Received:18September2018 | Revised:16January2019 | Accepted:16February2019

DOI: 10.1002/fsn3.990

SchoolofPharmacy,JiangxiUniversityof TraditionalChineseMedicine,Nanchang, China

Correspondence Ling‐yunZhong,SchoolofPharmacy,Jiangxi UniversityofTraditionalChineseMedicine, Nanchang,China. Email:[email protected]

Funding information NationalNaturalScienceFoundationof China,Grant/AwardNumber:81260643, 81460605and81660663

Objective: Patientswithgastrointestinaldisorderscommonlysufferfrompoortreatmentoutcomesandadverseeffectsoftraditionalpharmacologicaltherapy.Herbal medicineisafavorablealternativeduetothelowriskofsideeffects.Thisstudywas performedtoexploretheantiemeticeffectsandtheimprovementeffectongastrointestinalfunctionofcomponentsofthreegingerjuiceexcipients. Methods: Thecompositionswereanalyzedbyliquidchromatographmassspectrometer(LC‐MS),especiallythegingerolsofdriedgingerjuice(DGJ),freshgingerjuice (FGJ),andfreshgingerboiledjuice(FGBJ).Furthermore,therespectivegastrointestinaleffectsonratmodelswithfunctionaldyspepsia(FD)werecompared. Results: The6‐keto‐PGF1α levelsintheserumofthetreatedgroupsweresignificantlyreduced(p < 0.05),ascomparedwiththecontrolgroup.Comparedwiththe cisplatingroup,therewasanapparentreductioninkaolinintakeforDGJ,FGJ,and FGBJ(p < 0.01; p < 0.01; p < 0.05).Theintestinalpropulsiverateoftheratsinthe treatedgroupwassignificantlyhigherthanthatinthecontrolgroup(p < 0.05).Ginger juicessignificantlyimprovedgastrointestinalfunctioninrats.EightcommoncomponentswerefoundinDGJ,FGJ,andFGBJ,amongwhich6‐paradol,10‐gingerol,and 12‐shogaolledtoinhibitedgastricmucosaldamagefunctioneffectaccordingtothe Pearsoncorrelationanalysis.Only6‐shogaolwasfoundtohaveapositivecorrelation withgastrointestinalfunctioneffectthroughPearsoncorrelationanalysis. Conclusion: Gingerjuiceshouldberecommendedforthemedicinalmaterialsusedin thetreatmentofconcurrentsymptomsofFD.

K E Y W O R D S

driedgingerjuice,freshgingerboiledjuice,freshgingerjuice,gastrointestinaleffects,gingerol

GingerrhizomeofZingiber officinale Roscoe,awidelyusedherbalmedicine,ispungentandwarmaccordingtotraditionalChinesemedicine

(TCM)theory.Thechemicalcompositionofbiologicalpropertiesof gingerprimarilyincludesnonvolatilepungentcompounds,6‐,8‐,10‐ gingerolsand6‐,8‐,10‐shogaols(Ho&Chang,2018).Gingerhasbeen knowntopreventorarrestnauseaandvomiting,diastoleandprotect

ThisisanopenaccessarticleunderthetermsoftheCreativeCommonsAttributionLicense,whichpermitsuse,distributionandreproductioninanymedium, providedtheoriginalworkisproperlycited. ©2019TheAuthors.Food Science & NutritionpublishedbyWileyPeriodicals,Inc.

Food Sci Nutr. 2019;7:2205–2213. www.foodscience-nutrition.com | 2205

thecoronaryartery,inhibitthecontractionofsmallintestineandmuch more(Chatturong,Kajsongkram,Tunsophon,Chanasong,&Chootip, 2018;Thamlikitkuletal.,2017;Wuetal.,2018).InTCM,gingerisregularlyusedasaMinisterialdrugtopotentiatetheMonarchdruginthe treatmentofthemaindiseaseortotreattheconcurrentsymptoms.

