A comparative study using HPLC and packed column supercritical fluid chromatography for the assay of three anti-psychotic dosage forms

A reproducible and selective method was developed for the analysis of three anti-pschycotics, i.e. haloperidol, trifluoperazine and trihexyphenidyl in bulk and dosage forms using packed column supercritical fluid chromatography (SFC). The analytes were resolved by elution with supercritical fluid carbon dioxide doped with 16.67% (v/v) methanol containing 0.8% isopropylamine. Parallel studies were performed by HPLC using ion pairing reagent and a comparison is discussed. The method was successfully used for the assay of three formulations containing a combination of: (1) haloperidol-trihexyphenidyl; (2) haloperidol-trifluoperazine; (3) trifluoperazine-trihexyphenidyl.     

A simultaneous packed column supercritical fluid chromatographic method for ibuprofen, chlorzoxazone and acetaminophen in bulk and dosage forms

A reproducible and efficient method for the separation and estimation of ibuprofen, chlorzoxazone and acetaminophen has been developed using packed column supercritical fluid chromatography (SFC). The separations were performed on an ODS-RP JASCO column employing methanol modified supercritical fluid CO(2) as the mobile phase. The densities and polarities of the mobile phase were optimised from the effects of pressure, temperature and modifier concentration on retention times. In addition a flow programming of the mobile phase helped to obtain better resolution and a faster elution for acetaminophen. The analytes were detected using a uv detector at 254 nm. The study includes a successful attempt at quantitation of the 3 drugs. Chromatographic figures of merit, linear dynamic range, limit of quantitation (LOQ), precision and accuracy etc. were determined to assess the viability of the method. The method has been extended to commercial dosage forms containing all 3 drugs.     

Performance comparison between packed column supercritical fluid chromatography (SFC) and HPLC using cyclandelate as the model analyte

A reproducible and fast method has been developed for the assay of cyclandelate in bulk and drug forms using packed column supercritical fluid chromatography using dicyclohexyl phthalate (DCHP) as internal standard. The drug and the internal standard were resolved by elution with supercritical fluid carbon dioxide doped with 14.29% (v/v) methanol on an RP-C₁₈ column and detected spectrophotometrically at 228 nm. Chromatographic figures of merit using C₈, C₁₈, cyano and phenyl columns have been assessed. Parallel experiments have been performed by HPLC and the data have been compared. Supercritical fluid extraction using CO₂ modified with a small amount of methanol was found to give quantitative analytical recoveries of cyclandelate from a dosage form. SFC has been shown to be a viable, faster alternative technique to HPLC generating less disposable waste.     

Performance comparison between packed column supercritical fluid chromatography (SFC) and HPLC using cyclandelate as the model analyte

A reproducible and fast method has been developed for the assay of cyclandelate in bulk and drug forms using packed column supercritical fluid chromatography using dicyclohexyl phthalate (DCHP) as internal standard. The drug and the internal standard were resolved by elution with supercritical fluid carbon dioxide doped with 14.29% (v/v) methanol on an RP-C₁₈ column and detected spectrophotometrically at 228 nm. Chromatographic figures of merit using C₈, C₁₈, cyano and phenyl columns have been assessed. Parallel experiments have been performed by HPLC and the data have been compared. Supercritical fluid extraction using CO₂ modified with a small amount of methanol was found to give quantitative analytical recoveries of cyclandelate from a dosage form. SFC has been shown to be a viable, faster alternative technique to HPLC generating less disposable waste. Received: 20 June 1997 / Revised: 20 October 1997 / Accepted: 26 October 1997     

Application of packed column supercritical fluid chromatography to the simultaneous determination of seven anticonvulsant drugs

