Novel spectrofluorimetric method for boldine alkaloid determination in herbal drugs and phytopharmaceuticals.

A new green on-line method for Boldine determination (BOL) in herbal drugs and phytopharmaceuticals, using its native fluorescence in acid media (λex=282nm; λem=373nm) has been developed. The presented methodology involves for the first time, a flow injection (FI) strategy using a mini-column of multiwalled carbon nanotubes as retention agent coupled with molecular fluorescence. Different parameters influence as sample pH and flow rate, eluent flow rate and composition; on BOL sensitivity and elution time was investigated by multifactorial techniques. Adequate dynamic calibration range (r2=0.9993) was obtained over a concentration interval of 0.029-27.0µgmL-1 BOL. The limits of detection (LOD) and quantification (LOQ) were 0.008 and 0.029µgmL-1, respectively. The average recoveries in explored samples ranged from 95% to 103%. Under optimized conditions, the throughput sample as high as 30h-1 was achieved with high repeatability performance (99%). The proposed development represents a useful and valuable tool emulating the analytical efficiency of the official methodologies for quality control of herbal and phytopharmaceutical drugs containing BOL. Moreover, this approach shows advantages respect to low cost, simplicity and environmental and analyst friendly.

Liquid chromatography coupled to molecular fluorescence with postcolumn UV sensitization for thimerosal and derivative compounds monitoring in environmental samples.

A HPLC coupled with molecular fluorescence (MF) spectrometry method for determination of thimerosal (THM, sodium ethylmercurythiosalicylate, C9 H9 HgNaO2 S), and derivatives is proposed. A sensitization of MF was provoked by UV irradiation of analytes in a home-made photoreactor that served as interface between the LC column and MF spectrometer. This method is applied to determination of THM, ethyl mercury, and thiosalicylic acid in samples of pharmaceutical industry effluents, and waters of La Carolina and Jáchal rivers situated in the center-west side of San Luis city and in the east of San Juan city (Middle West, Argentine) where the effluents are dumped. The LODs calculated on basis of 3σ criterion were 1.8, 5, and 0.05 µmol/L for THM, ethyl mercury, and for thiosalicylic acid, respectively.

Determination of thimerosal in pharmaceutical industry effluents and river waters by HPLC coupled to atomic fluorescence spectrometry through post-column UV-assisted vapor generation.

A high performance liquid chromatography coupled with atomic fluorescence spectrometry method for the determination of thimerosal (sodium ethylmercury thiosalicylate, C9H9HgNaO2S), ethylmercury, and inorganic mercury is proposed. Mercury vapor is generated by the post-column reduction of mercury species in formic acid media using UV-radiation. Thimerosal is quantitatively converted to Hg(II) followed by the reduction of Hg(II) to Hg(0). This method is applied to the determination of thimerosal (THM), ethylmercury (EtHg) and inorganic Hg in samples of a pharmaceutical industry effluent, and in waters of the San Luis River situated in the west side of San Luis city (Middle West, Argentine) where the effluents are dumped. The limit of detections, calculated on the basis of the 3σ criterion, where 0.09, 0.09 and 0.07 µg L(-1) for THM, EtHg(II) and for Hg(II), respectively. Linearity was attained from levels close to the detection limit up to at least 100 µg L(-1).

Fluorescence method using on-line sodium cholate coacervate surfactant mediated extraction for the flow injections analysis of Rhodamine B.

An on-line surfactant mediated extraction method in a flow injection analysis format with fluorescence detection was developed for the determination of Rhodamine B (RhB) in food condiments. The sample was extracted using the phase separation behavior exhibited by the bile salt surfactant, sodium cholate (NaC), upon addition of sodium dodecylsulfate (SDS) in the presence of acid at room temperature. The RhB in the sample was incorporated into the NaC/SDS coacervate phase which was then collected on a glass-wool packed mini column from which it was subsequently eluted using a 1.00 mol L(-1) HCl solution. The inherent fluorescence (λex=555 nm; λem=575 nm) of RhB was employed for detection. Good linearity (r(2)=0.9933) was obtained over the concentration range 0.4-4794-479.0 µg L(-1) RhB. The detection (LOD) and quantification (LOQ) limits were 0.12 and 0.40 µg L(-1), respectively. The method was successfully applied for analysis of RhB in food condiments and spiked samples. The average recoveries ranged from 95.3% to 118.9% at spiked concentration levels of 1.19 and 2.39 µg L(-1). Under optimized conditions, a throughput of 50 samples per hour was achieved. The proposed method may be a valuable tool not only for quality control of food condiments and similar food confectioneries but for the analysis of a variety of other RhB-containing samples as well.

