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Comp ehensi e quali a i e and quan i a i e analysis o
la onoids in dandelion (Ta axacum o icinale) lowe s and
ood p oduc s
Jou nal:
Jou nal o Ag icul u al and Food Chemis y
Manusc ip ID
D a
Manusc ip Type:
A icle
Da e Submi ed by he
Au ho :
n/a
Comple e Lis o Au ho s:
Jed ejek, Da iusz; Ins y u Up awy Nawozenia i Gleboznaws wa,
Depa men o Biochemis y and C op Quali y
Pawelec, Sylwia; Ins y u Up awy Nawozenia i Gleboznaws wa,
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1Comp ehensi e quali a i e and quan i a i e analysis
2o la onoids in dandelion (Ta axacum o icinale)
3 lowe s and ood p oduc s
4Da iusz Jed ejek,* Sylwia Pawelec
5
6†Depa men o Biochemis y and C op Quali y, Ins i u e o Soil Science and Plan Cul i a ion, S a e
7Resea ch Ins i u e, Cza o yskich 8 S ., 24-100 Puławy, Poland
8
9*Co esponding au ho : E-mail: [email p o ec ed]; Phone: 0048 814786886
10
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12 ABSTRACT
13 Ta axacum o icinale, a cosmopoli an pe ennial, is widely used as edible, o age,
14 honey and medicinal plan . Unsu p isingly, de ailed LC-QTOF-MS p o iling o lowe
15 ex ac e ealed he p esence o se e al la onoid signals mos likely lacking li e a u e
16 con i ma ion. Ta ge ed isola ion, including DoE planning based ex ac ion, led o he
17 isola ion o 11 la onoids, o which h ee we e new (16, 18 and 33). Thei s uc u es
18 we e es ablished by he use o NMR echnique. Bo h bi la ones (16 and 18) we e
19 s uc u ed om wo lu eolin molecules, linked by a C-C bond h ough IB (C-6′) and IIA
20 (C-6″ o C-8″) ings. No el la onolignan (33) composed o icin and unusual,
21 con aining a ca boxyl unc ion, lignan moie y. The con en s o 28 la onoids
22 (glycosides, aglycones, bi la ones and la onolignans) was de e mined, based on LC-
23 MS/MS me hod, in aw lowe s and dandelion sy ups and inc u e. Mul i a ia e
24 analyses showed he sepa a ion o sp ing and au umn lowe s and a high di e si y o
25 ood p oduc s, and helped iden i y me aboli es co ela ed wi h he samples.
26
27 KEYWORDS: dandelion, la onoids, bi la ones, la onolignans, NMR, UHPLC-MS/MS
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29 INTRODUCTION
30 The genus Ta axacum, commonly known as dandelion, is one o he be e -known and
31 commonly used by humans ep esen a i es o As e aceae plan amily. Ta axacum
32 o icinale F.H.Wigg. (common dandelion) is a yellow- lowe ing he baceous plan na i e
33 o empe a e clima es, usually sp eading in pas u es and was eland.1 O e he
34 cen u ies, common dandelion and o he local Ta axacum species (e.g. T. mongolicum
35 and T. co eanum) ha e been used in he Eu opean, Chinese and No h Ame ican olk
36 medicines as a emedy in he ea men o a ious ailmen s, such as li e and diges i e
37 diso de s, diabe es, skin in lamma ions, a h i ic and heuma ic diseases.2 Nowadays,
38 he whole plan (including oo , lea es, and lowe s) has “gene ally ecognized as sa e”
39 s a us app o ed by U.S. Food and D ug Adminis a ion, and is commonly p ocessed
40 in o di e en edible p oduc s – salads, eas, inc u es, sy ups, jellies, cakes, wines,
41 honeys and co ee subs i u es.3 In addi ion, T. o icinale he b is a Pha macopeial
42 ma e ial ha is app o ed o use as an he bal supplemen , in he o m o inc u e and
43 ea p epa a ions, by Eu opean Medicines Agency.4,5
44 Ta axacum he b ex ac s (wa e and alcoholic) ha e been shown o exe a wide ange
45 o in i o and in i o biological ac ions, such as an ioxidan , an i-in lamma o y,
46 an ica cinogenic, an i-hype glycemic, an i- h ombo ic, an imic obial, an i i al, and
47 hepa op o ec i e, in a numbe o scien i ic s udies.5–11 Polyphenols, mainly
48 hyd oxycinnamic acids and la onoids, as well as e penoids and sesqui e pene
49 lac ones ha e been ound as majo bioac i e me aboli es o dandelion plan
50 ma e ial.12,13 Albei , despi e he gene ally well- ecognized chemical p o ile o dandelion
51 he b and lowe s, new me aboli es (o me aboli es no ye epo ed in he axon) a e
52 occasionally documen ed, as exempli ied by he ecen ly cha ac e ized la onoids o
53 T. mongolicum o T. o icinale ui s.14,15 In addi ion, despi e he ai ly widesp ead use
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54 o dandelion he b, he amoun o de ailed esea ch de o ed o he quan i a i e con en
55 o bioac i e me aboli es (beyond colo ime ic measu emen o o al polyphenols and
56 la onoids) seems o be negligible and insu icien .15–20 I should be no ed ha he as
57 majo i y o quan i a i e wo ks on dandelion he b la onoids include only a ew majo
58 componen s (such as que ce in and lu eolin and hei glycosides) in a single
59 analysis.16–20
60 The wo k is phy ochemically explo a o y, wi h se e al speci ic objec i es, including
61 de ailed HR-QTOF-MS quali a i e analysis o Ta axaci Flos, a ge ed isola ion and
62 iden i ica ion o la onoid oligome s and la onolignans, and measu emen o he
63 quan i a i e con en o nea ly 30 la onoid compounds in aw plan ma e ial (sp ing and
64 au umn lowe s) and ood p oduc s based on dandelion lowe s (sy ups and inc u e).
