Contact Toxicity of Various Stemona Collinsiae Root Extracts Against Periplaneta Americana (Dictyoptera: Blattodea) And Detection of Didehydrostemofoline Distribution in Tissue Using MALDI Imaging Mass Spectrometry

Contact toxicity against Periplaneta americana has never been tested with S. collinsiae root extract. Hexane, dichloromethane, ethanol and water extracts were tested in final-instar nymphs and adult P. americana by topical application method. The dichloromethane extract showed the high-est potency of contact toxicity against the final-instar nymphs (41-100% corrected mortality at 48 hours), lowest LC50 (1.5±0.2 %w/v at 48 hours), and lowest LT50 (36.1±0.8 hours at 10%w/v) while the water crude extract lacked the contact toxicity (0-0% corrected mortality at 48 hours). Signs of toxicity, such as excited movement, trembling body, motionlessness, and swollen abdomen segment including irregularly extended foregut were found at the both stages of P. americana dropping with solutions of dichloromethane extract. Detection of didehydrostemofoline distri-bution using IMS revealed that didehydrostemofoline distributed in the tissue of the dead fi-nal-instar nymph and adult P. americana contacting with dichloromethane extract, but it was not found in tissue of euthanized P. americana which exposed to the water extract. Didehydrostemo-foline in the extract was a cause of toxicity signs and death of P. americana via a contact route. Thus, dichloromethane extract and didehydrostemofoline could be used as an active ingredient and chemical marker in aerosol and spray formulations for cockroach control.

HPLC Method for Separation of Cannabidiol Hemp Seed Oil with Skin Lipids and Tandem HRMS Technology for Characterization of a Chemical Marker

Cannabidiol (CBD) hemp seed oil is a commercial raw material with antioxidant and anti-inflammatory benefits that has been formulated into body wash and skin care products. The biggest analytical challenge is how to simultaneously quantify CBD and hemp seed oil as they deposited on the skin surface. CBD is easily separated and quantified from skin surface extracts via a HPLC-mass spectrometry methodology. However, the structural skeleton of triacylglycerides (TAGs) in hemp seed oil is same as those from the skin surface sebum. The strong hydrophobicity with subtle structural difference challenges their separation. In this project, a new reverse phase HPLC-high resolution mass spectrometry methodology was developed with a strong mobile phase normal propanol. The separated hemp seed oil TAGs in the chromatogram were identified and characterized using data-dependent acquisition (DDA) technology. Based on the daughter ion characterization, the separated peak with an ammonium adduct at 890.7226 [M + NH4]+ was confirmed as the parent ion of glycerol with three omega-3 fatty acid chains. This is the first time TAG structure with direct HPLC-tandem mass spectrometry technology has been elucidated without a hydrolysis reaction. The confirmed TAG structure with an ammonium adduct at 890.7226 ± 0.0020 can be used as a representative chemical marker for the hemp seed oil quantification.

Organ Specific Differences in Alteration of Aquaporin Expression in Rats Treated with Sennoside A, Senna Anthraquinones and Rhubarb Anthraquinones

Senna and rhubarb are often used as routine laxatives, but there are differences in mechanism of action and potential side effects. Here, we studied metabolites of senna anthraquinones (SAQ), rhubarb anthraquinones (RAQ) and their chemical marker, sennoside A (SA), in a rat diarrhea model. In in vitro biotransformation experiments, SAQ, RAQ and SA were incubated with rat fecal flora solution and the metabolites produced were analyzed using HPLC. In in vivo studies, the same compounds were investigated for purgation induction, with measurement of histopathology and Aqps gene expression in six organs. The results indicated that SAQ and RAQ had similar principal constituents but could be degraded into different metabolites. A similar profile of Aqps down-regulation for all compounds was seen in the colon, suggesting a similar mechanism of action for purgation. However, in the kidneys and livers of the diarrhea-rats, down-regulation of Aqps was found in the RAQ-rats whereas up-regulation of Aqps was seen in the SAQ-rats. Furthermore, the RAQ-rats showed lower Aqp2 protein expression in the kidneys, whilst the SA-rats and SAQ-rats had higher Aqp2 protein expression in the kidneys. This may have implications for side effects of SAQ or RAQ in patients with chronic kidney or liver diseases.