On-line enantioseparation of chlorpheniramine using ß-cyclodextrin and carbon nanotubes after multivariate optimization.

Multiwalled carbon nanotubes are evaluated here as solid phase extraction (SPE) sorbent aiming to (±)-chlorpheniramine (CPA) enantioresolution with fluorimetric detection. ß-cyclodextrin (CD) was added to the racemate and solutions with HCl and sodium dodecyl sulfate (SDS) in different proportions were assayed as eluents to achieve the separation between both enantiomers. The overall methodology involved a flow injection (FI) strategy enabling high sample throughput and low reagents consumption making it suitable for drug routine quality control. An adequate enantioresolution (2.08) with satisfactory responses for both (R)-CPA (peak area=285) and (S)-CPA (peak area=380) was achieved applying the proposed FI-SPE strategy under the optimized conditions [ß-CD] = 1.0 mmol L(-1), [HCl] = 1.0 × 10(-2) mol L(-1), [SDS] = 4.0 × 10(-4) mol L(-1) and eluent flow rate = 8.0 rpm.

Monitoring of phenolic compounds for the quality control of Melissa officinalis products by capillary electrophoresis.

INTRODUCTION: Official assays for the quality control of Melissa officinalis L. (Lamiaceae) leaves establish the quantification of total hydroxycinnamic derivatives expressed as rosmarinic acid. OBJECTIVE: The goal of this work was to develop a simple, fast and reliable method for monitoring the phenolic composition in herbs from the Lamiaceae family and for rapidly detecting M. officinalis adulteration or substitution in commercial medicinal samples in Argentina. METHODOLOGY: A capillary zone electrophoresis (CZE) method was performed under the following conditions: the background electrolyte (BGE) consisted of 20 m m sodium tetraborate buffer, pH 9.2; the applied voltage was 25 kV; the capillary and sample temperatures were kept at 25 °C; the hydrodynamic mode was selected for the sample injection (3.45 kPa during 5 s). RESULTS: A CZE method that achieved the separation and simultaneous determination of eight related phenolic compounds in less than 11 min was optimised for application to control quality analysis of M. officinalis-based products. The method was validated according to the US Federal Drug Agency requirements and offers advantages in terms of analysis time, cost and operation. CONCLUSIONS: The proposed methodology can be applied to the standardisation and quality control of plant material and phytopharmaceutical products derived from the Lamiaceae family, as indicated by the results obtained in the analysis of commercial medicinal products in Argentina.

Analysis of potential adulteration in herbal medicines and dietary supplements for the weight control by capillary electrophoresis.

Four different phytopharmaceutical dosage forms for use in weight control programs were analyzed. Two different ground herbal blends and their correspondent infusions, a capsule and a tincture were investigated for the presence of compounds used as adulterants in these products. A capillary electrophoresis (CE) method was developed and validated. The optimized experimental conditions were: BGE, sodium tetraborate buffer 20mM, pH 9.2, voltage applied 30kV, capillary temperature 25°C, injection sample at 0.5Psi during 5s. Ephedrine, norephedrine, caffeine and furosemide were baseline separated in less than 7min; the migration times were found to be 2.65, 2.90, 3.75 and 6.58min, respectively. The analysis showed in sample 3 concentrations of 0.45±0.03mgg(-1) (ephedrine), 0.33±0.02mgg(-1) (norephedrine), 1.09±0.41mgg(-1) (caffeine) and 0.80±0.17mgg(-1) (furosemide). Caffeine content in samples 1, 2 and 4 was 0.61±0.06mgg(-1), 15.66±1.05mgg(-1) and 2.27±0.13mgml(-1), respectively. Linearity was obtained in the concentration range of 1-1000µgml(-1). Limits of detection (LOD) and quantification (LOQ) were determined as 0.42µgml(-1) and 1.40µgml(-1) (ephedrine), 0.47µgml(-1) and 1.40µgml(-1) (norephedrine), 0.12µgml(-1) and 0.48µgml(-1) (caffeine), 0.22µgml(-1) and 0.73µgml(-1) (furosemide). The common constituents of the samples did not interfere with the potential adulterants. Repeatability was better than 0.24% RSD for the retention time and 1.43% for the peak area. Intermediate precision was tested by changing the capillary, the day of operation and the operator, in all the cases the %RSD was better than 3.06.