Simultaneous analysis of 35 specific antihypertensive adulterants in dietary supplements using LC/MS/MS

A sensitive and specific LC/MS/MS method was developed for the simultaneous analysis of 35 compounds used for treating hypertension as adulterants in dietary supplements. The method was validated for specificity, linearity, accuracy, precision, limit of detection, limit of quantitation, stability and recovery. The limit of detection and limit of quantitation ranged from 0.20 to 20.0 and 0.50 to 60.0 ng/g, respectively. The linearity was good (r ² > 0.999), with intra‐ and interday precision levels of 0.43–7.87% and 0.65–9.95% and the intra‐ and interday accuracies of 84.36–115.82% and 83.78–118.69%, respectively. The stability (relative standard deviation) was <14.75%. The mean recovery was 80.81–117.86% (relative standard deviation <10.00%). Ninety‐seven commercial dietary supplements available in South Korea were analyzed. While none contained detectable amounts of the 35 antihypertensive compounds, the developed LC/MS/MS procedure can be used for routine analysis to monitor illegal adulteration in various forms of dietary supplements.     

Simultaneous determination of anti‐diabetes/anti‐obesity drugs by LC/PDA,and targeted analysis of sibutramine analog in dietary supplements by LC/MS/MS

The safety of dietary supplements is questionable as there have been occasional reports of products contaminated with illegal adulterants. The present study was carried out to develop trustworthy methodologies to screen for six anti‐diabetic drugs (phenformin, rosiglitazone, glipizide, glimepiride, glybenclamide and gliclazide) and six anti‐obesity drugs (ephedrine, fenfluramine, T3, T4, fluoxetine and sibutramine) in dietary supplements. A simultaneous determination method of the 12 drugs by liquid chromatography coupled with a photodiode array (LC/PDA) was established and was validated for linearity (r² > 0.99), precision (RSD <13.3%), recoveries (88.8–115.9%) and reproducibility. Sibutramine and its analogs, N‐desmethylsibutramine, were subject to further investigation by LC/MS/MS because they were one of the major illegal adulterants. Our proposed method to monitor illegal drug adulterations in dietary supplements using LC/PDA is a simple and reliable, and therefore applicable to routine drug‐adulteration screening. Copyright © 2009 John Wiley & Sons, Ltd.     

Total and inorganic arsenic in dietary supplements based on herbs, other botanicals and algae—a possible contributor to inorganic arsenic exposure

The content of total and inorganic arsenic was determined in 16 dietary supplements based on herbs, other botanicals and algae purchased on the Danish market. The dietary supplements originated from various regions, including Asia, Europe and USA. The contents of total and inorganic arsenic was determined by inductively coupled plasma mass spectrometry (ICP-MS) and anion exchange HPLC-ICP-MS, respectively, were in the range of 0.58 to 5.0 mgkg^?1 and 0.03 to 3.2 mg?kg^?1, respectively, with a ratio between inorganic arsenic and total arsenic ranging between 5 and 100 %. Consumption of the recommended dose of the individual dietary supplement would lead to an exposure to inorganic arsenic within the range of 0.07 to 13 μg?day^?1. Such exposure from dietary supplements would in worst case constitute 62.4 % of the range of benchmark dose lower confidence limit values (BMDL₀₁ at 0.3 to 8 μg kg bw^?1 kg^?1 day^?1) put down by European Food Safety Authority (EFSA) in 2009, for cancers of the lung, skin and bladder, as well as skin lesions. Hence, the results demonstrate that consumption of certain dietary supplements could contribute significantly to the dietary exposure to inorganic arsenic at levels close to the toxicological limits established by EFSA.     

Determination of lecithin and soybean oil in dietary supplements using partial least squares-Fourier transform infrared spectroscopy

Lecithin and soybean oil in dietary supplements were determined by Fourier transform infrared spectrometry transmission measurements in dichloromethane in combination with a partial least squares (PLS) regression. Two different PLS models were developed, using 16 synthetic mixtures of analytes in dichloromethane, making measurements in the spectral range from 931.8 to 1252.3 cm⁻¹ for lecithin and from 911.4 to 1246.9 cm⁻¹ and 1695.3 to 1774.5 cm⁻¹ for soybean oil. Seven products from the Spanish market with lecithin concentrations between 21.1% and 99.1% and soybean oil concentrations between 0% and 37.2% were analyzed by the proposed method and the data was compared to a chromatographic reference procedure obtaining accurate results. For samples spiked with amounts between 50 and 250 mg of lecithin and soybean oil recovery percentages between 98.0% and 102.1% and between 93.6% and 102.0% with an average precision of 0.35% and 0.41% were achieved for lecithin and soybean oil, respectively. This method can be applied for the quality control of dietary supplements.     

