RP Conditions Research Articles

In‐line coupling of a reversed‐phase column to cope with limited chemoselectivity of a quinine carbamate‐based anion‐exchange type chiral stationary phase

A chiral stationary phase based on tert-butylcarbamoyl quinine has shown remarkable enantiomer separation capability for the thyroid hormone thyroxine (T(4)) and its structural analogue triiodothyronine (T(3)) employing hydroorganic buffered mobile phases (typical RP conditions). To overcome the problem of a somehow limited chemoselectivity for the critical peak pair between adjacent L-thyroxine (L-T(4)) and D-thyroxine (D-T(4)) peaks on the chiral anion-exchanger CSP when all four compounds need to be analysed simultaneously like in impurity profiling of L-T(4 )drug products, an RP column (Gemini C18) was serially coupled with the chiral anion-exchanger column to add a hydrophobic selectivity increment and to improve thereby the critical resolution between L-T(3) and D-T(4). Various parameters such as organic modifier content, pH, buffer concentration and type, type of achiral column as well as sequence of achiral and chiral column have been investigated with individual and tandem columns. With the optimized conditions and use of the tandem column a significantly improved separation, as compared to the chiral anion-exchanger column alone, with a critical resolution as high as 3.7 and an almost equal band spacing of the four components of the test mixture could be obtained. The sequence of the columns (achiral-chiral or chiral-achiral) had no significant effect on the separation performance.

Preparation and chromatographic properties of a multimodal chiral stationary phase based on phenyl‐carbamate‐propyl‐β‐CD for HPLC

A phenylcarbamate derivative of 2-hydroxypropyl-beta-CD bonded stationary phase was prepared by a previously described method. Its enantiomeric recognition abilities were evaluated as chiral stationary phase (CSP) in normal, polar organic and RP conditions by HPLC. The relevant structural features of the prepared stationary phase which make it an effective chiral selector are discussed. This material seems to have an excellent enantioselectivity for a variety of racemic analytes in the three modes. Hence it can be considered a highly effective multimodal column. Retention factor (k), selectivity (alpha) and resolution (R(s)) were the chosen parameters to describe the column performance. Optimization of these separations was discussed in terms of mobile phase composition, flow rate and structural patterns of the injected analytes.

Enantiomeric separation of chiral pesticides by high performance liquid chromatography on cellulose tris‐3,5‐dimethyl carbamate stationary phase under reversed phase conditions

Twenty chiral pesticides were tested, of which seven samples were directly separated by HPLC using cellulose tris-3,5-dimethyl carbamate (CDMPC) chiral stationary phase under RP conditions. The influence of mobile phase composition and column temperatures from 0 degrees C to 40 degrees C on the separations were investigated. The mobile phases were methanol/water or ACN/water at a flow rate of 0.8 mL/min with UV detection at 230 or 210 nm. Epoxiconazole, terallethrin, benalaxyl, and diclofopmethyl were observed to obtain the baseline separation under suitable conditions and other pesticides pyriproxyfen, lactofen, and quizalofop-ethyl were separated partially. The retention factors (k) and selectivity factor (alpha) for the enantiomers of most investigated pesticides decreased upon increasing the temperature except for the selectivity factors (alpha) of pyriproxyfen in methanol/water. The ln alpha - 1/T plots for racemic chiral pesticides were linear at the range of 0-40 except for that of pyriproxyfen enantiomers in methanol/water and the chiral separations were controlled by enthalpy. Better separations were not always at low temperature. The elution orders of the eluting enantiomers were determined by a circular dichroism (CD) detector.

Preparation of particle‐fixed silica monoliths used in capillary electrochromatography

Fused-silica capillarieswere packed with porous 1 microm bare silica microspheres and immobilized by potassium silicate-formamide in order to obtain columns with silica-based monolithic packing. After curing, the particle-fixed monolithic columns were octadecylated insitu with dimethyloctadecylchlorosilane. The columns were mechanically strong and permeable. No noticeable loss in efficiency was found after using a column continuously for 1 month. The performances of the particle-fixed silica monolithic columns were evaluated for CEC under RP conditions. High separation efficiency (about125 200 plates/m) was obtained by using these new types of columns.

