反相高效液相色谱分离果酸的研究

水果和蔬菜中含有的有机酸,称谓果酸,果酸的定量方法很多,气相色谱法需要将果酸衍生后才能分析。本文阐述用反相高效液相色谱及紫外与示差折光检测器,流动相为0.01M(NH_4)_2HPO_4,用H_3PO_4调节pH值为2.5,使果酸得到分离。用保留时间定性,用标准酸样的直线方程进行定量。并对橙子中的柠檬酸、健力宝饮料中的酒石酸作了定量测定。回收率达90%以上。此法快速,分离效果较好,是食品生产与分析发展的趋势。     

RPLC分离果酸的研究

本文用反相高压液相色谱(RHLC)研究了果酸的分离,找到了用紫外与示差折光检测器同时检测分离果酸的最佳条件,并以0.01M(NH_4)_2HPO_4 用H_3PO_4调节PH2.2～2.5。在上述条件下使果酸得到分离,用保留时间定性,用标准酸样的直线方程来定量。并对果实橙子中的柠檬酸和健力宝饮料中的酒石酸作了定量。     

Separation of racemic 2,4-dinitrophenyl amino acids on carboxymethyl-β-cyclodextrin coated zirconia in RPLC

We report preparation and use of carboxymethyl-β-cyclodextrin-coated zirconia as a chiral stationary phase (CSP) for separation of enantiomers of 2,4-dinitrophenyl (DNP) amino acids in reversed-phase high performance liquid chromatography. The CSP showed good enantioselectivity for some of the amino acids studied. Effects of pH and amount of methanol in mobile phases on retention and enantioselectivity for the analytes were examined.     

唾液中残存消毒剂TCS的检测

TCS(2，4，4’-三氯-2’-羟基二苯醚，triclosan，中文名称玉洁纯、玉洁新、三氯新等)，是一种非离子型的高效广谱抗微生物剂，对于一般常见的细菌、真菌及酵母等病原菌都具有杀灭功效及抑制作用，广泛用于牙膏、洗漱液和洗发液中。国内外学者长期的临床观察证实含有TCS的牙膏和漱口水能有效地控制菌斑和缓解牙龈炎症状。     

Separation and determination of amitriptyline and nortriptyline by dispersive liquid-liquid microextraction combined with gas chromatography flame ionization detection

Dispersive liquid–liquid microextraction (DLLME) coupled with gas chromatography–flame ionization detection (GC–FID) was applied for the determination of two tricyclic antidepressant drugs (TCAs), amitriptyline and nortriptyline, from water samples. This method is a very simple and rapid method for the extraction and preconcentration of these drugs from environmental sample solutions. In this method, the appropriate mixture of extraction solvent (18 μL Carbon tetrachloride) and disperser solvent (1 mL methanol) are injected rapidly into the aqueous sample (5.0 mL) by syringe. Therefore, cloudy solution is formed. In fact, it is consisted of fine particles of extraction solvent which is dispersed entirely into aqueous phase. The mixture was centrifuged and the extraction solvent is sedimented on the bottom of the conical test tube. 2.0 μL of the sedimented phase is injected into the GC for separation and determination of TCAs. Some important parameters, such as kind of extraction and disperser solvent and volume of them, extraction time, pH and ionic strength of the aqueous feed solution were optimized. Under the optimal conditions, the enrichment factors and extraction recoveries were between 740.04–1000.25 and 54.76–74.02%, respectively. The linear range was (0.005–16 μg mL⁻¹) and limits of detection were between 0.005 and 0.01 μg mL⁻¹ for each of the analytes. The relative standard deviations (R.S.D.) for 4 μg mL⁻¹ of TCAs in water were in the range of 5.6–6.4 (n = 6). The performance of the proposed technique was evaluated for determination of TCAs in blood plasma.     

