Pharmacokinetic Study of Ipriflavone in Healthy Volunteers by HPLC and Determination of the Relative Bioavailability of Its Tablet Preparation

Two kinds of Ipriflavone tablets (IP) were orally cross administered to 8 healthy volunteers. The serum level of IP at different time was determined by HPLC, 3P87 pharmacokinetic package was used in the analysis of the time course of serum concentration and it had shown that the data well fitted a 2 compartment model. Pharmacokinetic parameters and area under curve (AUC) were calculated. The relative bioavailability of Tabellae IP made in China vs that made in Japan (as a reference) was 1.003( P >0.05). The values of AUC T max and C max of the two preparations were also comparable by the statistical analysis, so they were bioequivalent.

Simultaneous Determination of Four Pesticide Residues in Fruit Juice by HPLC

[Objective] This study was conducted to develop a system for simultaneous determination of imidacloprid, diflubenzuron, thiabendazole and carbendazim in fruit juice by HPLC. [Method] Using acetonitrile as the extraction solvent, the pesticides in fruit juice were purified through a NH2 solid phase extraction （SPE） cartridge, then detected by HPLC. [Result] There was a good linear relationship between the peak area and the concentrations of imidacloprid, diflubenzuron, thiabendazole and carbendazim in a range of 0.05-5.0 μg/ml, and the linear correlation coefficient varied in a range of 0.999 0-0.999 8; the limit of detection for imidacloprid, diflubenzuron, thiabendazole and carbendazim was 0.003, 0.005, 0.003 and 0.007 mg/kg, respectively. The recovery rate of imidacloprid, diflubenzuron, thiabendazole and carbendazim standards added at three levels （0.1, 0.5 and 1.0 mg/kg） ranged from 82% to 107%, with RSD less than 4.5%. [Conclusion] The sensitivity, accuracy and precision of this method were able to meet the requirements for pesticide residue analysis.

Simultaneous Determination of Ticlopidine Hydrochloride and Nitrendipine in a New Tablet Formulation by HPLC

An accurate, simple and sensitive high performance liquid chromarographic (HPLC) method for simultaneous determination of ticlopidine hydrochloride and nitrendipine in a new tablet formulation is described. Chromatographic separation of ticlopidine hydrochloride and nitrendipine was achieved on a Hypersil BDS C18 column using a mobile phase consisting of acetonitrile-methanol-10 mmol/L ammonium acetate (60∶10∶30(V/V), pH 6.5) at a flow rate of 1.0 mL/min. Absorbance was monitored at 236 nm where both drugs have significant absorption. The proposed method was validated with respect to linearity, selectivity, accuracy, precision, and limits of detection and quantitation. The linear ranges for ticlopidine hydrochloride and nitrendipine were found to be 75-750 μg/mL and 1-10 μg/mL, respectively. The mean recoveries were 100.1%(S R=0.6%,n=9) for ticlopidine hydrochloride and 99.9%(S R=0.7%,n=9) for nitrendipine. The within-day precision and between-day precision for ticlopidine hydrochloride and nitrendipine were 0.63% and 0.89%, and 0.74% and 1.0%, respectively. The proposed HPLC method can be used for the simultaneous determination of both drugs in pharmaceutical preparations.

SIMULTANEOUS DETERMINATION OF VITAMIN B COMPLEX (THIAMINE, RIBOFLAVIN, NIACIN, PYRIDOXINE AND FOLIC ACID) IN FOODS BY HPLC

