Primary constituents of blue cohosh: quantification in dietary supplements and potential for toxicity

Dietary supplements containing dried roots or extracts of the roots and/or rhizomes of blue cohosh (Caulophyllum thalictroides) are widely available. This botanical has a long history of use by Native Americans and its use continues to the present day. The primary constituents of blue cohosh are its alkaloids and saponins. The structures of the alkaloids magnoflorine, baptifoline, anagyrine, and N-methylcytisine have been known for many years. The last 10 years have seen a great increase in isolation and identification of the large number of saponins present in blue cohosh. Important developments in nuclear magnetic resonance techniques have contributed substantially to the increase in elucidation of the structures of the complex saponins. Several authors have described quantitative methods for both the alkaloids and saponins in blue cohosh. Such methods have made it possible to quantify these constituents in dietary supplements containing this botanical ingredient. Concentrations of both alkaloids and saponins vary substantially in dietary supplements of blue cohosh. The nicotinic alkaloid, N-methylcytisine, a potent toxicant, has been found in all dietary supplements of blue cohosh analyzed. The teratogenic alkaloid anagyrine has been found in some but not all dietary supplements.     

Promising Antimicrobial Properties of Bioactive Compounds from Different Honeybee Products

Bee products have been known for centuries for their versatile healing properties. In recent decades they have become the subject of documented scientific research. This review aims to present and compare the impact of bee products and their components as antimicrobial agents. Honey, propolis, royal jelly and bee venom are bee products that have antibacterial properties. Sensitivity of bacteria to these products varies considerably between products and varieties of the same product depending on their origin. According to the type of bee product, different degrees of activity were observed against Gram-positive and Gram-negative bacteria, yeasts, molds and dermatophytes, as well as biofilm-forming microorganisms. Pseudomonas aeruginosa turned out to be the most resistant to bee products. An analysis of average minimum inhibitory concentration values for bee products showed that bee venom has the strongest bacterial effectiveness, while royal jelly showed the weakest antibacterial activity. The most challenging problems associated with using bee products for medical purposes are dosage and safety. The complexity and variability in composition of these products raise the need for their standardization before safe and predictable clinical uses can be achieved.     

Status of methodology for the determination of fat-soluble vitamins in foods, dietary supplements, and vitamin premixes

Fat-soluble vitamins (FSVs) include vitamin A, carotenoids, vitamins D, E, and K. New legislation is being introduced in many countries to reinforce regulatory compliance of declared concentrations of vitamins and other micronutrients in food products and dietary supplements. The levels of FSVs are likely to be more closely scrutinized due to their potential health risks associated with overdosing, in particular of vitamin D. However, a proviso of stricter regulatory compliance is that analytical methods must be fit-for-purpose, providing adequate accuracy and precision. Official methods have been published by organizations such as AOAC INTERNATIONAL, European Committee for Standardization, International Dairy Federation, U.S. Pharmacopeia, and International Organization for Standardization. The methods available for foods, dietary supplements, and vitamin premixes are evaluated in this review. In general, these methods show adequate precision for regulatory compliance; however, the field of application has not often been evaluated for a sufficiently large range of food matrixes. Gaps have been noted in the range of published official procedures, particularly for carotenoids and vitamin premixes. The potential of some recent developments in sample preparation and chromatographic techniques were evaluated to provide improved procedures for FSV analysis the future.     

A flow-injection mass spectrometry fingerprinting method for authentication and quality assessment of <Emphasis Type="Italic">Scutellaria lateriflora</Emphasis>-based dietary supplements

Scutellaria lateriflora, commonly known as skullcap, is used as an ingredient in numerous herbal products. However, it has been occasionally adulterated/contaminated with Teucrium canadense and/or Teucrium chamaedrys, commonly known as germander, due to the morphological similarities between the two genera. The latter contains hepatotoxic diterpenes. Despite the potential hepatotoxicity introduced by germander contamination, analytical methodologies for the authentication and quality assessment of S. lateriflora-based dietary supplements have not been reported. In this study, a flow-injection/mass spectrometry fingerprinting method in combination with principal component analysis was used to survey S. lateriflora-based dietary supplements sold in the USA.     