65
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66 MATERIALS AND METHODS
67 Plan Ma e ial. Dandelion lowe s we e collec ed om a na u al si e in sou heas e n
68 Poland (Kazimie ski Landscape Pa k, 51.378605 N, 22.072647 E) in 2021 (Oc obe ),
69 2022 (Ap il and Oc obe ) and 2023 (Ap il). Plan ma e ial was p ese ed by eeze-
70 d ying, hen was inely g ound wi h elec ic mill and sie ed (1 mm), and inally s o ed
71 a 4 °C un il use. A ouche specimens a e deposi ed in ou labo a o y. In addi ion,
72 h ee di e en comme cially a ailable dandelion lowe -based sy ups (sy up I (48%
73 lowe decoc ion), II (60% lowe decoc ion) and III (40% lowe decoc ion), pu chased
74 on he Polish ma ke in May 2023) and a handmade e hanol lowe inc u e ( o mula ion
75 acco ding o he medicinal plan ex books21–23) we e used in he s udy. De ailed
76 in o ma ion on sy up composi ions and a desc ip ion o he inc u e p epa a ion a e
77 gi en in he Supplemen a y Ma e ials (Desc ip ions S1-S2).
78 Ins umen s and Ma e ials. 1H- and 13C-NMR spec a (including one- and wo-
79 dimensional) we e eco ded on a B uke A ance III HD Ascend-500 spec ome e
80 (B uke BioSpin, Rheins e en, Ge many), equipped wi h 3 mm b oad-band in e se
81 (BBI) p obe. HRESIMS spec a, exac masses o compounds, MS/MS agmen a ion
82 pa e ns, and molecula o mulas we e ob ained using a B uke Impac II HD (B uke ,
83 Bille ica, MA, USA) quad upole ime-o - ligh (Q-TOF) mass spec ome e coupled wi h
84 a The mo Scien i ic Ul ima e 3000 RS ch oma og aphic sys em. HPLC sepa a ions
85 owa ds ex ac ac iona ion and compounds isola ion we e pe o med wi h a Dionex
86 (Sunny ale, CA, USA) sys em equipped wi h a pho odiode a ay de ec o PDA-100
87 and ac ion collec o FC 204 (Gilson, Middle on, WI, USA) using a Sephadex LH-20
88 (Me ck, Wa saw, Poland) column (80 × 2.8 cm i.d.) and e e sed-phase (RP) HPLC
89 columns (A lan is T3, 250 × 19 mm, 10 µm, and 250 × 10 mm, 5 µm, Wa e s, Mil o d,
90 MA, USA; and Cosmosil πNAP, 250 × 10 mm, 5 µm, Nacalai Tesque Inc., Kyo o,
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91 Japan). Quan i a i e UHPLC analyses we e pe o med using a Wa e s ACQUITY
92 ch oma og aphic sys em equipped wi h PDA and Xe o TQ-S mic o T iple Quad upole
93 (TQD) mass de ec o and RP column (BEH C18, 100 × 2.1 mm, 1.7 µm, Wa e s). Plan
94 ma e ial, ex ac s and isola ed compounds we e d ied using Gamma 2–16 LSC eeze
95 d ye (Ch is , Os e ode am Ha z, Ge many).