Separation of ternary systems of hydrophilic ionic liquid with miscible organic compounds by RPLC with refractive index detection

A rapid and simple analytical method was developed for the simultaneous and quantitative determination and separation of hydrophilic imidazolium ionic liquids (ILs) (1-butyl-3-methylimidazolium chloride, [C(4)mim]Cl; 1-hexyl-3-methylimidazolium chloride, [C(6)mim]Cl; 1-octyl-3-methylimidazolium chloride, [C(8)mim]Cl; 1-allyl-3-methylimidazolium chloride, [Amim]Cl; or 1-allyl-3-methylimidazolium bromide, [Amim]Br) with miscible ethyl acetate and EtOH and their mixtures using reverse phase liquid chromatography coupled with refractive index detection (RPLC-RI). The influence of 60 to 100% (volume percentage) methanol in the mobile phase on the IL systems ([C(4)mim]Cl, [C(6)mim]Cl, [C(8)mim]Cl, [Amim]Br, or [Amim]Cl)-ethyl acetate-EtOH was investigated. The optimum mobile phase for the system [C(8)mim]Cl-ethyl acetate-EtOH, [C(4)mim]Cl-ethyl acetate-EtOH, [Amim]Br-ethyl acetate-EtOH and [Amim]Cl-ethyl acetate-EtOH was methanol/water (60:40, v/v), and methanol/water (70:30, v/v) for [C(6)mim]Cl-ethyl acetate-EtOH. Under optimum mobile phase conditions for each system, the RSD of the retention time ranged from 0.02 to 0.04%, and the RSDs of the peak area percent ranged from 0.23 to 1.85%, which showed good reproducibility of the RPLC-RI method. The RPLC-RI method can determine IL, ethyl acetate, and EtOH simultaneously in 5 min, and the analytes, especially IL, can be eluted completely. The results show that the RPLC-RI method can be used to separate and determine ILs in mixtures with organic compounds simultaneously and quantitatively.     

New thermodynamically consistent competitive adsorption isotherm in RPLC

A new equation of competitive isotherms was derived in the framework of the ideal adsorbed solution (IAS) that predicts multisolute adsorption isotherms from single-solute isotherms. The IAS theory makes this new isotherm thermodynamically consistent, whatever the saturation capacities of these single-component isotherms. On a Kromasil-C(18) column, with methanol-water (80/20 v/v) as the mobile phase, the best single-solute adsorption isotherm of both toluene and ethylbenzene is the liquid-solid extended multilayer BET isotherm. Despite a significant difference between the monolayer capacities of toluene (370 g/l) and ethylbenzene (170 g/l), the experimental adsorption data fit very well to single-component isotherms exhibiting the same capacities (200 g/l). The new competitive model was used for the modeling of the elution band profiles of mixtures of the two compounds. Excellent agreement between experimental and calculated profiles was observed, suggesting that the behavior of the toluene-ethylbenzene adsorbed phase on the stationary phase is close to ideal. For example, the concentrations measured for the intermediate plateau obtained in frontal analysis differ by less than 2% from those predicted by the IAS model.     

Properties of the Adsorption Equilibrium Isotherms Used and Measured in RPLC

Monitoring anabolic steroids in meat-producing animals is a challenging task. It implies very specific and sensitive analytical methods able to detect and identify sub-μg kg^?1 residue levels in complex biological matrices such as meat, urine, or hair. Gas and liquid chromatography coupled to mass spectrometry are the most efficient means of achieving these objectives. In this paper we review how developments in mass spectrometry have been rapidly applied to this problem, how efficient analysis of anabolic steroids in urine, edible tissue, and hair has been achieved, and, later, how measurement of conjugate steroids and determination of the origin of natural steroid hormones has been achieved. The performance characteristics of different mass spectrometers (quadrupole, ion-trap, electromagnetic, isotope-ratio, tandem, and hybrid instruments), the efficiency of different acquisition techniques (LR-SIM, HR-SIM, MRM), and, finally, sample introduction (gas chromatography and liquid chromatography, with discussion of alternative interfaces) are discussed, with numerous applications.     

Comparison of the Separation Characteristics of the Organic–Inorganic Hybrid Octadecyl Stationary Phases XTerra MS C18 and XBridge C18 and Shield RP18 in RPLC