A high performance liquid chromatographic assay has been developed for the simultaneous determination of five water soluble vitamins:thimine, riboflavin, niacin, pyridoxine and folic acid in cereal products and fresh vegetables. Food samples were hydrolyzed in 0.4 mol/L HCl, autoclaved at 120℃ 15 psi for 20 minutes, using a μ-Bondapak column (3.9×300mm, Waters Co.), a mobile phase of methanol-water (30:70) (0.005 mol/L heptanesulfonic acid) and a flow rate of 1.0 ml/min gave the most satisfactory separation of the five water soluble vitamins. A double channel deteetion was used: four vitamins (B_5, folic acid, B_2, B_1) were detected hy UV spectrophotometry (254 nm) first, pyridoxine (B_6) was detected by fluoromelry (EX 290nm, EM 395 nm) afterwards. Detection limits were 2, 5, 5, 2and 5ng, linear ranges were 5-10ng, 5-50ng, 5-40ng, 5-50ng and 10-50ng for B_1, B_2, B_5, B_6 and folic acid respectively. Recoveries were 92-100%(B_5), 51-52% (folio acid), 103—105% (B_2), 99—100% (B_6) and 91.3—102% (B_1) respectively. In comparison with a reference method and checking with food composition tables, very satisfactory results were obtained by this method.

Simultaneous determination of ten components in Dianxiankang Capsules by HPLC QAMS

Objective:To establish an high performance liquid chromatography-quantitative analysis of multi components by single marker(HPLC-QAMS)method for simultaneous determination of gastrodin,parishin E,parishin B,parishin,tenuifolin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andαasarone in Dianxiankang Capsules.Methods:Waters Symmetry C_(18)column was used with acetonitrile-0.05%phosphoric acid solution as mobile phase for gradient elution.Multiwavelength switching detection.The contents of gastrodin,parishin E,parishin B,parishin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andα-asarone were calculated by relative correction factor.At the same time,the contents of 10 components in 12 batches of Dianxiankang Capsules were determined by external standard method(ESM).Results:An HPLC-QAMS method was established tenuifolin as the internal reference substance was established.The relative correction factors of gastrodin,parishin E,parishin B,parishin,onjisaponin B,methylophiopogonanone A,methylophiopogonanone B,β-asarone andα-asarone were 0.8238,0.7239,1.0229,1.1881,0.7272,1.3108,0.9314,0.6549 and 1.0572,respectively.The relative correction factors had good repeatability and no significant difference with ESM(P>0.05).Conclusion:HPLC-QAMS can be used for simultaneous determination of multi-index components in Dianxiankang Capsules.

SIMULTANEOUS DETERMINATION OF FAT SOLUBLE VITAMINS A, D_3, E AND K_1 IN FORTIFIED MILK POWDERS BY HPLC

A high performance liquid chromatographic method for the simultaneous determination of four fat soluble vitamins: retinol (vitamin A), cholecalciferol(vitamin D), tocopherol(vitamin E) and phylloquinone(vitamin K_1) in fortified milk powders and egg yolk has been developed. The method requires enzymatic hydrolysis of lipid component of the sample as a pretreatment. Several factors which influence the enzymatic hydrolysis were studied-Separation was achieved using μ-Bondapak C-18 column(3.9×300mm), 98% methanol as mobile phase, a double channel detection was selected; vitamins D_3 E_1 K_1 were detected by UV spectrophotometry (265 nm) first, then vitamin A by fluorometry (EX 325nm, EM 480mm). The retention times of vitamin A_1 D_3, E and K_1 4.87, 9.00, 10.58 and 15.45 min respectively. Detection limit were 0.64, 0.25, 0.50 and 0.07 ng; and the recoveries were 90.5%～103.6% 90.0%～95.6%, 91.7%～98.8%, 91.5%～98.6%, respectively. The vitamins A, D_0, E, K_1 contents in foods were determined satisfactorily.

Development of RP-HPLC method for simultaneous determination of docetaxel and curcumin in rat plasma: Validation and stability

The purpose of the present research was to develop a suitable, simple, precise, accurate,robust, and reproducible RP-HPLC method for a reliable simultaneous quantification of docetaxel(DTX) and curcumin(CCM) in rat plasma samples using paclitaxel(PTX) as an internal standard. The samples were assayed by the Agilent 1260 Infinity HPLC instrument using a Capcell Pak C8 column(4.6 mm × 150 mm, 5 μm) under isocratic conditions. The mobile phase consisted of acetonitrile and triple distilled water(40/60, v/v) with a flow rate of 1.0 ml/min. The eluent was monitored at 230 nm for simultaneous measurement of curcumin and docetaxel. The method was validated by determining system suitability, selectivity, sensitivity, linearity, inter-day and intra-day precision, accuracy, robustness, and stability in accordance with the guidelines of the United States Food and Drug Administration(FDA).The developed chromatographic method proved to be simple, precise, accurate, robust and reproducible. Moreover, the samples showed stability at room temperature over a period of 48 h. Thus, this method would be employed for routine simultaneous quantification of docetaxel and curcumin in rat plasma samples.