96 Ex ac ion and Isola ion o Fla onoids. The powde ed plan ma e ial (750 g) was
97 de a ed wi h n-hexane (6 L) o e 10 h using Soxle appa a us. The esidue (600 g)
98 was wice ex ac ed wi h 90% me hanol (MeOH) (2 × 6 L) a 65 °C and 180 pm o 55
99 min (Inno a 44R shake , Eppendo , Hambu g, Ge many), based on he op imiza ion
100 esul s o Design o Expe imen (DoE) design (Desc ip ion S3, Tables S1-S4, Fig. S1-
101 S2). The aw ex ac (E1) was il e ed and concen a ed unde educed p essu e,
102 ollowed by liquid-liquid ex ac ion wi h e hyl ace a e (3 × 1 L). A e e apo a ion o he
103 sol en , he esidue was suspended in 50% -BuOH ( / ) and eeze-d ied, yielding
104 10.4 g o la onoid ex ac (E2). The E2 was u he ac iona ed on a Sephadex LH-20
105 (Me ck) column (100 × 2.8 cm), which was elu ed wi h 95% MeOH ( / ) a a low a e
106 o 2.4 mL/min. The sepa a ion was unde he con ol o HPLC-UV350nm de ec ion and
107 addi ionally moni o ed by LC-MS analyses. The se en pooled ac ions (A–G) we e
108 collec ed, concen a ed, and eeze-d ied o gi e 1.23 g o ac ion A, 0.95 g o ac ion
109 B, 1.21 g o ac ion C, 483 mg o ac ion D, 51 mg o ac ion E, 49 mg o ac ion F,
110 and 31 mg o ac ion G. F ac ions p omising wi h new me aboli es, including A, and
111 D-G, we e u he sepa a ed o indi idual compounds using e e se-phase HPLC
112 (Dionex HPLC-PDA sys em). F ac ion A was ch oma og aphed in 3 s eps using
113 A lan is T3 (250 × 19 mm, Wa e s; g adien 20–55% o ace oni ile (MeCN), low a e
114 7 mL/min, empe a u e 30 °C) and Cosmosil πNAP (Nacalai Tesque Inc.; g adien 25–
115 30% o MeCN, low a e 3.5 mL/min, empe a u e 30 °C; and g adien 60–64% o
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116 MeOH, low a e 3 mL/min, empe a u e 25 °C) columns. F ac ions D-G we e pu i ied
117 using A lan is T3 (250 × 10 mm, Wa e s; g adien 26–29% o MeCN, low a e 3
118 mL/min, empe a u e 25 °C) column. A o emen ioned ch oma og aphic me hods
119 esul ed in 11 pu e la onoid me aboli es, including compound 16 (17 mg) was
120 eco e ed om ac ion E, compound 18 (14 mg) om ac ion G, and compound 33 (9
121 mg) om ac ion A.
122 Liquid Ch oma og aphy High-Resolu ion Mass Spec ome ic Analysis. HR-
123 MS/MS da a we e ob ained using a B uke Impac II HD Q-TOF mass spec ome e
124 coupled wi h a Ul ima e 3000 RS (The mo Scien i ic) UHPLC sys em. The MS ope a ed
125 in elec osp ay ioniza ion and bo h pola i y modes, wi h he ollowing se ings: mass
126 scan ange 50–1400 m/z; capilla y ol age 4.0 kV (ESI+) o 3.0 kV (ESI–); nebulize
127 and d ying gas (N2) 2.0 ba and 10 L/min, espec i ely; d y gas empe a u e 220 °C.
128 The MS/MS spec a we e egis e ed using a collision ene gy (CE) o 50 eV wi h
129 s epping be ween 50–125% o CE. The acqui ed da a we e calib a ed in e nally wi h
130 sodium o ma e (10 mM solu ion in 50% 2-p opanol) which was injec ed o he ion
131 sou ce be o e he sample analysis. Da a we e p ocessed using Da aAnalysis 4.4
132 so wa e (B uke ). Ch oma og aphic sepa a ion was ca ied ou on an HSS C18
133 column (100 × 2.1 mm, 1.7 µm, Wa e s) equipped wi h a p e-column. A 28 min linea
134 g adien (10→35%) o ace oni ile–wa e (bo h acidi ied wi h 0.1% o mic acid), wi h a
135 low a e o 0.4 mL/min, was applied. The column was held a 45 °C. Injec ion olume
136 was 2 µL.
137 NMR Measu emen s. 1D and 2D NMR spec a we e eco ded on B uke A ance III
138 HD Ascend-500 spec ome e (B uke BioSpin, Rheins e en, Ge many) equipped wi h
139 5 mm b oad-band in e se (BBI) p obe. The 1H and 13C NMR spec a (a 500 and 125
140 MHz, espec i ely) we e measu ed in DMSO-d6 a 30 °C. Chemical shi s a e gi en on
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141 he δ scale. Da a p ocessing was pe o med wi h he Topspin so wa e ( e sion 3.5pl2,
142 B uke BioSpin).
143 Quan i a i e UHPLC Analysis o Fla onoids. Fo plan ma e ial (au umn 2021-
144 sp ing 2023), a 250 mg o g ound lowe s we e ex ac ed wice (2 × 4 mL) using 80%
145 MeOH in wo s eps: I – he mo-shaking (20 min, 60 °C, 800 pm; The momixe ,
146 Eppendo ), and II – ul asonic ba h (10 min, 40 °C, 35 kHz). A e cen i uga ion (10
147 min, 5 °C, 12,000 × g), he supe na an s we e combined and d ied in a he mo-block
148 (40 °C) unde N2 and e-dissol ed in 2 mL o 80% MeOH. Samples o sy ups (4 mL)
149 and inc u e (15 mL a e concen a ion o 3 mL) we e pu i ied using SPE echnique
150 (Oasis HLB 500 mg column (Wa e s)). Pola componen s we e elu ed wi h 2% MeOH,
151 while la onoids using 95% MeOH (con aining 1% ammonia). Pu i ied samples we e
152 d ied simila ly o he ex ac s and e-dissol ed in 1 mL o 80% MeOH. All samples we e
153 s o ed a -20 °C un il analysis. Fi e independen ex ac s we e p epa ed o g ound
154 lowe s and ou pu i ied samples o sy ups and inc u e. P io o analyses, he
155 samples we e app op ia ely dilu ed wi h 80% MeOH and il e ed (0.22 µm).
