Rapid and robust analysis of aristolochic acid I in Chinese medicinal herbal preparations by surface-enhanced Raman spectroscopy.

The use of Chinese herbs containing aristolochic acid can induce the exchange of adenine and thymine in gene mutations and even cause liver cancer. To eliminate the harm of aristolochic acids (AAs) to humans, a rapid and robust method of AAs screening is a prerequisite. In this work, a facile and robust Surface-enhanced Raman spectroscopy (SERS) method was used for the qualitative and quantitative detection of AAs in Chinese medicinal herbal preparations based on the mandelic acid modified Ag nanoparticles SERS substrate. Qualitative and quantitative SERS detection of Aristolochic acid I (AAI) was achieved with a good linear relationship ranging from 0.2 - 120.0 µM and a limit of detection (LOD) of 0.06 µM. The proposed method demonstrates a refined strategy for sensitivity analysis of AAs with the advantages of easy operation, time-saving, high sensitivity, and molecular specificity, making it a preferred platform for the screening of AAI in regular inspections of herbal products and regulatory supervision of the supply chain.

Thermal resistance and high-performance microwave decontamination assessment of Bacillus endospores isolated from food-grade herbal extracts.

Generally, endospore contamination can occur from different sources during product manufacturing in many industries and therefore lower its quality by affecting physicochemical properties and shelf-life. Bacterial endospores can germinate inside the product and produce several enzymes, which can cause several undesirable changes. This study assessed the spores thermal resistance and applied a microwave decontamination technique toward herbal extracts (Tilia tomentosa and Centella asiatica) containing ethanol or glycerol. Based on 16S rRNA analysis, the detected contaminant endospores belonged to different Bacillus species, namely B. subtilis, B. zhangzhouensis, and B. pumilus. The thermal resistance assessment using inoculated endospores in the actual products revealed B. pumilus T2 as the most resistant endospore to the heat treatments tested in both T. tomentosa and C. asiatica extracts. Finally, a high-performance microwave technique was used to decontaminate T. tomentosa extract against the mixture of Bacillus spores. Results from the microwave technique indicate that the increase of temperature from 100°C to 105°C not only decontaminated the product but also could dramatically decrease the effective thermal treatment time (10 times), which can benefit the product quality. The results provided in this study considerably contribute to improving an original decontamination method for products containing glycerol and ethanol with the most negligible effect on product quality.

A Graded Approach for Evaluating Health Claims about Plant-Based Food Supplements: Application of a Case Study Methodology.

The implementation of REGULATION (EC) No 1924/2006 has led to the formation of a list of health claims that can be used in food supplements (EU 432/2012). However, such supplements are often composed of plant preparations with claims omitted from this list. The peculiarity of plants is related to their long history of use, that could allow claims based on traditionally recognized health effects. In addition, the scientific literature has been enriched over the years through clinical studies that have assessed the bioavailability and efficacy of bioactive components, and investigated their mechanisms of action. Based on existing recognized models which aim to classify research according to the level of scientific evidence, Synadiet developed a three-grade model (A, B or C) for assessing plants health claims. In this paper, the applicability of the model is illustrated through an example for which a Grade B health claim attesting the possible contribution of red clover isoflavones to the improvement of blood lipid levels in postmenopausal women has been attributed. The model appears able to be easily extrapolated to claims pertaining to other plants. If adopted by consensus at European level, this model could initiate the implementation of a positive list of health claims on plant preparations.

Effect-directed analysis as a method for quality and authenticity estimation of Rhodiola rosea L. preparations.