RP-HPLC Method for the Simultaneous Determination of Lisinopril and NSAIDs in API, Pharmaceutical Formulations and Human Serum

High performance liquid chromatographic method was developed valdated and applied for the simultaneous determi- nation of lisinopril and NSAIDs in bulk, pharmaceuticals formulations and human serum. A Purospher star C18 (5 μm, 25 × 0.46 cm) column was used with mobile phase consisting of methanol: water: acetonitrile (80:17.5:2.5 v/v, pH 3.0) and quantitative evaluation was performed at 225 nm with a flow rate of 1.0 mL?min–1. The retention time of lisinopril was 2.2 min while naproxen, flurbiprofen, diclofenac sodium and mefenamic acid were found to be 4.0, 4.5, 5.0 and 6.7 min respectively. Suitability of this method for the quantitative determination of the drugs was proved by validation in accordance with the requirements laid down by International Conference on Harmonization (ICH) guidelines. The method is selective, precise, accurate and can be used for analysis of pharmaceutical preparations in quality control and clinical laboratories.

Content Determination of Astragaloside Ⅳ in Astragalus from Three Different Regions by HPLC

[Objectives] The content of astragaloside in Astragalus membranaceus(Fisch.) Bge.var.mongholicus(Bge.) Hisao from three different regions was determined.[Methods] Referring to the method recorded in the Chinese Pharmacopoeia(2015 edition),the content of astragaloside IV in A.membranaceus was determined by HPLC.[Results] There were great differences in the astragaloside IV content of A.membranaceus among different regions.The content of astragaloside IV in A.membranaceus cultivated in Inner Mongolia was highest(0.155%),followed by that(0.143%) in A.membranaceus cultivated in Gansu,and the content of astragaloside IV in A.membranaceus cultivated in Shanxi was lowest(0.080%).The contents of astragaloside IV in A.membranaceus from different regions were all in line with the standard(not less than 0.040%) of Chinese Pharmacopoeia(2015 edition).[Conclusions]The content of astragaloside IV in A.membranaceus cultivated in three different regions met the medicinal standards.

Study on HPLC Fingerprint and Simultaneous Determination of Three Active Components of Dipsaci Radix Collected in Different Time

[Objectives] To establish methods of HPLC fingerprint and simultaneous determination of the three bioactive components(namely, asperosaponin VI, loganin and sweroside) of Dipsaci Radix, and explore the quality situation of this crude medicine collected in various months in autumn. [Methods] The analysis was performed on Thermo BDS Hypersil C_(18)(4.6 mm×250 mm, 5 μm) column with a mixture of acetonitrile and 0.1% phosphoric acid as mobile phase in gradient mood at a flow rate of 1.0 mL/min, the column temperature was set at 30 ℃, and the detection wavelength was 220 nm. [Results] The analysis methods for HPLC fingerprint and determination of the three components in Dipsaci Radix had been established. The results showed that 12 batches of samples, which were collected in four different places from September to November, possessed the similarities of greater than 0.976. Through the quantitative analysis of asperosaponin VI, loganin and sweroside in Dipsaci Radix, it was found that the quality variation of this herbal medicine and the different collected months of autumn showed a low correlation. [Conclusions] The established methods of HPLC characteristic fingerprint and simultaneous determination of three glycosides provided a new way for quality analysis of Dipsaci Radix. It was preliminarily indicated that collecting this plant medicine in different months of autumn would not affect its quality.

