Novel reusable and switchable deep eutectic solvent for extraction and determination of curcumin in water and food samples.

For the first time, a fast and easy extraction method based on a unique reusable and switchable deep eutectic solvent (made of octylamine, succinic acid, and water as precursors) was presented and utilized for the microextraction and determination of curcumin as a model analyte. The main factors used to induce a phase transition in the as-prepared deep eutectic solvent were solutions of NaOH and HCl. Among the standout characteristics of the suggested deep eutectic solvent are the removal of toxic organic solvents like THF, the lack of a need for centrifugation, and the ability to be reused in subsequent extractions. The influence of effective parameters (i.e., proportions of deep eutectic solvent structure components, volume of prepared deep eutectic solvent, volume and concertation of NaOH, volume of HCl, and salt effect) on the extraction procedure were investigated. The calibration curve also was linear in the range of 35-500 µg L-1 with coefficients of determination (R2) of 0.9976. Limit of detection (S/N = 3) 10.0 µg L-1, the limit of quantification (LOQ) of 35.0 µg L-1, the relative standard deviations (RSDs %) composed of intra-day RSD (4.7) and inter-day RSD (6.4), preconcentration factor of 40.0, enrichment factor of 38.68, and relative recovery of 92.6%-100.3 % were achieved. The reusable and switchable deep eutectic solvent based-dispersive liquid-liquid microextraction technique was proficiently employed to expedite easy and fast extraction of curcumin from water and food samples.

A wearable colorimetric sweat pH sensor-based smart textile for health state diagnosis.

Sweat pH is an important indicator for diagnosing disease states, such as cystic fibrosis. However, conventional pH sensors are composed of large brittle mechanical parts and need additional instruments to read signals. These pH sensors have limitations for practical wearable applications. In this study, we propose wearable colorimetric sweat pH sensors based on curcumin and thermoplastic-polyurethane (C-TPU) electrospun-fibers to diagnose disease states by sweat pH monitoring. This sensor aids in pH monitoring by changing color in response to chemical structure variation from enol to di-keto form via H-atom separation. Its chemical structure variation changes the visible color due to light absorbance and reflectance changes. Furthermore, it can rapidly and sensitively detect sweat pH due to its superior permeability and wettability. By O2 plasma activation and thermal pressing, this colorimetric pH sensor can be easily attached to various fabric substrates such as swaddling and patient clothing via surface modification and mechanical interlocking of C-TPU. Furthermore, the diagnosable clothing is durable and reusable enough to neutral washing conditions due to the reversible pH colorimetric sensing performance by restoring the enol form of curcumin. This study contributes to the development of smart diagnostic clothing for cystic fibrosis patients who require continuous sweat pH monitoring.

Separation and identification of terpene-conjugated curcuminoids based on liquid chromatography-tandem mass spectrometry and their in vitro anti-inflammatory activities.

Terpene-conjugated curcuminoids are conjugates of curcuminoids and bisabolanes in the rhizomes of Curcuma longa L. The fragmentation pathways of known three terpene-conjugated curcuminoids (bisabolocurcumin-ether, bisabocurcumin, and demethoxybisabolocurcumin ether) and curcumin, demethoxycurcumin, and bisdemethoxycurcumin were investigated using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry in negative mode to rapidly search and discover similar unknown compounds of the acetone fraction of turmeric. Subsequently, compounds 1-3 were founded in the acetone fraction based on molecular weight and above fragmentation pathways (the characteristic fragment ions, the most and second most abundant fragment ions produced in MS2 spectra). Terpecurcumin X (1) and terpecurcumin Y (3) were further separated by liquid chromatography-tandem mass spectrometry guided isolation technique to verify their structures by nuclear magnetic resonance, electrospray ionization high-resolution mass spectroscopy, ultraviolet and visible spectra and infrared spectra. Interestingly, 1 and 3 were new compounds. The results indicate the feasibility and significant advantages of liquid chromatography-tandem mass spectrometry for the rapid discovery and analysis of new constituents in traditional Chinese medicine. In vitro, Terpene-conjugated curcuminoids had better nitric oxide inhibitory activity than the other seven curcuminoids (demethoxycurcumin, bisdemethoxycurcumin, curdione, curcumenone, bisacurone, curcumenol, and germacron).