156 Quan i a i e UHPLC-MS/MS (Wa e s) measu emen s we e based on mul iple eac ion
157 moni o ing (MRM) de ec ion mode and ex e nal calib a ion o he 16 e e ence
158 la onoid subs ances. De ailed in o ma ion ega ding de ec ion: capilla y ol age,
159 p ecu so → agmen ions’ ansi ions, and collision ene gy is shown in Table S6; while
160 calib a ion pa ame e s a e placed in Table S7. The gene al ch oma og aphic
161 sepa a ion condi ions we e as ollows: column BEH C18 (100 × 2.1 mm, 1.7 µm,
162 Wa e s), column empe a u e 45 °C, a 14 min linea g adien (15→35%) o ace oni ile–
163 wa e (bo h acidi ied wi h 0.1% o mic acid), low a e o 0.4 mL/min, and injec ion
164 olume 2 µL. The MS de ec o was ope a ed in posi i e ion mode wi h he ollowing
165 se ings: capilla y ol age (3.1 kV), sou ce and desol a ion gas empe a u es (150 °C
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314 dis ance and Wa d's algo i hm) analyses highligh ed he g ea dis inc ness o sp ing
315 and au umn lowe s, as well as a g ea e a ia ion in sp ing ma e ial (sepa a ion o
316 sp ing lowe s 2022 and 2023 along PC2) compa ed o au umn samples was obse ed
317 (Fig. 4A-B). None heless, he sepa a ion o au umn lowe s om he wo ha es yea s
318 was la e e iden along PC3 (Fig. S23B). The PCA biplo s (Fig. 4C and S23C) and
319 hea map (Fig. 4B) con i med he seasonal clus e ing o he lowe s wi h espec o
320 chemical composi ion, and among he posi i e co ela ions eco ded we e: au umn
321 2021 – compounds 5, 24 and 27; au umn 2022 – compounds 21 and 40-43; sp ing
322 2022 – compounds 15, 18+19, 22, 25-26, 28, 33 and 37-39; sp ing 2023 – compounds
323 3, 8, 14, 16 and 20. The cu en wo k is he i s o a emp o iden i y seasonal
324 phy ochemical a ia ion in dandelion lowe s.
325 The quan i a i e la onoid analysis o dandelion ood p oduc s e ealed hei high
326 a iabili y (6.5-30.9 µg/mL o o al la onoids, Table 3), and hey we e gene ally
327 a anged in he ollowing o de : sy up I ≥ inc u e > sy up II > sy up III. The p opo ion
328 o he speci ied la onoid g oups in he samples was also he e ogeneous: glycosides
329 (70% o o al la onoids) and aglycones (27%) p edomina ed in sy up I, while in sy ups
330 II-III he p opo ion was e e sed (aglycones – abou 70% and glycosides – abou
331 25%), simila ly, aglycones (83%) de ini ely p edomina ed in he inc u e, bu
332 la onolignans (11%) we e second. In each p oduc , o al bi la ones we e a mino
333 ac ion (less han 2% o o al la onoids), bu in he inc u e (0.35 µg/mL) hei le el
334 was se e al imes (3-15) highe han hose in sy ups (0.02–0.1 µg/mL) (Table 3).
335 Su p isingly, he h ee comme cial sy ups di e ed signi ican ly in hei quan i a i e
336 la onoid p o ile, which, in addi ion, was no simply ela ed o he manu ac u e ’s
337 decla ed dandelion decoc ion con en (sy ups I-III – 48%, 60% and 40%, espec i ely).
338 Acco ding o Table 3, he dis inguishing ea u e o sy up III was he lowes eco ded
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339 concen a ions o all la onoids, o sy up I he highes le els o he h ee lu eolin
340 glycosides, and o he inc u e he highes concen a ions o mos o he compounds
341 (aglycones, bi la ones and la onolignans). I can be no ed ha he numbe o
342 publica ions de o ed o quan i a i e chemical analysis o dandelion ood p oduc s, like
343 lowe s, is negligible. O e all, he la onoid le els in comme cial sy ups, especially I-II,
344 a e in ai ly close ag eemen wi h he esul ob ained o he handmade sy up (22 µg
345 lu eolin eq/mL) in he pape by Majewski e al. 2022.11 On he o he hand, ou alcoholic
346 lowe inc u e (85 g/1000 mL) had a signi ican ly highe la onoid con en (269 µg/g
347 dw) compa ed o he b inc u e (50 g/500 mL, 90 µg/g dw) de eloped by Epu e e al.