Cartwright's method as a physico-chemical marker of antimonium crudum biological effect

Mice bearing Leishmania (L) amazonensis infection and treated with Antimonium crudum (AC) 30cH presented significant reduction of the monocyte migration to the site of infection with clinical improvement. In vitro, the treatment of infected macrophages with AC 30cH produced inhibition of the parasite-induced peaks of CCL2 (a chemokine for monocytes migration) and inhibition of lysosome activity, explaining the results obtained previously in vivo. In the following studies, physical-chemical parameters of the remedy and respective controls were evaluated, to search for a correlation with the former described biological effects. The study of polarity changes in different water-based dilutions of AC using Cartwright´s method, revealed dilution-dependent variations in the absorbance of three solvatochromic dyes ET 33, BDN and methylene Violet (MV), used as “probes” to evaluate the dipole features of the medicine. The electrical activity of the homeopathic preparations appears to be dilution-dependent and related to their biological effects. Further experiments were performed using samples of the supernatant of infected macrophages after 96 hours of incubation with AC in different dilutions. These samples were processed using the same procedures as used for the original medicines and were analyzed by MV method. All tested potencies presented a sharp increase of absorbance at 580 nm, in relation to all controls (supernatant from untreated cells and cells treated with succussed water), as performed by one-way ANOVA, being F = 176.208; p = 0.001 and ?2 = 0.988. This results strongly suggest that biological systems could amplify the electric signal and the following changes in the medium polarity.

Effect of Scopoletin and Carotenoids on Postharvest Physiological Deterioration (PPD) of Transgenic High Beta Carotene Cassava

Cassava tubers suffer from postharvest physiological deterioration (PPD) which normally sets in within 72 hours of harvest. This study examines the role of scopoletin and carotenoids in the onset or delay in PPD in two transgenic varieties EC20-7 and EC20-8 compared to a wild variety TME-7. Scopoletin and carotenoids were quantified by liquid chromatography-mass spectrometry. The scopoletin content (0.10 – 0.20 nmol/g) in the fresh varieties was not significantly (P>0.05) different from the amount in stored cassava roots (12.58 – 14.90 nmol/g). The carotenoid content values in EC20-7 variety were 6.66 µg/g (α-carotene), 80.45 µg/g (β-carotene) and 5.98 µg/g (lutein). As for EC20-8, α-carotene, β-carotene and lutein values were 6.19 µg/g, 69.11 µg/g and 3.12 µg/g, respectively. There was no significant (P>0.05) difference between the varieties in α-carotene content but in their lutein content. The results indicate that carotenoids are more relevant in the delay of PPD and scopoletin content is not a major factor in PPD vascular streaking or discolouration. Hence scopoletin content of cassava varieties may not be considered as a chemical marker for determining the potential of PPD in cassava tubers.

Electrochemical Determination of the “Furanic Index” in Honey

5-(hydroxymethyl)furan-2-carbaldehyde, better known as hydroxymethylfurfural (HMF), is a well-known freshness parameter of honey: although mostly absent in fresh samples, its concentration tends to increase naturally with aging. However, high quantities of HMF are also found in fresh but adulterated samples or honey subjected to thermal or photochemical stresses. In addition, HMF deserves further consideration due to its potential toxic effects on human health. The processes at the origin of HMF formation in honey and in other foods, containing saccharides and proteins—mainly non-enzymatic browning reactions—can also produce other furanic compounds. Among others, 2-furaldehyde (2F) and 2-furoic acid (2FA) are the most abundant in honey, but also their isomers (i.e., 3-furaldehyde, 3F, and 3-furoic acid, 3FA) have been found in it, although in small quantities. A preliminary characterization of HMF, 2F, 2FA, 3F, and 3FA by cyclic voltammetry (CV) led to hypothesizing the possibility of a comprehensive quantitative determination of all these compounds using a simple and accurate square wave voltammetry (SWV) method. Therefore, a new parameter able to provide indications on quality of honey, named “Furanic Index” (FI), was proposed in this contribution, which is based on the simultaneous reduction of all analytes on an Hg electrode to ca. −1.50 V vs. Saturated Calomel Electrode (SCE). The proposed method, validated, and tested on 10 samples of honeys of different botanical origin and age, is fast and accurate, and, in the case of strawberry tree honey (Arbutus unedo), it highlighted the contribution to the FI of the homogentisic acid (HA), i.e., the chemical marker of the floral origin of this honey, which was quantitatively reduced in the working conditions. Excellent agreement between the SWV and Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) data was observed in all samples considered.

Determination of astilbin in dietary supplements containing Smilax glabra using high-performance liquid chromatography

Smilax glabra rhizoma (SGR) has been used in traditional medicine remedies for the treatment of joint pain and inflammation, and recently, has been presented in dietary supplements for supporting arthritis and gout treatment. In the 5th Vietnamese Pharmacopoeia, astilbin was chosen as a chemical marker for the quality control of Smilax glabra as herbal medicine. A high performance liquid chromatographic (HPLC) method was developed for the qualitative and quantitative determination of astilbin in dietary supplements containing Smilax glabra. Ultrasonic extraction with 60% methanol was used for astilbin extraction from herbal products. HPLC analysis was performed with a C18 column and a mobile phase consisting of methanol and water in gradient mode, with UV detection at 291 nm. The method was validated according to AOAC International’s requirements with high specificity and precision.