Stable Isotope Ratio Analysis for the Authentication of Natural Antioxidant Curcuminoids from Curcuma longa (Turmeric).

Curcuminoid complex, a mixture of curcumin, demethoxycurcumin and didemethoxycurcumin, is one of the most popular antioxidants of natural origin, and it has a multitude of other health benefits. It is threatened by the proliferation of counterfeit products on the market containing synthetic curcuminoids whose addition is difficult to identify as they present the three curcuminoid forms in the correct ratios. Consequently, the necessity to detect this fraudulent practice is escalating. Carbon-14 analysis is the most effective available method, but it is also expensive and difficult to implement. This paper describes the first attempt to characterize natural curcuminoids and their synthetic form, used as an adulterant, through the analysis of stable isotope ratios of carbon and hydrogen (expressed as δ13C and δ2H). Carbon values greater than -28.6‱ and hydrogen values greater than -71‱ may indicate the addition of synthetic curcuminoids to the natural ones.

The phytoequivalence of herbal extracts: A critical evaluation.

The concept of phytoequivalence was developed in Germany in mid-1990 to indicate that a herbal extract is equivalent to another one, more specifically, to a clinically validated one. The pharmacological/physiological activity of an extract is related to its composition, but, due to the presence of many constituents, reliable technique are necessary to compare extracts. While the concept of equivalence among pure chemicals or isolated substances is feasible using the modern chromatographic and spectroscopic technology, specific guideline for herbal extracts or botanicals are still missing. This is mainly due to the multicomponent nature of the herbal extracts and to the natural variability of their constituents. Nevertheless, by using mathematical and/or chemometric approaches, the concept of phytoequivalence can be precisely addressed, as exemplified by a series of examples that are discussed in detail. Helpful practical recommendations based on literature data and personal experience are presented.

The use of stable isotope ratio analysis to characterise saw palmetto (Serenoa Repens) extract.

Saw palmetto extract (SPE) has many pharmacological effects. Thus, its demand and value has increased steadily, along with the presence of counterfeit SPEs on the market. In this work bulk δ13C, δ2H, δ18O and fatty acid δ13C, δ2H analysis was performed in 20 authentic and 9 commercial SPEs, 12 meat fats and 4 pure fatty acids. Authentic SPEs are characterised by bulk values from -31.0‰ to -29.7‰ for δ13C, -176‰ to -165‰ for δ2H, 27.2‰ to 40.7‰ for δ18O, and values of capric, caprylic, lauric, myristic, palmitic and oleic acids from -37.4‰ to -30.5‰ for δ13C and -187‰ to -136‰ for δ2H. The isotopic values of all the commercial SPEs were out of these ranges and more similar to those of meat fat and pure fatty acids. Stable isotope ratio analysis can therefore be proposed as a suitable tool for detecting adulteration in SPEs.

Combined use of isotopic fingerprint and metabolomics analysis for the authentication of saw palmetto (Serenoa repens) extracts.

Saw palmetto (Serenoa repens, SP) is the most expensive oil source of the pharmaceutical and healthfood market, and its high cost and recurrent shortages have spurred the development of designer blends of fatty acids to mimic its phytochemical profile and fraudulently comply with the current authentication assays. To detect this adulteration, the combined use of isotopic fingerprint and omic analysis has been investigated, using Principal Component Analysis (PCA) to handle the complex databases generated by these techniques and to identify the possible source of the adulterants. Surprisingly, the presence of fatty acids of animal origin turned out to be widespread in commercial samples of saw palmetto oil.

Topical anti-inflammatory activity of extracts and compounds from Hypericum perforatum L.