Curcumin quantification in skin and mucosa: Optimization of extraction and chromatographic method validation.

Curcumin is a natural phenol found in the rhizome of Curcuma longa. It has been studied to treat several human carcinomas, such as melanomas and breast, head and neck, prostate, and ovary cancers. Here, we develop and validate a method for recovering curcumin from the skin layers and mucosa and selectively quantifying it, aiming to support permeation studies in developing topical formulations containing the natural compound. Recovery of curcumin from the stratum corneum, remaining skin, and mucosa was performed using ethanol, DMSO/ethanol, and DMSO, respectively, under mild stirring for specific periods. The separation of curcumin from the other curcuminoids, skin, and mucosa interferents was obtained using a C18 column as a stationary phase. The mobile phase was composed of pH 3.0 phosphoric acid at 1.0 mmol/L and acetonitrile (47:53, v/v), which flowed at 1 mL min-1. UV-Vis detection of curcumin was at 424 nm. The chromatographic method was selective, linear (r > 0.999), with a regression curve in the concentration range from 1.0 to 30.0 µg mL-1, robust, precise, and accurate, with curcumin recovery rates higher than 95 ± 7 % from the mucosa, 82 ± 2 % from the stratum corneum, and 65 ± 10 % from the remaining skin. Finally, the method was successfully used in a skin permeation test performed with porcine skin and mucosa. The validated method is, therefore, suitable for the recovery and quantification of curcumin from the skin layers and mucosa, favoring the development of new topical formulations destined for these sites of administration.

Effect of curcumin on the formation of polycyclic aromatic hydrocarbons in grilled chicken wings.

Organic macromolecules form carcinogenic and toxic substances such as polycyclic aromatic hydrocarbons (PAHs) under high temperature baking. Thus, this study investigated the effects and inhibition pathways of different curcumin concentrations (0.01, 0.05, 0.25, 0.3 mg/g) on seven PAHs in grilled chicken wings. The results demonstrated that curcumin concentrations displayed positive effects in inhibiting the formation of PAHs (16%-72%), increasing the total phenolic content (397.5-1934.4 mg/g) and free radical scavenging activity, and reducing TBARS values (31.15%-47.76%) and fatty acid content. Additionally, PCA and Pearson correlation analyses indicated that lipid oxidation (r = 0.42) and unsaturated fatty acids (r = 0.55) could promote the production of PAHs, while DPPH, ABTS and TPC could counteract their facilitation of PAHs. In conclusion, the addition of appropriate amounts of curcumin before grilling is a feasible strategy to reduce fat oxidation levels and the number of free radicals for the purpose of limiting PAHs content.

Development and Validation of a Green Stability-Indicating HPTLC Method for Estimation of Curcumin, Gallic Acid, and Ursolic Acid From Polyherbal Formulation Jatyadi Taila.

BACKGROUND: Jatyadi taila (JT) is a well-known Ayurvedic wound-healing product, comprising 16 different medicinally important plants, including Curcuma longa, Terminalia chebula, and Jasminum officinale. OBJECTIVE: The proposed work discusses the development and validation of the green and economical stability-indicating HPTLC method for quantification of the key marker phytoconstituents, curcumin (CUR), gallic acid (GA), and ursolic acid (UA), from JT. METHOD: Quality standard parameters for JT were determined following standard procedures. The marker constituents CUR, GA, and UA were resolved from JT using toluene-ethyl acetate-formic acid (6:2:1, v/v/v) as the mobile phase and subsequently derivatized to estimate UA. The developed plates were subjected to HPTLC-MS analysis. All constituents were subjected to forced degradation to determine the proposed technique's stability-indicating property and the accelerated stability studies of marketed formulation and marker constituents. Greenness evaluation of the method was aided by the AGREE methodology. RESULTS: The Rf values of CUR, GA, and UA were found to be 0.60 and 0.60; 0.27 and 0.28; and 0.74 and 0.77 from reference standard and oil samples respectively, when analyzed at 366 nm, 290 nm, and 366 nm, respectively. HPTLC-MS was carried out to verify the active constituents present in JT. The constituents followed first-order degradation kinetics. The quantity of CUR, GA, and UA in JT was reduced at the end of accelerated stability studies. The developed approach was validated in compliance with the International Conference on Harmonization (ICH) Q2 (R2) guideline. CONCLUSIONS: Among the chosen key markers, GA was highly unstable during forced degradation. JT should be stored at a controlled temperature using more protective packaging material to ensure its quality and efficacy. HIGHLIGHTS: The developed method can be used as a quality control tool for JT as it can be used to determine the stability of the key marker compounds the herbal formulation.