Enantiomer separation on monolithic silica HPLC columns using chemically bonded methylated and methylated/acetylated 6-O-tert-butyldimethyl- silylated β-cyclodextrin

A new 2,3-methylated 3*-monoacetylated 6-O-tert-butyldimethylsilylated beta-CD derivative was synthesized and chemically bonded onto aminopropyl derivatized monolithic silica HPLC columns. In this CD derivative, only one of seven methyl groups in 3-position was substituted by an acetyl group. Its applicability as a chiral stationary phase for HPLC was tested and compared with exclusively 2,3-methylated 6-O-tert-butyldimethylsilylated beta-CD immobilized onto aminopropyl-modified monoliths. Thirty-two chiral compounds from different chemical classes and different functionalities were tested under RP conditions. Fourteen compounds were resolved into their enantiomers by methylated 6-O-tert-butyldimethylsilylated beta-CD. By use of methylated/acetylated 6-O-tert-butyldimethylsilylated beta-CD as the chiral stationary phase 7 analytes were successfully stereodifferentiated.

A (4R)‐Hydroxy‐L‐proline‐derived chiral scaffold and its oligomers as chiral selectors in liquid chromatography chiral stationary phases for enantioseparation

The chromatographic behaviour of a poly-L-proline-derived chiral stationary phase (CSP) is compared to the corresponding single proline-derived CSP. Structurally diverse racemic test compounds and mobile phases, including normal- and RP conditions, were used. Although the application domain of the poly-L-proline-derived CSP (CSP-3) was considerably restricted, this CSP showed a higher retention and a slightly broader application domain than the monomeric analogue (CSP-1) when heptane/2-PrOH was used as mobile phase. The presence of an alcohol in the mobile phase was essential for enantioseparation in the poly-L-proline-derived CSP when normal-phase conditions were applied.

Stationary phase-based two-dimensional chromatography combining both covalent and noncovalent interactions on a single HPLC column

A new type of 2-D separation material was synthesized and studied. The material is suitable for 2-D chromatography utilizing both covalent and noncovalent interactions. The first dimension is boronate affinity chromatography, and the second dimension is RP chromatography (or vice versa). The polymeric media were prepared using p-vinylphenylboronic acid as the functional monomer. This monomer was selected due to the presence of the boronic acid group for the cis-diol/boronate interaction in boronate chromatography. Two crosslinkers were evaluated, namely ethylene glycol dimethacrylate and divinylbenzene. The crosslinker content was varied to maximize the polymer strength and the RP performance of the packed column. Several parameters were evaluated to define the optimum for polymer strength and column performance including crosslinker, porogen, initiator, and column-packing parameters. The polymer-based HPLC columns were successful in separating phenol, catechol, dimethylphthalate, and hydroquinone under RP conditions, and thus can be used as an RP HPLC column. The columns were also successful in separating catechol and adenosine under boronate chromatography conditions, and thus can be used as a boronate affinity column. Moreover, the two types of chromatography can be performed consecutively on the same column during one complete chromatographic run, making it a 2-D chromatography. Under these 2-D conditions, the catechol was separated from a mixture of phenol, catechol, dimethylphthalate, and hydroquinone; the adenosine ribonucleoside was separated from a mixture of adenosine ribonucleoside, adenosine deoxyribonucleoside, and uridine deoxyribonucleoside. This type of single-column 2-D HPLC eliminates the requirement of a complex and expensive multidimensional HPLC instrument and provides increased peak capacity for separation.

How Revealing is Revealed Preference?

This lecture address the following two key criticisms of the empirical application of revealed preference theory: When the RP conditions do not reject, they do not provide precise predictions; and when they do reject, they do not help characterize the nature of irrationality or the degree/direction of changing tastes. Recent developments in the application of RP theory are shown to have rendered these criticisms unfounded. A powerful test of rationality is available that also provides a natural characterization of changing tastes. Tight bounds on demand responses and on the welfare costs of relative price and tax changes are also available and are shown to work well in practice.

<I>Presidential Address</I>: How Revealing Is Revealed Preference?

This lecture address the following two key criticisms of the empirical application of revealed preference theory: When the RP conditions do not reject, they do not provide precise predictions; and when they do reject, they do not help characterize the nature of irrationality or the degree/direction of changing tastes. Recent developments in the application of RP theory are shown to have rendered these criticisms unfounded. A powerful test of rationality is available that also provides a natural characterization of changing tastes. Tight bounds on demand responses and on the welfare costs of relative price and tax changes are also available and are shown to work well in practice. (JEL: )

Dual chamber arrhythmia detection in the implantable cardioverter defibrillator.