Three preparations of Hypericum perforatum L. (a hydroalcoholic extract, a lipophilic extract and an ethylacetic fraction) and the pure compounds hypericin, adhyperforin, amentoflavone, hyperoside, isoquercitrin, hyperforin dicyclohexylammonium (DHCA) salt and dicyclohexylamine were evaluated for their topical anti-inflammatory activity. H. perforatum preparations provoked a dose-dependent reduction of Croton-oil-induced ear oedema in mice, showing the following rank order of activity: lipophilic extract > ethylacetic fraction > hydroalcoholic extract (ID50 (dose that inhibited oedema by 50%) 220, 267 and >1000 microg cm(-2), respectively). Amentoflavone (ID50 0.16 micromol cm(-2)), hypericin (ID50 0.25 micromol cm(-2)), hyperforin DHCA salt (ID50 0.25 micromol cm(-2)) and adhyperofrin (ID50 0.30 micromol cm(-2)) had anti-inflammatory activity that was more potent or comparable to that of indometacin (ID50 0.26 micromol cm(-2)), whereas isoquercitrin and hyperoside were less active (ID50 about 1 micromol cm(-2)). As dicyclohexylamine alone was inactive, the effect of hyperforin DHCA salt can be attributed completely to the phloroglucinol moiety. The pharmacological activity and phytochemical profile of the tested extracts and fraction suggest that different constituents are involved in the topical antiphlogistic property of H. perforatum in-vivo.

Single-laboratory validation for the determination of terpene lactones in Ginkgo biloba dietary supplement crude materials and finished products by high-performance liquid chromatography with evaporative light-scattering detection.

A single-laboratory validation was completed for a method to determine total terpene lactones in Ginkgo biloba products. The method determines terpene lactones on the basis of the main terpene lactones (Bilobalide, Ginkgolide A, Ginkgolide B, Ginkgolide C, and Ginkgolide J) by high-performance liquid chromatography with evaporative light-scattering detection after extraction. Nine matrixes were chosen for study, including crude leaf material, standardized dry powder extract, single- and multiple-entity finished products, and alcohol and glycerin tinctures. The sample purification with prepacked columns allows selective extraction of the terpene lactones with no interferences from any matrix under study. A Youden ruggedness trial testing 7 instrumental and preparation factors with the potential to affect quantitative results showed that 2 factors (volume of the column elution solvent and pH of the diluent) were the most important parameters to control during sample preparation. The method performed well in terms of precision; 4 matrixes tested in triplicate over a 3-day period showed an overall repeatability relative standard deviation (RSD) of about 3%. HorRat values were within the limits for performance acceptability, ranging from 0.5 to 1.0. Analysis of variance testing at a = 0.05 showed no significant differences among the within-or between-group sources of variation, although comparison of within-day, between-day, and total precision showed that most of the RSD came from within-day determinations except those for the Ginkgo dry extract (Gb-SLV-2). Accuracy testing at 4 concentration levels of terpene lactones obtained by spiking a negative control matrix at approximately 300, 750, 1500, and 2250 microg/mL gave recoveries of about 91% for the 300 microg/mL level, about 98% for the 750 microg/mL level, about 99% for the 1500 microg/mL level, and 97% for the 2250 microg/mL level with an overall recovery of 96% and an RSD of 3.2%.

New liquid chromatography method with ultraviolet detection for analysis of anthocyanins and anthocyanidins in Vaccinium myrtillus fruit dry extracts and commercial preparations.

The Vaccinium myrtillus fruits (bilberry) are a well-known anthocyanins source, and their extracts are widely used in dietary botanicals and pharmaceutical products for the treatment of vascular and vision disorders. Different analytical methods used for standardization of the bilberry extracts and their preparations are available from pharmacopeias and from the literature. However, the methods reported in the literature do not allow the detection of free anthocyanidins, which are markers of poor product quality. A new liquid chromatography method was developed and validated for the identification and quantification of both anthocyanins and anthocyanidins present in bilberry extracts and products. The method shows a good reproducibility and, due to its high specificity, is suitable to identify unequivocally the botanical raw materials used for manufacturing and to evaluate the extract composition, thus ensuring a high degree of product consistency and quality. Forty typical bilberry preparations belonging to 24 different brands were purchased in the marketplace and evaluated for their quality by using the developed method. Results revealed marked differences among the brands despite a common origin and labeling.

Metabolomic evaluation of ginsenosides distribution in Panax genus (Panax ginseng and Panax quinquefolius) using multivariate statistical analysis.