Effect of dentin biomodification using natural collagen cross-linkers on the durability of the resin-dentin bond and demineralized dentin stiffness.

OBJECTIVE: The purpose of this study was to evaluate the effect of using natural cross-linkers as sumac and curcumin on the durability of the resin-dentin bond and stiffness of demineralized dentin matrix. METHODS: Thirty sound molars were divided into 5 groups: Control (CO), Grape Seed extract (GSE), Cacao seed extract (CSE), Sumac extract (SE) and Curcumin extract (CE). The teeth had their coronal dentin exposed, etched, and pre-treated for 1 min with the extracts. Teeth were then bonded using Single-Bond II adhesive and 4 mm composite was built up on dentin surface. Teeth were sectioned into 1 × 1 × 8mm beams and their micro-tensile bond strength (µTBS) was tested after 24 h and 6 months of water storage. For stiffness testing, 15 teeth were sectioned to obtain dentin beams (1 × 1 × 6.5 mm), the beams were demineralized in 10% phosphoric acid then rinsed and divided into 5 groups. Beams were then immersed in their respective extract solution for 1 min after which they were subjected to a 3- point loading test using a universal testing machine to calculate their modulus of elasticity. RESULTS: After 24 h, no significant difference in µTBS was shown between all groups. After 6 Months, GSE, CE, and SE showed significantly higher µTBS compared to CO (p ≥ 0.05). For the modulus of elasticity; only GSE showed a significantly higher modulus compared to other groups. CLINICAL RELEVANCE: The application of grape seed extract, curcumin and sumac extract as dentin pre-treatments appear to be a promising approach to enhance the durability of the resin-dentin bond in a clinically relevant application time.

Visualizing the distribution of curcumin in the root of Curcuma longa via VUV-postionization mass spectrometric imaging.

Curcumin is a dietary spice and coloring agent widely used in food and herbal medicine. Herein, we visualized the distribution of curcumin in fresh Curcuma longa (turmeric) root sections using the state-of-the-art vacuum-ultraviolet (VUV, 118 nm) single photon-postionization mass spectrometric imaging method. Compared with other mass spectrometric imaging methods, the proposed method does not require any sample pre-treatment. The proposed approach could be more conducive to in situ detection of small molecules. The mass spectroscopic imaging (MSI) images of curcumin sections with a lateral resolution of 100 µm indicated that the concentrations of curcumin decreased from the phloem to the xylem of the root. We also show MS imaging of curcumin in the turmeric root at different maturity periods, revealing the transformation of this endogenous species. The result of quantitative analysis indicates that the total curcumin content of the mature turmeric root is estimated to be 3.43%, which is consistent with the previous report that the content of curcumin in the turmeric root is estimated between 3% and 5%. The report indicated that the proposed method of VUV single photon postionization MSI can be used to explore the metabolic process of plants, which is critical for herbal farming, harvest, and its ingredient extraction.

Colorimetric ELISA based on urease catalysis curcumin as a ratiometric indicator for the sensitive determination of aflatoxin B1 in grain products.