Adulterations of food and pharmaceutical preparations are the important global problem. On the one hand, fraud practices are becoming more and more sophisticated while on the other, monitoring and uncovering falsifications are insufficient. One of the most common consumer concern is the quality and authenticity of the purchased products, related to the confidence that they have composition and properties in accordance with the manufacturer's declaration on the label. This refers also to pharmaceuticals potentially delivering great health benefits such as Rhodiola rosea L. supplements. The aim of this study was defining authenticity and possible adulterations of two R. rosea preparations basing on their TLC-bioprofiles and the presence of biomarker compounds characteristic for this plant. The effect-directed analysis (EDA), i.e. TLC hyphenated with micro-chemical and biological assays performed directly on TLC plates followed by HPLC-ESI-MS was used for the bioprofiling of antioxidants, antibacterials, and inhibitors of lipase, acetylcholine, α-glucosidase and tyrosinase as well as for the identification of the biomarkers. The results pointed to the possible adulteration of one of the tested products related to the absence of two rosavins, the most important quality markers of R. rosea.

Phytochemical profile and ethnopharmacological applications of Lecythidaceae: An overview.

ETHNOPHARMACOLOGICAL RELEVANCE: The family Lecythidaceae has about 25 genera and 400 identified species, which are distributed especially in the pantropical region, mostly found in the tropics of Central and South America, Southeast Asia and Africa. The third most abundant family in Amazonian forests and the genus Eschweilera, with the large number of species in Lecythidaceae. AIM OF THE REVIEW: The present review compiles information since the 1934s about of ethnopharmacology, and chemical constituents of species of Lecythidaceae, as well as a summary of the bioactivities shown by the extracts, fractions, and secondary metabolites. MATERIALS AND METHODS: All relevant information on ethnopharmacology, and chemical constituents of species of Lecythidaceae were gathered from electronic databases including Web of Science, Science Direct, Elsevier, ResearchGate, and Google Scholar. Information was also obtained from local books, PhD. and MSc. Dissertations. RESULTS: The phytochemical diversity of the family was demonstrated with 180 different metabolites that have been reported from 25 species, most of them being triterpenes or flavonoids. The pharmacological studies carried out with the extracts, fractions and compounds showed promising antibacterial, antifungal, anti-inflammatory, antinociceptive, cytotoxic and antioxidant activities. CONCLUSION: The present review provides an insight into ethnopharmacology, phytochemistry and pharmacology of species of Lecythidaceae. Based on the pharmacological studies it has been found that different plant species of Lecythidaceae possess a wide range of bioactivities such as anti-arthritic, anti-inflammatory, antileishmanial, antibacterial and antifungal. These activities are due to the presence of bioactive compounds including triterpenoids and their glycosides derivatives, flavonoids, steroids, sesquiterpenoids, alkaloids, and other compounds. However, there are many plants, which have not been assessed pharmacologically and hence warrant further studies.

Risk Associated with the Use of Selected Ingredients in Food Supplements.

This review focuses on four new product categories of food supplements: pre-workout, fat burner/thermogenic, brain/cognitive booster, and hormone/testosterone booster. Many food supplements have been shown to be contaminated with unauthorized substances. In some cases, the ingredients in the new categories of dietary supplements were medicinal products or new synthetic compounds added without performing clinical trials. Some of the new ingredients in dietary supplements are plant materials that are registered in the pharmacopoeia as herbal medicines. In other cases, dietary supplements may contain plant materials that have no history of human use and are often used as materials to 'camouflage' stimulants. In the European Union, new ingredients of dietary supplements, according to European Food Safety Authority or unauthorized novel food. Furthermore, selected ingredients in dietary supplements may be prohibited in sports and are recognized as doping agents by World Anti-Doping Agency.

Quality evaluation of Pinellia tuber by LC-TOF/MS targeted to ephedrine.