Simultaneous Determination of Six Constituents in Cangbaiqutong Capsule by HPLC Wavelength Switching Technique

[Objectives] To establish a high performance liquid chromatography (HPLC) wavelength switching method for the simultaneous determination of content of six constituents (phellodendrine chloride, gentiopicrin, paeoniflorin, tetrandrine, berberine hydrochloride and paeonol) in Cangbaiqutong capsules, and provide a scientific basis for the comprehensive evaluation of the quality of Cangbaiqutong capsule.[Methods] The chromatographic column of Waters XSELECT CSH-C 18 (4.6 mm × 150 mm, 5 μm), the mobile phase of acetonitrile-0.1% phosphate solution, gradient elution (0-15 min,10%-18% A;15-30 min,18%-50% A;30-35 min, 50%-10% A);the flow rate of 1.0 mL/min, wavelength switching of 284 (0-7 min, phellodendrine), 274 (7-10 min, gentiopicrin), 230 (10-14 min, paeoniflorin) and 274 nm (14-35 min, tetrandrine, berberine hydrochloride, paeonol), the injection volume of 10 μL.[Results] There was a good linear relationship between the area of chromatographic peak and the injection volume of phellodendrine chloride, gentiopicrin, paeoniflorin, tetrandrine, berberine hydrochloride and paeonol in the range of 0.150-1.504, 0.768-7.680, 1.096-10.960, 0.220-2.200, 0.296-2.956, 0.0345-0.345 μg, respectively;the average recovery rates ( n =6) were 98.3%, 99.2%, 98.8%, 98.8%, 99.1% and 98.2%, respectively;the RSD value was 1.32%, 1.46%, 1.08%, 1.31%, 1.26% and 1.21%, respectively.[Conclusions] The method can be used to determine many kinds of constituents at the same time, and the operation is simple, accurate and reproducible, and can be used for the quality control of compound Cangbaiqutong capsules.

Development of a novel stability indicating RP-HPLC method for quantification of Connexin43 mimetic peptide and determination of its degradation kinetics in biological fluids

Connexin43 mimetic peptide （Cx43MP） has been intensively investigated for its therapeutic effect in the management of inflammatory eye conditions, spinal cord injury, wound healing and iscbemia-induced brain damage. Here, we report on a validated stability-indicating reversed-phase high performance liquid chromatography （RP-HPLC） method for the quantification of Cx43MP under stress conditions. These included exposure to acid/base, light, oxidation and high temperature. In addition, the degradation kinetics of the peptide were evaluated in bovine vitreous and drug-free human plasma at 37℃. Detection of Cx43MP was carried out at 214 nm with a retention time of 7.5 min. The method showed excellent linearity over the concentration range of 0.9-250μg/mL （R2a0.998）, and the limits of detection （LOD） and quantification （LOQ） were found to be 0.90 and 2.98 μg/mL, respectively. The accuracy of the method determined by the mean percentage recovery at 7.8, 62.5 and 250μg/mL was 96.79%, 98.25% and 99.06% with a RSD of〈 2.2%. Accelerated stability studies revealed that Cx43MP was more sensitive to basic conditions and completely degraded within 24 h at 37℃（0% recovery） and within 12 h at 80℃ （0.34% recovery）. Cx43MP was found to be more stable in bovine vitreous （t1/2 slow= 171.8 min） compared to human plasma （t1/2slow = 39.3 min） at 37℃ according to the two phase degradation kinetic model. These findings are important for further pre-clinical development of Cx43MP.

RP-HPLC method for the simultaneous determination of daidzein,genistein and formonetin in Solanum Lyratum Thunb

A rapid method for the simultaneous determination of daidzein,genistein and formonetin in solanum Lyratum Thunb by high performance liquid chromatography(HPLC)was developed.Separation was achieved on a Diamonsil C18 column(250 mm×4.6 mm,5 μm)with isocratic elution,using a mobile phase of methanol-tetrahydrofuran-water(44∶3∶53,v/v).The wavelength was set at 260 nm and column was maintained at 35 ℃.The linear ranges of daidzein,genistein and formonetin were 1.0-40.0,0.1-4.0 and 0.1-4.0 μg/mL,respectively.The average recoveries were between 98.4% and 101.3%.This method could be used for the quality control of Solanum lyratum Thunb due to its simplification,reliability,rapidity and excellent precision.

