Marzanna Kurzawa

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 quercetin and rutin in selected herbs and pharmaceutical preparations.

Optimization of the extraction time of rutin and quercetin from herbal plant (Inflorescentia tiliae, Violae tricoloris herba, Anthodium arnica, Flos sambuci, Herba rutae, and Herba hyperici) is described. Determination of aforementioned flavonols was performed by pharmacopoeial, spectrophotometric, and chromatographic methods. The highest quercetin concentration was obtained for Inflorescentia tiliae whereas Herba hyperici contains the highest amount of rutin. The methods were tested on pharmaceutical preparations. The obtained results were compared by statistical test.

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.

Application of isotachophoretic and conductometric methods for neomycin trisulphate determination

Determination of neomycin trisulphate (NMS) in a dosage form (Neox and Neosol) was carried out by capillary isotachophoresis (cITP) with conductometric detection. The following electrolytes: leading: 10 mmol dm−3 sodium acetate + 0.08 % hydroxyethylcelulose (HEC) and acetic acid to pH = 5.5, and terminating: 10 mmol dm−3β-alanine were tested for isotachophoretic separation of NMS. The calibration curve was linear over the range of 10.00 mg dm−3 to 100.00 mg dm−3 with LOD = 5.69 mg dm−3 and LOQ = 18.96 mg dm−3. The results were compared to the conductometric determination of NMS with: ammonium molybdate (VI), silver nitrate (V) and Reinecke salt. Good accuracy was obtained from conductometric titration of NMS with Reinecke salt, the recoveries being as follows: 100 % (RSD = 1.99 %); 96.17 % (RSD = 2.10 %) and 95.22 % (RSD = 1.55 %) for NMS in pure form, Neosol and Neox, respectively.

Determination of benzoate, sorbate, citrate and orthophosphate ions in beverage samples using two-dimensional isotachophoretic method

A two-dimensional capillary isotachophoretic method (cITP-cITP) using electrolyte system consisting of leading electrolytes (LE1): [10 mM HCl + β-alanine (pH 3.9) + 0.1% hydroxyethylcellulose (HEC)] and (LE2): [10 mM HCl + aminocaproic acid (pH 5.00) + 0.1% HEC], and 5 mM caproic acid as terminating electrolyte (TE) was studied. Two methods of detection, conductometric and UV-Vis, were applied to the determination of selected food preservatives and additives. Practical applicability was demonstrated by simultaneous determination of benzoates, sorbates, citrates and orthophosphates in 12 samples of beverages. The proposed method revealed linearity with R2 between 0.9992 and 0.9999 for the concentration ranges: 10–100 mg/L (orthophosphate and citrate ions), 20–100 mg/L (sorbates) and 40–120 mg/L for benzoates. The detection limits for all studied ions were from 0.85 to 3.1 mg/L whereas the quantification ones were from 2.8 to 10 mg/L. The variation coefficients for five-fold analysis of all ions ranged between 0.4 and 9.1%. Obtained recoveries (from 97 to 104%) confirmed satisfactory accuracy of the proposed cITP-cITP method for the determination of tested food additives.

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.

Determination of metoprolol tartrate by capillary isotachophoresis

This paper describes two isotachophoretic methods of metoprolol tartrate (MT) determination in pure and dosage forms. The first method was used for direct analysis where the following electrolyte system was applied: 10 mmol dm−3 3-morpholino-2-hydroxypropanesulfonic acid, 10 mmol dm−3 NaCl, 2 % hydroxyethylocelulose as leading (LE) and 10 mmol dm−3 glycyl-glycine as terminating (TE) electrolytes. The second method was used for indirect analysis of MT as tartrate ions. In this case, the leading electrolyte consisted of 10 mmol dm−3 HCl, β-alanine (BALA), pH 4-5, and the terminating one of 5 mmol dm−3 glutamic acid, 10 mmol dm−3β-alanine. Calibration curves were calculated as follows: for system A: y = (0.52 ± 0.05)x − (0.9 ± 0.2) (LOD = 13.0 mg dm−3, LOQ = 31.7 mg dm−3); and for system B: y = (0.240 +- 0.001)x + (0.18 ± 0.06) (LOD = 1.8 mg dm−3, LOQ = 4.4 mg dm−3). The isotachophoretic method was compared with the pharmacopoeial one by statistical tests.