Ginseng is any one of the eleven species belonging to the genus Panax of the family Araliaceae and is found in North America and in eastern Asia. Ginseng is characterized by the presence of ginsenosides. Principally Panax ginseng and Panax quinquefolius are the adaptogenic herbs and are commonly distributed as health food markets. In the present study high performance liquid chromatography has been used to identify and quantify ginsenosides in the two subject species and the different parts of the plant (roots, neck, leaves, flowers, fruits). The power of this chromatographic technique to evaluate the identity of botanical material and to distinguishing different part of the plants has been investigated with metabolomic technique such as principal component analysis. Metabolomics provide a good opportunity for mining useful chemical information from the chromatographic data set resulting an important tool for quality evaluation of medicinal plants in the authenticity, consistency and efficacy.

Metabolomics study of Saw palmetto extracts based on 1H NMR spectroscopy.

Preparations containing Saw palmetto extracts are used in traditional medicine to treat benign prostatic hyperplasia. According to the European and the American Pharmacopoeias, the extract is obtained from comminuted Saw palmetto berries by a suitable extracting procedure using ethanol or supercritical carbon dioxide or a mixture of n-hexane and methylpentanes. In the present study an approach to metabolomics profiling using nuclear magnetic resonance (NMR) has been used as a finger-printing tool to assess the overall composition of the extracts. The phytochemical analysis coupled with principal component analysis (PCA) showed the same composition of the Saw palmetto extracts obtained with carbon dioxide and hexane with minor not significant differences for extracts obtained with ethanol. In fact these differences are anyhow lower than the batch-to-batch variability ascribable to the natural-occurring variability in the Saw palmetto fruits' phytochemical composition. The fingerprinting analysis combined with chemometric method, is a technique, which would provide a tool to comprehensively assess the quality control of Saw palmetto extracts.

Inverse genetic predisposition to colon versus lung carcinogenesis in mouse lines selected based on acute inflammatory responsiveness.

Mouse lines produced by bidirectional selection on the basis of maximum (AIRmax) or minimum (AIRmin) acute inflammatory reactions were examined for the development of chemically induced acute colitis and colon tumors and the development of lung tumors. AIRmax mice were more susceptible than AIRmin to acute colitis induced by ingestion of dextran sodium sulfate showing a 3-fold higher disease activity index and presenting an intense inflammatory infiltrate in the base of colon crypts as well as elevated expression of IL-1beta, TNFalpha, IFNgamma and IL-6 mRNA in colon tissue. AIRmax were also more susceptible than AIRmin to colon cancer induced by 2 or 7 weekly doses of 1,2-dimethylhydrazine (DMH), showing significantly higher numbers of colonic aberrant crypt foci (ACF) at 150 days after DMH treatment (P = 0.01) and significantly higher numbers of tumors affecting larger intestinal areas at 300-475 days. At the latter time point, however, multiple lung adenomas and large adenocarcinomas were found in AIRmin but not in AIRmax mice. Treatment of mice with nimesulide for 60 days beginning 24 h before the first of two DMH doses almost completely inhibited the appearance of ACF in both lines. Furthermore, ACF numbers and the degree of acute inflammation directly co-segregated in an F2 (AIRmax x AIRmin) intercross population. The results demonstrate that genetic determinants of the inflammatory response differentially influence susceptibility to colon and lung carcinogenesis in the AIRmax and AIRmin mouse model.

Genetic selection for high acute inflammatory response confers resistance to lung carcinogenesis in the mouse.

Mice selected for a high acute inflammatory response (AIRmax) are resistant to chemically induced lung tumorigenesis, whereas the low responders (AIRmin) are susceptible. In urethane-treated mice, anti-inflammatory drugs increased the tumor incidence in AIRmax but not AIRmin mice, and an inverse correlation (P<.001) between the degree of acute inflammatory response (AIR) and lung tumorigenesis was found in an F2 (AIRmax x AIRmin) intercross population. The results provide evidence for the involvement of lung tumor modifier loci in AIR regulation and implicate AIR quantitative trait loci in the inherited predisposition to lung cancer.

Hamartomas pulmonares: 18 casos operados / Hamartoma of the lung: 18 cases operated

Se presentan 18 casos de hamartomas pulmonares, tratados quirúrgicamente, 12 enucleaciones, 3 resecciones atípicas y 3 lobectomías. No hubo mortalidad