静脉注射格尔德霉素,于给药后不同时间点采集血样。以 N-苯基-1-萘胺作为内标,Oasis HLB 同相萃取柱优化提取富集,HPLC 法测定其浓度;用药代动力学计算程序3P97拟合药物浓度-时间曲线,求算药动学参数。结果:格尔德霉素在0.01-1.0μg·mL^(-1)线性范围内,低、中、高3种不同浓度质控样品(0.01,0.1,1.0μg·mL^(-1))日内 RSD 分别为5.6%,4.6%,1.9%(n=5);日间 RSD 分别为6.9%,3.2%,6.7%(n=5)。绝对回收率分别为85.0%±3.0%,91.1%±2.2%,94.0%±0.7%(n=5);低、中、高剂量组的 T_(1/2β)分别为89.2,116.6,101.6 min;AUC 分别为6.9,12.7,24.9 min·μg·mL^(-1);Vc 分别为858.0,964.4,981.3 mL·kg^(-1);CL 分别为35.9,39.4,40.1mL·min^(-1)·kg^(-1)。结论:固相萃取-HPLC 法测定格尔德霉素准确,快速,回收率高。格尔德霉素在 Beagle 犬体内药代动力学符合二室静脉注射模型。这对以后的临床应用指导具有十分重要的意义。

离子对SPE-HPLC法检测生物检材中的百草枯

目的建立生物检材中百草枯的简便、快速、灵敏、可靠的高效液相色谱分析方法。方法生物检材酶解后,用以十二烷基三甲基溴化铵和十二烷基硫酸钠预处理过的C18固相柱萃取,HPLC/DAD进行分析。结果回收率81%～94%,检出限为1ng·mL-1,线性范围50ng·mL-1～1mg·mL-1,结论此方法适用于中毒生物检材中百草枯的检测。

Optimization of Extraction Conditions of Some Phenolic Compounds from White Horehound (<i>Marrubium vulgare</i>L.) Leaves

This research was aimed to optimize the extraction conditions of three phenolic compounds: total phenolics, flavonoids and condensed tannins, from White Horehound’s leaves (Marrubium vulgare L.). Distilled water and different organic solvents such as: methanol, ethanol and acetone, were used, with various concentrations (20% - 80%, v/v), temperatures (20&deg;C - 60&deg;C) and extraction times (30 - 450 min). Results showed that the maximum total phenolics amounts (293.34 ± 14.60 mg gallic acid equivalent/g dry weigh), were obtained with 60% aqueous methanol at 25&deg;C for 180 min;total flavonoids (79.52 ± 0.55 mg catechin equivalent/g dry weigh) with 80% aqueous methanol at 20&deg;C for 450 min, and condensed tannins (28.15 ± 0.80 mg catechin equivalent/g dry weigh) with 60% aqueous acetone at 50&deg;C and for 180 min. ANOVA test showed the significant effect (***P < 0.001) of the extraction conditions tested on phenolic compounds. The Principal Component Analysis (PCA) exhibited the positive effect of low temperatures on total phenolics and flavonoids extraction, and the effect of high temperatures on the condensed tannins extraction. The Response Surface Methodology (RSM) provided predicted values of extraction conditions and maximum polyphenols amounts similar to those obtained experimentally.

蒲公英的乙酸乙酯部位HPLC指纹图谱定性分析和咖啡酸的定量测定

目的 运用指纹图谱技术建立蒲公英的色谱定性和定量方法。方法 采用RP -HPLC方法 ,HypersilBDSC18分析柱 (5μm ,2 5 0mm× 4 .6mm) ;流动相A为乙腈 ,流动相B为磷酸盐缓冲液 (磷酸调pH为 3.7) ;梯度洗脱时间为 0、38、4 0min时 ,A(% )分别为 5 %、75 %、75 % ;柱温 35℃ ;流速 0 .7ml·min-l;检测波长 32 3nm。对蒲公英的乙酸乙酯部位进行指纹图谱定性分析 ,并对有效成分咖啡酸进行定量测定。结果 在色谱条件下 ,11份不同蒲公英药材中 ,乙酸乙酯部位的色谱指纹图谱中可检出 11个相对位置稳定的共有峰 ,通过与标准品的保留时间及紫外光谱比较 ,3号峰和 6号峰分别鉴定为咖啡酸和阿魏酸 ,蒲公英药材中咖啡酸的含量为 0 .0 15 %～ 0 .0 4 0 %。结论 蒲公英乙酸乙酯部位HPLC指纹图谱定性和咖啡酸的定量分析方法具有较强的针对性和准确性 。

HPLC法同时测定健腰密骨片浸膏粉中5种指标成分

目的:建立健腰密骨片浸膏粉的多成分定量分析方法,为评价浸膏粉中间体的质量提供依据。方法:采用HPLC法,以乙腈-磷酸盐缓冲液为流动相,梯度洗脱,测定健腰密骨片浸膏粉中5种成分的含量。结果:木兰花碱、丹酚酸B、朝藿定C、淫羊藿苷和宝藿苷Ⅰ5种成分在各自质量浓度范围内呈良好线性关系,r≥0.9997;5种化合物的平均回收率分别为100.47%、104.37%、95.38%、95.93%、96.66%。该方法的专属性、重复性均良好。结论:该分析方法准确、重复性好,可作为健腰密骨片浸膏粉的质量控制方法。