Studies of speed, resolution, and selectivity have shown that packed column supercritical fluid chromatography (PCSFC) is a viable technique for the isocratic, isothermal and isobaric separation of seven anticonvulsants, viz., phenobarbitone, phenytoin sodium, phethenylate sodium, nitrazepam, clonazepam, carbamazepine, and primidone, and their simultaneous estimation. The drugs were eluted from a JASCO, RP-C₁₈ (250×4.6 mm) 10 μ packed column with a binary mobile phase of carbon dioxide and methanol, using ibuprofen as the internal standard. The effect of pressure, temperature, modifier concentration, and the rate of flow of CO₂ on retention and selectivity of all the analytes were studied and the parameters optimised. Without methanol in the mobile phase none of the solutes eluted. Changing modifier concentration was the most effective physical parameter for changing retention and selectivity. The analytes were detected using a UV detector at 215 nm. An arbitrary mixture of eight components was baseline resolved in 7 min. The study includes a successful attempt at quantification of the drugs. Chromatographic and analytical figures of merit have been listed. The present work holds promise for a possible replacement of HPLC with SFC for the separation and assay of drugs of different families.     

Using subcritical/supercritical fluid chromatography to separate acidic,basic, and neutral compounds over an ionic liquid-functionalized stationary phase

We packed an ionic liquid (IL)-functionalized stationary phase – based on 1-octyl-3-propylimidazolium chloride covalently bounded to silica gel – into a 3.2 mm × 250 mm column for the simultaneous separation of acidic, basic, and neutral compounds using carbon dioxide subcritical/supercritical fluid chromatography (SFC), and examined the effects of the pressure, temperature, co-solvents, and additives on the retention behavior of the analytes. The model compounds tested for SFC separation are acetaminophen, metoprolol, fenoprofen, ibuprofen, naphthalene, and testosterone. The data indicate that hydrogen-bonding and hydrophobic interactions between the analytes and the IL-modified stationary phase seem to involve in the separation process. Simultaneous separation of acidic, basic, and neutral compounds via SFC was successful at a co-solvent content of 20% MeOH, a pressure of 110 bar, and a column temperature of 35 °C. The relative standard deviations of the retention times and peak areas at 50 ppm were all less than 4 and 8% (n = 6), respectively.     

基于巯基-烯点击化学法的β-环糊精固定相多模式色谱保留行为研究

基于巯基硅胶与单取代-6A-烯丙氨基-β-环糊精的巯基-烯点击化学反应,制备了β-环糊精(Click TE-CD)共价键合固定相。元素分析结果表明β-环糊精被成功键合到硅胶表面。以黄酮苷类化合物为模型,考察了Click TE-CD固定相在亲水、反相和超临界流体色谱等分离模式下的色谱保留行为。黄酮苷类化合物保留时间随流动相中乙腈含量的变化呈现典型的U型曲线,表明Click TE-CD固定相具有亲水/反相的双重保留特性。应用几何学方法测得Click TE-CD固定相在反相/亲水、亲水/超临界、反相/超临界混合模式下的正交性分别为69.8%、50.8%、50.8%。对比复杂中药样品降香提取物在反相、亲水、超临界等模式下的分离情况,结果表明Click TE-CD固定相在分离中药复杂样品方面具有极大潜力,可以在一根色谱柱上通过分离模式的改变,实现二维液相色谱的分离。Click TE-CD固定相不同分离模式的分离性能和较好的正交性表明该固定相具有在液相色谱方法发展和二维液相色谱分离方面应用的潜力。     

Characterization and classification of stationary phases in HPLC and SFC – a review

Packed column supercritical fluid chromatography (pSFC) is an attractive technique in drug discovery related analysis because it offers several advantages over the more commonly used high-performance liquid chromatography (HPLC) technique. The environmental-friendly CO₂ mobile phase, the high-throughput capacity, the increased efficiency and the lower operational costs give SFC additional benefits over HPLC in analysis related to drug development. The latter technique is well established and has been used for decades in the pharmaceutical industry. On the other hand, SFC is still in its infancy, even though the technique has been known for decades and researchers are still discovering the possibilities and limitations of this technique.     