Macroscopic and microscopic authentication of Chinese and North American species of Ephedra

Ephedra sinica Stapf or Ma Huang has been used in traditional Chinese medicine for over 5000 years as a bronchodilating and stimulatory agent. In the West, it is popularly used in dietary supplements for weight loss and to enhance athletic performance. Adverse events have been reported following consumption of dietary supplements containing ephedrine alkaloids. There are about 50 known species of Ephedra. The ratio of ephedrine to other alkaloids varies from species to species; all North American species lack alkaloids. The method commonly used in the dietary supplement industry for botanical authentication is to analyze the product for the presence of chemical markers known to be present in the specific herb. However, this method does not ensure that the product contains authentic herb, especially if it has been spiked with chemical marker compounds. In the trade and raw drug market, Ephedra is available in the form of stem cuttings or powders, without any vouchers, thus making identification of the species difficult. Using light microscopy, we can detect the presence of Ephedra herb, even in powder form, and identify within certain limits its geographical origin. Identification of Chinese and North American species of Ephedra has been made easier by developing a key using leaf and internode length as key identification characters.     

Multielemental Analysis of Bee Pollen,Propolis, and Royal Jelly Collected in West-Central Poland

Beehive products possess nutritional value and health-promoting properties and are recommended as so-called “superfoods”. However, because of their natural origin, they may contain relevant elemental contaminants. Therefore, to assess the quality of bee products, we examined concentrations of a broad range of 24 selected elements in propolis, bee pollen, and royal jelly. The quantitative analyses were performed with inductively coupled plasma-mass spectrometry (ICP-MS) and inductively coupled plasma optical emission spectrometry (ICP-OES) techniques. The results of our research indicate that bee products contain essential macronutrients (i.e., K, P, and S) and micronutrients (i.e., Zn and Fe) in concentrations depending on the products’ type. However, the presence of toxic heavy metals makes it necessary to test the quality of bee products before using them as dietary supplements. Bearing in mind that bee products are highly heterogenous and, depending on the environmental factors, differ in their elemental content, it is necessary to develop standards regulating the acceptable levels of inorganic pollutants. Furthermore, since bees and their products are considered to be an effective biomonitoring tool, our results may reflect the environment’s condition in west-central Poland, affecting the health and well-being of both humans and bees.     

Chromatographic and mass spectrometric fingerprinting analyses of Angelica sinensis (Oliv.) Diels-derived dietary supplements

Angelica sinensis (Oliv.) Diels (“Danggui” in Chinese) is one of the most commonly used traditional Chinese medicines. It has been used to invigorate blood circulation for the treatment of anemia, hypertension, chronic bronchitis, asthma, rheumatism, and cardiovascular diseases. There are a number of A. sinensis-derived dietary supplements in the US markets. However, no study have been conducted to investigate the quality of these dietary supplements. In this paper, high-performance liquid chromatographic and flow-injection mass spectrometric fingerprints were both evaluated to assess the consistency of A. sinensis-derived dietary supplements. Similarity analysis was carried out on the high-performance liquid chromatographic (HPLC) fingerprints. Meanwhile, principal component analysis (PCA) was performed on the data obtained from flow-injection mass spectrometric (FIMS) fingerprints, which can analyze each sample in 2 min, compared with 30 min required for the chromatographic fingerprint. Both methods show significant chemical differences between samples that may be due to differences in growing locations, growing conditions, harvesting times, and/or botanical processing. The loading plots obtained from PCA singled out the discriminatory ions that were responsible for chemical differences of A. sinensis-derived dietary supplements.