Gingerolsandshogaolsareactiveconstituentsofginger,which aredifficulttopurifyduetoitsgreatvarietyandstructuralsimilarity.Thus,itisnecessarytodeveloparapidanalysisandidentificationmethodforgingeranditssubtypes.Analysismethodshave been developed, including nuclear magnetic resonance (NMR), high‐performanceliquidchromatography(HPLC),andultrahigh‐performanceliquidchromatography–massspectrometry(UHPLC‐MS) (Ghasemzadeh, Jaafar, Baghdadi, & Tayebi‐Meigooni, 2018; Li et al.,2018;Park&Jung,2016;Raietal.,2015).However,theHPLC technologyneedstobeappliedincombinationwiththestandard forqualitativeanalysisofunknowncompounds.GC‐MShasbeen widelyappliedingingeranalysis,butgingerolishardtobedetected becauseofitslowvolatility.HPLC‐MScouldintegratethestrong separation ability of liquid chromatography with high sensitivity andhighselectivity,providingstructuralinformation.Inparticular, high‐resolutiontime‐of‐flightmassspectrometrycanprovideaccuratemolecularweightinformationanddeducetheformulaofthe specificcompound.Park&Jungseparatedandquantified8major gingerolsandshogaolsfromgingerusingtheultrahigh‐performance liquid‐phasechromatography–electrosprayionization–tandemmass spectrometry(HPLC‐ESI‐MS/MS)technology(Park&Jung,2016).

NumerousTCMscanpreventgastricmucosaldamage,vomiting, andgastrointestinalmovement.Theinhibitioneffectsofaqueous extractfromArtemisiacapillariswereinvestigatedintermsofROS andNF‐kBofacutegastricmucosalinjuryinducedbyethanol(Yeo, Hwang,Kim,Youn,&Lee,2018).Strikingly,increased6‐keto‐PGF1α waspreviouslydemonstratedtoreducethedegreeofgastricmucosalinjury(Kouetal.,2018).Thegingerextractcanpreventthe picabehaviorofratsinducedbycisplatin(Huetal.,2016).Tangweian decoction,aChineseherbalmedicine,cansignificantlypromoteintestinalpropulsionrateindiabeticmice(Tianetal.,2017).

BasedontheChinesePharmacopoeia(2015version),thereare twosubtypesofginger,namelyfreshgingeranddriedginger.Both formscanbeusedtoobtaingingerjuice.AccordingtotheNational ProcessingStandardofTraditionalChineseMedicine—1988,ginger includesfreshgingerjuice(FGJ),freshgingerboiledjuice(FGBJ), and dried ginger juice (DGJ). However, no clear distinction was identifiedamongthegingerjuices.Thesegingerjuicesareoften interchangeablewhenpreparingaTCMformulation.Inthecurrent study, we aimed to analyze the nonvolatile components in FGJ, FGBJ,andDGJbyLC‐MS.Aftertheratswereadministeredwith gingerjuices,theethanol‐inducedgastricmucosaldamageindex andthelevelsofIL‐8,TNFα,6‐keto‐PGF1α,cisplatin‐inducedemesis,andtheintestinalpropulsionrateweredeterminedtoevaluate the effect on gastroenteric motivity. The chemical compositions of these three ginger juices were compared to determine which oneisthemosteffectiveinthepreparationofChinesetraditional medicine.

2 | MATERIALS AND METHODS

2.1 | Materials

A liquid chromatography coupled with a mass selective detector wasemployedtoanalyzethenonvolatilecomponents.Inaddition, ahigh‐speedrefrigeratedcentrifugeandfreeinstrumentwerealso used.Allinvolvedchemicalswereofanalyticalgrade,includinganhydrousethanol,petroleumether,andchloroform,whichwereseparatelypurchasedfromXilongChemicalCo.,Ltd,andTianjinDamao ChemicalReagentFactory.Nutritionalsemi‐pasteswerealsosupplied.Kaolinwaspreparedaspreviouslydescribed.Briefly,kaolin andarabicgumweremixedataratioof99:1,followedbytheadditionofdistilledwater(volume)toformathickpaste.Thepaste wasfurthercutintopiecesresemblingregularratchowpellets.After beingcut,thepelletsweredriedatroomtemperaturefor48hr(Han etal.,2014).