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

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.     

An Analysis of Oxidative Changes and the Fatty Acid Profile in Stored Poultry Sausages with Liquid and Microencapsulated Fish Oil Additives

This study deals with the fatty acid profile and oxidative changes (TBARS) in vacuum-packed (VP) or modified-atmosphere-packed (MAP) finely-comminuted poultry sausages with liquid fish oil and microencapsulated fish oil (MC) additives. An analysis of omega-3 fatty acids (EPA and DHA) showed that their content in the samples with the fish oil additive decreased from the initial value of 0.22 g∙100 g⁻¹ of the product to 0.18 g∙100 g⁻¹ (MAP) and 0.17 g∙100 g⁻¹ (VP), respectively. After in vitro digestion, the total EPA and DHA content in the sample with microencapsulated oil amounted to 0.17 g∙100 g⁻¹ of the product. The TBARS values showed the VP samples with both forms of the fish oil additive had the lowest values on the first day of storage. Storage of the samples for 21 days caused a slight increase in the degree of lipid oxidation. The research indicated that the forms of the oil additive did not have a negative influence on the sensory features or the physicochemical properties of the sausages. The EPA and DHA levels in samples with liquid fish oil and those with oil microcapsules were sufficient for the sausage producer to declare high content of these fatty acids in accordance with the current EC regulation.     

Antioxidant activity of clove oil – A powerful antioxidant source

The number of methods to measure the antioxidants in botanicals, foods, nutraceuticals and other dietary supplements has increased considerably in the last decade. Clove oil is obtained by distillation of the flowers, stems and leaves of the clove tree. In the present paper, clove oil was evaluated by employing various in vitro antioxidant assay such as α,α-diphenyl-β-picryl-hydrazyl free radical (DPPH) scavenging, 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, total antioxidant activity determination by ferric thiocyanate, total reducing ability determination by Fe³⁺–Fe²⁺ transformation method, superoxide anion radical scavenging by riboflavin/methionine/illuminate system, hydrogen peroxide scavenging and ferrous ions (Fe²⁺) chelating activities. Clove oil inhibited 97.3% lipid peroxidation of linoleic acid emulsion at 15 μg/mL concentration. However, under the same conditions, the standard antioxidant compounds such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), α-tocopherol and trolox demonstrated inhibition of 95.4, 99.7, 84.6 and 95.6% on peroxidation of linoleic acid emulsion at 45 μg/mL concentration, respectively. In addition, clove oil had an effective DPPH scavenging, ABTS⁺ scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, ferric ions (Fe³⁺) reducing power and ferrous ions (Fe²⁺) chelating activities. Also, these various antioxidant activities were compared to BHA, BHT, α-tocopherol and trolox as reference antioxidant compounds.     

Detection of refined olive oil adulteration with refined hazelnut oil by employing NMR spectroscopy and multivariate statistical analysis

NMR spectroscopy was employed for the detection of adulteration of refined olive oil with refined hazelnut oil. Fatty acids and iodine number were determined by ¹H NMR, whereas ³¹P NMR was used for the quantification of minor compounds including phenolic compounds, diacylglycerols, sterols, and free fatty acids (free acidity). Classification of the refined oils based on their fatty acids content and the concentration of their minor compounds was achieved by using the forward stepwise canonical discriminant analysis (CDA) and the classification binary trees (CBTs). Both methods provided good discrimination between the refined hazelnut and olive oils. Different admixtures of refined olive oils with refined hazelnut oils were prepared and analyzed by ¹H NMR and ³¹P NMR spectroscopy. Subsequent application of CDA to the NMR data allowed the detection of the presence of refined hazelnut oils in refined olive oils at percentages higher than 5%. Application of the non-linear classification method of the binary trees offered better possibilities of measuring adulteration of the refined olive oils at a lower limit of detection than that obtained by the CDA method.     