An LC-MS screening method with library identification for the detection of steroids in dietary supplements

For many years anabolic-androgenic steroids (AAS) are by far the most frequently detected pharmacological substances in doping control. In order to improve their performances, professional sportsmen are often tempted to take dietary supplements. However, due to the frequent and widespread occurrence of contaminated supplements, the use of such products is not without risk for the athletes involved. In order to minimize the chances of an unattended positive doping test or serious health problems, fast and reliable screening methods for the detection of anabolic steroids in dietary supplements are needed. A general screening procedure requires the fast and unambiguous detection of a large range of steroids. Gas chromatography-mass spectrometry (GC-MS) has been used intensively in the detection of doping substances for the past 40 years. Over time, many laboratories have delivered spectra to be included in standard reference databases, one of which is maintained by the National Institute of Standards and Technology (NIST) (Gaithersburg, MD, USA). In recent years, however, liquid chromatography coupled to mass spectrometry (LC-MS) has gained popularity. Unfortunately, existing GC-MS libraries are not applicable to LC-MS analysis. In the present study, a new mass spectral library of 88 steroids was developed, along with a fast UPLC-MS method. For the construction of this mass spectral library, three different mass spectra were measured for each steroid, with a sample cone voltage of 30, 60 and 100 V, respectively. This method was then successfully tested on contaminated dietary supplements which had previously been tested by means of a targeted LC-MS/MS method. Overall, the library search was shown to identify the same compounds as the MRM method.     

Analysis of Citrus Bioflavonoid Content and Dipeptidyl Peptidase-4 Inhibitory Potential of Commercially Available Supplements

Citrus bioflavonoids are polyphenolic plant-derived pigments found in high levels in oranges, lemons, grapefruits and other citrus fruits. The three most abundant types of citrus bioflavonoids are hesperidin, naringenin and eriocitrin. Citrus bioflavonoids have long been known to possess powerful free radical-scavenging properties and cardioprotective effects. The study involved the analysis of 10 commercially available citrus bioflavonoid supplements from three different countries: Australia, the United States and Canada. The supplements were tested for their citrus bioflavonoid content which varied from 0.8 to 33.3% w/w. The daily bioflavonoid dose varied from 19 mg to 560 mg. Hesperidin was the major citrus bioflavonoid in nine out of ten supplements. One supplement was found to contain less than 10% of the quantity of rutin claimed to have been added. The DPP-4 inhibitory potential, compared through an estimation of rutin equivalence, ranged from 1.9 mg to 400 mg per day. This data highlights the variability between the supplements in their potential to inhibit DPP-4 for subsequent health benefits.     

Simultaneous determination of vitamin A and beta-carotene in dietary supplements by liquid chromatography

Several liquid chromatography (LC) methods for analysis of vitamin A in foods and feeds have been previously reported but only a few have been applied in non-food matrixes. A validated LC method is needed for determination of vitamin A and beta-carotene in the various matrixes presented by dietary supplements. The performance of a reversed-phase method with methanol-isopropanol gradient elution was evaluated with standard retinyl derivatives and beta-carotene. The reversed-phase method is capable of separating retinol from other derivatives such as retinyl acetate, retinyl palmitate, and beta-carotene. Two types of extraction were used to extract the analytes from the dietary supplements: a hexane-methylene chloride extraction for soft-gel capsules containing beta-carotene, and a direct solvent extraction for dietary supplements in tablet form. The direct solvent extraction consisted of treatment with ethanol and methylene chloride following addition of hot water (55 degrees C). Results with the reversed-phase method for vitamin A and beta-carotene in the products examined (n = 8) indicated excellent method performance. The main form of vitamin A or beta-carotene in dietary supplements was the all-trans isomer. The reversed-phase method avoids saponification and is rapid, accurate, precise, and suitable for simultaneous determination of retinyl derivatives and beta-carotene in dietary supplements.     