348 2023.20
349 Subsequen ly, mul i a ia e s a is ical me hods we e used o g aphically illus a e he
350 di e ences/simila i ies be ween dandelion ood p oduc s. The esul s o PCA (95%
351 a iabili y co e ed by he i s wo componen s, Fig. S24) and HCA (Wa d's algo i hm
352 and Euclidean dis ance) analyses con i med he high a iabili y o he p oduc s
353 (sepa a e g ouping o samples, Fig. 4D-E). Simila ly, he PCA-biplo s (Fig. 4F and S24)
354 and hea map (Fig. 4E) con i med he sample clus e ing wi h espec o chemical
355 composi ion, which was p e iously highligh ed a he le el o Table 3 (e.g., he posi i e
356 co ela ion o sy up I wi h glycosides, as well as inc u e wi h la one aglycones).
357 Dandelion is a well-known use ul plan wi h many documen ed biological p ope ies.
358 HR-QTOF-MS/MS analysis o he lowe ex ac o his plan e ealed he p esence o
359 nume ous la onoids, among which we e well-known and bioac i e me aboli es such
360 as lu eolin and i s glycosides, ch ysoe iol, icin, apigenin. In addi ion, mo e han 20
361 o he la onoid compounds we e de ec ed, including se e al oligome s and mo e han
362 a dozen la onolignans (lignan de i a i es o icin). Ta ge ed isola ion o me aboli es
363 using ex ac ion op imized by DoE planning and a ious ch oma og aphic me hods led
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364 o he isola ion o 11 me aboli es whose chemical s uc u es we e de e mined by
365 spec oscopic da a. Among hem we e ou bi la ones, including philono is la one
366 (p e iously epo ed in he axon) and dic anolomin ( epo ed o he i s ime in he
367 axon), as well as wo new compounds, 6',8''-bilu eolin (16) and 6',6''-bilu eolin (18).
368 On he o he hand, among he se en la onolignans isola ed, he e was one new
369 compound, a axalignan A (33), as well as six al eady desc ibed me aboli es, excep
370 ha wo me hoxycalquiquelignan de i a i es (40-41) ha e no ye been epo ed in he
371 Ta axacum. The quan i a i e con en o mo e han 30 la onoids was de e mined,
372 based on UHPLC-ESI-MS/MS analysis, in aw lowe s and dandelion ood p oduc s.
373 Compa a i e and chemome ic analysis (including PCA, HCA and PLS-DA) using
374 quan i a i e la onoid da a showed seasonal clus e ing o lowe samples (sp ing s.
375 au umn), wi h compounds 27, 24, 34, 29, 35, 16, 5, 32 and 25 among he ai s wi h
376 he g ea es impac . A simila app oach o ood p oduc s ( h ee comme cial sy ups
377 and a handmade inc u e) showed hei high a ia ion in phy ochemical composi ion.
378 Ou wo k is he i s such comp ehensi e s udy o la onoids in dandelion lowe s, and
379 ills many phy ochemical knowledge gaps o his plan and he ood p oduc s made
380 om i .
381
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382 ACKNOWLEDGEMENTS
383 We hank Lukasz Pecio o pe o ming NMR expe imen s, Aga a Soluch o echnical
384 assis ance and Anna Rybicka o hos ing he UHPLC-MS ins umen .
385
386 FOUNDING SOURCES
387 The wo k was pa ially suppo ed om he Polish Na ional Science Cen e p ojec
388 PRELUDIUM 2022/45/N/NZ9/03440.
389
390 SUPPORTING INFORMATION
391 Supplemen a y Ma e ials include Desc ip ions S1-S3, Tables S1-S7 and Figu es S1-
392 S24. The ma e ial is a ailable ee o cha ge ia In e ne a h p:// pubs.acs.o g.
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394 REFERENCES
395 (1) WFO (2024): Ta axacum o icinale F.H.Wigg. Published on he In e ne ;
396 h ps://www.wo ld lo aonline.o g/ axon/w o-0000062154. Accessed on: 30 Jan 2024.
397 (2) Ma inez, M.; Poi ie , P.; Chamy, R.; P ü e , D.; Schulze-G ono e , C.; Jo que a,
398 L.; Ruiz, G. Ta axacum O icinale and Rela ed Species—An E hnopha macological
399 Re iew and I s Po en ial as a Comme cial Medicinal Plan . J. E hnopha macol. 2015,
400 169, 244–262. h ps://doi.o g/10.1016/j.jep.2015.03.067.
401 (3) González-Cas ejón, M.; Visioli, F.; Rod iguez-Casado, A. Di e se Biological
402 Ac i i ies o Dandelion. Nu . Re . 2012, 70 (9), 534–547.
403 h ps://doi.o g/10.1111/j.1753-4887.2012.00509.x.