Bioelectrochemical evaluation of the total phenols content in olive oil mill wastewaters using a tyrosinase–colloidal gold–graphite–Teflon biosensor

The use of a robust tyrosinase biosensor, fabricated from graphite–Teflon rigid electrode matrices modified with gold nanoparticles, for the estimation of the total phenols content in olive oil mill wastewaters (OMW), is proposed. The performance of this bioelectrode using both continuous stirring and flow-injection amperometry was studied. A potential value of ?0.10?V was selected for the sensitive and stable detection of various phenolic compounds present in OMW samples: catechol, 3,4-dihydroxycinnamic acid (caffeic acid), 3,4-dihydroxyphenylacetic acid (DOPAC), 4-hydroxyphenylacetic acid, 4-hydroxyphenylethanol (tyrosol), and 4-hydroxyphenylpropionic acid. Using catechol as the target phenol, linear calibration graphs were obtained in the 1.0?×?10^?8???8.0?×?10^?6?mol?L^?1 (batch) and 1.0?×?10^?7???1.0?×?10^?5?mol?L^?1 (FI) concentration ranges, with slope values of 750?mA?L?mol^?1 and 103?mA?L?mol^?1, respectively. Batch amperometry was chosen for the analysis of real samples because of its higher sensitivity. For example, the limit of detection for caffeic acid was 80?nM. The ‘pool’ of phenolic compounds was estimated in OMW obtained from different extraction systems and containing phenols at diverse levels of concentration. A comparison of these results with those obtained by applying the Folin–Ciocalteau spectrophotometric reference method was carried out.     

Development of a methodology based on headspace-gas chromatography-ion mobility spectrometry for the rapid detection and determination of patin fish oil adulterated with palm oil

The present research intends to develop a new method based on headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) for the detection and determination of patin fish oil adulterated with different percentages of palm oil, because it is a cheaper vegetable oil. Five variables that affect headspace generation (incubation time and temperature, amount of sample, injection volume, and pre-heating time) have been optimized by means of a Box Behnken design in combination with Response Surface Methodology. Then, repeatability and intermediate precision have been studied where coefficients of variation lower than 10% were obtained. This new method has been applied to several samples of patin fish oil adulterated with palm oil at 20 different levels (5–50% palm oil content). The results have confirmed the suitability of the HS-GC-IMS for a rapid, easy, and reliable detection and discrimination of adulterated oil samples since a characteristic fingerprint that allows 100% successful discrimination between adulterated and unadulterated patin oil samples was achieved. Finally, a regression model has been developed to determine each sample’s adulteration level with an error lower than 10% and a coefficient of determination greater than 0.95.     

A New Polymer Flooding Agent Prepared Through Intermacromolecular Complexation

Abstract

This paper reports a new polymer flooding agent used for enhanced oil recovery (EOR), poly(acrylamide-acrylic acid) [P(AM-AA)]/poly(acrylamide-dimethyldiallylammonium chloride) [P(AM-DMDAAC)] polyelectrolyte complex. The solution viscosity of prepared P(AM-AA)/P(AM-DMDAAC) complex is enhanced due to the strong interaction between the two oppositely charged copolymers, i.e., P(AM-AA) and P(AM-DMDAAC), which were prepared through radical copolymerization. The ionic content could be controlled by changing the reaction conditions. The structures of the two copolymers were characterized through FT-IR, ¹H NMR, and acidic and precipitation titration. The formation as well as the factors affecting the P(AM-AA)/P(AM-DMDAAC) polyelectrolyte complex were investigated by means of viscosity measuring and light transmittance testing. The experimental results show that the composition of the copolymers, the pH value, and the concentration of the polymer solutions have remarkable effects on the formation of P(AM-AA)/P(AM-DMDAAC) polyelectrolyte complex and the solution viscosity. When DMDAAC content in P(AM-DMDAAC) is 3.2 mol%, AA content in P(AM-AA) is 48–58 mol%, the weight ratio of P(AM-AA) to P(AM-DMDAAC) is 70/30–30/70, the pH value of the solution is 6–10, and the concentration of solution is 1000–3500 ppm, then a homogeneous solution of P(AM-AA)/P(AM-DMDAAC) poly-electrolyte complex could be obtained which exhibits a much higher solution viscosity compared with its components.     

Detection of antioxidant butylated hydroxytoluene (BHT) in Antarctic krill (Euphausia superba Dana)

In this study, butylated hydroxytoluene (BHT) was separated and prepared from Antarctic krill. BHT was separated from the volatile oil of Antarctic krill using high performance thin layer chromatography (HPTLC) with petroleum ether/ethyl acetate/hexane (4:1:0.5, v/v/v) was used as the developing solvent. The content of BHT in volatile oil was 9.20?mg/g and the content of BHT in dried Antarctic krill was 0.35?mg/g (0.070?mg/g in frozen whole Antarctic krill). The linearity, accuracy, stability, and recovery of the analysis showed that HPTLC is the most suitable method for the determination of BHT in Antarctic krill. The BHT crude sample was obtained by scraping the separated spot in the thin layer chromatography plate, which was then analyzed using high performance liquid chromatography (HPLC). The resulting sample was identified with 96.05% purity based on the HPLC analysis. The structure of BHT was determined using gas chromatography–mass spectrometry (GC–MS). The results of the HPLC and GC–MS analysis validated the HPTLC method. BHT is a widely used antioxidant in food, pharmaceuticals, and in industrial production. The exploitation and utilization of BHT in Antarctic krill is of great economic value.     

