Herbal Food Supplements Research Articles

Analysis of phosphodiesterase type 5 inhibitors and analogues as adulterants in herbal dietary food supplements in Saudi market using green UPLC-MS/MS method: Assessment of the greenness and whiteness profiles

Herbal dietary supplements are popular alternatives to oral phosphodiesterase type 5 inhibitors (PDE5Is) for treating Erectile Dysfunction (ED), yet concerns arise regarding their adulteration with undeclared synthetic PDE5Is or analogues, posing health risks. The aim of this study was to develop and validate a green UPLC-MS/MS method for sensitive and rapid detection of PDE5Is and analogues in sexual enhancement herbal dietary supplements marketed in Saudi Arabia. Method validation revealed good linearity with determination coefficients (r2) ≥ 0.9990, and LODs and LOQs in the range of 0.4 – 32 ng g−1 and 1––96 ng g−1, respectively. The accuracy ranged from 80.8 % to 103.4 % while inter-day precision was ≤ 8.6 % and intra-day precision was ≤ 8.4 %. The method was successfully utilized to analyze 32 herbal dietary supplements. The results revealed that 18 samples (56.3 %) tested positive for at least one of the target analytes. Sildenafil emerged as the most frequently detected analyte, found in 15 samples (20.1 µg g−1 to 37,492 µg g−1). Tadalafil was detected in 7 samples, with concentrations ranging from 4.46 µg g−1 to 19,986 µg g−1. Yohimbine was detected in only 4 samples, while icariin was detected at lower concentrations in 3 samples. Furthermore, the method greenness and whiteness profiles were evaluated using four metrics including the Analytical Eco-Scale, Green Analytical Procedure Index (GAPI), Analytical GREEnness (AGREE) metric, and Red-Green-Blue 12 (RGB 12) metric.This study provides foundational data on the adulteration of dietary supplements in Saudi Arabia and offers insights for future interventions to promote consumer safety and awareness regarding herbal dietary supplement consumption.

Sea hedgehog-inspired surface-enhanced Raman scattering biosensor probe for ultrasensitive determination of Staphylococcus aureus in food supplements

Staphylococcus aureus contamination in food supplements poses substantial challenges to public health and large-scale production but the sensitive detection in a timely manner remains a bottleneck. Drawing inspiration from the sea hedgehog, gold nanostars (AuNSs) were leveraged to design an ultrasensitive surface-enhanced Raman scattering (SERS) biosensor for the determination of Staphylococcus aureus in food supplements. Besides the surface enhancement furnished by the AuNSs, Raman reporter molecules and specific aptamers sequentially self-assembled onto these AuNSs to construct the “three-in-one” SERS biosensor probe for label-based quantitation of Staphylococcus aureus. Following incubation with contaminated health product samples, the gold nanostars@Raman reporter-aptamer specifically recognize and assemble around Staphylococcus aureus cells, forming a distinctive sea hedgehog structure. This unique configuration results in an amplified Raman signal at 1338 cm−1 and an enhancement factor of up to 6.71 × 107. The entire quantitative detection process can be completed within 30 min, boasting an exceptional limit of detection as low as 1.0 CFU mL−1. The method exhibits a broad working range for the determination of Staphylococcus aureus, with concentrations spanning 2.15 CFU mL−1 to 2.15 × 105 CFU mL−1. Furthermore, it demonstrates outstanding precision, with relative standard deviation values consistently below 5.0%. As a showcase to validate the practicality of the SERS method, we conducted tests on determining Staphylococcus aureus in a herbal food supplement, i.e., Ginkgo Biloba extract (GBE); the results align closely with those obtained through the conventional lysogeny broth agar plate method, pointing to the potential applicability in real-world scenarios.

A Pilot Prospective Study Evaluating the Effect of Curcuma-Based Herbal Food Supplement on the Outcome of In Vitro Fertilization in Patients Testing Positive for Four Immunological Biomarkers.