Freshforms(FoshanCity,GuangdongProvince)anddriedforms of ginger(QianweiCity,SichuangProvince)werepurchasedfrom the Tian Qi Hall medicine material crude slices limited company (ZhangshuCity,JiangxiProvince,China).

Theaforementioneddriedgingerwaspreparedforusebycuttingintopieces(0.3cm×0.3cm×0.3cm),followedbybeingbaked at55°Cfor18hr.

MaleSpragueDawley(SD)rats(weighing180and220g)were purchasedfromJi'nanPengYueExperimentalAnimalBreedingCo., Ltd(CertificateNo.SCXK(Lu)2014‐0007).Additionally,adultmale WistarratsandSpragueDawleyrats(ShanDong,CertificateNo. SCXK20130001),weighingbetween200and230g,wereincluded inthecurrentstudy.Allanimalexperimentswerecarriedoutin accordancewithguidelinesevaluatedandapprovedbytheEthics CommitteeofJiangxiUniversityofTraditionalChineseMedicine.

Firstly, 150g of fresh ginger was washed and squeezed as juice. Afterfilteringthejuice,theobtainedresidueswereresuspendedin theappropriateamountofwaterandsqueezedagain.Subsequently, theobtainedfiltrateswerecombined.

Next,wesoaked150goffreshgingerand50gofdriedginger,respectively,in1Lwaterfor1hr.Afterboilingfor30minandfiltration,theobtainedresidueswereresuspendedinwaterandboiled foranother30min.Thefiltratewascombinedandconcentratedto 150ml.Thedosageratiooffreshanddriedgingeris3:1.

Sampleswerepreparedbydissolving10mlofgingerjuicein20ml ofmethanol.Next,thesamplesunderwentultrasonicfor10minto breakthecells.Thesolutionswerefiltered,andthefiltratewasfurtherstrainedthrougha0.22‐μmMilliporefilter.

1. 2.2 | Methods

1. 2.2.1 | LC‐MS analytical condition
2. 2.2.2 | Grouping and administration Ethanol‐induced gastric mucosal damage Theratswererandomlydividedintofivegroups:thenormalgroup, the model group, the FGJ group, the FGBJ group, and the DGJ group.FGJ,FGBJ,andDGJwereadministeredtoratsonceaday foradurationof7daysexcludingtheratsinthenormalandmodel groups,whichweregiven0.9%physiologicalsalineinsteadofFGJ, FGBJ,orDGJ.Alltheratsexceptthoseinthenormalgroupwere given 1ml absolute ethanol 1hr after the last administration of drugs.Subsequently,allanimalswereanesthetized.Next,thestomachswereremovedandopenedalongthelongcurvaturetoobserve thelesionsmacroscopically1hrafterbeinginjectedwith6mlof 4%paraformaldehyde.Scoringstandardwasasfollows:Intactgastricmucosawasscoredas0;point‐shapedbleeding,point‐shaped erosion, and strip‐shaped lesion of no more than 1mm were all scored as 1; strip‐shaped lesion with more than 1mm was doublyscored.Totalscoresofeveryratwereconsideredasthelesion index. Inhibition ratio of lesion formation (%)=(A−B)/A×100% (A,Bwerethelesionindicesofthemodelgroupanddrug‐treated group, respectively). Gastric tissues in the same location were fixed with 4% formalin, embedded with paraffin, and stained by HE.Inaddition,bloodsampleswereobtainedfromtheabdominal aorta.IL‐8,TNFα,and6‐keto‐PGF1αinplasmawereexaminedby radioimmunoassay.