There is an urgent need to measure aflatoxin B1 (AFB1) in food to prevent contaminated food consumption. In this work, a novel colorimetric enzyme-linked immunosorbent assay (ELISA) was developed for the detection of AFB1 using curcumin as a colorimetric indicator. An indirect competitive enzyme-label immunoassay was developed using urease and rabbit anti-mouse immunoglobulin G labeled with gold nanoparticles as the signal-transduction tag. Urease catalyzed the hydrolysis of urea to produce ammonia, which increased the pH of the solution. The phenolic hydroxyl group of curcumin ionized into phenolic oxygen anions under alkaline conditions, which strengthened the synergistic effect of electron supply and absorption in curcumin. As a result, the color of curcumin changed from yellow to reddish-brown, producing a visible color change. Under optimal conditions, AFB1 could be qualitatively determined with the naked eye, and quantitatively assessed by measuring the ratio of absorbance at wavelengths of 550 and 428 nm. The change in the ratio of absorbance Δ550/Δ428 decreased linearly in a range of 0.01-5 ng mL-1, and the limit of detection was 67 pg mL-1. Therefore, the selectivity and reliability of this proposed method were well validated. This method was also successfully used for the quantitation of AFB1 in spiked rice flour and wheat flour samples. This approach may broaden the application field of colorimetric ELISA for aflatoxin, providing a promising platform for the rapid screening of aflatoxin in food.

Quality assessment of Curcuma dietary supplements: Complementary data from LC-MS and 1H NMR.

Due to the numerous potential health benefits of Curcuma, turmeric dietary supplements (DS) are among the top selling products. To assess the quality of these formulations, thirty Curcuma DS along with five standard Curcuma rhizomes were analyzed with UHPLC-MS and 1H NMR. The chemometric treatment of the UHPLC-MS spectra showed a significant variability of their chemical composition that was confirmed by 1H NMR which allowed the absolute quantification of the Curcuma major bioactive components, i.e. curcuminoids (curcumin, demethoxycurcumin and bisdemethoxycurcumin), and turmerones (aryl-, α- and ß-) as well as piperine, a commonly associated curcumin bioavailability enhancer: respectively 3.5-556, 0-8.6, 0.18-8.1 mg/capsule or tablet. The comparison of the actual and claimed quantities of curcuminoids and piperine showed that 58% of the DS contained the expected amounts of actives.

Analytical strategies to determine the labelling accuracy and economically-motivated adulteration of "natural" dietary supplements in the marketplace: Turmeric case study.

Turmeric has faced authenticity issues as instances of economic-adulterations to reduce the cost. We used carbon-14 and HPLC analyses as complementary methods to verify "all-natural" label claims of commercial dietary supplements containing turmeric ingredients. A high percentage of curcumin-to-curcuminoids value was used as an indicator to imply the presence of synthetic curcumin. However, using the HPLC method alone did not provide direct evidence of curcuminoids' natural origin, whereas using only the carbon-14 method cannot test for potency label claims and determine which constituent(s) contain 14C radiocarbon. By analyzing results from both methods, a significant correlation between the percentage of curcumin-to-curcuminoids and % biobased carbon (Pearson's r = -0.875, p < 0.001) indicated that synthetic curcumin was greatly attributed to determined synthetic ingredients. Only four out of the 14 samples analyzed supported authentic label claims. This orthogonal testing strategy showed its potential for the quality control of turmeric products.

Evaluation of curcuminoids, physiological adaptation, and growth of Curcuma longa under water deficit and controlled temperature.

Turmeric (Curcuma longa L.; Zingiberaceae), an economically important crop and a major spice in Indian cuisine, produces natural yellow color (curcumin) as well as curcuminoids which are widely utilized in traditional and modern medicinal practices. During the turmeric culture, the fluctuations of precipitation and seasonal changes in the whole life cycle play a major role, especially water shortage and decreasing temperature (in winter season), leading to rhizome dormancy under extreme weather conditions. The objective of this investigation was to understand how the water deficit and reduced temperature affect turmeric growth, physiological adaptation, quantity, and quality of turmeric rhizomes. Four-month-old turmeric plants were subjected to four treatments, namely normal temperature and well-watered (RT-WW), or water-deficit (RT-WD) conditions in the greenhouse, 25 °C controlled temperature and well-watered (CT-WW), or water-deficit (CT-WD) conditions in glasshouse. Leaf osmotic potential considerably declined in 30 days CT-WD treatment, leading to chlorophyll degradation by 26.04%, diminution of maximum quantum yield of PSII (Fv/Fm) by 23.50%, photon yield of PSII (ΦPSII) by 29.01%, and reduction of net photosynthetic rate (Pn) by 89.39% over CT-WW (control). After 30 days water withholding, fresh- and dry-weights of rhizomes of turmeric plants grown under CT-WD declined by 30-50% when compared with RT-WW conditions. Subsequently, curcuminoid content was reduced by 40% over RT-WW plants (control), whereas transcriptional expression levels of curcuminoids-related genes (CURS1, CURS2, CURS3, and DCS) were upregulated in CT-WD conditions. In summary, the water withholding and controlled temperature (constant at 25 °C day/night) negatively affected turmeric plants as abiotic stresses tend to limit overall plant growth performances and curcuminoid yield.