Supercritical fluid chromatographic solvatochromic modifier polarity studies

To understand the chromatographic process as a whole, whether it be for gas chromatography (GC), liquid chromatography (LC), or supercritical fluid chromatography (SFC), one needs to know the chemical and physical nature of the mobile and stationary phases and also the interactions that take place between analytes (solutes) and the two phases. An approach towards Investigating the ways that stationary and mobile phases contribute to chromatographic retention Involves exploring the effects of solvent polarity on the strength of the mobile phase. In SFC this could involve determining the polarity of several different modifier/carbon dioxide mobile phases. In this paper, the use of a solvatochromic indicator to learn more about the effects of SFC modifier/mobile phase polarity will be investigated and discussed using several different modifiers and a diolmodified silica column.     

填充柱超临界流体色谱系统中的溶剂效应

 考察了填充柱超临界流体色谱法 (SFC)中的样品溶剂及连续进样等因素对化合物保留行为变化的影响规律。以超临界 CO2 或含低体积分数甲醇的 CO2 为流动相时 ,氨基柱上组分的保留时间随着样品溶剂的极性增大而增大 ,而溶剂对 C1 8柱上组分的保留时间影响不大 ;在 C1 8柱上 ,溶剂对连续进样的后续效应不强 ;而在氨基柱上 ,甲醇溶液的后续效应比丙酮、氯仿溶液的后续效应强。当甲醇的体积分数大于 1 .0 %时 ,溶剂的效应明显减弱。这种变化规律对填充柱 SFC的合理进样并获得重现性良好的色谱数据具有实际意义。     

High-performance liquid chromatography and capillary supercritical-fluid chromatography separation of vegetable carotenoids and carotenoid isomers

Carotenoids from carrots and tomatoes were separated with high-performance liquid chromatography (HPLC) and capillary supercritical fluid chromatography (SFC). All trans alpha- and beta-carotene were separated from their respective cis-isomers with capillary SFC. Carotenoids extracted from tomatoes included xanthophyll, lycopene and beta-carotene, while alpha- and beta-carotene were extracted from carrots. The HPLC separations were accomplished isocratically with a 25-cm column containing 5-microns ODS and methanol-acetonitrile-chloroform (47:47:6) or acetonitrile-dichloromethane (80:20). beta-Carotene cis-isomers were separated with SFC with a SB-cyanopropyl-25-polymethylsiloxane column, while alpha-carotene isomers were separated with two SB-cyanopropyl-50-polymethylsiloxane columns. Carotenoids from carrots and tomatoes were separated with a SB-phenyl-50-polymethylsiloxane column. Carbon dioxide with 1% ethanol was the SFC mobile phase. The eluent was monitored at 461 nm for HPLC and either 453 or 461 nm for SFC.     

Comparative study of the enantioselective separation of several antiulcer drugs by high-performance liquid chromatography and supercritical fluid chromatography

A comparative study of the enantiomeric separation of several antiulcer drugs such as omeprazole, lansoprazole, rabeprazole and pantoprazole using HPLC and supercritical fluid chromatography (SFC) on the Chrialpak AD column is presented in this work. The results show that employing the above mentioned column only two compounds (omeprazole and pantoprazole) could be enantiomerically resolved using HPLC, on the contrary SFC allowed the enantiomeric separation of all the compounds studied with higher resolutions and lower analysis times.     

Synthesis and application of mono-6-(3-methylimidazolium)-6-deoxyperphenylcarbamoyl-beta-cyclodextrin chloride as chiral stationary phases for high-performance liquid chromatography and supercritical fluid chromatography

The synthesis of mono-6-(3-methylimidazolium)-6-deoxyperphenylcarbamoyl-beta-cyclodextrin chloride (MPCCD) and its application in chiral stationary phases (CSPs) for high-performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) are being reported. This chiral selector is coated onto silica gel in different weight percentages (15, 20 and 35%, w/w) to obtain CSPs having different loading content. These new chiral stationary phases are tested using normal-phase HPLC for enantioseparation of racemic aromatic alcohols. Indeed, the enantiodiscrimination abilities of these CSPs are found to be influenced by the loading content of the chiral selector. Among the three columns (MPCCD-C15, MPCCD-C20 and MPCCD-C35), the best enantioseparation results are obtained using a column containing 20% (w/w) of MPCCD (MPCCD-C20). The resolution (R(s)) obtained for p-fluorophenylethanol, p-chlorophenylethanol, p-bromophenylethanol, p-iodophenylethanol and p-fluorophenyl-3-buten-1-ol using MPCCD-C20 ranges from 3.83 to 5.65. Good enantioseparation results are obtained for these analytes under SFC separation conditions using the MPCCD-C20 column.     