After acclimatization to the laboratory environment for 3days, all animals were randomly divided into five groups, namely control, cisplatin,FGJ,FGBJ,andDGJgroups.Themiceinthecontrolgroup wereadministeredwithperitonealinjectionsandgastriclavagewith normalsaline.Theanimalsinthecisplatingroupreceivedperitoneal injectionsofcisplatin(5mg/kg)andgastriclavagewithnormalsaline,whileanimalsinthethalidomideandgranisetrongroupsreceived peritonealinjectionsofcisplatin(5mg/kg)andgastriclavagewith,respectively,FGJ(10ml/kg),FGBJ(10ml/kg),orDGJ(10ml/kg).

Miceweredividedinto4groups,namelythenormal,FGJ,FGBJand DGJgroups.Themicewerefastenedfor12hrpriortotheexperiment.Fifteenminutesafterthelastadministrationofdrugs,themice weregivensemi‐solidnutritioninavolumeof1ml.Theanimalswere thensacrificedbycervicaldislocation20minafterthemeal,andthe wholegastrointestinaltractwasremoved.Theentirelengthofthe intestineswasstretchedoutonpaper,andthedistancetheinkhad travelledfromthepyloruswasexpressedasthepercentageofthe totallengthofsmallintestinebetweenpylorusandcecum.Atotalof 10animalswereusedforeachdoseortreatment,respectively.

Allexperimentswereperformedatleastsixtimes.Theresultswere presentedasmeanvalue±standarderror(SE).Statisticalanalyses wereperformedusingtheSPSS19softwarewithindependent‐samplesttest(ref).Avalueofp < 0.05wasconsideredtobestatistically significant.Pearsoncorrelationanalysiswasappliedfortherelative analysisofcommoningredientsinallgingerjuicesandtheirpharmacodynamicsindicesusingtheSPSS19software.

3 | RESULTS

Typicaltotalionchromatograms(TICs)ofthenonvolatilecomponent fractionextractedfromFGJ,FGBJ,andDGJareshowninFigure1. AlthoughtheTICswerecomplicated,mostchromatographicpeaks werenotedtobewellseparated.PositiveionmodetotalionchromatogramswereanalyzedaccordingtoPeakviewTM1.7.viamethods oftargetscreeningandnontargetscreening.Thecommonchemical compositions of FGJ, FGBJ, and DGJ were initially identified (Table1)(Denniff,Macleod,&Whiting,1981;Hu,Guo,etal.,2011; Hu,Rayner,etal.,2011;Jiang,Sólyom,Timmermann,&Gang,2010; Kikuzaki,Kawasaki,&Nakatani,1994;Zhan,Wang,Xu,&Yin,2008; Zhang,Gan,&Hong,2005).

Evidentlargebandlikehemorrhagicerosionsintheglandularstomachwerenotedinthemodegroup(Figures2and3,Table2).In the FGJ, FGBJ, and DGJ groups, gastric mucosal damages were preventedinadose‐dependentmanner,whencomparedwiththe modelgroup.

Comparedwiththecontrolgroup,thegastricmucosaofrats inthemodelgroupwasfoundtobesignificantlydamaged.Inaddition, shedding, bleeding, and infiltration of inflammatory cells were apparent in the mucosa. Relative to the model group, the gastric mucosa was observed to be lighter and presented with lessinflammatorycellinfiltrationinthegingerboiledjuicegroup and ginger juice group, while slightly alleviated gastric mucosal

1. (a)
2. (b)
3. (c)

1. FIGURE 1 LC‐MSTIC(FGJ‐A,FGBJ‐B,DGJ‐C)

lesionandinflammatorycellinfiltrationwerenotedintheginger boiled juice group. However, the gastric was severely damaged inthedriedgingerboiledjuicegroupandgingerjuicegroup.The aforementionedresultsindicatethatgingerjuicecaninhibitthe inflammationofgastricmucosaandcanfurthermorepromotethe repairingprocessofgastricmucosa.