Pinellia tuber (PTE, , , , , , , , ) is derived from the tuber of Pinellia ternata Breitenbach (Araceae), which is a crude drug used in traditional Japanese Kampo medicine for the purpose of antiemesis and expectoration. Since the separation of ephedrine from PTE in 1978, it has been listed as a PTE component in textbooks and internet information. Therefore, there are harmful effects on appropriate use in clinical practice because PTE is dealt with as a crude drug for doping target, and traditional Japanese Kampo medicine containing PTE must be carefully administered to the elderly. However, since the 1978 published report, there has not been any report on the isolation of ephedrine from PTE and the interpretation of biosynthesis remains questionable. In the present study, we analyzed the PTE samples in market distribution products by LC-TOF/MS. From the analysis of the result of ephedrine's m/z 148.113 [M + H-H2O]+, PTE was not detected (n = 55, detection limit: 0.5 ppb). Additionally, the tuber of P. tripartite (PTR, ), the tuber of P. pedatisecta (PPE, ), Arisaema Tuber (ART, ), and the tuber of Typhonium flagelliforme (TFI, ) that have a similar description to PTE were also not detected. Moreover, the genetic analysis of experimental samples showed that PTE is derived from P. ternata. Furthermore, our attempt to isolate ephedrine from PTE based on the past literature was unsuccessful. These results suggest that PTE in market distribution products may not contain ephedrine as a component.

Rapid identification and determination of pyrrolizidine alkaloids in herbal and food samples via direct analysis in real-time mass spectrometry.

Pyrrolizidine alkaloids (PAs) are naturally occurring plant toxins associated with severe liver damage if excessive ingestion. Herein, a novel analytical strategy on utilizing direct analysis in real-time mass spectrometry (DART-MS) was developed, and applied in analysis of six representative PAs. The calibration curves in the range of 10-1000 ng·mL-1 were established, and relative standard deviations (RSDs) were less than 10%. The limits of detection (LODs) and limits of quantitation (LOQs) were 0.55-0.85 ng·mL-1 and 1.83-2.82 ng·mL-1, respectively. The feasibility of method was indicated by analysing real samples including Gynura japonica, drug tablets, granules, and fresh cow's milk. Moreover, the results of DART-MS were in good agreement with those observed by high performance liquid chromatography mass spectrometry (HPLC-MS), but consumed less time without chromatographic separation. This research provides a facile fashion for safety assessment of herbal and food products containing PAs and presents promising applications in food, pharmaceutical and clinical analysis.

Quantitative determination of aloin, antioxidant activity, and toxicity of Aloe vera leaf gel products from Greece.

BACKGROUND: Aloe vera is a popular medicinal plant used widely by the cosmetic, pharmaceutical, and food industries. The A. vera leaf gel, which is used mostly for its positive effects on human health, contains over 75 different bioactive compounds, including aloin. Aloin is a toxic compound, and its content in A. vera leaf gel products depends on the different cultivation conditions and especially on leaf processing. RESULTS: In this study, A. vera leaf gel products, varied in terms of leaf processing, were analyzed using liquid chromatography for their aloin content, their antioxidant activity by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) radical cation (ABTS·+ ) and the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH· ) antioxidant activity assays and their toxicity against Aliivibrio fisheri and SH-SY5Y cells. In the samples processed with industrial methods and in those filtered in the lab, the content of aloin was found below the limit (0.1 mg L-1 ) of the EU legislation however, the unprocessed and unfiltered samples were found to contain more than 10 mg L-1 . Antioxidant activity was estimated to vary from 1.64 to 9.21 µmol Trolox mL-1 for DPPH· and from 0.73 to 5.14 µmol Trolox mL-1 for ABTS·+ . Toxicity values on A. fisheri, expressed as the concentration at 50% loss of initial luminescence, ranged from 0.03 to 0.09 mg mL-1 . The cytotoxic study indicated that aloin A at low concentrations (1 and 10 µg mL-1 ) protects SH-SY5Y cells from toxicity induced by hydrogen peroxide. CONCLUSIONS: Consequently, the filtration process of A. vera leaf gels, either laboratory or industrial, resulted in aloin A content below the EU legislation detection limits. © 2020 Society of Chemical Industry.

Suspected-target and non-targeted screenings of phosphodiesterase 5 inhibitors in herbal remedies using liquid chromatography-quadrupole time-of-flight-mass spectrometry.

