Enantiospecific binding of Rotigotine and its antipode to serum albumins: Investigation of binding constants and binding sites by partial-filling ACE.

Enantiospecific binding of antiparkinsonian medication Rotigotine (S-enantiomer) and its antipode to HSA or BSA was demonstrated employing partial-filling ACE (PF-ACE) under near-physiological conditions (50 mM phosphate, pH 7.4, 37 degrees C). The enantioseparation of the enantiomers was achieved by PF-ACE. Subsequently, the binding characteristics of the enantiomers to the serum albumins were investigated. Based on the PF-ACE data, the following binding constants were obtained: K(b,HSA,S)=8884+/-255 M(-1), K(b,HSA,R)=17648+/-587 M(-1), K(b,BSA,S)=7348+/-237 M(-1), K(b,BSA,R)=9353+/-352 M(-1). It revealed that Rotigotine had weaker affinity for the two serum albumins, and both enantiomers showed stronger affinity for HSA than BSA. The presence of either site marker (warfarin or ketoprofen) had adverse effect on the enantioseparation due to the competitive binding, or even eliminated the enantioselective binding of the enantiomers to the albumin when the molar ratio of the site marker to the albumin was at certain level. Although there might be a synergistic binding between the drug and the albumin, it was suggested that site II and I were the preferential binding site of the drug on HSA and BSA, respectively.

Enantioseparation of esbiothrin by cyclodextrin-modified microemulsion and micellar electrokinetic chromatography.

A comparison between chiral cyclodextrin-modified microemulsion electrokinetic chromatography (CD-MEEKC) and cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) for the enantiomeric separation of esbiothrin was carried out. For both methods, the separation conditions were optimized by varying CD types and concentration, running buffer pH and compositions, organic modifiers, and temperature. The optimal CD-MEEKC conditions were 0.8% n-heptane, 2.3% SDS, 6.6% n-butanol, 90.3% 10 mM sodium tetraborate containing 3% (w/v, the ratio of CD mass to microemulsion volume) methyl-beta-cyclodextrin, pH 10, 25 degrees C. The optimized CD-MEKC conditions were 3.3% SDS, 96.7% 10 mM sodium tetraborate containing 5% (w/v) beta-CD, pH 10, 25 degrees C. The difference in physicochemical properties of the buffer and CDs resulted in different optimal CD type. The competitive distribution between the microemulsion (or micelle) and chiral CD contributed to the chiral separation. Both methods provided excellent separation (R(s) approximately 3) with similar migration time (ca. 15 min). CD-MEEKC provided higher separation efficiencies (>300000) than CD-MEKC (>200000). The LODs for CD-MEEKC and CD-MEKC were 4.7 microg/mL and 3.2 microg/mL, respectively. The RSDs of migration time and peak area for CD-MEEKC were slightly higher than for CD-MEKC. Both the demonstrated CD-MEEKC and CD-MEKC methods provided high efficiencies, low LODs, and reproducible enantioseparations of esbiothrin.

Synthesis and binding site characteristics of 2,4,6-trichlorophenol-imprinted polymers.

2,4,6-Trichlorophenol (2,4,6-TCP)-imprinted micro- and submicrospheres prepared by precipitation polymerization were compared with templated materials obtained by conventional bulk polymerization. The influence of the type and amount of functional monomer, the type and amount of cross-linker, polymerization temperature, porogen, and the ratio of template molecule and functional monomer to cross-linker on the size of the obtained particles were investigated. UV-Vis spectrophotometer experiments revealed that the microsphere polymers provided higher affinity to the template in contrast to imprinted polymers prepared by bulk polymerization. The binding properties of the microspheres, including binding isotherms and affinity distribution, were studied via Freundlich isotherm affinity distribution (FIAD) analysis. The obtained results indicated that microspheres prepared by precipitation polymerization provided superior rebinding properties during equilibrium binding in contrast to bulk polymers and submicrosphere polymers. Moreover, release experiments showed that 80% of rebound 2,4,6-TCP was released from the imprinted microspheres within the first 2 h, while more intimately bound 2,4,6-TCP molecules were released in the following 40 h. The morphologies and porosities of the resulting imprinted materials were characterized by scanning electron microscopy (SEM) and Brunauer-Emmett-Teller (BET) analysis, respectively. The microsphere polymers exhibited a regular spherical shape with a high degree of monodispersity to the corresponding bulk polymers. Furthermore, the micro- and submicrospheres were characterized by narrow distribution of pores in contrast to a heterogeneity index of m = 0.6647 for the microsphere imprinted polymer.

Simultaneous enantioseparation of antiparkinsonian medication Rotigotine and related chiral impurities by capillary zone electrophoresis using dual cyclodextrin system.

A dual cyclodextrin (CD) system consisting of sulfated beta-CD (S-beta-CD) and methyl-beta-CD (M-beta-CD) modified capillary zone electrophoresis (CZE) method was proposed to separate the antiparkinsonian drug Rotigotine ((-)-(S)-2-(N-propyl-N-2-thienylethylamino)-5-hydroxytetralin) and related chiral impurities (2-(N-propylamino)-5-hydroxytetralin, 2-(N-propylamino)-5-methoxytetralin). The method was optimized by varying the CD type, the buffer pH, individual CD concentration of the dual system and the ionic strength of background electrolyte. Under the optimum conditions, i.e. 2% (w/v) S-beta-CD and 2% (w/v) M-beta-CD in 100mM sodium phosphate (pH 2.5) as the running buffer, separation voltage -20 kV, detected at 200 nm and temperature controlled at 20 degrees C, a satisfactory separation of the six analytes was accomplished. The optimized method was validated for specificity, precision, linearity, accuracy and stability using sodium benzenesulfonate as the internal standard. The relative standard deviation for migration time was less than 0.58%, and 3.78% for peak area ratio. The linearity ranged from 0.005 to 0.25 mM. The recovery ranged from 95.9% to 108.3%. The limits of detection and limits of quantification for each enantiomer were 0.003 and 0.01 mM, respectively. This method was utilized for evaluating the chiral purity of five batches of Rotigotine.

Studies on degradation of imazalil enantiomers in soil using capillary electrophoresis.

A scheme was demonstrated to elucidate the degradation behaviors of the two enantiomers of the fungicide imazalil in soil using cyclodextrin-modified capillary zone electrophoresis. The separation buffer was 50 mmol/L NaH2PO4, 5 mmol/L (NH4)H2PO4, and 5 mmol/L beta-cyclodextrin (pH 3.0). The limits of detection of this CE method were 0.24 and 0.26 microg/mL for (-)- and (+)-imazalil, respectively. Five different soil conditions were investigated in laboratory microcosms: under sunlight; in darkness; under UV irradiation; in sterilized soil; and in soil with wheat planted in it. Radiation, microorganisms, and uptake by wheat benefited the degradation of imazalil in this study. The half-lives (t1/2) of imazalil in soil under the above conditions were 20, 30.5, 11, 27.5, and 21.5 days, respectively. The degradation rate of imazalil in soil under the five different sets of conditions decreased in the order: UV irradiation > sunlight > soil with wheat planted in it > sterilized soil > soil kept in darkness. Imazalil in soil (pH 8.2, slightly alkaline) collected in the suburbs showed non-enantioselective degradation.