Comparative analysis of the oil and supercritical CO2 extract of Ridolfia segetum (L.) Moris

Supercritical carbon dioxide extraction allowed to obtain the volatile oil of different aerial parts of Ridolfia segetum (L.) Moris. Extraction conditions were as follows: pressure, 90 bar; temperature, 50°C and carbon dioxide flow, Φ?=?1.0?kg?h^?1. Waxes were entrapped in the first separator set at 90?bar and ?10°C. The oil was recovered in the second separator working at 15?bar and 10°C. The main components of the flower oil were α-phellandrene (19.4%), terpinolene (20.5%), piperitenone oxide (11.6%), β-phellandrene (8.2%), (Z)-β-ocimene (7.8%), myristicin (7.5%) and p-cymene (4.4%). The comparison with the hydrodistilled (HD) oil reveal that the significative difference was the content of sesquiterpenes which are higher in the supercritical fluid extraction (SFE) products. Collection of samples at different extraction times during supercritical extraction, allowed to monitor the change of the oil composition. Lighter compounds, as hydrocarbon monoterpenes, were extracted in shorter times than the heavier hydrocarbon and oxygenated sesquiterpenes. The oil from the steams was characterized by a high content of α-phellandrene (12.9%), terpinolene (11.6%), myristicin (11.0%), p-cymene (9.9%), β-phellandrene (8.2%) and (Z)-β-ocimene (6.0%) while the main components of the fruits were found to be myristicin (70.8%), piperitenone oxide (19.9%) and dill apiole (4.2%).     

Quality of Fish-Oil-Based Dietary Supplements Available on the Italian Market: A Preliminary Study

The global market of food supplements is growing rapidly with a large turnover. Fish oil supplements represent a significant part of this turnover as they are believed to have important health benefits. Conversely, there are few papers in the literature about the quality control of fish oil capsules. As prior studies illustrate, a perfect agreement with the label is rarely found, and in some isolated cases, large amounts of soybean oil are also detected, indicating a true adulteration rather than a non-compliance with the label. None of the available studies refer to the Italian market, which ranks first in Europe in the consumption of food supplements. In this present communication, a quality control of fish-oil-based supplements from the Italian market was carried out for the first time. With minor deviations, all results showed substantial agreement with the label. However, the most important conclusion from this research is that compliance with the label is not enough to judge a product of good quality. The analysis of the overall fatty acid composition showed that some supplements have a high level of saturated fatty acids, and therefore they did not undergo a proper purification process. This may represent a safety issue since the purification process also allows the removal of toxic contaminants.     

Thermodynamic interactions and characterisation of naphthenic oil by inverse gas chromatography

The thermodynamic properties of naphthenic oil, a plasticiser, were investigated by means of inverse gas chromatography (IGC) using 10 different kinds of solvents as probes. Some thermodynamic parameters, such as specific retention volume, weight fraction activity coefficient, Flory–Huggins interaction parameter, partial molar heats of mixing and solubility parameter were obtained to judge the interactions between oil and solvents and the solubility of oil in these solvents. The results indicated that n-heptane, n-hexane, cyclohexane, chloroform, benzene and diethyl ether are good solvents for oil at experimental temperatures. The solubility parameters of oil varied from 13.94 to 13.21?(J?cm^?3)^1/2 at temperature range 323–353?K. The solubility parameter of oil was calculated to be 14.38?(J?cm^?3)^1/2 at room temperature, which is consistent with that obtained using surface tension–solubility parameter relation method.     