The lucrative market of herbal remedies spurs rampant adulteration, particularly with pharmaceutical drugs and their unapproved analogues. A comprehensive screening strategy is, therefore, warranted to detect these adulterants and, accordingly, to safeguard public health. This study uses the data-dependent acquisition of liquid chromatography-quadrupole time-of-flight-mass spectrometry (LC-QTOF-MS) to screen phosphodiesterase 5 (PDE5) inhibitors in herbal remedies using suspected-target and non-targeted strategies. For the suspected-target screening, we used a library comprising 95 PDE5 inhibitors. For the non-targeted screening, we adopted top-down and bottom-up approaches to flag novel PDE5 inhibitor analogues based on common fragmentation patterns. LC-QTOF-MS was optimised and validated for capsule and tablet dosage forms using 23 target analytes, selected to represent different groups of PDE5 inhibitors. The method exhibited excellent specificity and linearity with limit of detection and limit of quantification of <40 and 80 ng/mL, respectively. The accuracy ranged from 79.0% to 124.7% with a precision of <14.9% relative standard deviation. The modified, quick, easy, cheap, effective, rugged, and safe extraction provided insignificant matrix effect within -9.1%-8.0% and satisfactory extraction recovery of 71.5%-105.8%. These strategies were used to screen 52 herbal remedy samples that claimed to enhance male sexual performance. The suspected-target screening resulted in 33 positive samples, revealing 10 target analytes and 2 suspected analytes. Systematic MS and tandem MS interrogations using the non-targeted screening returned insignificant signals, indicating the absence of potentially novel analogues. The target analytes were quantified from 0.03 to 121.31 mg per dose of each sample. The proposed strategies ensure that all PDE5 inhibitors are comprehensively screened, providing a useful tool to curb the widespread adulteration of herbal remedies.

The colloidal gold nanoparticle-based lateral flow immunoassay for fast and simple detection of plant-derived doping agent, higenamine.

Higenamine (HM), an alkaloid found in various plant species, is obtained when norcoclaurine synthase selectively condenses dopamine and 4-hydroxyphenylacetaldehyde to give (S)-higenamine ((S)-HM). The World Anti-doping Agency has listed HM as a prohibited agent in athletics. As a result, many commercial, academic, and regulatory bodies across the globe are invested in finding a rapid method for (S)-HM detection. In the current study, a lateral flow immunoassay (LFA) was developed in which the relevant biosensor was generated as a conjugate of the monoclonal antibody against (S)-HM (namely, MAb E8) and colloidal gold nanoparticles. The HM-γ-globulin conjugates and rabbit anti-mouse IgG antibodies were placed in the test and control zones, respectively. The free (S)-HM molecules in the samples and the immobilized HM-γ-globulin conjugates competitively reacted with the developed biosensor in the LFA. An inverse relationship existed between the biosensors' visible response, which was noted by the variation in the intensity of a pinkish spot in the test zone, and the content of the free (S)-HM. The limit of detection of the developed LFA was 156 ng/mL. Various validation methods confirmed that the LFA exhibited sufficient sensitivity, selectivity, repeatability, and reliability, making it ideal for (S)-HM detection in plant samples and plant-containing products. The developed system required only a small sample volume (20 µL) and a concise sample preparation time compared with conventional LFAs. Thus, the LFA reported in this study could serve as a rapid response kit for the detection of (S)-HM in plant samples.

Rapid Quantification and Validation of Biomarker Scopoletin in Paederia foetida by qNMR and UV-Vis for Herbal Preparation.

Scopoletin has previously been reported as a biomarker for the standardization of Paederia foetida twigs. This study is the first report on the determination and quantification of scopoletin using quantitative nuclear magnetic resonance (qNMR) in the different extracts of Paederia foetida twigs. The validated qNMR method showed a good linearity (r2 = 0.9999), limit of detection (LOD) (0.009 mg/mL), and quantification (LOQ) (0.029 mg/mL), together with high stability (relative standard deviation (RSD) = 0.022%), high precision (RSD < 1%), and good recovery (94.08-108.45%). The quantification results of scopoletin concentration in chloroform extract using qNMR and microplate ultraviolet-visible (UV-vis) spectrophotometer was almost comparable. Therefore, the qNMR method is deemed accurate and reliable for quality control of Paederia foetida and other medicinal plants without extensive sample preparation.