The ulcer index (UI) of the groups following administration wasfoundtobesignificantlylowerthanthatinthemodelgroup (p < 0.01).ComparedwiththeFGBJgroup,asignificantreduction intheratewasrevealedintheFGJandDGJgroups(p < 0.05).The significantreductionssuggestthatadministrationofgingerjuiceled todecreasedintestinalpropulsiverate(p < 0.05)(Tables2and3).

AsshowninTable3,themodelgroupshowednosignificantdifferencesrelativetothecontrolgroupinthecontentofIL‐8(p > 0.05).

ThecontentofTNFα waselevated,buttherewerenosignificant differences within any groups. And the content of 6‐keto‐PGF1α inserumsignificantlyincreasedcomparedwiththatinthecontrol group (p < 0.05).Thisdemonstratesthattheacuteinjurystimulates thebody'sinflammatoryresponse.Thecontentof6‐keto‐PGF1α in serumofginger‐treatedgroupwassignificantlydecreasedcompared withthemodelgroup(p < 0.01,p < 0.05).Furthermore,thecontent ofFGJgroupwaslowerthantheFGBJgroup,inwhichithasbasicallyreturnedtonormallevels.

Twenty‐fourhoursafteradministeringperitonealinjectionsofcisplatin, kaolin intake was shown to be significantly increased in

1. TABLE 1 PossiblecommonchemicalcompositionofthesamplesbyLC‐MS

N Name TR/min Formula MS Error (ppm)

Area% DGJ FGJ FGBJ

1. 1 4‐shogaol 27.01 C15H20O3 249.1484[M+H]+ −0.5 1.75 0.89 1.83
2. 2 6‐gingerdiol 40.84 C17H28O4 317.1717[M+Na]+ −1.9 1.62 1.60 5.27
3. 3 6‐gingerol 44.56 C17H26O4 295.1896[M+H]+ −1.9 7.54 19.59 13.83 317.1717[M+Na]+ −2.7
4. 4 6‐shogaol 44.57 C17H24O3 277.1792[M+H]+ −2.2 0.90 0.57 1.24
5. 5 methyl‐6‐gingerol 50.61 C18H28O4 331.1875[M+Na]+ −1.3 0.82 1.61 1.91
6. 6 3‐or5‐acetoxy‐6‐gingerdiol 51.81 C19H30O5 361.1982[M+Na]+ −1.1 0.63 0.65 0.89
7. 7 diacetoxy‐4‐gingerdiol 52.69 C19H28O6 375.1771[M+Na]+ −1.9 0.29 0.21 0.45
8. 8 8‐shogaol 54.13 C19H28O3 305.2105[M+H]+ −2.2 1.59 4.01 0
9. 9 8‐gingerol 54.13 C19H30O4 345.2033[M+Na]+ −1 1.59 5.34 2.65
10. 10 methyl‐3‐or
11. 11 10-gingerdione 55.67 C21H32O4 371.2184[M+Na]+ −2.5 0.08 0 0
12. 12 6‐paradol 56.1 C17H26O3 301.1771[M+H]+ −1 0 4.27 0
13. 13 1‐dehydro‐6‐gingerdione 56.33 C17H22O4 313.1408[M+Na]+ −0.9 0 0.12 0.52
14. 14 acetoxy‐8‐gingerol 56.94 C21H32O5 387.2124[M+Na]+ −4.7 0.13 3.82 0
15. 15 10‐gingerol 56.96 C21H34O4 373.234[M+Na]+ −2.5 0.13 3.82 0
16. 16 acetoxy‐10‐gingerol 58.99 C23H36O5 415.2441[M+Na]+ −3.3 0 1.50 0
17. 17 12‐shogaol 59.12 C23H36O3 361.2727[M+H]+ −3 0 0.87 0
18. 18 10‐shogaol 59.65 C21H32O3 355.2228[M+Na]+ −4.4 0.14 0.72 0
19. 19 1‐dehydro‐10‐gingerdione 60.51 C21H30O4 347.2204[M+H]+ −3.8 0 1.15 0 369.2019[M+Na]+ −4.6

Note.DGJ:driedgingerjuice;FGJ:freshgingerjuice;FGBJ:freshgingerboiledjuice;LC‐MS:LiquidChromatograph‐MassSpectrometer.