Aquaculture Production of the Brown Seaweeds Laminaria digitata and Macrocystis pyrifera: Applications in Food and Pharmaceuticals

Seaweeds have a long history of use as food, as flavouring agents, and find use in traditional folk medicine. Seaweed products range from food, feed, and dietary supplements to pharmaceuticals, and from bioenergy intermediates to materials. At present, 98% of the seaweed required by the seaweed industry is provided by five genera and only ten species. The two brown kelp seaweeds Laminaria digitata, a native Irish species, and Macrocystis pyrifera, a native New Zealand species, are not included in these eleven species, although they have been used as dietary supplements and as animal and fish feed. The properties associated with the polysaccharides and proteins from these two species have resulted in increased interest in them, enabling their use as functional foods. Improvements and optimisations in aquaculture methods and bioproduct extractions are essential to realise the commercial potential of these seaweeds. Recent advances in optimising these processes are outlined in this review, as well as potential future applications of L. digitata and, to a greater extent, M. pyrifera which, to date, has been predominately only wild-harvested. These include bio-refinery processing to produce ingredients for nutricosmetics, functional foods, cosmeceuticals, and bioplastics. Areas that currently limit the commercial potential of these two species are highlighted.     

LC/MS/MS analysis of α‐tocopherol and coenzyme Q10 content in lyophilized royal jelly,beebread and drone homogenate

This study shows the results of application liquid chromatography‐tandem mass spectrometry (LC/MS/MS) for assay of the content of α‐tocopherol and coenzyme Q₁₀ in bee products of animal origin, i.e. royal jelly, beebread and drone homogenate. The biological matrix was removed using extraction with n‐hexane. It was found that drone homogenate is a rich source of coenzyme Q₁₀. It contains only 8 ± 1 µg/g of α‐tocopherol and 20 ± 2 µg/g of coenzyme Q₁₀. The contents of assayed compounds in royal jelly were 16 ± 3 and 8 ± 0.2 µg/g of α‐tocopherol and coenzyme Q₁₀, respectively. Beebread appeared to be the richest of α‐tocopherol. Its level was 80 ± 30 µg/g, while the level of coenzyme Q₁₀ was only 11.5 ± 0.3 µg/g. Copyright © 2016 John Wiley & Sons, Ltd.     

High amounts of 17-methylated anabolic-androgenic steroids in effervescent tablets on the dietary supplement market

In numerous studies it has been demonstrated that several nutritional supplements contain prohormones not declared on the label. In the current study two products (effervescent tablets) containing high amounts of the 17-methylated anabolic androgenic steroids metandienone (product 1: 16.8 mg/tablet) and stanozolol (product 2: 14.5 mg/tablet) were identified. Additionally in both products norandrostenedione was detected, in product 2 with minor amounts of several other steroids. The substances identified can cause enormous health risks. In addition, the use of the analyzed tablets can lead to positive doping results for metabolites of the respective steroids in sports. This study again shows the insufficient surveillance of the production and trade of dietary supplements. Consumers should be aware of the enormous health and doping risks connected with the use of such products. For GC-MS identification of the analytes the trimethylsilyl derivatives of the steroids and the mixed N-t-butyldimethylsilyl,O-trimethylsilyl derivatives were used. The quantitation of metandienone, norandrostenedione, and stanozolol was performed using HPLC-DAD.     

Extension of AOAC official method 999.14 (choline in infant formula and milk) to the determination of choline in dietary supplements

AOAC Official Method 999.14 is applicable for the determination of choline in milk and infant formulas. To date, its use has not been extended beyond these matrixes. We modified Official Method 999.14 and applied it to the determination of choline in a range of choline-containing dietary supplements. Dietary supplement tablets, capsules, wafers, softgels, liquid products, and drink powders were included. We found that the standard curve could be extended to cover a wider range of choline concentrations and defined a procedure for the use of Norit for samples in which the vitamin C content was high enough to interfere with the analysis. Recoveries of choline added to infant formula powders and to representative dietary supplement tablets, capsules, powdered drink mix, and wafer products were 85-114%. The use of Norit during the procedure did not affect the recovery of choline added to infant formula powders or to dietary supplements. An alkaline digestion was included for use with a product containing lecithin as the sole source of choline. Ten of 11 dietary supplement products analyzed by the modified method contained amounts of choline at or above declarations found on the product labels. The remaining product contained about 40% of the label-declared amount of choline.     

