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Chemical Analysis of Lepidium Meyenii (Maca) and Its Effects On molecules Article Chemical Analysis of Lepidium meyenii (Maca) and Its Effects on Redox Status and on Reproductive Biology y in Stallions 1, 1, , 2 3 Simona Tafuri z, Natascia Cocchia * z, Domenico Carotenuto , Anastasia Vassetti , Alessia Staropoli 3,4, Vincenzo Mastellone 1, Vincenzo Peretti 1, Francesca Ciotola 1, Sara Albarella 1 , Chiara Del Prete 1, Veronica Palumbo 1, Luigi Esposito 1 , Francesco Vinale 3,4 and Francesca Ciani 1 1 Department of Veterinary Medicine and Animal Production, University of Naples Federico II, 80137 Naples, Italy; [email protected] (S.T.); [email protected] (V.M.); [email protected] (V.P.); [email protected] (F.C.); [email protected] (S.A.); [email protected] (C.D.P.); [email protected] (V.P.); [email protected] (L.E.); [email protected] (F.C.) 2 UNMSM, Universidad Nacional Mayor San Marcos, Lima 11-0058, Peru; [email protected] 3 Institute for Sustainable Plant Protection, National Research Council, 80055 Portici (Na), Italy; [email protected] (A.V.); [email protected] (A.S.); [email protected] (F.V.) 4 Department of Agricultural Sciences, University of Naples Federico II, 80055 Naples, Italy * Correspondence: [email protected]; Tel.: +39-81-253-6017; Fax: +39-81-253-6042 Running title: Maca: Chemical Analysis and Biological Activity. y The authors contributed equally to this work. z Academic Editor: Francesca Giampieri Received: 15 March 2019; Accepted: 21 May 2019; Published: 23 May 2019 Abstract: The present study was conducted to assess the chemical composition of Yellow Maca (Lepidium meyenii) and its biological activity on stallions following oral administration of hypocotyl powder. Maca was subjected to methanolic extraction and the chemical analysis was carried out by LC-MS-QTOF (liquid chromatography-mass spectrometry). Our results showed that Maca contains some effective antioxidants, a high percentage of glucosinolates, and other important components with a high antioxidant capacity. To evaluate the plant biological activity in stallions fed with Maca powder for 60 days, the redox status and some reproductive parameters were investigated. Blood and semen samples were collected at 0, 30, 60, and 90 days from the beginning of this study. Blood samples showed a decrease of the reactive oxygen metabolites, evaluated by d-ROMs test, and an increase of the antioxidant barrier in terms of biological antioxidant potential (BAP test), powerful oxidant capacity (OXY-Adsorbent test), and thiols evaluation (-SHp test). Furthermore, semen samples showed a positive trend during Maca administration in the following parameters: ejaculate volumes and sperm concentrations, total and progressive motility, and acrosome integrity. Keywords: Lepidium meyenii (Maca); stallion semen; oxidative stress; antioxidants; chemical analysis 1. Introduction Maca (Lepidium meyenii Walp), belonging to the Brassicaceae family, was discovered more than 2000 years ago in the Andes highlands of Peru, where it grows exclusively between 3500 and 4500 m above sea level [1]. The hypocotyls have been widely used as a nutritional supplement and in folk medicine to increase fertility and sexual function [2]. The dried hypocotyls of Maca are rich in high nutritional value elements, such as carbohydrates, proteins, lipids, essential amino acids, and free fatty acids. Furthermore, Maca contains several secondary metabolites, such as macamides, macaridine, alkaloids, and glucosinolates [3]. The most abundant glucosinolates detected in Maca are aromatic Molecules 2019, 24, 1981; doi:10.3390/molecules24101981 www.mdpi.com/journal/molecules Molecules 2019, 24, x 2 of 11 medicine to increase fertility and sexual function [2]. The dried hypocotyls of Maca are rich in high nutritional value elements, such as carbohydrates, proteins, lipids, essential amino acids, and free Molecules 2019, 24, 1981 2 of 12 fatty acids. Furthermore, Maca contains several secondary metabolites, such as macamides, macaridine, alkaloids, and glucosinolates [3]. The most abundant glucosinolates detected in Maca glucosinolates,are aromatic glucosinolates, namely benzyl namely glucosinolate(glucotropaeolin) benzyl glucosinolate(glucotropaeolin) and m-methoxybenzyl and m-methoxybenzyl glucosinolate (glucolimnanthin).glucosinolate (glucolimnanthin). Maca has been Maca mainly has been classified mainly in threeclassified ecotypes in three according ecotypes to according the color ofto the hypocotyls:color of the hypocotyls: red, yellow, andred, black.yellow, These and black. showed Th diesefferent showed biological different activity biological depending activity on depending the type of cultivation,on the type processing,of cultivation, and processing, extraction, and onextraction, the concentration and on the of concentration different bioactive of different metabolites bioactive [3,4]. Themetabolites main chemical [3,4]. The components main chemical of Maca components were considered of Maca to were be responsible, considered among to be re othersponsible, things, among for its aphrodisiac,other things, immunostimulant,for its aphrodisiac, andimmunostimulant, energizing properties and energizing [5,6]. properties [5,6]. Previous studies have demonstrated the eeffectsffects of MacaMaca onon spermatogenesis,spermatogenesis, sperm count, and sperm motility [[1,7].1,7]. Furthermore, in a rat model Maca restored the spermatogenesis decrease when altitudealtitude increasesincreases [ 8[8].]