Background and Objectives: Inflammation and oxidative stress have been described to reduce the chance for pregnancy instauration and maintenance. NOFLAMOX, a recently developed herbal preparation with recognized antioxidant and anti-inflammatory properties, can represent an interesting treatment to increase the chance of pregnancy, both physiological or after in vitro fertilization (IVF). The aim of this study was to assess NOFLAMOX's effect; a population with unexplained infertility was screened for the recently described IMMUNOX panel based on four immunological biomarkers with a prospective study approach. Materials and Methods: Patients with unexplained infertility and positive for at least one of the biomarkers of the IMMUNOX panel were included in this study and treated with NOFLAMOX for three months prior to an IVF cycle. Results: Eighty-six patients (n = 86) were screened with the IMMUNOX panel and the forty-seven (54.5%) found positive were included in this study. In more detail, 11 were positive for TNFα (23.4%), 18 (38.3%) for glycodelin (GLY), 29 (61.7%) for Total Oxidative Status (TOS), and 32 (68.1%) for Complement Activity Toxic Factor (CATF). After three months of treatment, a significant reduction in the number of IMMUNOX-positive patients was observable, with 26 patients who turned IMMUNOX-negative displaying a quantitative statistically significant variation of 100% (11/11), 38.9% (7/18), 65.5% (18/29), and 75% (24/32), for TNFα, glycodelin, TOS, and CATF, respectively. Followed in the subsequent IVF cycle, this NOFLAMOX-treated population showed a pregnancy rate of 42.3% compared to the 4.7% of the IMMUNOX-positive group of patients. Conclusions: Taken together, the results of this study suggest that NOFLAMOX could represent an interesting option for those patients with unexplained infertility of inflammatory/oxidative origin. Further studies are needed to confirm these results and explore possible strategies to restore fertility in women with immune-mediated sterility.

A review of botany, traditional applications, phytochemistry, pharmacological applications, and toxicology of Rubus ellipticus Smith fruits.

Rubus ellipticus Smith. (Family Rosaceae), often known as the yellow Himalayan raspberry (Yellow Hissar), is one of the most widely used edible fruits in Indian folk medicinal systems. The current review aims to identify the gap between research and existing applications of this fruit to help scientists explore the current trends and opportunities for future development. Fruits of R. ellipticus are the source of several classes of compounds. Fruits of R. ellipticus are also rich in nutrients such as carbohydrates, vitamins, and minerals. It has been shown to have significant medical value in a variety of studies, including as an anti-diabetic, nephroprotective, anti-inflammatory, analgesic, antipyretic, antitumor, wound healing, antifertility, oviposition deterrent, antibacterial, and antioxidant. Fruits of R. ellipticus have been the subject of several in vitro and in vivo investigations, all of which have corroborated their wide range of biological activities and demonstrated their potential for the identification of new therapeutic candidates and the development of innovative herbal food supplements. Additional mechanism-based pharmacological evaluation and clinical research shouldprovide an adequate scientific basis for the traditional usage of R. ellipticus fruits, which is currently not sufficiently supported by the available research on its active components and molecular mechanisms.

Herbal food supplements usage awareness of university students: Example of Echinacea and St. John's Wort

In recent years, usage of food supplements (Fs) has increased in order to maintain healthy living, have well-being, and be protected from the diseases. There are many medicinal plants used as herbal food supplements (HFs). Within the scope of this study, Echinacea and St. John’s Wort were selected among the plants that are frequently encountered. This descriptive, cross-sectional study was conducted through completing a self-administered online questionnaire by health students. The questionnaire was filled out by 211 students studying at the faculties of Medicine, Dentistry, Pharmacy and Health Sciences at Istanbul Medipol University. The greatest participation was achieved with Pharmacy students (47.4%), whereas the lowest participation was reached with Dentistry students (5.7%). The most commonly used products as Fs were vitamin D (21.3%) and multivitamins (16.1%), while the use of HFs was 8.5%. St. John’s Wort is commonly preferred for wound and burn treatment and Echinacea is used to boost immunity. In parallel with this use, St. John’s Wort is preferred as olive oil maceration and Echinacea as herbal infusion. While the use of HFs was 58.3%, that of the Fs were 44.5%. Echinacea use was found to be 14.4% and St. John’s Wort was 31.3%. The relationship between the presence of chronic disease and the use of Fs or HFs was not statistically significant. This study is significant to detect the opinions and knowledge levels of health students about Fs, especially HFs, St. John’s Wort and Echinacea, which are available in the market.