Simultaneous Determination of Four Ingredients in Guanxin Danshen Tablets by RP-HPLC

[Objectives]This study aimed to establish a method for simultaneous determination of four ingredients in Guanxin Danshen tablets by RP-HPLC.[Methods]HPLC chromatography was adopted with column of Thermo Hypersil-Keystone C 18 column(4.6 mm×250 mm,5μm),mobile phase of acetonitrile(A)-0.03 mol/L ammonium acetate(pH adjusted to 2.4 with formic acid)(B),gradient elution(0-5 min,5%A;5-10 min,5%A→19%A;10-40 min,19%A;40-68 min,19%A→36%A;68-90 min,36%A→95%A),flow rate of 1.0 mL/min and column temperature of 25℃.[Results]The content of salvianic acid A sodium,protocatechuic aldehyde,salvianolic acid B and tanshinone II A showed good linear relationship with chromatographic peak area in the range of 3.310-18.66,0.03950-0.2370,0.7500-4.500,0.05920-0.3550μg,respectively.The recovery rate(n=6)was 101.75%,96.86%,104.15%and 99.03%,respectively,and the RSD was 1.52%,2.81%,1.80%,and 1.37%respectively.The established method has good precision,reproducibility and stability.[Conclusions]This method can be used for the quality control of multiple ingredients of Guanxin Danshen tablets.

Simultaneous determination of five diterpenoid alkaloids in Herba Delphinii by HPLC/ELSD

A HPLC-ELSD method was developed and validated for simultaneous determination of five Hetisane-type diterpenoid alkaloids in a Tibetan traditional herbal medicine, ＂Gebu Dilu＂ （Herba Delphinii）, using a Kromasil C18 column （250 mmx 4.6 mm, 5 gm） with the mobile phase consisting of acetonitrile and 0.1% triethylamine in gradient （detected by evaporative light scattering detector）. The linear ranges of five compounds were determined and method validation was evaluated completely. The established method is rapid and accurate with high repeatability, and can be applied for the quality control of Herba Delphinii.

Determination of Phytoene Content in Tomato Ketchup by HPLC

[Objective] The aim was to establish an HPLC method for determination of phytoene content in tomato ketchup. [Method] The chromatographic conditions were as follows： column, Agilent AT C18 （250 mm×4.6 mm, 5 μm）; mobile phase, methanol-THF （75：25）; detection wavelength, 287 nm; flow rate, 1.0 ml/min; column temperature, 30 ℃; sample size, 50 μl. [Result] There was a good linear relation- ship in the phytoene content range of 0.186-1.116μg. The average recovery rate was 103.8% with RSD of 1.47%. The phytoene content in the tomato ketchup sample was determined as 10.7 rag/100 g simple, accurate, rapid and reliable, and phytoene content in tomato ketchup. [Conclusion] The established method is it can be used for the determination of

HPLC法同时测定脑脉泰胶囊中12种成分的含量

目的建立HPLC法同时测定脑脉泰胶囊中3′-羟基葛根素、2-羟基红花黄色素A、葛根素、葛根素芹菜糖苷、3′-甲氧基葛根素、大豆苷、二苯乙烯苷、木犀草苷、染料木苷、木犀草素、丹酚酸B、迷迭香酸的含量。方法分析采用KromasilC_(18)色谱柱(250mm×4.6mm,5μm);流动相乙腈-水(含1.0g/L磷酸),梯度洗脱;体积流量0.8mL/min;柱温30℃;检测波长280nm。再进行聚类分析、主成分分析。结果12种成分在各自范围内线性关系良好(r>0.9990),平均加样回收率97.07%~98.30%,RSD0.86%~1.82%。10批样品聚为4类,4种主成分累积方差贡献率达86.262%。结论该方法简便稳定,可靠准确,可为脑脉泰胶囊的质量控制提供科学依据。