. Additionally, Additionally, diet diet supplementation supplementation of Macaof Maca showed showed a beneficial a beneficial effect oneffect semen on qualitysemen quality of hypofertile of hypofertile and fertile and stallions fertile stallions [9], as well [9], as well on the as quantityon the qua ofntity fresh of semen fresh andsemen the and quality the ofquality semen of aftersemen storage after storage at 5 ◦C upat 5 to °C 72 up h [to10 ].72 h [10]. The use ofof artificialartificial inseminationinsemination (AI)(AI) hashas greatlygreatly increasedincreased inin recentrecent decades.decades. AI has severalseveral advantages comparedcompared to naturalto natural service, service, including including higher pregnancyhigher pregnancy rates and reduced rates transportationand reduced coststransportation of a mare costs to the of stallion, a mare with,to the consequently, stallion, with, lower consequently, stress and lower prevention stress ofand a stallionprevention overuse. of a Semenstallion used overuse. for AI Semen should used be preserved for AI should from damagebe preserved occurring from due damage to oxidative occurring stress due [10 ].to Aoxidative strategy tostress prevent [10]. oxidative A strategy damage to toprevent spermatozoa oxidative may da bemage found to in foodspermatozoa supplementation may be with found antioxidants. in food Thesupplementation presence of compounds,with antioxidants. such as The macamides presence and of glucosinolates, compounds, whichsuch a ares macamides hydrolisated and to isothiocyanatesglucosinolates, which by myrosinase, are hydrolisated allows Maca to isothiocyanates to scavenge free by radicals myrosinase, and protect allows cells Maca from to oxidativescavenge stressfree radicals (OS). Reactive and protect oxygen cells species from (ROS) oxida aretive produced stress (OS). in diff erentReactive sites oxygen in mammalian species bodies(ROS) [11are], andproduced were foundin different to accumulate sites in duringmammalian the normal bodies testicular [11], and spermatogenesis were found to andaccumulate steroidogenesis during [12the]. Physiologicalnormal testicular low levelsspermatogenesis of ROS play and an important steroidogenesi role ins processes [12]. Physiological such as capacitation, low levelshyperactivation, of ROS play an acrosomeimportant reaction,role in andprocesses sperm-oocyte such fusion,as capacitation, thus ensuring hyperactivation, appropriate fertilization,acrosome reaction, whereas highand levelssperm-oocyte of ROS mayfusion, determine thus ensuring sperm pathologies,appropriate suchfertiliza as ATPtion, depletion whereas andhigh loss levels of sperm of ROS motility may anddetermine viability sperm [13]. pathologies, ROS activity such is of majoras ATP concern depletion for spermand loss quality of sperm both inmotility vivo and and during viabilityin vitro[13]. incubation,ROS activity as is well of major as during concern its cool for storagesperm [quality14]. The both cellular in vivo antioxidant and during defense in vitro systems incubation, are able as to controlwell as theduring deleterious its cool estorageffects of [14]. ROS, The but cellular if an imbalance antioxidant between defense oxidant systems and are antioxidant able to control systems the determinesdeleterious ROSeffects accumulation, of ROS, but if OS an may imbalance occur [ 15between–17]. oxidant and antioxidant systems determines ROS Inaccumulation, the present study OS may the occur chemical [15–17]. composition of Yellow Maca (Lepidium meyenii) was investigated by usingIn the LC-MS- present QTOF. study Moreover, the chemical the biological composition effects onof stallionsYellow Ma orallyca fed(Lepidium during meyenii the breeding) was seasoninvestigated with Maca by using hypocotyl LC-MS- powder QTOF. was Moreover, evaluated the in termsbiological of antioxidant effects on activity stallions and orally semen fed quantity during andthe breeding quality. season with Maca hypocotyl powder was evaluated in terms of antioxidant activity and semen quantity and
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