1. FIGURE 2 Protectionofgingerjuicesongastricmucosalinjuryinducedbyethanolinrats.(a)Normalgroup;(b)Modelgroup;(c)FGBJ group;(d)FGJgroup;(e)DGJgroup
2. FIGURE 3 Gastrictissues.(a)Normalgroup;(b)Modelgroup;(c)FGBJgroup;(d)FGJgroup;(e)DGJgroup

1. TABLE 2 Effectofethylacetateextractfromgingerjuiceon gastriclesionsinducedbyethanolinrats

Groups Number UI Normal 6 – Model 6 64.17±6.49 FGBJ 6 48.83±3.31* FGJ 6 27.33±2.80*,† DGJ 6 26.17±3.60*,†

Note.DGJ:driedgingerjuice;FGJ:freshgingerjuice;FGBJ:freshginger boiledjuice;UI:ulcerindex.

*p < 0.01versusmodelgroup;†p < 0.05versusFGBJgroup.

TABLE 3 EffectofgingerjuicesonIL‐8,TNFα,and6‐keto‐PGF1α activityofgastricmucosadamagedbyethanolinrats

Groups IL‐8 (pg/ml) TNFα (pg/ml) 6‐keto‐PGF1α (pg/ml) Normal 47.21±6.73 57.70±7.77 82.42±7.34 Model 47.01±2.34 63.90±6.42 95.67±7.95* FGJ 46.27±5.22 55.75±8.14 81.89±5.16‡ FGBJ 46.40±5.05 61.35±5.87 83.58±10.318† DGJ 47.11±2.27 67.04±6.27 84.55±8.00†

Note.DGJ:driedgingerjuice;FGJ:freshgingerjuice;FGBJ:freshginger boiledjuice.

*p < 0.01versusnormalgroup;†p < 0.05versusmodelgroup;‡p < 0.01 versusmodelgroup.

thecisplatinandadministeredgroups(p < 0.01,vs.controlgroup; Table4).However,thedifferenceinkaolinintakebetweentheFGJ groupandDGJgroupreflectednostatisticallysignificantdifference. After24hr,graduallydecreasingkaolinintakewasobservedinthe administeredandcisplatingroups.

TheresultsshowedthatFGBJ,FGJ,andDGJgroupsweresignificantlydifferentfromthenormalgroup(p < 0.01).Comparedwith theFGBJgroup,theFGJgroupwassignificantlyreduced(p < 0.05). Boiledgingerjuiceexhibitedthestrongesteffectongastrointestinal propulsion,followedbygingerboiledjuiceandgingerjuice.Among these,FGBJpresentedwiththestrongesteffectonintestinepropulsion(asshowninTable5).

accountedfor10%,45%,and45%,respectively,forthetotalscores ofthegastricmucosaldamagefunction.Kaolinintakeandintestinal propulsiverateaccountedfor50%and50%,respectively,forthe totalscoresofthegastrointestinalfunction.

Theingredients,including4‐shogaol,6‐gingerol,6‐shogaol,8‐ gingerol,6‐paradol,1‐dehydro‐6‐gingerdione,10‐gingerol,and12‐ shogaol,wereanalyzedaccordingly(Table6).Theresultsrevealed that6‐shogaol,6‐paradol,10‐gingerol,and12‐shogaolshowedsignificantpositivecorrelationwithgastrointestinalfunctionandinhibitedgastricmucosaldamagefunction(p < 0.05)(Table7).

