Anna Filipiak-Szok

Determination of toxic metals by ICP-MS in Asiatic and European medicinal plants and dietary supplements.

The potentially toxic metals content was determined in selected plants, used in Traditional Chinese Medicine (Angelica sinensis, Bacopa monnieri, Bupleurum sinensis, Curcuma longa, Cola accuminata, Emblica officinalis, Garcinia cambogia, Mucuna pruriens, Ocimum sanctum, Panax ginseng, Pueraria lobata, Salvia miltiorrhiza, Schisandra sinensis, Scutellaria baicalensis, Siraitia grosvenorii, Terminalia arjuna and Terminalia chebula), and some European herbs (Echinacea purpurea, Hypericum perforatum, Vitis vinifera). Samples were mineralized in a closed microwave system using HNO3 and the concentrations of Cd, Pb, Al, As, Ba, Ni and Sb were determined by ICP-MS method. Some relevant aspects of potential toxicity of metallic elements and their compounds were also discussed. Results of metal content analysis in dietary supplements available on Polish market, containing studied plants, are presented as well. The results were analyzed by principal component analysis (PCA) and cluster analysis.

Determination of anti-oxidant capacity and content of phenols, phenolic acids, and flavonols in Indian and European gooseberry

Phenolic acids and derivatives of quercetin in Indian (amla) and European gooseberry were determined by high-performance liquid chromatography with diode array detector. The calibration curves were constructed using phenolic compounds standards (the coefficient of determination (R2) was 0.9990–0.9997 for phenolic acids and 0.9989–0.9994 for flavonols, respectively). The lowest detection limit was 0.28 mg L−1 and 0.35 mg L−1 for hyperoside and gallic acid, respectively, whereas the highest was 1.80 mg L−1 and 7.98 mg L−1 for quercetin and chlorogenic acid, respectively. The quantification limits calculated were 0.85–24.04 mg L−1 for hyperoside and chlorogenic acid, respectively. The predominant phenolic acid in amla and gooseberry is gallic acid: (5.37 ± 0.04) mg per 100 g of dry mass (d.m.) and (3.21 ± 0.03) mg per 100 g of d.m., respectively. The next one was caffeic acid, 0.65–1.22 mg per 100 g of d.m., followed by p-coumaric acid, 0.84–1.17 mg per 100 g of d.m. Out of the flavonols, rutin is predominant: (3.11 ± 0.13) mg per 100 g of d.m. and (2.12 ± 0.03) mg per 100 g of d.m., respectively. Anti-oxidant activity was also determined.

Determination of phytochemicals, antioxidant activity and total phenolic content in Andrographis paniculata using chromatographic methods

Antioxidant activity, total phenolics content and selected phytochemicals (alkaloids and andrographolides) were determined in Andrographis paniculata and in dietary supplements containing this plant. Antioxidant activity was measured by FRAP, CUPRAC and DPPH procedures and ranged from 503.36 to 6164.09μmol TE/100g d.m. depending on methods, part of plant and kind of dietary supplement. The total phenolics (175.13-1723.79mg GAE/100g) and andrographolides content (19.44-85.13mg/g) in the studied samples were correlated with antioxidant activities determined by CUPRAC, FRAP and DPPH (r>0.95, p<0.05 level). Purine alkaloids: caffeine, theobromine, theophylline and indole alkaloids: harmine, harmane, harmol, yohimbine, brucine and strychnine were detected in the studied samples by different chromatographic techniques (HPLC-DAD, LC-MS/MS, GC-MS). The total alkaloids content in APs-roots and APs-leaves varies from 50.71±0.36mg/g d.m. to 78.71±0.48mg/g d.m., respectively, whereas for dietary supplements (Pn and DK) TAC was found between 19.52±0.15mg/g and 22.18±0.15mg/g d.m.. The highest concentration of andrographolides was found in A. paniculata leaves, whereas the lowest in dietary supplement Pn. Moreover principal component analysis, cluster analysis and one-way ANOVA follow by Duncans tests were also performed.