Risk assessment of aflatoxin B1 in herbal medicines and plant food supplements marketed in Malaysia using margin of exposure and RISK21 approaches

Aflatoxin B1 (AFB1) is a mycotoxin produced by several species of Aspergillus fungi which can cause liver cancer in animals and humans. This study aims to perform the risk assessment of AFB1 in herbal medicines and plant food supplements (PFS) in Malaysian market. A total of 31 herbal medicines and PFS were purchased through online platforms and over the counter using a targeted sampling strategy. Of 31 samples analysed using the ELISA method, 25 (80.6%) were contaminated with AFB1 at levels ranged from 0.275 to 13.941 μg/kg. The Benchmark Dose Lower Confidence level of 10 (BMDL10) of 63.46 ng/kg bw/day and the estimated dietary intake of the adult population ranged from 0.006 to 10.456 ng/kg bw/day were used to calculate the Margin of Exposure (MOE). The MOEs for 24 (96%) out of the 25 positive samples were lower than 10,000. The RISK21 matrix revealed that AFB1 exposure levels from herbal medicines and PFS differed greatly over the world. The calculated population risk of acquiring liver cancer from AFB1 exposure ranged from 0 to 0.261 cancers/100,000 populations/year and accounted for an estimated percentage of liver cancer incidence ranged from 0.002 to 4.149%. This study revealed a moderate risk of liver cancer attributable to AFB1 from herbal medicine and PFS among Malaysian populations and emphasised an urgency for risk management actions.

Natural Product – Drug Interactions and Their Evaluation Using a Caco-2 Cell Culture Model

Natural products, such as herbal medicinal products, food supplements, and functional food, are widely used to support well-being and for promoting health. In general, the effects of using these products are desired and beneficial, but unexpected adverse effects might also occur, especially when natural products are used with medication. Consumers do not often even know that they are exposed to bioactive compounds that might interact with the body and have effects on their well-being. One of the objectives of the World Health Organization is to promote herb–drug interaction monitoring, and it is important to know how marketed preparations and compounds from common foods interact when they are absorbed. In this review, we describe how a Caco-2 cell absorption model has been used to study how natural products, such as flaxseed, rapeseed, purple loosestrife, pine, echinacea, certain berries and herbs, anthranoid laxatives, and traditional medicinal plants, affect the absorption of co-administered drugs. We discuss the types of interactions and adverse effects that might occur and their possible reasons. Overall, we conclude that the Caco-2 cell absorption model is a useful tool for studying the absorption of natural products with drugs; and that to enable the safe use of natural products with medicines, concomitant use should be studied.

Human Health Risk Assessment of Arsenic and Other Metals in Herbal Products Containing St. John's Wort in the Metropolitan Area of Mexico City.

Consumption of St. John's wort plant is high worldwide due to its various medicinal properties. However, herbal products containing St. John's wort may be contaminated with toxic metals. This is often related to contamination of both water and the atmosphere, lack of proper cultivation methods, and inadequate plant storage conditions, as well as a lack of stricter sanitary supervision. A safety assessment of copper (Cu), lead (Pb), cadmium (Cd) and arsenic (As) content in 23 products containing St. John's wort (pharmaceutical herbal products, food supplements and traditional herbal remedies) sold in the metropolitan area of Mexico City was conducted. The analysis of metals was determined using a graphite-furnace atomic absorption spectrometer. All herbal products were contaminated with Cu, Pb, Cd and As. The pharmaceutical herbal items showed less contamination by metals. The daily human intake (DHI) values for Pb exceeded the permissible limits in the group of traditional herbal remedies. The DHI calculation for As exceeded the permitted intake values for all items in the group of traditional herbal remedies, five food supplements and one pharmaceutical herbal product. The hazard indicator calculation of the non-carcinogenic cumulative risk values for traditional herbal remedies was greater than 1, suggesting a risk to human health.