引火汤冻干粉HPLC指纹图谱建立及8种成分含量测定

目的建立引火汤冻干粉HPLC指纹图谱,并测定地黄苷D、麦冬甲基黄烷酮A、毛蕊花糖苷、原儿茶酸、梓醇、五味子醇甲、五味子乙素、五味子甲素的含量。方法指纹图谱建立采用Supersil AQ18色谱柱(4.6 mm×250 mm,5μm);流动相乙腈-0.1%磷酸,梯度洗脱;体积流量1.0 mL/min;柱温30℃;检测波长230 nm。UPLC-MS/MS含量测定采用Alphasil VC-C_(18)色谱柱(2.1 mm×100 mm,2.5μm);流动相乙腈-0.1%甲酸,梯度洗脱;体积流量0.3 mL/min;柱温30℃;电喷雾离子源;正负离子扫描;多反应监测模式。结果10批冻干粉指纹图谱中有17个共有峰,相似度大于0.90,指认出毛蕊花糖苷、水晶兰苷、去乙酰车叶草苷酸、五味子醇甲。8种成分在各自范围内线性关系良好(R^(2)>0.9906),平均加样回收率98.7%~101.1%,RSD 2.0%~5.2%。结论指纹图谱结合含量测定能完整表征引火汤基准样品质量,从而为其关键化学属性质量评价提供参考。

基于HPLC和UHPLC-MS/MS方法探究植物生长调节剂对白术品质的影响

目的:通过比较白术内在成分含量的变化及检测农药残留,探究植物生长调节剂的使用对白术品质产生的影响。方法:通过田间试验分别喷施不同浓度多效唑(A组)、矮壮·甲哌鎓(B组),并设计空白对照组(C组),收集样品;建立HPLC方法,同时测定白术中白术内酯Ⅰ、Ⅱ、Ⅲ及苍术酮的含量,比较A组、B组与C组的成分含量差异;建立UHPLC-MS/MS方法,同时检测白术中多效唑、矮壮素等8种植物生长调节剂的残留量,分析白术样品的残留情况。结果:HPLC测定白术内酯Ⅰ、Ⅱ、Ⅲ和苍术酮的含量,结果显示A组和B组的白术内酯Ⅰ、Ⅱ、Ⅲ含量均降低,且部分样品含量降低差异有统计学意义(P<0.01),A组的苍术酮含量先升高后降低,当多效唑浓度最高时,样品A5的苍术酮含量最低,且与C组比较,差异有统计学意义(P<0.01);B组的苍术酮含量变化程度不一。UHPLC-MS/MS检测结果显示,样品共检测出3种植物生长调节剂,分别为矮壮素、多效唑和烯效唑。白术A组多效唑残留量在2.81~246.67μg/kg;白术B组中矮壮素残留量在1.36~16.13μg/kg。白术A组多效唑的残留量和B组中矮壮素的残留量显著高于C组,且随着浓度的增加,残留量呈增大趋势。结论:白术种植过程中使用植物生长调节剂不但降低白术内在质量,还会使药材出现一定程度的有害残留,降低白术药材的安全性、有效性,进而可能会影响其临床用药,因此建议科学监管植物生长调节剂在白术种植过程中的使用。

HPLC-UV法同时测定赤血康脉胶囊中五种成分

本研究旨在建立HPLC-UV法同时测定赤血康脉胶囊中芍药苷、毛蕊异黄酮葡萄糖苷、β-蜕皮甾酮、阿魏酸和党参炔苷的含量。采用DiamonsilR C_(18)(250 mm×4.6 mm,5μm)色谱柱;流动相为乙腈-0.1%磷酸(梯度洗脱);流速为1.0 mL/min;柱温为35℃;检测波长为268 nm,进行分析。结果显示,5种化学成分在各自范围内线性关系良好(r>0.9995);精密度、稳定性、重复性试验结果的RSD均小于2.0%,平均加样回收率99.35%~101.59%,RSD 0.95%~2.69%。该含量测定方法简单可靠、重复性好,操作便捷,适用于同时测定赤血康脉胶囊中五种成分的含量。