Elemental Analysis of Medicinal Herbs and Dietary Supplements

Selected plants used in Traditional Chinese Medicine (Garcinia cambogia, Emblica officinalis, Angelica sinensis, Schisandra sinensis, Scutellaria baicalensis, Salvia miltiorrhiza, and Ocimum sanctum), European herbs (Echinacea purpurea, Cichorium intybus, and Vitis vinifera), and supplements derived from these products were analyzed for their macroelements (K, Na, Ca, and Mg) and microelements (Zn, Cu, Cr, Se, Mn, Mo, and Fe) concentrations by wavelength-dispersion x-ray fluorescence (WD-XRF), scanning electron microscopy–energy dispersive x-ray spectrometry (SEM–EDX), ion chromatography (IC), and inductively coupled plasma–mass spectrometry (ICP–MS). In addition, relevant aspects of heavy metal toxicity are discussed.

Evaluation of antioxidants in Dong quai (Angelica sinensis) and its dietary supplements

The antioxidant activity (AA), total phenolic content (TPC) and total flavonoids content (TFC) in Dong quai (DQ, Angelica sinensis) raw materials and dietary supplements (DS) containing this plant were determined using the CUPRAC, FRAP and fluorescence methods. The antioxidant activity for DQ aqueous extracts revealed by CUPRAC was (1330.45 ± 1.30) μmol Trolox equivalent (TE) per 100 g of dry mass (DM), whereas the antioxidant activity as determined by FRAP was (1813.9 ± 2.0) μmol of TE per 100 g of DM. Lower values were noted for the fluorescence method than for CUPRAC and FRAP (ranging from (35.96 ± 0.3) to (304.6 ± 1.4) μmol of TE per 100 g of DM). The highest TPC values were determined for an aqueous extract of DQ ((3330.3 ± 2.3) μmol of TE per 100 g of DM), while TFC for ethanolic extracts of DQ was ((146.50 ± 0.5) mg of quercetin equivalent (QE) per 100 g of DM). Cinnamic acid, isomers of benzoic acid and derivatives of quercetin were analysed by HPLC-PDA. The ferulic acid concentration in an ethanolic extract of DQ was (21.83 ± 0.07) mg per 100 g of DM. Of the flavonols detected, rutin exhibited the highest concentration in ethanolic extract of DQ ((3.32 ± 0.13) mg of QE per 100 g of DM). Other phytochemicals (alkaloids, saponins, flavonoids, anthraquinones, tannins, steroids, etc.) were identified by phytoscreening colour reaction. The results were analysed by principal component analysis (PCA), cluster analysis and one-way ANOVA tests.

Optimization of Capillary Isotachophoretic Method for Histidine Determination in Protein Matrices

The capillary isotachophoretic method was optimized and used for histidine determination in food samples. The optimum conditions for histidine separation and determination were found on the experimental conditions such as: selectivity, separation speed, pH, concentration of the leading and terminating electrolytes, and electroosmotic flow additives. The optimum electrolytes composition [leading electrolyte: 7 mM NH4OH + 15 mM 2-(N-morpholino)ethanesulfonic acid + 1% hydroxyethylcellulose; pH = 6.10 and terminating electrolyte: 15 mM aminocaproic acid +5 mM acetic acid +40% methanol; pH = 5.10] and conditions of analysis were adopted for histidine determination in food samples (meat and fish products). The proposed electrolyte system was characterized by linearity (10–100 and 100–430 mg · L−1 with R2 = 0.9976 and 0.9991), accuracy (99.5% and 98%), intra-assay of the relative step height (1.40% for standard and 3.20% for food samples analysis), inter-assay of the relative step height (3.65% and 6.30%) and satisfactory quantification and detection limits. The obtained results were compared to a chromatographic method (reversed-phase (RP)-HPLC) for determination of histidine. The average concentrations of histidine in the samples assayed by both methods were statistically comparable. It should be noted that the proposed histidine determination method can be considered as a contribution to Green Analytical Chemistry.

Optimization of extraction procedure and determination by high performance liquid chromatography of flavonols and phenolic acids from Hypericum Perforatum L.