Differences in the system constants of the solvation parameter model, discontinuities in retention factor plots (log k against volume fraction of organic solvent) and retention factor correlation plots are used to study the retention mechanism on XTerra MS C₁₈, XBridge C₁₈ and XBridge Shield RP₁₈ stationary phases with acetonitrile–water and methanol–water mobile phases containing from 10 to 70% (v/v) organic solvent. Wetting of XBridge C₁₈ at 10 and 20% (v/v) acetonitrile is incomplete and is responsible for small changes in the retention mechanism. The intermolecular interactions responsible for retention on XTerra MS C₁₈ and XBridge C₁₈ are similar with minor differences in cavity formation and hydrogen-bonding interactions responsible for small selectivity differences. On the other hand, for bulky solutes there are large changes in retention at low volume fractions of organic solvent (<40% v/v) associated with steric repulsion on the XTerra MS C₁₈ stationary phases that is absent for XBridge C₁₈. Selectivity differences are more apparent for XBridge C₁₈ and XBridge Shield RP₁₈. For acetonitrile-water mobile phases cavity formation in the solvated XBridge Shield RP₁₈ is slightly more difficult and hydrogen-bond acid and base interactions are more important than for XBridge C₁₈. With methanol–water mobile phases it becomes slightly easier to form a cavity in the solvated XBridge RP₁₈ compared with XBridge C₁₈. In addition, the methanol-water solvated XBridge RP₁₈ is a stronger hydrogen-bond base and more dipolar/polarizable than XBridge C₁₈. These variations in selectivity justify the use of XBridge C₁₈ and XBridge Shield RP₁₈ as complementary stationary phases for method development.     

Off‐line 2‐D RPLC/RPLC method for separation of components in Dalbergia odorifera T. Chen

An off‐line 2‐D RPLC/RPLC system incorporating a β‐CD based column and a BEH C18 column was developed in the separation of components in Dalbergia odorifera T. Chen. (Jiang Xiang). Both orthogonality and van Deemter curves of the two columns were investigated. The orthogonality was about 57%, which was evaluated with 40 standard solutes. Optimized column efficiency could be achieved under optimal linear velocity (0.04 cm/s) on the CD column or under high linear velocity (0.32 cm/s) on the BEH C18 column. With the 2‐D LC system, totally 637 peaks were separated in 114 fractions from the extraction of Jiang Xiang. Meanwhile, 19 flavanoids were tentatively identified from 114 fractions with Q‐TOF MS. The results demonstrated the separation power of this 2‐D LC system and further proved the high orthogonality between CD and C18 columns.     

Modelling chromatographic behaviour as a function of pH and solvent composition in RPLC

Summary In this work we establish the basic layout of IONICS, an expert system for optimizing the separation of ionogenic solutes in Reversed-Phase Liquid Chromatography, using the pH and the organic-modifier concentration of the mobile phase as parameters. We also present REMO, a front-end system that automates the retention modelling stage, based on a 9-parameter model. This system uses a scale transformation to suppress several numerical problems previously observed and features a strategy for automatic calculation of an initial approximation to the model optimum. The successful application of this system to a set of seven drugs is described. The final models are accurate and have smaller numerical problems. We also describe the use of a genetic algorithm instead of classical non-linear least-squares for fitting the model to the experimental data. Results indicate that genetic algorithms are a valuable, complementary tool for retention modelling.     

Column-switching RPLC for the trace-level determination of ethylenethiourea in aqueous samples

Summary Column-switching with two C18 separation columns combined with UV detection at 233 nm has been used to determine ETU in ground water samples. The direct assay of ETU (limit of detection, 1 ppb) takes less than 10 min. For the determination of ETU down to a level of 0.1 ppb, preconcentration of ETU is achieved by a liquid-liquid extraction procedure allowing the analysis of at least 12 water samples per day.For both methods (1-ppb and 0.1-ppb level), the overall recovery (96 and 82%), the repeatability (4.5 and 4.3%) and the reproducibility (5.0 and 7.9%) are satisfactory (n=18 and 28, respectively). More than one hundred samples of ground water were analysed during a 10-month period; over 0.1 ppb of ETU (range, 0.1–300 ppb) was found in 30% of the samples tested.     

Selectivity enhancement of diastereomer separation in RPLC using crystalline-organic phase-bonded silica

Summary Poly(octadecyl acrylate) exists in a crystalline state at room temperature. This crystalline state was formed even after being grafted onto silica through the terminal group of the polymer. When this polymer-grafted silica (Sil-ODA_n) was used as a reversed phase liquid chromatography packing material, better selectivity for diastereomerized phenylethylamines derivatized by (S)-(−)-(2,3-naphthalenedicarboximidyl)propionyl fluoride was observed at room temperature than was observed for simple octadecylated silica (ODS). On the other hand, increasing temperature reduced the selectivity to be close to that of ODS. Additional experiments showed the better selectivity derived from Sil-ODA_n was due to both the highly-oriented structure of the polymer and the carbonyl-π interaction with the diastereomer. A theoretical investigation of this carbonyl-π interaction is also described briefly.     