Curcuma longa extract protects against 5-fluorouracil-induced oral mucositis in hamsters

Abstract Curcumin, contained at Turmeric (Curcumalonga), can exert many beneficial pleiotropic activities in the gastrointestinal tract. This study evaluated the antioxidant and anti-inflammatory activity of C. longa on 5-fluorouracil (5-FU)-induced oral mucositis (OM) in hamsters. Phytochemical analysis of crude C. longa extract (CLE) was performed to detect the presence of curcumin by TLC and HPLC. Golden Syrian hamsters were orally pre-treated with CLE (5, 50, or 100mg/kg). Cheek pouch samples were subjected to macroscopic and histopathological evaluation. ELISA was performed to quantify the inflammatory cytokines IL-1ß and TNF-α. Superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) levels were assessed by ultraviolet-visible spectroscopy analysis. Behavior analysis was conducted by the open field test. Curcumin content in the CLE was 0.55%m/m ± 0.0161 (2.84%). The group treated with 5mg/kg CLE showed healing evidence with macroscopic absence of ulceration (p<0.05) and microscopic aspect of re-epithelialization, discrete inflammatory infiltrate and absence of edema. Treatment with 5mg/kg CLE significantly increased GSH levels, and reduced MDA levels and SOD activity (p˂0.05), and decreased IL-1ß (p˂0.05) and TNF-α (p˂0.01) levels. A significant reduction in walking distance, ambulation, speed, and rearing was observed for motor activity. Curcumin reduced oxidative stress, inflammation, and motor activity in hamsters with 5-FU-induced OM.

Curcumin solid dispersion based on three model acrylic polymers: formulation and release properties

Abstract To investigate structure-property relationship of polymer-based curcumin solid dispersion (SD), three acrylic polymers were used to formulate curcumin SD by solvent evaporation method. Curcumin Eudragit EPO SD (cur@EPO), curcumin Eudragit RS PO SD (cur@RSPO) and curcumin Eudragit RL PO SD (cur@RLPO) showed deep red, golden orange and reddish orange color, respectively. Cur@RSPO entrapped 15.42 wt% of curcumin followed by cur@RL PO and cur@EPO. FTIR spectra indicated that in cur@EPO, curcumin may transfer hydrogen to the dimethylaminoethyl methacrylate group and thus change its color to red. In contrast, curcumin may form hydrogen bonding with Eudragit RS PO and Eudragit RL. Curcumin exists in amorphous state in three SDs as proved by differential scanning calorimetry and X-Ray diffraction measurement. In vitro digestion presented that lower pH value in simulated gastric fluid (SGF) stimulates the curcumin release from cur@EPO while permeability influences the release profile in other two SDs. When in simulated intestinal fluid (SIF), first order release model governs the release behaviors of all three SDs which showed sustained release pattern. Our results are helpful to elucidate how structure of polymer may impact on the major properties of curcumin contained SD and will be promising to broaden its therapeutic applications.

Curcumin nanoliposomes mitigate wound tissue inflammatory response caused by tooth extraction

Abstract Curcumin is a plant-derived compound with polypharmacological properties that are hampered by its poor solubility, fast degradation, etc. Wound closure complications that follow tooth extraction are numerous, and relatively frequently additional treatment is needed to prevent unwanted process chronification. The present study aims to compare the effects of free and the nanoliposome-encapsulated curcumin on tooth extraction wound closure. The experiments were performed on Wistar rats where both forms of curcumin were applied topically on a tooth extraction wound for seven days. Changes in tissue oxidative stress (malondialdehyde and oxidized proteins concentrations, and catalase activity) and inflammation (nitric oxide levels and myeloperoxidase activity) related parameters were studied three and seven days following the tooth extraction. Also, the extent of pathohistological changes and osteopontin immunohistochemical expression were studied. The obtained results indicate that both forms of curcumin prevent an increase in oxidative stress and inflammation-related parameters in the studied samples at 3-and 7-day time points. Additionally, we found that curcumin diminished tissue inflammatory response and osteopontin expression, while at the same time it caused faster granulation tissue maturation. The encapsulation of curcumin in nanoliposomes proved to be better in improving the extraction wound healing process than the free curcumin, giving this formulation a potential in the pharmaceutical industry.