Quality Assessment of Sitopaladi Churna Using High-Performance Liquid Chromatography Coupled with Multivariate Analysis.

"Sitopaladi churna," a well-known formulation of Ayurveda, is prescribed to treat the disease like Bronchitis, Pneumonia, allergic conditions, viral infection of the respiratory tract and as a natural antioxidant. A novel method based on reversed-phase high-performance liquid chromatography (RP-HPLC) coupled to photodiode array detector was established and validated for sitopaladi churna. Here, HPLC fingerprints data obtained for 28 samples including standard sample were then treated to chemometric analysis like principal component analysis and hierarchical clustering analysis for further analysis to evaluate the differences in market samples of sitopaladi churna. Additionally, one major marker compound, piperine was quantified and it also facilitated for relative retention time. The simulative mean spectrum was also generated. The validation results showed that the developed method was simple, precise and stable. Thus, the developed chromatographic method adjoined with multivariate analysis can be used as an efficient and practical approach for quality assessment of sitopaladi churna.

A new type of tomato puree with high content of bioactive compounds from 100% whole fruit.

In this work, a new type of tomato puree was obtained from the whole fruit including seeds and skins without any waste. Currently, the food industry aims to develop products that both bring benefits to consumer health and are ecosustainable. Several efforts in fact have been carried out to revalue tomato by-products, such as seeds and skins, for human use. The physicochemical properties of the whole tomato puree immediately after production and after 6 months of storage were investigated and compared to the properties of a traditional tomato puree. Two tomato varieties were used in this study: Villa Literno and Piccadilly. pH, dry weight, water activity, titratable acidity, reducing sugars, soluble solid, lycopene, polyphenol, and volatile organic compound contents were evaluated. Tomato puree obtained from whole fruits mainly showed higher lycopene (90 to 150 mg/kg), flavonoid (39 to 41 mg/kg), and aldehyde (in particular hexanal) contents and a greater stability over time compared to those of the traditional puree. The proposed process allowed to obtain a tomato puree with a higher content of bioactive compounds, producing no waste. PRACTICAL APPLICATION: In this study, we proposed a new type of tomato puree made from the whole fruit. The whole tomato puree is richer in phenolic compounds and volatile organic compounds than the traditional puree that is made by removing skins and seeds. It can be an interesting product for the food industries because it is a source of bioactive compounds and reduces to zero the amount of waste.

Contents of key bioactive and detrimental compounds in health performance coffees compared to conventional types of coffees sold in the United States market.

The United States is the largest coffee consuming country worldwide. Recently, in addition to cup quality, the focus on health promotion has increased significantly in the country, with launching of many brands with health claims, mainly highlighting the antioxidative and stimulating properties of the beverage. On the other hand, mycotoxins and, to a lesser extent, acrylamide, have raised concerns among consumers and health authorities. This study investigated the contents of the main bioactive compounds (caffeine, chlorogenic acids and their 1,5-γ-quinolactones, and trigonelline) in health performance coffees and compared them to those of conventional roasted coffees available on the U.S. market. The following categories were compared by ANOVA at p ≤ 0.05, followed by Fisher's test: 1 - health performance, 2 - gourmet and 3 - traditional, totaling 127 samples. As complementary results, the contents of acrylamide and ochratoxin A were evaluated in part of the samples (n = 58). The mean contents (g per 100 g) of bioactive compounds for categories 1 to 3, respectively, were 1.09, 1.11 and 1.07 for caffeine; 1.75, 1.88 and 1.34 for chlorogenic acids/lactones, and 0.63, 0.64 and 0.56 for trigonelline. The mean contents (µg kg-1) of acrylamide for categories 1 to 3, respectively, were 82, 71 and 85. Only about 7% of the evaluated samples presented quantifiable amounts of OTA and all of them were within the maximum limits established by health authorities. In general, the contents of bioactive and potentially harmful compounds were not consistently different among categories, with high and low individual amounts in all of them. Most health claims on labels related to the amount of bioactive compounds in health performance coffees were unjustified, suggesting the need for improvement in coffee labeling regulation in the U.S.