Effect of ionic additives on the elution of sodium aryl sulfonates in supercritical fluid chromatography

Addition of a small amount of polar solvent (i.e., modifier) to CO2 in packed column supercritical fluid chromatography (SFC) has shown major improvements in both polar analyte solubility and interaction of the polar analyte with the stationary phase. Recently, the addition of an ionic component (i.e., additive) to the primary modifier by one of us has been shown to extend even further the application of SFC to polar analytes. In this work, the effect of various ionic additives on the elution of ionic compounds, such as sodium 4-dodecylbenzene sulfonate and sodium 4-octylbenene sulfonate, has been studied. The additives were lithium acetate, ammonium acetate, tetramethylammonium acetate, tetrabutylammonium acetate, and ammonium chloride dissolved in methanol. Three stationary phases with different degrees of deactivation were considered: conventional cyanopropyl, deltabond cyanopropyl, and bare silica. The effect of additive concentration and additive functionality on analyte retention was investigated. Sodium 4-dodecylbenzene sulfonate was successfully eluted using all the additives with good peak shape under isocratic/isobaric/isothermal conditions. Different additives, however, yielded different retention times and in some cases different peak shapes.     

Kinetic behaviour in supercritical fluid chromatography with modified mobile phase for 5 μm particle size and varied flow rates

After much development of stationary phase chemistry, in recent years the focus of many studies in HPLC has shifted to increase the efficiency and analysis speed. Ultra high pressure liquid chromatography (UHPLC) using sub-2 μm particles, and high temperature liquid chromatography (HTLC), using temperatures above 100°C have received much attention. These new approaches allow the use of flow rates higher than those classically used in HPLC, reducing the analysis duration. Due to the low viscosity of supercritical fluids, high velocities, i.e. high flow rates, can be achieved with classical pumping systems typically used in supercritical fluid chromatography (SFC). The effects of the flow rate increase with CO(2)/methanol mobile phase was studied on the inlet pressure, t(0), the retention factor of the compounds, and on the efficiency. Simple comparisons of efficiencies obtained at varied temperature between SFC and HPLC, with a packed column containing 5 μm particles, show the greater kinetic performances achieved with the CO(2)/methanol fluid, and underline specific behaviours of SFC, occurring for high flow rates and sub-ambient temperature. Some values (N/t(0)) are also compared to UHPLC data, showing that good performance can be achieved in SFC without applying drastic analytical conditions. Finally, simple kinetic plots (t(0) vs N) at constant column length are used to select combinations of temperature and flow rate necessary to achieve a required theoretical plate number.     

Development of an orthogonal method for mometasone furoate impurity analysis using supercritical fluid chromatography

While supercritical fluid chromatography (SFC) has received great popularity in chiral separation and purification, it has rarely been used for trace level pharmaceutical impurity analysis, partially due to the limitation of instrument sensitivity. In this study, a packed column SFC method has been developed for the quantitative analysis of mometasone furoate and its trace level impurities. The UV detection was optimized to improve the sensitivity by 2-4 fold. In combination with an increased sample concentration, this SFC method is capable of trace level (0.05% of the active) analysis of the impurities. The SFC method used a silica column and a mobile phase consisting of CO(2) and methanol. The new method provides an orthogonal selectivity complementary to the reversed phase HPLC (RP-HPLC) method. All of the impurities and the active were baseline separated within 12 min on SFC, which is less than one third of the RP-HPLC method run time. The method was also partially validated for linearity, accuracy, precision (repeatability), and limit of quantitation. This study demonstrated that the SFC method, with improved sensitivity, can be a valuable tool to provide orthogonal selectivity for trace level impurity separation. With further validation, the method may be suitable for release testing and stability testing for mometasone furoate drug substance.     