Development, validation, and application of a multimatrix UHPLC-MS/MS method for quantification of Datura-type alkaloids in food

A quantitative ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for the determination of tropane alkaloids (TAs), atropine and scopolamine, in a variety of food products. The sample preparation of cereal-based food, oilseeds, honey, and pulses consisted of a solid–liquid extraction with an acidified mixture of methanol and water, while an additional step of solid-phase extraction on a cation-exchange sorbent was introduced in the treatment of teas and herbal infusions, aromatic herbs, spices and food supplements. The limits of quantification of the method varied from 0.5 to 2.5 µg kg−1. Apparent recovery was in the range of 70–120%, and repeatability and intermediate precision were below 20%. The method was successfully applied in a proficiency testing exercise as well as in the analysis of various commercial foods. Only 26% of the analysed food samples contained one or both TAs. The mean concentrations for atropine and scopolamine amounted to 21.9 and 6.5 µg kg−1, respectively, while the maximum concentrations were 523.3 and 131.4 µg kg−1, respectively. Overall, the highest levels of TA sum were found in an herbal infusion of fennel and a spice mix containing fennel and anise seeds.

Disclosing Frauds in Herbal Food Supplements Labeling: A Simple LC-MS/MS Approach to Detect Alkaloids and Biogenic Amines

Food supplements are a category of products perceived safe and therefore commonly used by different categories of consumers without any particular attention or precaution. However, health risks associated with the consumption of supplements containing undeclared substances cannot be excluded. A variety of analytical methods are used to control supplement quality composition, but usually, these procedures are complex and time-consuming. Here, we report the results of a simple and fast liquid chromatography-tandem mass spectrometry (LC–MS/MS) method, to detect and quantify simultaneously different categories of active molecules, such as biogenic ammines and natural alkaloids that at high doses can produce negative health effect in consumers. Three categories of products intended for body weight loss, energy boosting, and erectile dysfunction treatment, purchased through e-commerce sites and from local supermarkets, were analyzed (n = 91). The caffeine, synephrine, agmatine sulfate, yohimbine, phenethylamine, and icariin were correctly separated and identified with good precision (RSD < 20%) and recovery (89–109%). The identification and quantification of the analytes in real samples highlighted that the 26% of the samples were not compliant with labeling, confirming that frauds are very common also in the natural supplements market. This LC-MS/MS method could be easily used to test natural supplements in order to check the correct labeling and to protect consumers from potential health risks and food frauds.

Safety Assessment of Herbal Food Supplements: Elemental Profiling and Associated Risk.

Increased usage of herbal food supplements by both the paediatric and adult populations prompted a health risk assessment study. The elemental profiles of 52 supplements collected in Serbia were obtained using ICP-MS. The calculated contribution to permitted daily exposure was in compliance with the guidelines for elemental impurities: up to 46.8% for Pb (infants), 67.2% for Cu (toddlers), 6.8% for As (preschool children), and 8.0% for Hg (adolescents). Hazard quotients indicated high exposure of toddlers to Cu (100.9%) and Zn (112.7%), although by only one supplement, for which hazard indices ranged from 229 to 105% (aligning from toddlers to adolescents). Based on the margin of exposure, As and Pb were not of safety concern. Conversely, the proportion of supplements exceeding the acceptable level of lifetime cancer risk due to As exposure varied from 41.9% in adolescents to 54.3% in adults. Simultaneous use of multiple supplements over a prolonged period of time could further deepen health concerns.

