Ultra-High-Performance Liquid Chromatography with Photodiode Array and High-Resolution Time-of-Flight Mass Spectrometry Detectors (UHPLC-PDA-ESI-ToF/HRMS) for the Tentative Structural Characterization of Bioactive Compounds of Salvia verbenaca Extracts in Relation to Their Biological Activities.

BACKGROUND: Salvia verbenaca of the Lamiaceae family is a Mediterranean plant widely used in the Moroccan traditional folk medicine. The aim of this work was to explore the phytochemical composition of Salvia verbenaca extracts and its antioxidant activity. METHODS: Separation and identification of the major phytochemicals present in the two hexane and ethyl acetate explored extracts have been achieved through ultra-high-performance liquid chromatography separation technique coupled to photodiode array and high-resolution time-of-flight mass spectrometry detectors. Antioxidant activity of the obtained extracts was evaluated through DPPH• (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azinobis(3-ethylbenzothiazoline6-sulfonic acid) diammonium salt) assays and the obtained results were compared to TROLOX (6-hydroxy2,5,7,8-tetramethylchroman-2-carboxylic acid). RESULTS: Using the analysis technique indicated above, eighteen phytochemicals belonging to phenolic acids, phenolic diterpenes and flavonoids have been characterized on the basis of the obtained UV and mass spectroscopic properties and on the subsequent fragmentations. The antioxidant activity of the explored extracts showed potential scavenging activities compared to TROLOX. A detailed discussion of the attained results has been presented considering the activities observed of each extract. CONCLUSIONS: The research herein presented an analysis technique allowing to screen Salvia verbenaca phytochemicals. The explored plant could be considered as a source of functional phenolic compounds. These could be useful for further pharmacological studies such as new drugs design after clinic and its safety evaluation. It is thus hoped that the information presented here might prompt further studies that will possibly lead to development of therapeutic agents from this plant.

The Challenge of Measuring Sweet Taste in Food Ingredients and Products for Regulatory Compliance: A Scientific Opinion.

The Codex Alimentarius Commission, a central part of the joint Food and Agricultural Organization/World Health Organizations Food Standards Program, adopts internationally recognized standards, guidelines, and code of practices that help ensure safety, quality, and fairness of food trade globally. Although Codex standards are not regulations per se, regulatory authorities around the world may benchmark against these standards or introduce them into regulations within their countries. Recently, the Codex Committee on Nutrition and Foods for Special Dietary Uses (CCNFSDU) initiated a draft revision to the Codex standard for follow-up formula (FUF), a drink/product (with added nutrients) for young children, to include requirements for limiting or measuring the amount of sweet taste contributed by carbohydrates in a product. Stakeholders from multiple food and beverage manufacturers expressed concern about the subjectivity of sweetness and challenges with objective measurement for verifying regulatory compliance. It is a requirement that Codex standards include a reference to a suitable method of analysis for verifying compliance with the standard. In response, AOAC INTERNATIONAL formed the Ad Hoc Expert Panel on Sweetness in November 2020 to review human perception of sweet taste, assess the landscape of internationally recognized analytical and sensory methods for measuring sweet taste in food ingredients and products, deliver recommendations to Codex regarding verification of sweet taste requirements for FUF, and develop a scientific opinion on measuring sweet taste in food and beverage products beyond FUF. Findings showed an abundance of official analytical methods for determining quantities of carbohydrates and other sweet-tasting molecules in food products and beverages, but no analytical methods capable of determining sweet taste. Furthermore, sweet taste can be determined by standard sensory analysis methods. However, it is impossible to define a sensory intensity reference value for sweetness, making them unfit to verify regulatory compliance for the purpose of international food trade. Based on these findings and recommendations, the Codex Committee on Methods of Analysis and Sampling agreed during its 41st session in May 2021 to inform CCNFSDU that there are no known validated methods to measure sweetness of carbohydrate sources; therefore, no way to determine compliance for such a requirement for FUF.

Single-Laboratory Validation to Extend the Scope of AOAC Official MethodSM 2015.06 to Indian Infant, Child, and Adult Nutritional Matrixes.

BACKGROUND: AOAC Method 2015.06 is a Final Action Official MethodSM for the determination of 12 elements (Na, Mg, P, K, Ca, Cr, Mn, Fe, Cu, Zn, Se, and Mo) in infant formula and adult nutritional products, based on inductively coupled plasma (ICP)-MS. Currently, its scope does not include certain kinds of formulations used in India. The method would likely be used more in Indian laboratories if its performance were characterized on Indian matrixes. OBJECTIVE: In this study we describe a typical single-laboratory validation (SLV) exercise designed to characterize the precision and accuracy of AOAC Method 2015.06 for common Indian nutritional matrixes so that the scope of the method can be extended to include them. METHOD: Six matrixes specific to the Indian market were carried through an SLV and the Standard Method Performance Requirements (SMPRs®) previously published for this method were used to evaluate the results. RESULTS: The method demonstrated typical repeatability (<5% RSD), and intermediate precision (5-8% RSD) on the Indian matrixes, with very few exceptions. Accuracy was demonstrated by overspike recoveries in the range of 90-110% over 3 days for the Indian matrixes, as well as excellent agreement with previously published results for three additional matrixes tested. Some of the new Indian matrixes required alternate sample preparation procedures versus the usual reconstitution prescribed by the SMPRs. CONCLUSIONS: The SLV results showed that AOAC Method 2015.06 can be extended to include these Indian matrices. HIGHLIGHTS: The two special sample preparation procedures can now be considered validated.

Structural characterization of bioactive compounds in Cotula cinerea extracts by ultra-high-performance liquid chromatography with photodiode array and high-resolution time-of-flight mass spectrometry detectors.

RATIONALE: Cotula cinerea of the Asteraceae family is a traditional Moroccan plant with various biological activities such as analgesic, cytotoxic and antioxidant effects which are often related to the presence of secondary metabolites. The present work aims to screen and identify the main phytochemicals compounds of Cotula cinerea extracts. METHODS: A method was developed that coupled a rapid and simple ultra-high-performance liquid chromatography system with both photodiode array and high-resolution time-of-flight mass spectrometry detectors (UHPLC-PDA/TOF-HRMS) for the identification of the main secondary metabolites of three investigated extracts (hexane, AcOEt and n-BuOH). RESULTS: A total of 30 phytocomponents pertaining to phenolic compounds and terpenoids have been detected, identified and quantified. Among these were previously reported free and conjugated coumaric and caffeic acids along with free and conjugated flavones and flavonols with kaempferol, quercetin, luteolin and apigenin aglycones. In addition, sulfated flavonoids were identified in the investigated extracts and are reported in this work for the first time in Cotula cinerea. The obtained results have been discussed in relation to the biological activities of the corresponding extracts. CONCLUSIONS: This study proposed a practical strategy for rapidly screening and identifying secondary metabolites of Cotula cinerea and provided new information on the phytochemicals of this Saharan plant. This work has therefore provided useful results for further pharmacological studies and the design of new drugs based on this species and will facilitate the utilization of Cotula cinerea in the clinic and its safety evaluation. It is also hoped that the information presented here might stimulate further studies that will possibly lead to development of therapeutic agents from this plant.