TheDGJgroupwiththelowestUIwasindexedas100points.The FGJ group with the lowest concentration of TNFα in serum was taken as 100 points. The FGJ group with the lowest concentrationof6‐keto‐PGF1αinserumwastakenas100points.TheFGBJ groupwiththelowestkaolinintakewasalsoindexedas100points. TheFGBJgroupwiththehighestintestinalpropulsiveratewasindexedas100points.Inaddition,theaveragevaluesoftheother groupsweresequentiallycalculated.UI,TNFα,and6‐keto‐PGF1α

TABLE 4 Changesinkaolinintake

Kaolin 24 hr 48 hr

Groups

Normal 0.1731±0.028 0.1556±0.058 Cisplatin 3.66±0.75 0.54±0.18 FGJ 2.86±0.68* 0.50±0.14 FGBJ 1.58±0.22* 0.40±0.29 DGJ 2.23±0.78* 0.77±0.21

Note.DGJ:driedgingerjuice;FGJ:freshgingerjuice;FGBJ:freshginger boiledjuice.

*p < 0.01versuscisplatingroup.

Thechiefpungentcompoundofgingerisgingerol.Gingerols,unstableinheat,arepronetodehydrationreactionandproducingthe correspondingshogaols(Jungetal.,2017),whichexertantioxidant, anti‐inflammatory,antiallergic,anticancer,andantimicrobialeffects (Semwal,Semwal,Combrinck,&Viljoen,2015).

Interestingly,thegastrointestinaleffectisoneofitsmajorfunctions.Inthecurrentstudy,weanalyzedgingerolsbyLC‐MSandtheir gastrointestinaleffects.Thegingerolschangedingingerjuiceswhatevermaybetheircomposition,andthegastrointestinaleffectsof gingerjuiceswereextremelydifferent.Atotalof8gingerolswere foundinDGJ,FGJ,andFGBJ,amongwhichthree,namely6‐paradol,10‐gingerol,and12‐shogaol,sharedapositivecorrelationwith

TABLE 5 Intestinalpropulsiverateofgroups

Intestinal propulsive distance (cm)

Intestinal propulsive rat%

Group n

Normal 10 15.19±1.09 28.48±2.48 FGBJ 10 24.59±2.18 52.01±3.18* FGJ 10 23.25±1.98 48.15±3.50* DGJ 10 26.07±2.54 50.66±4.14*

Note.DGJ:driedgingerjuice;FGJ:freshgingerjuice;FGBJ:freshginger boiledjuice.

*p < 0.01versusnormalgroup.

1. TABLE 6 Commoningredients(relativecontents)X1 to X8andgastrointestinalcomprehensivevalueY
2. TABLE 7 Pearsoncorrelationanalysis

X1 X2 X3 X4 X5 X6 X7 X8 Y1 Pearson −0.983 0.905 −0.801 0.986 0.994 0.994 −0.191 0.990 1

Y Relevance 0.059 0.140 0.204 0.054 0.035 0.035 0.439 0.044 1 Significant * * * N 3 3 3 3 3 3 3 3

X1 X2 X3 X4 X5 X6 X7 X8 Y2 Pearson 0.955 −0.600 0.988 −0.793 −0.929 −0.929 0.628 −0.940 1

Y Relevance 0.096 0.295 0.049 0.209 0.120 0.120 0.284 0.111 1 Significant * N 3 3 3 3 3 3 3 3

*Correlationissignificantatthe0.05level(1‐tailed).

theinhibitoryeffectsongastricmucosaldamage,onthebasisofa Pearsoncorrelationanalysis.Notably,only6‐shogaolpresentedwith a positive correlation with the gastrointestinal effect. Among the fourcompounds,6‐paradoland12‐shogaolwerepresentinFGJonly. 10‐gingerolwasfoundinDGJandFGJ.6‐shogaolwasfoundinDGJ, FGJ,andFGBJ.Importantly,thecontentof6‐shogaolineachginger juicesfromhightolowwasFGBJ,DGJ,andFGJ.The6‐shogaolof boiledgingerjuiceishigherthanthatofunboiledgingerjuice,itmay beattributedtothatgingerolinboiledgingerjuiceistransformedinto shogaolafterheating,andthecontentofgingerolsinfreshgingermay behigherthanthatinfriedginger.