A novel botanical formula improves eye fatigue and dry eye: a randomized, double-blind, placebo-controlled study.

BACKGROUND: With the frequent use of video display units, eye fatigue is becoming more common globally. An alternative nutritional strategy is needed to prevent the aggravation of eye fatigue symptoms. OBJECTIVES: The objective was to evaluate the protective effect of a novel botanical combination of lutein ester, zeaxanthin, and extracts of blackcurrant, chrysanthemum, and goji berry on adults with eye fatigue in a randomized, double-blind, placebo-controlled clinical trial. METHODS: We randomly allocated 360 participants into 4 groups to receive placebo and 3 doses of our formula (chewable tablets, containing 6 mg, 10 mg, or 14 mg of lutein) once daily for 90 d. Each participant had 3 visits at baseline (V1), 45 d (V2), and 90 d (V3) during the study. RESULTS: Intervention with the formula improved individual scores of eye fatigue symptoms, including eye soreness, blurred vision, dry eye, foreign body sensation, and tearing. Compared with placebo, the formula at all 3 doses significantly decreased the total score of eye fatigue symptoms and increased the visuognosis persistence time at both V2 and V3. According to the Schirmer test, both 10-mg and 14-mg lutein formula groups had improved tear secretion at V3 compared with the placebo. The keratography results indicated that the first tear break-up time, average tear break-up time, and tear meniscus height were significantly increased after formula intervention. The formula at all 3 doses significantly increased the macular pigment optical density at V2 and V3 compared with the placebo, whereas optical coherence tomography showed no significant difference in retinal thickness and retinal volume across all groups at both visits. CONCLUSIONS: Our botanical formula improves eye fatigue, dry eye, and macular function without changing the retinal structure, and thus it could serve as an effective nutritional strategy in improving eye fatigue without causing serious side effects.Clinical Trial Registry: chictr.org.cn (ChiCTR1800018987).

Optimization of pectinase-assisted extraction of Annona muricata L. juice and the effect of liquefaction on its pectin structure.

BACKGROUND: Soursop (Annona muricata L.) is an underutilized tropical and subtropical fruit with high nutritional and therapeutic benefits. This fruit is faced with enormous post-harvest losses due to its high perishability. This work was aimed to optimize the pectinase-assisted extraction conditions of soursop juice using Doehlert design and to study the effect of pectinase on its pectin structure. RESULTS: The predicted models were validated for all the responses studied and the regression coefficients ranged from 0.905 to 0.987 (P ≤ 0.05). An incubation time of 172 min, enzyme concentration of 0.04% (w/w) and incubation temperature at 42.9 °C were found to be the optimal conditions for soursop juice extraction, which resulted in 75.20%, 3.74, 7.35 °Brix, 87.06%T, and 0.44% MAE for soursop juice yield (%), pH, total soluble solids (TSS) (°Brix), clarity (%T) and titratable acidity (% malic acid equivalent, MAE), respectively. Morphologically, untreated soursop pulp presented a non-uniform spherical surface; enzyme hydrolyzed soursop exhibited ruptured and wrinkled surface; meanwhile for the different pectin obtained, untreated soursop pectin depicted porous surface and enzyme hydrolyzed soursop pectin showed whirling rough surface. Fourier-transform infrared (FTIR) confirmed the presence of similar chemical group stretching and vibrations in commercial pectin and soursop pectin. CONCLUSION: Under the optimum conditions, the numerical predictions were similar to the experimental data obtained, thus confirming the validity of the models. Application of enzyme treatment caused the breakdown of pectin structure as illustrated by scanning electron microscopy (SEM) and FTIR analyses.

Investigation of Chocolate Matrix Interference on Cannabinoid Analytes.