404 (4) The Eu opean Pha macopoeia Ten h Edi ion (2019); Council o Eu ope,
405 S asbou g, F ance; ISBN: 978-92-871-8912-7.
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478
479
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Table 1. UHPLC-HR-QTOF-MS/MS cha ac e iza ion o me aboli es de ec ed in dandelion lowe la onoid ex ac .
No
RT
(min)
UV λmax
(nm)
Obse ed [M+H]+,
m/z
Fo mula
E o
(ppm)
MS/MS agmen s
Iden i y
1
1.69
323, 295sh
163.0393/181.0498
C9H9O4
-1.2
89.0384, 117.0336, 136.0516
ca eic acid*
2
2.73
308
147.0445/165.0547
C9H7O2
-0.6
-
couma ic acid*
3
3.78
254, 336
449.1084
C21H21O11
-1.1
287.0564
lu eolin-7-O-glucoside*
4
4.83
326, 295sh
499.1241/517.1351
C25H25O12
-1.4
163.0397
3,5-di-ca eoylquinic acid*
5
5.55
267, 337
449.1086
C21H21O11
-1.8
287.0555
lu eolin-4'-O-glucoside*
6
6.36
326, 295sh
499.1241/517.1348
C25H25O12
-1.5
163.0395
di-ca eoylquinic acid
7
6.59
250, 269, 345
463.1240
C22H23O11
-1.1
301.0712
ch ysoe iol-O-hexoside
8
6.84
267, 340
449.1083
C21H21O11
-0.9
287.0558
lu eolin-3'-O-glucoside*
9
7.01
-
442.2071 [M+NH4]+
C21H32NO9
0.0
171.1172, 217.1227
a axinic acid-1-O-glucoside*
10
8.49
254, 341
855.1186
C45H27O18
0.7
285.0400, 419.0771, 703.1097
la onoid ( i-lu eolin)
11
8.98
254, 344
855.1187
C45H27O18
0.6
285.0401, 153.0188, 703.1081
la onoid ( i-lu eolin)
12
9.31
256, 344
855.1183
C45H27O18
1.0
285.0403, 153.0186, 703.1076
la onoid ( i-lu eolin)
13
9.80
327, 295sh
513.1395/531.1500
C26H27O12
-0.5
163.0395
ca eoyl- e uloylquinic acid
14
10.16
252, 346
287.0557
C15H11O6
-2.3
153.0188, 135.0446
lu eolin*
15
11.85
253, 341
571.0882
C30H19O12
-1.9
419.0775, 153.0192, 285.0389
philono is la one#
16
12.53
254, 344
571.0881
C30H19O12
-1.8
419.0778, 153.0195, 445.0560
a axabilu eolin A#
17
13.23
254, 341
855.1200
C45H27O18
-0.9
153.0186, 551.0983, 703.1094
la onoid ( i-lu eolin)
18
13.66
254, 342
571.0885
C30H19O12
-2.5
419.0778, 445.0562
a axabilu eolin B#
19
13.68
254, 340
571.0887
C30H19O12
-2.6
419.0778, 445.0563
dic anolomin#
20
14.09
265, 338
271.0609
C15H11O5
-3.1
153.0183
apigenin*
21
15.52
250, 269, 346
301.0710
C16H13O6
-1.0
258.0529, 286.0478
ch ysoe iol*
22
15.67
250, 269, 351
331.0816
C17H15O7
-1.1
315.0501, 203.0338
icin*
23
16.25
255, 342
853.1036
C45H25O18
-0.1
153.0188, 269.0439, 497.0499
la onoid oligome
24
17.34
268, 339
497.1441
C26H25O10
0.2
331.0815, 315.0503
calquiquelignan D#
25
18.00
269, 339
527.1550
C27H27O11
-0.5
331.0824, 315.0506
salcolin B#
26
18.12
268, 330
331.0816
C17H15O7
-1.0
315.0507, 270.0528
apome zge in*
27
18.74
268, 337
497.1447
C26H25O10
-0.9
331.0818, 315.0504
calquiquelignan E#
28
19.58
269, 337
527.1549
C27H27O11
-0.2
331.0822, 315.0507
salcolin A#
29
20.25
269, 342
675.2076
C36H35O13
-0.5
331.0820, 315.0510, 270.0533
la onolignan ( icin de i a i e)
30
20.69
270, 337
495.1286
C26H23O10
-2.0
284.0687, 331.0821, 255.0661
la onolignan ( icin de i a i e)
31
20.86
297, 310
787.3688
C45H55O12
-1.9
147.0444, 204.1030, 641.3354
N-couma oyl-spe mine de i a i e
32
21.16
269, 341
705.2183
C37H37O14
-0.7
331.0830, 315.0512
la onolignan ( icin de i a i e)
33
21.46
270, 321
525.1399
C27H25O11
-1.5
331.0826, 284.0693, 255.0664
a axalignan A#
34
21.51
269, 340
675.2080
C36H35O13
-1.2
331.0822, 315.0511
la onolignan ( icin de i a i e)
35
22.03
269, 340
675.2079
C36H35O13
-1.0
331.0823, 315.0512
la onolignan ( icin de i a i e)
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36
22.35
268, 339
479.0979
C25H19O10
-1.4
257.0456, 435.1101, 287.0557
la onoid
37
22.56
269, 341
705.2185
C37H37O14
-1.1
331.0830, 315.0512
la onolignan ( icin de i a i e)
38
23.14
269, 341
705.2186
C37H37O14
-1.1
331.0832, 315.0513
la onolignan ( icin de i a i e)
39
23.89
269, 341
495.1299
C26H23O10
-2.8
287.0560, 315.0509
la onolignan ( icin de i a i e)
40
24.03
268, 340
511.1614
C27H27O10
-3.0
331.0828, 315.0513
7''-me hoxycalquiquelignan D#
41
24.58
268, 339
511.1610
C27H27O10
-2.2
331.0826, 315.0515
7''-me hoxycalquiquelignan E#
42
25.10
269, 340
541.1715
C28H29O11
-1.9
331.0821, 315.0509
la onolignan (me hoxysalcolin B)
43
25.13
269, 339
541.1713
C28H29O11
-1.7
331.0822, 315.0507
la onolignan (me hoxysalcolin A)
* he iden i y o he compound was con i med by compa ison wi h e e ence subs ance; # compound was isola ed and iden i ied in his s udy
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Table 2. 1H and 13C NMR da a o new la onoid compounds 16, 18 and 33 (in DMSO-d6).