Safety assessment of herbal food supplements: Ethanol and residual solvents associated risk

Availability of herbal food supplements and belief in their efficacy and safety contributed to their increased use, causing exposure to ethanol and residues of extraction solvents. Liquid herbal food supplements (103) were purchased from pharmacies (Novi Sad, Serbia) during 2021, and analysed for presence of ethanol and selected residual solvents using HSS-GC-MS method. Ethanol was not quantified in only one of 18 products for infants or toddlers less than two years old. WHO proposed requirement for ethanol content in products intended for children less than 6 years old (<0.5%) was not met by 26 samples, in products for children 6–12 years (<5%) by four samples and in products for children over 12 years (<10%) by two samples. Calculated blood alcohol concentrations exceeded the relevant toxicological levels for 37 samples following a single dose, and for 31 samples in case of accidental poisoning (entire package). Methanol was quantified in 38 samples, in one of them above the limit of 0.05%, (Ph. Eur. 9), causing exceedance of permitted daily exposure. Other residual solvents were quantified rarely and in low concentrations. Study findings confirmed a need for raising awareness of professionals and public and establishment of strict guidelines for ethanol content and labelling.

Target screening method for the quantitative determination of 118 pyrrolizidine alkaloids in food supplements, herbal infusions, honey and teas by liquid chromatography coupled to quadrupole orbitrap mass spectrometry

An analytical procedure for the screening of 118 pyrrolizidine alkaloids (PAs) was successfully validated and applied to their quantitative determination in food supplements, herbal infusions, honey, and teas. It provides the reliable analyte identification by high-resolution tandem mass spectrometry (HRMS/MS), the accurate determination of 21 regulated PAs, and broad contamination profiles. 10% of 281 analyzed samples resulted contaminated at levels above the maximum levels (MLs) of European legislation. The contamination of herbal infusions of mixed plants can represent a possible health concern (23%; mean of PA sum above ML). A high number of PAs not included in the regulation was detected in honey and herbal food supplements, but their contribution was only relevant to the overall level in honey. The results indicate the need to continue collecting contamination data in food supplements and infusions of mixed herbs and to expand the PA-pool to be monitored in honey and related products.

Subcritical Water Extraction of Rosmarinic Acid from Lemon Balm (Melissa officinalis L.) and Its Effect on Plant Cell Wall Constituents.

Rosmarinic acid (RA), an ester of caffeic acid and 3,4-dihydroxyphenyllactic acid, is a potent radical scavenger, a chelator of prooxidant ions, and an inhibitor of lipid peroxidation. RA-containing extracts are widely used natural antioxidants in food products, and many herbal preparations and food supplements, containing RA, are marketed with claims of beneficial health effects. The current study investigated the effectiveness of subcritical water extraction (SWE) for the recovery of RA from lemon balm (Melissa officinalis), as a "green" alternative to conventional hydro-alcoholic extraction. Different durations (10 min and 20 min) and extraction temperatures (100 °C and 150 °C) were applied. Subcritical water applied at a temperature of 100 °C was equally efficient as 50% ethanol in extracting RA. However, the further elevation of temperature to 150 °C decreased RA content by up to 20% due to thermal degradation. The content of RA in dried extracts was between 2.36% and 5.55% and the higher temperature of SWE increased extract yield by up to 41%. The higher extraction yield resulted from the degradation of plant material by subcritical water as evidenced by the increased extraction and degradation of proteins, pectin, and cellulose. These results reveal that SWE is an efficient technology for the extraction of RA and other antioxidants from lemon balm at reduced extraction time and without the use of toxic organic solvents. Furthermore, by modification of SWE conditions, dry extracts with different purity and content of RA are obtained. These extracts could be used in the food industry as food antioxidants, or in the development of food supplements and functional foods.

Tragocephala nobilis Fabricius (Coleoptera:Cerambycidae): A new insect pest threatening the domestication of Tetrapleura tetraptera (Schum and Thonn.) in Ghana

AbstractTragocephala nobilis (Fabricius) (Coleoptera:Cerambycidae), an insect pest (herbivorous insect), was recently found on a multipurpose tropical tree species, Tetrapleura tetraptera (Schum and Thonn.) Taub, in Ghana. Although the pest has been previously reported on other tree crops such as cocoa and Coffea arabica, this is the first report on T. tetraptera. T. tetraptera is a deciduous tree crop with numerous nutritional and medicinal benefits. The pest, a long‐horned beetle is found to attack T. tetraptera tree and damage is first observed as die‐back of twigs and branches leading to the formation of a witches broom in the crown. This is followed by a series of tunnels created by the larvae on the main stem resulting in branch death and/or tree mortality. In view of the threat posed by the pest to T. tetraptera, a broader study is needed urgently to develop an integrated pest management strategy for T. nobilis pest in T. tetraptera plantations as the interest in its domestication as a tree crop is burgeoning in response to increasing herbal medicine and food supplements demand.