Supercritical fluid chromatography and some of its applications: A review

The general topic of supercritical fluid chromatography (SFC) is introduced, and historical aspects of its development are discussed. The physical properties of supercritical fluids, gases and liquids are tabulated. SFC is compared and contrasted with the classical forms of chromatography - gas chromatography (GC) and high performance liquid chromatography (HPLC). The selectivity of SFC, GC, and HPLC are discussed and compared. Instrumentation employed for supercritical fluid chromatography is depicted. A wide variety of SFC applications are introduced. New examples of the use of SFC for analysis of a variety of complex oligomeric mixtures indcluding polypropylene glycol, polysiloxanes, fluorocarbon oligomers (i.e. -3M's fluoro-chemical surfactant Fluorad 171, and Kel-F) and high molecular weight normal alcohols are shown. The use of SFC for separation of mono-, di-, and triglycerides at low operating temperatures is described. Lastly, the use of SFC for separations of complex hydrocarbon mixtures from liquid fuels, polycyclic aromatic hydrocarbons, synthetic alpha-olefins, and petroleum functional group separations are depicted.     

Chiral high‐performance liquid and supercritical fluid chromatographic enantioseparations of limonene‐based bicyclic aminoalcohols and aminodiols on polysaccharide‐based chiral stationary phases

Enantioseparation of limonene‐based bicyclic 1,3‐aminoalcohols and 1,3,5‐ and 1,3,6‐aminodiols was performed by normal‐phase high‐performance liquid chromatographic and supercritical fluid chromatographic (SFC) methods on polysaccharide‐based chiral stationary phases. The effects of the composition of the mobile phase, the column temperature and the structures of the analytes and chiral selectors on retention and selectivity were investigated by normal‐phase LC and SFC technique. Thermodynamic parameters derived from selectivity–temperature‐dependence studies were found to be dependent on the chromatographic method applied, the nature of the chiral selector and the structural details of the analytes. Enantiorecognition in most cases was enthalpically driven but an unusual temperature behavior was also observed: decreased retention times were accompanied by improved separation factors with increasing temperature, i.e. some entropically driven separations were also observed. The elution sequence was determined in all cases. The separation of the stereoisomers was optimized in both chromatographic modalities.     

超临界流体色谱流程设计及其应用

本文设计了多功能超临界流体色谱流程,流程中包括毛细管/微填充柱SFC,GC,计算机控制温度、压力、密度及信号采集、处理,配置有超临界流体萃取池,解决了超临界流体色谱分流口易堵问题。利用该流程,将石腊、DC-200气相色谱固定相、黄油、蜂蜡、救心油、红花油等样品进行超临界流体色谱分离。     

A study of polyethoxylated alkylphenols by packed column supercritical fluid chromatography

Alkylphenol polyethoxylates (APEs) are a widely used group of nonionic surfactants in commercial production. Characterization of the composition of APE mixtures can be exploited for the determination of their most effective uses. In this study sample mixtures contain nonylphenol polyethoxylates and octylphenol polyethoxylates. The separation of individual alkylphenols by ethoxylate units is performed by supercritical fluid chromatography (SFC)-UV as well as normal-phase high-performance liquid chromatographic (HPLC)-UV employing packed columns. The stationary phase and column length are varied in the SFC setup to produce the most favorable separation conditions. Additionally, combinations of packed columns of different stationary phases are tested. The combination of a diol and a cyano column is found to produce optimal results. An advantage of using packed columns instead of capillary columns is the ability to inject large amounts of sample and thus collect eluted fractions. In this regard, fractions from SFC runs are collected and analyzed by flow injection analysis-electrospray ionization-mass spectroscopy in order to positively identify the composition of the fractions. In comparing the separation of APE mixtures by SFC and HPLC, it is found that SFC provides shorter retention times with similar resolution. In addition, less solvent waste is produced using SFC.