Dual chamber implantable cardioverter defibrillator (ICD) technology extended ICD therapy to more than termination of hemodynamically unstable ventricular tachyarrhythmias. It created the basis for dual chamber arrhythmia management in which dependable detection is important for treatment and prevention of both ventricular and atrial arrhythmias. Dual chamber detection algorithms were investigated in two Medtronic dual chamber ICDs: the 7250 Jewel AF (33 patients) and the 7271 Gem DR (31 patients). Both ICDs use the same PR Logic algorithm to interpret tachycardia as ventricular tachycardia (VT), supraventricular tachycardia (SVT), or dual (VT+ SVT). The accuracy of dual chamber detection was studied in 310 of 1,367 spontaneously occurring tachycardias in which rate criterion only was not sufficient for arrhythmia diagnosis. In 78 episodes there was a double tachycardia, in 223 episodes SVT was detected in the VT or ventricular fibrillation zone, and in 9 episodes arrhythmia was detected outside the boundaries of the PR Logic functioning. In 100% of double tachycardias the VT was correctly diagnosed and received priority treatment. SVT was seen in 59 (19%) episodes diagnosed as VT. The causes of inappropriate detection were (1) algorithm failure (inability to fulfill the PR<RP condition in atrial tachyarrhythmias with 1:1 AV conduction, and far-field R wave sensing intermittently present during sinus tachycardia); (2) programming settings (atrial fibrillation/atrial flutter with ventricular rate above the SVT limit); and (3) algorithm limitations (atrial tachycardia with ventricular rate around the shortest programmable SVT limit and SVT redetection following VT therapy). Programming measures improved detection ability in 13 of 59 of inappropriately detected arrhythmias. Dual chamber detection algorithms evaluated in a subset of diagnostically difficult arrhythmias allow safe detection of double tachycardias but require further extension and programmability to improve VT:SVT discrimination rules.

Comparison of D,L-Mandelic Acid Resolution on Zeolite and Silica Supported Pirkle-Type Chiral Stationary Phases

Zeolite A, a material of crystalline character, and Hypersil silica have been used as support for the preparation of chiral stationary phases. On the amorphous silica support surface the silanol groups are randomly dispersed. The crystalline zeolite secondary building units consisting primarily of SiO4, AlO4− tetrahedra determine the regularity of surface silanol groups. Owing to the crystal lattice structure, the location of silanols is well determined and hence the dispersion of chiral selector molecules chemically bonded onto the zeolite surface silanol groups is fundamentally arranged. Amides of DNB-L-Leu, DNB-L-Phe, B-L-Leu chiral selector molecules were anchored onto the zeolite silanols and B-L-Leu onto the silica support silanols. Lipophilic buffer in RP conditions has dynamically modified the residual silanols of each support. The enantioseparation of ion paired D,L-mandelic acid from aqueous solution on the zeolite and silica supported chiral stationary phases prove a superior enantioseparation on the zeolite supported phases.

Separation and identification of various carotenoids by C30 reversed-phase high-performance liquid chromatography coupled to UV and atmospheric pressure chemical ionization mass spectrometric detection

In this paper the application of on-line HPLC–UV–APCI (atmospheric pressure chemical ionization) mass spectrometry (MS) coupling for the separation and determination of different carotenoids as well as cis/trans isomers of β-carotene is reported. All HPLC separations were carried out under RP conditions on self-synthesized polymeric C30 phases. The analysis of a carotenoid mixture containing astaxanthin, canthaxanthin, zeaxanthin, echinenone and β-carotene by HPLC–APCI-MS was achieved by scanning the mass range from m/z 200 to 700. For the characterization of a sample containing cis/trans isomers of β-carotene as well as their oxidation products, a photodiode-array UV–visible absorbance detector was used in addition between the column and the mass spectrometer for structural elucidation of the geometrical isomers. The detection limit for β-carotene in positive-ion APCI–MS was determined to be 1 pmol. In addition, an extract of non-polar substances in vegetable juice has been analyzed by HPLC–APCI-MS. The included carotenoids could be identified by their masses and their retention times.

Treating adult marijuana dependence: A test of the relapse prevention model.

Men (n = 161) and women (n = 51) seeking treatment for marijuana use were randomly assigned to either a relapse prevention (RP; G.A. Marlatt & J.R. Gordon, 1985) or a social support (SSP) group discussion intervention. Data collected for 12 months posttreatment revealed substantial reductions in frequency of marijuana use and associated problems. There were no significant differences between the cognitive-behavioral RP intervention and the SSP group discussion conditions on measures of days of marijuana use, related problems, or abstinence rates. Men in the RP condition were more likely than men in the SSP condition to report reduced use without problems at 3-month follow-up. Posttreatment increases in problems associated with alcohol did not appear to relate to reduced marijuana use. Results are discussed in terms of the need for further research with marijuana-dependent adults and the efficacy of RP.