Resonance light scattering determination of 6-mercaptopurine coupled with HPLC technique.

A simple, fast, costless, sensitive and selective method of resonance light scattering coupled with HPLC was established for the determination of 6-mercaptopurine in human urine sample. In a Britton-Robinson buffer solution of pH5.5, the formation of coordination complex between 6-mercaptopurine and metal palladium (II) led to enhance the RLS intensity of the system. The RLS signal was detected by fluorescence detector at λ(ex)=λ(em)=315 nm. The analytical parameters were provided by the coupled system, the linear of 6-mercaptopurine response from 0.0615 to 2.40 µg L(-1) and the limit of detection (S/N=3) was 0.05 µg L(-1). The presented method has been applied to determine 6-mercaptopurine in human urine samples which obtained satisfactory results. Moreover, the reaction mechanism and possible reasons for enhancement of RLS were fully discussed.

Rayleigh light scattering detection of three α1-adrenoceptor antagonists coupled with high performance liquid chromatograph.

Herein, a Rayleigh light-scattering (RLS) detection method combined with high performance liquid chromatograph (HPLC) without any post-column probe was developed for the separation and determination of three α1-adrenoceptor antagonists. The quantitative analysis is benefiting from RLS signal enhancement upon addition of methanol which induced molecular aggregation to form an hydrophobic interface between aggregates and water that produce a sort of superficial enhanced scattering effect. A good chromatographic separation among the compounds was achieved using a Gemini 5u C18 reversed phase column (250 mm × 4.6 mm; 4 µm) with a mobile phase consisting of methanol and ammonium acetate-formic acid buffer solution (25 mM; pH=3.0) at the flow rate of 0.7 mL min(-1). The RLS signal was monitored at λex=λem=354 nm. A limit of detection (LOD) of 0.065-0.70 µg L(-1) was reached and a linear range was found between peak height and concentration in the range of 0.75-15 µg L(-1) for doxazosin mesylate (DOX), 0.075-3.0 µg L(-1) for prazosin hydrochloride (PRH), and 0.25-5 µg L(-1) for terazosin hydrochloride (TEH), with linear regression coefficients all above 0.999. Recoveries from spiked urine samples were 88.4-99.0% which is within acceptable limits. The proposed method is convenient, reliable and sensitive which has been used successfully in human urine samples.

A reversed-phase high performance liquid chromatography coupled with resonance Rayleigh scattering detection for the determination of four tetracycline antibiotics.

A new reversed-phase high performance liquid chromatography with resonance Rayleigh scattering detection (HPLC-RRS) was developed for simultaneous separation and determination of four tetracycline antibiotics (TCs). A good chromatographic separation among the compounds was achieved using a Synergi Fusion-RP column (150 mm x 4.6 mm; 4 microm) and a mobile phase consisting of methanol-acetonitrile-oxalic acid (5mM) at the flow rate of 0.8 mLmin(-1). Column temperature was 30 degrees C. The RRS signal was detected at lambda(ex)=lambda(em)=370 nm. The recoveries of sample added standard ranged from 95.3% to 103.5%, and the relative standard deviation was below 2.79%. A detection limit of 2.12-5.12 microgmL(-1) was reached and a linear range was found between peak height and concentration in the range of 10.36-518.0 microgmL(-1) for oxytetracycline (OTC), 12.11-605.5 microgmL(-1) for tetracycline (TC), 11.79-589.5 microgmL(-1) for chlortetracycline (CTC) and 10.32-516.0 microgmL(-1) for doxycycline (DC). The linear regression coefficients were all above 0.999. The method has been applied successfully to the determination of OTC, TC, CTC, DC in pharmaceutical formulations and in honey. The method was simple, rapid and showed a better linear relation and high repeatability.

[Determination of theanine in tea leaves by HPLC-inhibited chemiluminescence with Co2+ catalyst].

A method for the separation and determination of theanine in tea leaves has been developed, based on inhibited chemiluminescence of cobalt-catalyzed luminol-H2O2 reaction by theanine. Separations were performed on a YWG-C18 column using a mobile phase of 0.01 mol.L-1 sodium acetate-acetic acid buffer at pH 5.5 at a flow rate of 0.8 mL.min-1. The optimum concentrations of Co2+, luminol, and H2O2 were 2 micrograms.L-1, 0.25 mmol.L-1, and 0.5 mmol.L-1, respectively. Under the conditions mentioned above, the relative peak area (Y) of inhibited chemiluminescence of theanine was linear with the concentration (X) of theanine in the range of 0.2 g.L-1-5.0 g.L-1 and its linear regression equation was Y = 33,862 x + 1.0605 (r = 0.9983). The average recovery was 97.82% with a RSD of 2.16%.