Quantitative Determination and Characterization of a Kashmir Saffron (Crocus sativus L.)-Based Botanical Supplement Using Single-Laboratory Validation Study by HPLC-PDA with LC–MS/MS and HPTLC Investigations

Abstract

Food ingredients hold a higher nutritional value as a botanical supplement playing a vital role in modifying and maintaining the physiological conditions that improve human health benefits. The Kashmir saffron ( Crocus sativus L; KCS) obtained from dried stigmas is known for its aroma precursors and apocarotenoid derivatives, imparting a wide range of medicinal values and therapeutic benefits. In the present study, a simultaneous determination of apocarotenoids and flavonoids in stigma-based botanical supplements was carried out using analytical investigations. The high-performance thin-layer chromatography-based qualitative analysis of the raw material (stigmas, stamens, and tepals) and stigma extract has been carried out to identify apocarotenoids and flavonoids. The rapid HPLC-PDA method for the simultaneous quantification of KCS apocarotenoids was robust, precise (<5.0%), linear ( R 2 > 0.99), and accurate (80–110%) as per the single-laboratory validation data. Furthermore, the combined-expanded uncertainty (95%; K = 2) was calculated and found as 0.0035–0.007% (<5.0%) as per the EURACHEM guide for this HPLC analysis. Additionally, an untargeted identification of 36 compounds in the botanical supplement was based on the elution order, UV–vis spectra, mass fragmentation pattern, and standards by ESI-MS/MS analysis with comprehensive chromatographic fingerprinting. Thus, these analytical approaches enable a composite profile of the stigma-based extract as a potential supplement for human health benefits.

Quantitative and rapid quality assessment methods for the multi‐class bioactive constituents of Tinospora cordifolia using high‐performance liquid and thin layer chromatography analysis with tandem mass spectrometry characterization

“…Further in this study, LC-MS/MS was used to identify, select and confirm analytical markers. Based on the existing literature [25] and the optimization trials, an HPLC-MS/MS method was developed [30]. The developed method was utilized to identify 10 phytometabolite in TCE.…”

“…The second solvent system for compounds (3-4) used as ethyl acetate: glacial acetic acid: formic acid: water (10:1:1:2.5), followed by derivatization with anisaldehyde sulfuric acid (method 2). The injection volume used was 5 μl with an 8 mm band length followed by a 12 min run time for method 1 and 20 min for method 2 respectively, with a run distance of 80 mm [30]. For HPTLC-MS analysis, bands were eluted with CAMAG TLC-MS interface 2 using methanol as a carrier solvent at a flow rate of 0.1 ml/min, followed by offline analysis by LC-ESI-MS/MS.…”

Turmeric ethanol extract (Curcuma longa L.) reduces apoptosis and promotes canine osteosarcoma cell proliferation

“…Curcuma longa L., popularly known as turmeric, belongs to the family Zingiberaceae of genus Curcuma; it is a food spice appreciated worldwide to color and flavor foods. Although the plant originates from the tropical forests of India, the biggest supplier worldwide (11,12) , it is also widely consumed and produced in Mara Rosa, a central municipality in Brazil. Turmeric contains a large fraction (6.8%) of curcumin, a highly therapeutic molecule (13) .…”

“…scores and semi-quantitative combined scoring system (SCSS) from Fedchenko & Reifenrath12 to assess slides with canine osteosarcoma cells of the lineage D-17 treated with the turmeric (Curcuma longa, L.) ethanolic extract. The cells were stained with anti-Bcl-2, -KI-67 or -mutant p53 antibody and stained with hematoxylin.…”

Extrato etanólico de açafrão (Curcuma longa L.) reduz apoptose e promove proliferação de células de osteossarcoma canino

“…É uma planta popularmente conhecida como açafrão, que se caracteriza como uma cultura de colheita abundante e floração perene. Trata-se de um condimento alimentar apreciado e amplamente utilizado no mundo por conferir cor e sabor aos alimentos, tendo sua origem no sudeste da Ásia, nas florestas tropicais da Índia, maior fornecedor mundial (11,12) . No Brasil, o Centro-Oeste, particularmente a região de Mara Rosa -GO, é a que mais se destaca na produção e consumo do açafrão, cujo teor de curcumina, a molécula com efeitos terapêuticos mais característica desta planta, apresenta o significativo teor médio de 6,8% (13) .…”