The first known findings of chocolate matrix interference on cannabinoid analytes is reported. Stock solutions of four biogenic cannabinoids (Δ9-tetrahydrocannabinol, cannabidiol, cannabinol, and cannabigerol) and one synthetic cannabinoid (cannabidiol dimethyl ether) are subjected to milk chocolate, dark chocolate, and cocoa powder. A clear trend of matrix interference is observed, which correlates to several chemical factors. The amount of chocolate present is directly proportional to the degree of matrix interference, which yields lower percent recovery rates for the cannabinoid analyte. Structural features on the cannabinoid analytes are shown to affect matrix interference, because cannabinoids with fewer phenolic -OH groups suffer from increased signal suppression. Additionally, aromatization of the p-menthyl moiety appears to correlate with enhanced matrix effects from chocolate products high in cocoa solids. These findings represent the first known documentation of chocolate matrix interference in cannabinoid analysis, which potentially has broad implications for complex matrix testing in the legal Cannabis industry.

Direct and Fast Screening of New Psychoactive Substances Using Medical Swabs and Atmospheric Solids Analysis Probe Triple Quadrupole with Data-Dependent Acquisition.

New psychoactive substances (NPS) have become a serious public health problem, as they are continuously changing their structures and modifying their potency and effects on humans, and therefore, novel compounds are unceasingly appearing. One of the major challenges in forensic analysis, particularly related to the problem of NPS, is the development of fast screening methodologies that allow the detection of a wide variety of compounds in a single analysis. In this study, a novel application of the atmospheric solids analysis probe (ASAP) using medical swabs has been developed. The swab-ASAP was coupled to a triple quadrupole mass analyzer working under a data-dependent acquisition mode in order to perform a suspect screening of NPS in different types of samples as well as on surfaces. The compounds were automatically identified based on the observed fragmentation spectra using an in-house built MS/MS spectra library. The developed methodology was applied for the identification of psychoactive substances in research chemicals and herbal blends. The sensitivity of the method, as well as its applicability for surface analysis, was also assessed by identifying down to 1 µg of compound impregnated onto a laboratory table. Another remarkable application was the identification of cathinones and synthetic cannabinoids on the fingers of potential consumers. Interestingly, our data showed that NPS could be identified on the fingers after being in contact with the product and even after cleaning their hands by shaking off with a cloth. The methodology proposed in this paper can be applied for routine analyses of NPS in different matrix samples without the need to establish a list of target compounds prior to analysis.

Application of an in vitro digestion model to study the metabolic profile changes of an herbal extract combination by UHPLC-HRMS.

BACKGROUND: STW 5 is a fixed herbal combination containing extracts from nine medicinal plants: bitter candytuft, greater celandine, garden angelica roots, lemon balm leaves, peppermint leaves, caraway fruits, licorice roots, chamomile flowers, and milk thistle fruit. STW 5 is a clinically proven treatment for functional dyspepsia and irritable bowel syndrome. PURPOSE: Using a static in vitro method, we simulated oral, gastric, and small intestinal digestion and analyzed the metabolic profile changes by UHPLC-HRMS to determine the impact of oro-gastro-intestinal digestion on STW 5 constituents. STUDY DESIGN AND METHODS: STW 5 was incubated according to the InfoGest consensus method. Samples of each digestive phase were analyzed by UHPLC-HRMS in ESI positive and negative modes. After data processing, background subtraction, and normalization, the peak areas of detectable compounds were compared to untreated reference samples and recovery ratios were calculated to monitor the metabolic profile of STW 5 during simulated digestion. RESULTS: Although the levels of some constituents were reduced, we did not observe complete degradation of any of the constituents of STW 5 upon in vitro digestion. We did not detect any new metabolites beyond increased levels of caffeic acid and liquiritigenin due to degradation of progenitor compounds. Changes observed in intestinal bioaccessibility ratios were mainly a result of isomerization, hydrolysis, protein binding, and low water solubility. CONCLUSION: The majority of STW 5 constituents are stable towards simulated in vitro digestion and can reach the colon to interact with gut microbiota if they remain unabsorbed in the upper intestinal tract.