16 ( a axabilu eolin A)
18 ( a axabilu eolin B)
33 ( a axalignan A)
No
δC
δH (J in Hz)
δC
δH (J in Hz)
δC
δH (J in Hz)
2
165.9
-
166.1
-
162.6
-
3
106.2
5.97, s
105.9
5.99, s
104.8
6.99, s
4
181.1
-
181.3
-
181.6
-
5
161.2
-
161.3
-
161.3
-
6
98.7
6.08, d (2.1)
98.8
6.11, d (2.0)
99.3
6.18, b s
7
164.1
-
164.2
-
165.6
-
8
93.2
5.74, d (2.0)
93.4
5.97, d (2.0)
94.4
6.51, b s
9
157.2
-
157.4
-
157.5
-
10
103.2
-
103.4
-
103.3
-
1'
123.0
-
122.8
-
125.5
-
2'
115.9
7.29, s
115.6
7.22, s
104.2
7.30, s
3'
145.1
-
144.9
-
152.2
-
4'
148.3
-
148.1
-
139.3
-
5'
119.9
6.82, s
119.9
6.74, s
152.2
-
6'
122.9
-
123.8
-
104.2
7.30, s
3',5'-OMe
-
-
-
-
56.3
3.77, s
1''
-
-
-
-
139.3
-
2''
163.8
-
163.8
-
111.7
7.51, d (2.0)
3''
102.4
6.59, s
102.9
6.69, s
147.5
-
4''
181.8
-
181.7
-
152.2
-
5''
160.2
-
158.3
-
114.9
6.88, d (8.3)
6''
98.5
6.31, s
111.8
-
123.6
7.58, dd (8.3,
2.0)
7''
161.4
-
161.5
-
83.3
5.52, (5.6)
8''
107.1
-
93.4
6.53, s
62.5
3.78, o
3.84, o
9''
153.8
-
156.0
-
194.0
-
10''
103.4
-
103.4
-
-
-
3''-OMe
-
-
-
-
55.6
3.81, s
1'''
121.3
-
121.4
-
-
-
2'''
113.6
7.04, d (2.3)
113.3
7.42, d (2.3)
-
-
3'''
145.7
-
145.8
-
-
-
4'''
149.8
-
149.9
-
-
-
5'''
115.6
6.76, d (8.3)
116.1
6.90, d (8.2)
-
-
6'''
118.6
7.02, dd (2.2, 8.3)
119.0
7.43, dd (2.3, 8.2)
-
-
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Table 3. Fla onoid con en o dandelion lowe s and ood p oduc s based on hem (sy ups and inc u e) (mean ± SD). Compound numbe ing is
consis en wi h Table 1.