Hypericum perforatum is a medicinal plant which has been known in traditional medicine as an antiinflammatory and healing agent. Nowadays the use of Hypericum extracts is concerned mainly with antidepressive applications. In the present work, HPLC – RP- C18 column chromatography with photodiode array detection was applied for the determination of the derivatives of cinnamic and benzoic acid (e.g., caffeic, chlorogenic, ferulic, sinapic, gallic acids) (Fig.1.) and flavonols - quercetine derivatives (quercetine, rhamnetine, quercitrin, mirycetine, keampferol and rutin) in Hypericum Perforatum. Phenolic compounds were extracted from the sample matrix with ethanol and ethanolwater mixture in different ratios solvent (3:7; 8:2; v/v) at 30°C and 60°C in water-bath shaker and by ultrasonic extraction and then analyzed before and after acid and basic hydrolysis. The total amount of studied flavonols and phenolic acids were compared with the total flavonoids content (TFC) and with total polyphenols content (TPC). UV-Vis spectrometry was used to investigate methods for qualitative and quantitative determination of these compounds.

Simultaneous Determination of Isoquinoline Alkaloids in Medicinal Asiatic Plants by Ultrasound-Assisted Extraction and High-Performance Liquid Chromatography – Mass Spectrometry with Principal Component Analysis

ABSTRACT Isoquinoline alkaloids (papaverine, noscapine, berberine, emetine, and quinine) were determined in medicinal plants and herbs used in Traditional Chinese Medicine and Ayurveda by liquid chromatography with tandem mass spectrometry detection (LC-MS/MS). The analyzed alkaloids were separated at gradient conditions using methanol and 2% acetic acid within 12 min. The validated method was successfully applied for 17 herbal samples: Ashwagandha, Astragalus membranaceus, Emblica officinalis, Mucuna pruriens, Pueraria lobata, Ocimum sanctum, Rehmannia glutinosa, Schisandra sinensis, Terminalia arjuna, Terminalia chebula, and dietary supplements. The highest concentration of studied alkaloids was observed for berberine in Puearia lobata (6.68 ± 0.62 mg 100 g−1 d.m.), while the lowest value was obtained for noscapine in a dietary supplement containing Terminalia arjuna (0.09 ± 0.01 mg 100 g−1 d.m.). Principal component analysis, cluster analysis, and one-way ANOVA tests were also performed. The results indicate the need to control plant materials and dietary supplements in terms of the content of alkaloids and toxic components.

Application of Capillary Isotachophoresis in Phenothiazines Determination

A capillary isotachophoretic method (cITP) using three electrolyte systems for determination of promethazine, thioridazine and chlorpromazine hydrochlorides in pharmaceutical preparations was demonstrated. Proposed systems were characterised by linearity range 5-100 mg/L with R2 in all cases were higher than 0.999. The elaborated method was tested on pharmaceutical preparations. The recovery values were from 96.5% to 101.3%. The proposed isotachophoretic method is simple with acceptable precision and accuracy and can be suitable for routine analysis of studied biological active compounds.

The influence of pH and selected cations on the spectrofluorometric determination of oxytetracycline hydrochloride

The spectrofluorometric method for the oxytetracycline hydrochloride in pure and in veterinary products Tetrox and Oxymed 50 determination is described. The influence of pH solution and presence selected cations on fluorescence intensity were studied too. It was ascertained that the highest fluorescence intensity take place at pH=9. Moreover, the quenching of fluorescence intensity was observed at presence Ca2+, Al3+ and Fe3+ in contrast to Mg2+ which caused increasing of intensity. The obtained recovery (97.23±0.12% for Tetrox and 95.21±0.10% for Oxymed 50) values meet the European Pharmacopoeia requirement. Moreover, the relative standard deviation (RSD) was below 0.23% confirmed high precise of the method. The statistical test (t-Student and F-Snedecor) used for comparison spectrofluorometric and chromatographic methods pointed that they are comparable in respect of precision but not of accuracy.