Prostaglandinsarerecognizedasdefensiverepairfactorsofthe gastricmucosa.6‐keto‐PGF1αisastablemetaboliteofPG,andthe contentof6‐keto‐PGF1αcanbeusedtoreflectthecontentofPG inplasma.Thecurrentstudyfoundthatallgingerjuicescansignificantlyreducethecontentof6‐keto‐PGF1αinserum.Furthermore, 6‐paradol,10‐gingerol,and12‐shogaolwerenotdetectedinFGBJ, whichwasconsistentwiththeUIresultsofgastricmucosalinjury inducedbyethanol,revealingFGJandDGJhavesignificanttherapeuticeffectscomparedwithFGBJ.After24hr,kaolinintakeinthe FGBJ,DGJ,andFGJgroupswasgraduallydecreasedandshowedno statisticallysignificantdifference,suggestingthatallkindsofgingers canpreventvomitingeffectively.Inaddition,allgingerscanpromote intestinepropulsion,andFGBJexhibitsthestrongesteffect.

6‐Shogaoland6‐paradolarecharacterizedbyantimicrobialactivity(Galal,2008),whichmaybehelpfulforinhibitingbacterialinfectionintheprocessofgastricmucosalinjury.Additionally,6‐shogaol and 10‐gingerol have effective antioxidant and anti‐inflammatory properties(Dugasanietal.,2010),whichcanpreventoxygenfree radicalsfromactingonsulfenyltodegenerateproteinsandinactivate

enzymes, thus alleviating gastric mucosal damage. Moreover, the aforementioned antioxidant and anti‐inflammatory activities can ameliorategastrointestinalinjurybysuppressingtheinflammatory cascadereaction(Zhang,Ma,Gao,Sun,&Zhang,2017).Currently, there are almost no specific pharmacological researches on 12‐ shogaol.However,accordingtotheresultofthecurrentstudy,we inferthatitspharmacologicaleffectsarerelatedtositeslocalized inthedigestivetractandgastrointestinalinjurytoacertainextent. Besides,6‐shagoalmaybethemostcloselyrelatedsubstancetogastrointestinalfunction.

In conclusion, our findings demonstrate that when the ginger juiceFGJorDGJcouldbeusedfortheinhibitionofgastricmucosalinjury.Additionally,FGBJmaybethebestchoiceforpromoting gastrointestinalpropulsionandallkindsofgingersaresuitablefor preventing vomiting.

Overall,evidencehasbeenpresenteddemonstratingabasicun-

derstandingofdifferentgingerjuicesandtheirtherapeuticfeatures. Furtherstudiesshouldaimtoexplorethecorrelationsbetweenthe componentsofgingerjuicesandotherpharmacodynamicindicesto revealtheentireeffectsofgingerjuicesfortherapeuticregimens.

Onbehalfof,andhavingobtainedpermissionfromalltheauthors, theauthordeclaresthat:

1. • Thestudy'sprotocolsandprocedureswereethicallyreviewedand approvedbyEthicsCommitteeonLaboratoryAnimalsofJiangxi UniversityofTraditionalChineseMedicine.Wecompliancewith LaboratoryAnimal‐Guidelineforethicalreviewofanimalwelfare andLaboratoryAnimal‐GeneralrequirementsforanimalexperimentofStateStandardofthePeople'sRepublicofChina.
2. • Humantestingisunnecessaryinourstudy

CONFLICT OF INTEREST Theauthorsdeclarethattheyhavenoconflictofinterest.

ORCID Heng‐li Tong https://orcid.org/0000‐0002‐0456‐0000