Botanical Ingredient Forensics: Detection of Attempts to Deceive Commonly Used Analytical Methods for Authenticating Herbal Dietary and Food Ingredients and Supplements

Botanical ingredients are used widely in phytomedicines, dietary/food supplements, functional foods, and cosmetics. Products containing botanical ingredients are popular among many consumers and, in the case of herbal medicines, health professionals worldwide. Government regulatory agencies have set standards (collectively referred to as current Good Manufacturing Practices, cGMPs) with which suppliers and manufacturers must comply. One of the basic requirements is the need to establish the proper identity of crude botanicals in whole, cut, or powdered form, as well as botanical extracts and essential oils. Despite the legal obligation to ensure their authenticity, published reports show that a portion of these botanical ingredients and products are adulterated. Most often, such adulteration is carried out for financial gain, where ingredients are intentionally substituted, diluted, or "fortified" with undisclosed lower-cost ingredients. While some of the adulteration is easily detected with simple laboratory assays, the adulterators frequently use sophisticated schemes to mimic the visual aspects and chemical composition of the labeled botanical ingredient in order to deceive the analytical methods that are used for authentication. This review surveys the commonly used approaches for botanical ingredient adulteration and discusses appropriate test methods for the detection of fraud based on publications by the ABC-AHP-NCNPR Botanical Adulterants Prevention Program, a large-scale international program to inform various stakeholders about ingredient and product adulteration. Botanical ingredients at risk of adulteration include, but are not limited to, the essential oils of lavender (Lavandula angustifolia, Lamiaceae), rose (Rosa damascena, Rosaceae), sandalwood (Santalum album, Santalaceae), and tea tree (Melaleuca alternifolia, Myrtaceae), plus the extracts of bilberry (Vaccinium myrtillus, Ericaceae) fruit, cranberry (Vaccinium macrocarpon, Ericaceae) fruit, elder (Sambucus nigra, Viburnaceae) berry, eleuthero (Eleutherococcus senticosus, Araliaceae) root, ginkgo (Ginkgo biloba, Ginkgoaceae) leaf, grape (Vitis vinifera, Vitaceae) seed, saw palmetto (Serenoa repens, Arecaceae) fruit, St. John's wort (Hypericum perforatum, Hypericaceae) herb, and turmeric (Curcuma longa, Zingiberaceae) root/rhizome, among numerous others.

Evaluation of stability and determination of shelf life of an herbal food supplement: Naturovita

Herbal formulations are easily prone to deterioration of the active compounds during storage over a period of time. The stability study helps in understanding the physical and phytochemical changes associated with natural product formulation. The present study was conducted to determine the shelf life of the herbal formula, Naturovita which is used as a food supplement to support respiratory health. The accelerated stability study (Temperature: 40 ºC ± 2 and relative humidity (RH): 75% ± 5) was conducted as per ICH guidelines. The change in the organoleptic, physico-chemical properties, proximate, active phytochemical content and microbial load were verified over a period of 6 months at an interval of 0th, 2nd, 4th and 6th month. The results from various analysis performed were taken in consideration for the determination of extrapolated shelf life of Naturovita. The extrapolated shelf-life value was calculated with 10% degradation of the physico-chemical parameters at an accelerated condition. The shelf life of Naturovita formula was found to be 24.08 months (2.01 years).