Determination of mycotoxins,alkaloids, phytochemicals,antioxidants and cytotoxicity in Asiatic ginseng (Ashwagandha,Dong quai, <Emphasis Type="Italic">Panax ginseng</Emphasis>)

Mycotoxins and selected hazardous alkaloids in the medicinal plants (Panax ginseng, Angelica sinensis, and Withania somnifera) and dietary supplements were determined. Purine alkaloids were found in majority of samples; however, isoquinoline alkaloids were less abundant than indole. The predominant alkaloids appear to be caffeine (purine group), harman (indole group) and berberine (isoquinoline). Examined medicinal plants and dietary supplements were contaminated by mycotoxins (especially ochratoxin A 1.72–5.83 µg kg^?1), and many species of mold (e.g. Cladosporium, Eurotium, Aspergillus, Rhizopus, Penicillium). MTT cytotoxicity tests revealed that plant and supplements extracts exhibited medium or high cytotoxicity (only Dong quai—low). Moreover, antioxidant activity, total phenolics content and selected phytochemicals were analyzed by spectrophotometric and chromatographic methods. Quercetin and rutin were predominant flavonols (1.94-9.51 and 2.20–7.28 mg 100 g^?1, respectively). Analysis of phenolic acids revealed—gallic acid, as the most abundant, except Panax ginseng, where ferulic acid was prevailing. The results were analyzed by chemometric methods (cluster analysis, ANOVA).     

Pomegranate Seed Oil and Bitter Melon Extract Affect Fatty Acids Composition and Metabolism in Hepatic Tissue in Rats

Pomegranate seed oil (PSO) and bitter melon dried fruits (BME) are used as natural remedies in folk medicine and as dietary supplements. However, the exact mechanism of their beneficial action is not known. The aim of study was to assess how the diet supplementation with PSO and/or with an aqueous solution of Momordica charantia affects the metabolism of fatty acids, fatty acids composition and the level of prostaglandin E₂ (PGE₂) in rat liver. Animals (Sprague-Dawley female rats, n = 48) were divide into four equinumerous groups and fed as a control diet or experimental diets supplemented with PSO, BME or both PSO and BME for 21 weeks. Fatty acids were determined using gas chromatography with flame ionization detection. PSO added to the diet increased the rumenic acid content (p < 0.0001) and increased accumulation of n-6 fatty acids (p = 0.0001) in hepatic tissue. Enrichment of the diet either with PSO or with BME reduced the activity of Δ⁶-desaturase (D6D) (p = 0.0019), whereas the combination of those dietary factors only slightly increased the effect. Applied dietary supplements significantly reduced the PGE₂ level (p = 0.0021). No significant intensification of the influence on the investigated parameters resulted from combined application of PSO and BME. PSO and BME have potential health-promoting properties because they influence fatty acids composition and exhibit an inhibiting effect on the activity of desaturases and thus they contribute to the reduction in the metabolites of arachidonic acid (especially PGE₂).     

A new approach to ion exchange chromatography with conductivity detection for adulterants investigation in dietary supplements

The extent of adulteration of dietary supplements has significantly increased in recent years. This situation worries health authorities and requires auxiliary analytical tools for the investigation of illegal substances purposely added. Ion exchange chromatography with conductivity detection is a consolidated analytical technique for the determination of inorganic compounds in various matrices. This technique has been applied to the pharmaceutical characterization of mainly impurities and degradation products. This work presents a new approach to ion exchange chromatography as a screening method to investigate the presence of amfepramone, femproporex, sibutramine, bisacodyl and amiloride in dietary supplements advertised for weight loss. The method was optimized and validated using a Metrosep C4 100/4.0 cation exchange column. The mobile phase consisted of 1.8 mm HNO₃ containing 2% acetonitrile (v/v), with a flow rate of 0.9 ml min^?1, and nonsuppressed conductivity detection was applied. The limits of detection and quantification varied from 1.01 to 3.62 mg L^?1 and from 1.48 to 8.72 mg L^?1, respectively. The proposed method was successful applied to 78 solid dietary supplement samples, in two of which adulterations were found. Moreover, ion exchange chromatography with conductivity detection could be easily used for quality control without prior complex sample pre‐treatment.     

A New Method for Olive Oil Screening Using Multivariate Analysis of Proton NMR Spectra

A new NMR-based method for the discrimination of olive oils of any grade from seed oils and mixtures thereof was developed with the aim of allowing the verification of olive oil authenticity. Ten seed oils and seven monovarietal and blended extra virgin olive oils were utilized to develop a principal component analysis (PCA) based analysis of ¹H NMR spectra to rapidly and accurately determine the authenticity of olive oils. Another twenty-eight olive oils were utilized to test the principal component analysis (PCA) based analysis. Detection of seed oil adulteration levels as low as 5% v/v has been shown using simple one-dimensional proton spectra obtained using a 400 MHz NMR spectrometer equipped with a room temperature inverse probe. The combination of simple sample preparation, rapid sample analysis, novel processing parameters, and easily interpreted results, makes this method an easily accessible tool for olive oil fraud detection by substitution or dilution compared to other methods already published.