Validation of an innovative analytical method for simultaneous quantification of curcumin and fluconazole using high-performance liquid chromatography from nanostructured lipid carriers.

Vulvovaginal candidiasis is a public health problem with a high incidence among female patients. Currently, there is an increase in the identification of Candida spp. resistant to current therapy, making it necessary to search for new therapeutic alternatives. The synergistic potential of curcumin with fluconazole is described in the literature. However, due to its high lipophilicity, it is necessary to use drug-delivery systems to adequately explore its potential, among which is the nanostructured lipid carrier. However, to date, there is no validated method of high-performance liquid chromatography for simultaneous determination of fluconazole and curcumin in the literature. Thus, the present work developed a high-performance liquid chromatography method for simultaneous determination of fluconazole and curcumin co-encapsulated in nanostructured lipid carrier which was validated according to the International Conference on Harmonization (Technical Requirements for Registration of Pharmaceuticals for Human Use) - Q2 (R1) and the Food and Drug Administration - Guidance for Bioanalytical Method. The method was applied to determine the encapsulation efficiency and drug-loading of curcumin and fluconazole in nanostructured lipid carriers. The developed method proved to be selective, precise, accurate, and robust for the simultaneous determination of both drugs, enabling the quantification of encapsulation efficiency and drug-loading of curcumin and fluconazole in nanostructured lipid carriers.

The Demethoxy Derivatives of Curcumin Exhibit Greater Differentiation Suppression in 3T3-L1 Adipocytes Than Curcumin: A Mechanistic Study of Adipogenesis and Molecular Docking.

Curcumin is a known anti-adipogenic agent for alleviating obesity and related disorders. Comprehensive comparisons of the anti-adipogenic activity of curcumin with other curcuminoids is minimal. This study compared adipogenesis inhibition with curcumin, demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC), and their underlying mechanisms. We differentiated 3T3-L1 cells in the presence of curcuminoids, to determine lipid accumulation and triglyceride (TG) production. The expression of adipogenic transcription factors and lipogenic proteins was analyzed by Western blot. A significant reduction in Oil red O (ORO) staining was observed in the cells treated with curcuminoids at 20 µM. Inhibition was increased in the order of curcumin < DMC < BDMC. A similar trend was observed in the detection of intracellular TG. Curcuminoids suppressed differentiation by downregulating the expression of peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer-binding protein α (C/EBPα), leading to the downregulation of the lipogenic enzymes acetyl-CoA carboxylase (ACC) and fatty acid synthase (FAS). AMP-activated protein kinase α (AMPKα) phosphorylation was also activated by BDMC. Curcuminoids reduced the release of proinflammatory cytokines and leptin in 3T3-L1 cells in a dose-dependent manner, with BDMC showing the greatest potency. BDMC at 20 µM significantly decreased leptin by 72% compared with differentiated controls. Molecular docking computation indicated that curcuminoids, despite having structural similarity, had different interaction positions to PPARγ, C/EBPα, and ACC. The docking profiles suggested a possible interaction of curcuminoids with C/EBPα and ACC, to directly inhibit their expression.

Colorectal Adenocarcinoma Cell Culture in a Microfluidically Controlled Environment with a Static Molecular Gradient of Polyphenol.