Mass spectrometric analysis of pharmaceutical adulterants in products labeled as botanical dietary supplements or herbal remedies: a review

The increased availability and use of botanical dietary supplements and herbal remedies among consumers has been accompanied by an increased frequency of adulteration of these products with synthetic pharmaceuticals. Unscrupulous producers may add drugs and analogues of various classes, such as phosphodiesterase type 5 (PDE-5) inhibitors, weight loss, hypoglycemic, antihypertensive and anti-inflammatory agents, or anabolic steroids, to develop or intensify biological effects of dietary supplements or herbal remedies. The presence of such adulterated products in the marketplace is a worldwide problem and their consumption poses health risks to consumers. Analytical methods that allow rapid and reliable testing of dietary supplements for the presence of synthetic drugs are needed to address such fraudulent practices. Mass spectrometry (MS) and hyphenated techniques such as liquid chromatography–mass spectrometry (LC–MS) and gas chromatography–mass spectrometry (GC–MS) have become primary tools in this endeavor. The present review critically assesses the role and summarizes the applications of MS in the analysis of pharmaceutical adulterants in botanical dietary supplements and herbal remedies. The uses of MS techniques in detection, confirmation, and quantification of known pharmaceutical adulterants as well as in screening for and structure elucidation of unexpected adulterants and novel designer drugs are discussed.     

Stingless Bee-Collected Pollen (Bee Bread): Chemical and Microbiology Properties and Health Benefits

Stingless bee-collected pollen (bee bread) is a mixture of bee pollen, bee salivary enzymes, and regurgitated honey, fermented by indigenous microbes during storage in the cerumen pot. Current literature data for bee bread is overshadowed by bee pollen, particularly of honeybee Apis. In regions such as South America, Australia, and Southeast Asia, information on stingless bee bee bread is mainly sought to promote the meliponiculture industry for socioeconomic development. This review aims to highlight the physicochemical properties and health benefits of bee bread from the stingless bee. In addition, it describes the current progress on identification of beneficial microbes associated with bee bread and its relation to the bee gut. This review provides the basis for promoting research on stingless bee bee bread, its nutrients, and microbes for application in the food and pharmaceutical industries.     

A flow-injection mass spectrometry fingerprinting method for authentication and quality assessment of Scutellaria lateriflora-based dietary supplements

Scutellaria lateriflora, commonly known as skullcap, is used as an ingredient in numerous herbal products. However, it has been occasionally adulterated/contaminated with Teucrium canadense and/or Teucrium chamaedrys, commonly known as germander, due to the morphological similarities between the two genera. The latter contains hepatotoxic diterpenes. Despite the potential hepatotoxicity introduced by germander contamination, analytical methodologies for the authentication and quality assessment of S. lateriflora-based dietary supplements have not been reported. In this study, a flow-injection/mass spectrometry fingerprinting method in combination with principal component analysis was used to survey S. lateriflora-based dietary supplements sold in the USA.     

Determination of cadmium residues in bee products using a ‘lab-made’ bismuth bulk electrode

Raw propolis and bee pollen has been widely used in different areas such as medicine, food and apitherapy because they have biological and medicinal properties. The beneficial effects of both bee products for human health include antibacterial, antiviral, antimicrobial and anti-inflammatory properties. However, hives exposure to heavy metals, due beekeeping practices and environment, is one of the problems facing beekeepers. The presence of cadmium in bee products should be controlled, because of its high toxicity. A lab-made Bismuth Bulk Electrode (BiBE) was developed for cadmium residue determination in Argentinean bee products such as raw propolis and bee pollen employing square wave anodic stripping voltammetry (SWASV) as electrochemical technique. The SWASV parameters were optimised based on a 2 ⁵ factorial design. The method presents a linear range from 2.00 to 100.0 µg L^?1 of Cd (II) with a limit of detection (LOD) and a limit of quantitation (LOQ) of 0.78 µg L^?1 and 2.61 µg L^?1, respectively. The repeatability was 8.03%, calculated as the relative standard deviation. Recovery experiments were performed using spiked raw propolis and bee pollen samples with standard deviation values from 1.16% to 7.20%. The BiBE is easy and fast to elaborate and the propose method is environmentally friendly and low cost. Then, it can be considered as a good alternative to the quality control of bee products due to its importance in the beekeeping industry.