Flowe s
Food p oduc s
No
2021-Au umn
2022-Sp ing
2022-Au umn
2023-Sp ing
Sy up I (48%
decoc ion)
Sy up II (60%
decoc ion)
Sy up III (40%
decoc ion)
Tinc u e (85
g/1000 mL)
µg/g ( lowe s) o ng/mL ood p oduc s
5
213.41 ± 44.9a
10.15 ± 0.50b
8.74 ± 0.99b
11.45 ± 0.19b
211.84 ± 8.05a
59.52 ± 0.85c
68.76 ± 0.79b
76.96 ± 2.03b
15
35.79 ± 1.54c
64.08 ± 2.89a
37.23 ± 5.84bc
45.23 ± 7.85b
38.79 ± 1.88c
91.34 ± 5.23b
24.06 ± 2.39d
317.65 ± 3.88a
16
2.53 ± 0.18b
3.08 ± 0.11b
1.89 ± 0.15c
4.30 ± 0.57a
acec
3.36 ± 0.11b
ace
11.96 ± 0.09a
18+19a
1.65 ± 0.08b
2.84 ± 0.14b
1.67 ± 0.24b
1.95 ± 0.34a
ace
2.85 ± 0.35b
ace
15.68 ± 0.61a
20
10.48 ± 0.86a
8.88 ± 0.87b
7.06 ± 0.35c
10.75 ± 0.43a
24.25 ± 0.69c
53.71 ± 0.34b
17.25 ± 0.17d
158.70 ± 3.01a
26
8.85 ± 0.35b
12.85 ± 0.89a
6.32 ± 0.6c
9.66 ± 0.34b
294.98 ± 6.74c
312.59 ± 7.87b
60.59 ± 0.52d
577.92 ± 7.86a
29b
1.17 ± 0.03a
0.98 ± 0.10b
1.08 ± 0.07ab
0.72 ± 0.02c
6.12 ± 0.19c
7.57 ± 0.30b
1.38 ± 0.02d
12.16 ± 0.07a
30b
ace
ace
ace
ace
ace
ace
ace
ace
32b
0.46 ± 0.02c
0.75 ± 0.06a
0.48 ± 0.02c
0.58 ± 0.03b
6.54 ± 0.20a
3.31 ± 0.06c
1.12 ± 0.01d
5.66 ± 0.05b
33
21.19 ± 0.64b
30.65 ± 6.26b
18.76 ± 1.17b
23.71 ± 1.94a
45.16 ± 0.82c
65.47 ± 2.14b
10.45 ± 0.57d
231.39 ± 1.12a
34b
0.78 ± 0.02a
0.65 ± 0.05b
0.72 ± 0.05a
0.49 ± 0.02c
2.29 ± 0.09c
3.41 ± 0.12b
ace
8.01 ± 0.18a
35b
0.86 ± 0.02a
0.73 ± 0.07b
0.81 ± 0.05ab
0.54 ± 0.02c
1.97 ± 0.07c
3.04 ± 0.08b
ace
9.76 ± 0.11a
36b
ace
ace
ace
ace
ace
ace
ace
ace
37b
0.31 ± 0.01c
0.51 ± 0.04a
0.32 ± 0.02c
0.37 ± 0.02b
2.42 ± 0.10b
1.59 ± 0.06c
ace
3.42 ± 0.06a
38b
0.40 ± 0.01c
0.70 ± 0.06a
0.42 ± 0.03c
0.49 ± 0.03b
2.31 ± 0.09b
1.67 ± 0.03c
ace
5.48 ± 0.13a
39b
4.91 ± 0.28c
7.64 ± 0.76a
6.23 ± 0.96b
5.24 ± 0.69bc
9.20 ± 0.21c
10.80 ± 0.42b
2.52 ± 0.03d
64.25 ± 0.28a
40b
0.43 ± 0.02b
0.35 ± 0.06b
0.57 ± 0.09a
0.41 ± 0.03b
0.49 ± 0.01c
0.94 ± 0.04a
ace
0.72 ± 0.06b
41b
0.55 ± 0.03ab
0.45 ± 0.09b
0.59 ± 0.09a
0.51 ± 0.04ab
0.69 ± 0.01c
1.35 ± 0.10a
ace
1.07 ± 0.08b
42+43b
5.01 ± 0.25bc
4.76 ± 0.34c
6.78 ± 1.59a
6.35 ± 0.47ab
1.59 ± 0.03c
2.61 ± 0.22b
0.72 ± 0.12d
6.15 ± 0.16a
mg/g ( lowe s) o µg/mL ood p oduc s
3
0.87 ± 0.05a
0.87 ± 0.05a
0.69 ± 0.04b
0.92 ± 0.12a
10.72 ± 0.38a
1.96 ± 0.05b
0.84 ± 0.01c
0.81 ± 0.03c
8
0.87 ± 0.05a
0.87 ± 0.05a
0.69 ± 0.04b
0.92 ± 0.12a
10.75 ± 0.39a
1.71 ± 0.04b
0.79 ± 0.01c
0.75 ± 0.02c
14
2.22 ± 0.08a
2.27 ± 0.20a
1.76 ± 0.06b
2.43 ± 0.12a
6.60 ± 0.07c
9.61 ± 0.16b
4.04 ± 0.03d
21.08 ± 0.33a
21
0.16 ± 0.00a
0.15 ± 0.02a
0.17 ± 0.02a
0.16 ± 0.00a
0.59 ± 0.01c
0.93 ± 0.01b
0.33 ± 0.00d
2.14 ± 0.03a
22
0.06 ± 0.00c
0.10 ± 0.01a
0.05 ± 0.00c
0.07 ± 0.01b
0.71 ± 0.01a
0.44 ± 0.00c
0.14 ± 0.00d
0.64 ± 0.01b
Page 24 o 30
ACS Pa agon Plus En i onmen
Jou nal o Ag icul u al and Food Chemis y