To study the simultaneous effect of the molecular gradient of polyphenols (curcumin, trans-resveratrol, and wogonin) and biological factors released from tumor cells on apoptosis of adjacent cells, a novel microfluidic system was designed and manufactured. The small height/volume of microfluidic culture chambers and static conditions allowed for establishing the local microenvironment and maintaining undisturbed concentration profiles of naturally secreted from cells biochemical factors. In all trials, we observe that these conditions significantly affect cell viability by stimulating cell apoptosis at lower concentrations of polyphenols than in traditional multiwell cultures. The observed difference varied between 20.4-87.8% for curcumin, 11.0-37.5% for resveratrol, and 21.7-62.2% for wogonin. At low concentrations of polyphenols, the proapoptotic substances released from adjacent cells, like protein degradation products, significantly influence cell viability. The mean increase in cell mortality was 38.3% for microfluidic cultures. Our research has also confirmed that the gradient microsystem is useful in routine laboratory tests in the same way as a multiwell plate and may be treated as its replacement in the future. We elaborated the new repetitive procedures for cell culture and tests in static gradient conditions, which may become a gold standard of new drug investigations in the future.

The Untargeted Capability of NMR Helps Recognizing Nefarious Adulteration in Natural Products.

Curcuma longa (turmeric) has an extensive history of ethnomedical use for common ailments, and "curcumin"-containing dietary supplements (CDS) are a highly visible portion of today's self-medication market. Owing to raw material cost pressure, CDS products are affected by economically motivated, nefarious adulteration with synthetic curcumin ("syncumin"), possibly leading to unexpected toxicological issues due to "residual" impurities. Using a combination of targeted and untargeted (phyto)chemical analysis, this study investigated the botanical integrity of two commercial "turmeric" CDS with vitamin and other additives that were associated with reported clinical cases of hepatotoxicity. Analyzing multisolvent extracts of the CDS by 100% quantitative 1H NMR (qHNMR), alone and in combination with countercurrent separation (CCS), provided chemical fingerprints that allowed both the targeted identification and quantification of declared components and the untargeted recognition of adulteration. While confirming the presence of curcumin as a major constituent, the universal detection capability of NMR spectroscopy identification of significant residual impurities, including potentially toxic components. While the loss-free nature of CCS captured a wide polarity range of declared and unwanted chemical components, and also increased the dynamic range of the analysis, (q)HNMR determined their mass proportions and chemical constitutions. The results demonstrate that NMR spectroscopy can recognize undeclared constituents even if they represent only a fraction of the mass balance of a dietary supplement product. The chemical information associated with the missing 4.8% and 7.4% (m/m) in the two commercial samples, exhibiting an otherwise adequate curcumin content of 95.2% and 92.6%, respectively, pointed to a product integrity issue and adulteration with undeclared synthetic curcumin. Impurities from synthesis are most plausibly the cause of the observed adverse clinical effects. The study exemplifies how the simultaneously targeted and untargeted analytical principle of the 100% qHNMR method, performed with entry-level high-field instrumentation (400 MHz), can enhance the safety of dietary supplements by identifying adulterated, non-natural "natural" products.

Radix Pseudostellariae protein-curcumin nanocomplex: Improvement on the stability, cellular uptake and antioxidant activity of curcumin.

Radix Pseudostellariae protein (RPP) with satisfactory antioxidant activity and self-assembled ability was extracted from dried Radix Pseudostellariae. In this study, RPP-curcumin nanocomplex (RPP-Cur) was fabricated, and its improvement on the stability, cellular uptake and antioxidant activity of curcumin was investigated. RPP-Cur with homogeneously spherical structure exhibited good stability, which could maintain the morphology against simulated gastrointestinal digestion and up to 300 mM ionic concentration. After RPP nanoparticles encapsulation, the retention of curcumin increased 1.45 times under UV irradiation for 6 h. Besides, RPP-Cur exhibited additive reducing power of curcumin and RPP. The transport efficiency of hydrophobic curcumin across Caco-2 cells monolayer was greatly improved by RPP nanoparticle by 3.7 folds. RPP-Cur was able to be internalized by Caco-2 cells dose-dependently via macropinocytosis and clathrin-mediated endocytosis. The cellular uptake efficiency of embedded curcumin in RPP nanoparticles by Caco-2 cells was significantly higher than that of free curcumin, which might contribute to the enhanced intracellular antioxidant activity of RPP-Cur. These findings suggest that the proteins from Radix Pseudostellariae have potential to be developed into novel delivery system with intrinsic antioxidant activity for the hydrophobic active molecules in healthy food field.