Controlling the quality of maca (Lepidium meyenii) dietary supplements: Development of compendial procedures for the determination of intact glucosinolates in maca root powder products.

The demand and sales of dietary supplements derived from maca (Lepidium meyenii) have skyrocketed in the last decade and a variety of related nutritional and healthcare products have mushroomed into a business with market prominence. However, the lack of standard testing protocols for quality control could jeopardize the immediate benefits of these products for public health. We describe herein the development of analytical procedures for the determination of glucosinolates (GLs), the biologically active ingredients in maca. Because of the high polarity and instability caused by enzymatic hydrolysis, GLs in maca have been exclusively analyzed using desulfated GLs. This indirect analysis requires additional sample preparation steps, which is labor-intensive, and may lose the original GLs and introduce artificial compounds. Furthermore, the reported GL profiles of maca are inconsistent and incomplete, some GLs may be structurally misidentified. In this context, we focused on direct analysis of intact GLs in maca without the enzymatic desulfation. Four GLs (sinalbin, glucolepigramin, glucolimnanthin, and glucotropaeolin) were identified as the major GLs in maca root powder. An HPLC method based on ion pair chromatography was developed to determine individual and total GLs; chromatographic separations were achieved on a Luna column (C18, 4.6 × 100 mm,3 mm) using 0.1 % TFA in water and in methanol as mobile phase in a gradient elution mode. The developed procedures were validated within the calibration range of 10-500 µg/mL. Inter- and intra-day precision were shown to be lower than 3% at all concentrations levels with recovery between 100.2 % and 103.3 %. The procedures were applied to a total of 42 maca root powder products from 11 manufacturers. Sample analysis revealed a consistent correlation of glucotropaeolin: glucolimnanthin (1: 0.19) across all products with a correlation coefficient of 0.994. The correlation in combination with total GL contents for each product could be used for authentication and GL content determination. Incorporation of the developed procedures into USP monographs will strengthen the public standards for maca powder dietary supplements.

Proanthocyanidin Characterization, Antioxidant and Cytotoxic Activities of Three Plants Commonly Used in Traditional Medicine in Costa Rica: Petiveria alliaceae L., Phyllanthus niruri L. and Senna reticulata Willd.

The phenolic composition of aerial parts from Petiveria alliaceae L., Phyllanthus niruri L. and Senna reticulata Willd., species commonly used in Costa Rica as traditional medicines, was studied using UPLC-ESI-TQ-MS on enriched-phenolic extracts. Comparatively, higher values of total phenolic content (TPC), as measured by the Folin-Ciocalteau method, were observed for P. niruri extracts (328.8 gallic acid equivalents/g) than for S. reticulata (79.30 gallic acid equivalents/g) whereas P. alliaceae extract showed the lowest value (13.45 gallic acid equivalents/g). A total of 20 phenolic acids and proanthocyanidins were identified in the extracts, including hydroxybenzoic acids (benzoic, 4-hydroxybenzoic, gallic, prochatechuic, salicylic, syringic and vanillic acids); hydroxycinnamic acids (caffeic, ferulic, and p-coumaric acids); and flavan-3-ols monomers [(+)-catechin and (-)-epicatechin)]. Regarding proanthocyanidin oligomers, five procyanidin dimers (B1, B2, B3, B4, and B5) and one trimer (T2) are reported for the first time in P. niruri, as well as two propelargonidin dimers in S. reticulata. Additionally, P. niruri showed the highest antioxidant DPPH and ORAC values (IC50 of 6.4 µg/mL and 6.5 mmol TE/g respectively), followed by S. reticulata (IC50 of 72.9 µg/mL and 2.68 mmol TE/g respectively) and P. alliaceae extract (IC50 >1000 µg/mL and 1.32 mmol TE/g respectively). Finally, cytotoxicity and selectivity on gastric AGS and colon SW20 adenocarcinoma cell lines were evaluated and the best values were also found for P. niruri (SI = 2.8), followed by S. reticulata (SI = 2.5). Therefore, these results suggest that extracts containing higher proanthocyanidin content also show higher bioactivities. Significant positive correlation was found between TPC and ORAC (R² = 0.996) as well as between phenolic content as measured by UPLC-DAD and ORAC (R² = 0.990). These findings show evidence for the first time of the diversity of phenolic acids in P. alliaceae and S. reticulata, and the presence of proanthocyanidins as minor components in latter species. Of particular relevance is the occurrence of proanthocyanidin oligomers in phenolic extracts from P. niruri and their potential bioactivity.

Phenolic Assesment of Uncaria tomentosa L. (Cat's Claw): Leaves, Stem, Bark and Wood Extracts.

The phenolic composition of extracts from Uncaria tomentosa L. from different regions of Costa Rica was studied using advanced analytical techniques such as UPLC/TQ-ESI-MS and (13)C-NMR. Samples from leaves, stems, bark and wood (n = 22) were subjected to extraction to obtain phenolic and alkaloid extracts, separately. Comparatively, higher values of total phenolic content were observed for leaves, stems and bark (225-494 gallic acid equivalents/g) than for wood extracts (40-167 gallic acid equivalents/g). A total of 32 non-flavonoid and flavonoid compounds were identified in the phenolic extracts: hydroxybenzoic acids (benzoic, salicylic, 4-hydroxybenzoic, prochatechuic, gallic, syringic and vanillic acids), hydroxycinnamic acids (p-coumaric, caffeic, ferulic and isoferulic acids), flavan-3-ols monomers [(+)-catechin and (-)-epicatechin)], procyanidin dimers (B1, B2, B3, B4, B5, B7 and two other of unknown structure) and trimers (C1, T2 and one of unknown structure), flavalignans (four unknown structures pertaining to the cinchonain family) and propelargonidin dimers (four unknown structures, reported for the first time in U. tomentosa). Additionally, alkaloid extracts obtained from the plant residue after phenolic extraction exhibited a content of tetracyclic and pentacyclic alkaloids ranging between 95 and 275 mg/100 g of dry material for bark extracts, and between 30 and 704 mg/100 g for leaves extracts. In addition, a minor alkaloid was isolated and characterized, namely 18,19-dehydrocorynoxinoic acid. Our results confirmed the feasibility of U. tomentosa as a suitable raw material for obtaining phenolic- and alkaloid-rich extracts of potential interest.

UHPLC-PDA-ESI/HRMSn profiling method to identify and quantify oligomeric proanthocyanidins in plant products.

Oligomeric proanthocyanidins were successfully identified by UHPLC-PDA-HRMS(n) in a selection of plant-derived materials (jujube fruit, Fuji apple, fruit pericarps of litchi and mangosteen, dark chocolate, and grape seed and cranberry extracts). The identities of 247 proanthocyanidins were theoretically predicted by computing high-accuracy masses based on the degree of polymerization, flavan-3-ol components, and the number of A type linkages and galloyls. MS(n) fragments allowed characterization on flavan-3-ol based on the monomer, connectivity, and location of A-type bonds. Identification of doubly or triply charged ions of 50 PAs was made on the basis of theoretical calculations. A single catechin standard and molar relative response factors (MRRFs) were used to quantify the well-separated PAs. The ratios of the SIM peak counts were used to quantify each of the unseparated isomers. This is the first report of direct determination of each of the proanthocyanidins in plant-derived foods and proanthocyanidins containing an epifisetinidol unit in grape seeds.