Adsorption isotherms of organic modifiers and the determination of the dead volume in RPLC

Summary In RPLC the dead volume can be defined as the difference between the maximum column hold-up volume and the volume of the adsorbed phase. The composition of the adsorbed phase depends on the composition of the mobile phase and therefore, the dead volume also varies with it. In this work, the alkyl bonded phase acetonitrile (ACN)-water mobile phase system is investigated. In the system, deuterated water (D₂O) and deuterated acetonitrile (D-ACN) are retained due to the isotopic dilution effect. By means of D₂O and D-ACN, the absolute adsorption isotherm of the organic modifier ACN is measured. Based on the isotherm, the chromatographic behaviour of ACN, D-ACN and D₂O, the variation of the dead volume with the composition of the mobile phase, and the approach to determine the maximum column hold-up volumn are explained. In addition, the various approaches to determine the dead volume are compared and the recommendations are given for the case of common unbuffered binary systems (MeOH/H₂O, THF/H₂O and ACN/H₂O).     

Release of nortriptyline hydrochloride from oil-water microemulsions

The release of nortritptyline hydrochloride from oil-in-water (o/w) microemulsions (isopropyl myristate as oil, propylene glycol as cosurfactant, polysorbate 80 as surfactant and phosphate buffer, pH 7.4, as the continuous phase) containing increasing concentrations of polyethylene glycol 400, used to facilitate the diffusion of a drug from the inner oily phase of the microemulsion to the outer aqueous phase of such a dispersion system, was studied by determining the permeability constants of the drug through hydrophilic and lipophilic membranes separating the o/w microemulsions from the receiving aqueous phase (phosphate buffer pH 7.4). The permeability of nortriptyline hydrochloride from microemulsions through the lipophilic membrane increased as the concentration of polyethylene glycol 400 in the disperse system increased. The apparent permeability constant for nortriptyline hydrochloride, from the microemulsion without polyethylene glycol, was 1.36 x 10(-3) cm x h(-1), it increased up to 7.80 x 10(-3) cm x h(-1) in the presence of polyethylene glycol at a concentration of 50% (v/v) of the initial volume of the aqueous phase.     

Separation mechanism of chiral compounds in chiral stationary phase liquid chromatography

In this paper, the concept of reversed- or normal-phase chiral stationary phase liquid chromatography has been put forward according to the polar strength of mobile and stationary phases. The statistical model developed in HPLC has been used to investigate the separation mechanism of D- and L-enantiomer in chiral stationary phase liquid chromatography. It has been observed that the variation of capacity factor of enantiomers with mobile phase composition in both reversed-phase and normal-phase chiral stationary phase liquid chromatography can be described by the fundamental elution equation lnk' = a + blnC_b + _cC_b. The effect of mobile phase composition on the selectivity of enantiomers D and L in normal-phase chiral stationary phase liquid chromatography can be described by the equation lnα = Δa + ΔblnC_b, but in reversed-phase chiral stationary phase liquid chromatography the selectivity is almost independant of the mobile phase composition.     

Determination of antioxidants in polyolefins using total dissolution methodology followed by RPLC

Antioxidants are added to polyolefins to improve the stability of the resin from oxidation and degradation during processing of the finished article and to increase product lifetime. Without antioxidants, polyolefins would quickly degrade during and after the extrusion or thermoforming process, which would cause inferior appearance and physical properties. The proper level must be added to protect the polymer and to minimize cost. Antioxidants are usually extracted from the resin and the extract is analyzed by RPLC, GC, or Fourier transform infrared spectroscopy. Unfortunately, many of these procedures require significant manual labor, time, and solvent, rendering them impractical for high-throughput work processes. In addition, they may not provide complete extraction of the additives depending upon the type of resin. A validated analytical method was needed for the determination of three common antioxidants, Irganox(?) 1010, Irganox(?) 1076, and Irgafos(?) 168 in polyolefin resins. This paper shows the determination of these antioxidants using dissolution followed by precipitation with o-xylene and methanol. Direct analysis of the solution is achieved in 8?min using RPLC.