Assessment of morphological pharmacognostic characteristics of the content and label information of dried herbs marketed as food supplements in Bulgaria

According to the current food legislation in Bulgaria the dried herbs are classified under the large group of food supplements and their trade is allowed in pharmacies, drugstores and grocery stores. The aim of this study is to assess the morphological pharmacognostic characteristics of the content and the additional information on the labels of food supplements containing dried herbs in Bulgaria, in the light of key standard and regulatory documents related to the quality of herbal substances and food supplements. 91 herbal substances of 10 main groups were studied, which included 103 commercial products from 8 companies. The authenticity of the herbal substances was confirmed with macro-and microscopic tests. The macroscopic morphological indicators which were used as elements of trade-commodity analysis were changes in color, presence of other parts of the same or different plants, organic and mineral impurities, evidences of diseases and pest infestations. The phenological phase in which the herbal substances had been collected was determined also. We found that very small part of the studied products meet the standards for all of the selected criteria according to the considered documents which implies the need of strengthening control.

Curcumin encapsulated polylactic acid nanoparticles embedded in alginate/gelatin bioinks for in situ immunoregulation: Characterization and biological assessment

Curcumin is a known naturally occurring anti-inflammatory agent derived from turmeric, and it is commonly used as a herbal food supplement. Here, in order to overcome the inherent hydrophobicity of curcumin (Cur), polylactic acid (PLA) nanoparticles (NPs) were synthesised using a solvent evaporation, and an oil-in-water emulsion method used to encapsulate curcumin. Polymeric NPs also offer the ability to control rate of drug release. The newly synthesised NPs were analysed using a scanning electron microscope (SEM), where results show the NPs range from 50 to 250 nm. NPs containing graded amounts of curcumin (0 %, 0.5 %, and 2 %) were added to cultures of NIH3T3 fibroblast cells for cytotoxicity evaluation using the Alamar Blue assay. Then, the curcumin NPs were incorporated into an alginate/gelatin solution, prior to crosslinking using a calcium chloride solution (200 nM). These hydrogels were then characterised with respect to their chemical, mechanical and rheological properties. Following hydrogel optimization, hydrogels loaded with NP containing 2 % curcumin were selected as a candidate as a bioink for three-dimensional (3D) printing. The biological assessment for these bioinks/hydrogels were conducted using THP-1 cells, a human monocytic cell line. Cell viability and immunomodulation were evaluated using lactate dehydrogenase (LHD) and a tumour necrosis factor alpha (TNF-α) enzyme-linked immunosorbent (ELISA) assay, respectively. Results show that the hydrogels were cytocompatible and supressed the production of TNF-α. These bioactive hydrogels are printable, supress immune cell activation and inflammation showing immense potential for the fabrication of tissue engineering constructs.

Characterization of phenolics and discovery of α-glucosidase inhibitors in Artemisia argyi leaves based on ultra-performance liquid chromatography-tandem mass spectrometry and relevance analysis

Artemisia argyi leaves (AAL) has been widely used as herbal medicine and food supplement and in China and other Asian countries. The aim of this work is to qualitative and quantitative characterization of phenolic compounds in AAL and screening of natural product inhibitors of α-glucosidase from AAL. Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) was employed to rapid and comprehensive identification of phenolic compounds in AAL, and a total of thirty-three phenolic compounds were identified. High performance liquid chromatography with diode array detection (HPLC-DAD) was established and validated to simultaneously determinate ten main bioactive phenolics compounds in different batches of AAL samples. Meanwhile, the inhibitory capacities of different batches of AAL samples on α-glucosidase were evaluated. Then, relevance analysis, including grey relational analysis and Pearson correlation analysis were employed to investigate the correlations between the contents of phenolic compounds and α-glucosidase inhibitory activities, and discover the α-glucosidase inhibitors in AAL. The relevance analysis results indicated that three phenolic compounds, 3-caffeoylquinic acid, 3,4-dicaffeoylquinic acid and 3,5-dicaffeoylquinic acid could be potential α-glucosidase inhibitors in AAL. Moreover, the α-glucosidase inhibitory activities of the three phenolic compounds were validated by in vitro and in vivo experiments. The possible inhibiting effect of the three phenolic compounds on α-glucosidase was also explored by molecular docking analysis, and the results indicated that the binding of the three α-glucosidase inhibitors to α-glucosidase mainly by hydrogen bonds, hydrophobic forces and ionic bonds. The present research provided a deep insight into phenolic compounds and α-glucosidase inhibitory activities of AAL, and discovered the α-glucosidase inhibitors in AAL.