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Floral origin markers for authenticating Lavandin honey (Lavandula angustifolia xlatifolia). Discrimination from Lavender honey (Lavandula latifolia)

L. Castro-Vzquez, V. Leon-Ruiz, M.E. Alaon, M.S. Prez-Coello, A.V. Gonzlez-

Porto

PII: S0956-7135(13)00438-6

DOI: 10.1016/j.foodcont.2013.09.003

Reference: JFCO 3445

To appear in: Food Control

Received Date: 7 May 2013

Revised Date: 31 August 2013

Accepted Date: 2 September 2013

Please cite this article as: Castro-VzquezL., Leon-RuizV., AlaonM.E., Prez-CoelloM.S. & Gonzlez-

PortoA.V., Floral origin markers for authenticating Lavandin honey (Lavandula angustifolia x latifolia).Discrimination from Lavender honey (Lavandula latifolia), Food Control(2013), doi: 10.1016/

j.foodcont.2013.09.003.

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http://dx.doi.org/10.1016/j.foodcont.2013.09.003


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Floral origin markers for authenticating Lavandin honey (Lavandula angustifolia x1

latifolia). Discrimination from Lavender honey (Lavandula latifolia)2

3

L. Castro-Vzqueza *

, V. Leon-Ruizb

, M. E. Alaonc

, M. S. Prez-Coelloc

, A.V.4

Gonzlez-Portob

5

6

7

8

aFood Technology Area, Faculty of Pharmacy, University of Castilla-La Mancha,9

Campus Universitario, Albacete, Spain10

11

b Beekeeping Research Centre of Marchamalo, Camino San Martn s/n. 19180.12

Marchamalo. Guadalajara, Spain.13

14

cFood Technology Area, Faculty of Chemistry, University of Castilla-La Mancha,15

Campus Universitario Ciudad Real, Spain16

17

18

19

AUTHOR INFORMATION20

* Corresponding author. Tel.: +34-926-295300, Fax: +34-926-295318.21

E-mail: [email protected]

23

24

25


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ABSTRACT26

The aim of this study was to identify the botanical origin of lavandin honey, a27

monofloral product of recent proliferation obtained from a hybrid of the species28

Lavandula angustifolia and L. latifolia. Lavandin was authenticated here in chemical,29

palynological and sensory terms, and discriminated from the more common Lavender30

honey (L. latifolia).31

A total of 14 physicochemical parameters, 26 pollen types, 13 sensory attributes32

and 80 volatile compounds were identified and/or quantified for this purpose.33

High concentrations of -nonalactone, farnesol and acetovanillone, which were for34

the first time identified as components of honey aroma in this study, together with35

several lactones, dehydrovomifoliol, 4-methoxyacetophenone and decanal are proposed36

as chemical markers for authenticating lavandin monofloral honey. In sensory terms,37

increased scores for caramel and peach compote sensations, together with an also38

increased Rubus pollen content, provide additional useful information for the accurate39

authentication of lavandin honey and its discrimination from lavender honey (L.40

latifolia).41

42

Results showed enough floral markers to authenticate the botanical origin of43

Lavandin honey allowing its marketing as monofloral rather than generic honey.44

45

Keywords: Lavandin-honey; volatile-markers, physicochemical-parameters,46

palynological-analysis, sensory-profile.47

48

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1. Introduction50

Lavender is a popular aromatic mediterranean herb belonging to Lamiaceae51

family. The genus Lavandula contains several species, although the more common52

species that is believed to have aromatherapy and pharmaceutical values areLavandula53

dentata (French lavender), Lavandula angustifolia, Lavandula latifolia, Lavandula54

stoechas andLavandula hybrida.55

56

In the beekeeping context, honey from Lavender (Lavandula latifolia), commonly57

known as spike Lavender, is currently among the most appreciated high-quality58

honeys by virtue of its pleasant aroma and taste. Although, in recent years cultivation59

of the hybrid Lavender species known as Lavandin, which is a cross betweenLavandula60

angustifoliaandLavandula latifolia,has proliferated widely at the expense of Lavender61

because it has major advantages for flavouring industries (Paschalina & Apostolos,62

2003). This has been especially so in Spain, where large cropping areas of Lavandin (L.63

angustifolia x L. latifolia) have be established lately.64

65

When Lavender honey is commercialized, no distinction between species66

(Lavender or Lavandin) is made, however honey producers know that honey from L.67

latifoliaexhibits substantial sensory differences from Lavandin honey. For this reason,68

the Spanish beekeeping industry, which is concentrated in the Alcarria region (central69

Spain), is highly interested in finding ways to differentiate these species with a view to70

characterizing and marketing Lavandin honey as a monofloral product with its own,71

identifiable features.72

73

The traditional approach to identifying the botanical origin of honey relies on74

examination of its pollen (Anklam, 1998; Louveaux, Maurizio, & Vorwohl, 1978;75


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Soria, Gonzlez, de Lorenzo, Martnez-Castro, & Sanz, 2004).This analysis requires a76

high skill and does not ensure reliable identification if the honey concerned contains77

little or no pollen. The minimum proportion ofL. latifoliapollen needed to characterize78

Lavender honey as monofloral is about 15% (La Serna Ramos, 2007; Sanz, Prada,79

Gmez-Ferreras, Lorenzo, & Jrez, 1980), which is the typical proportion for all other80

species of the family Labiatae. In honey from Lavandin, which contains few pollen81

grains, this botanical analysis is of little use. Therefore, authenticating Lavandin honey82

entails using additional analytical and physicochemical markers not identified to date.83

84

Since the chief factor in honey characterization is its aroma, the analysis of85

volatile components is a useful tool to authenticate unifloral honeys. Lavender honey86

has so far been characterized in terms of its hexanal, heptanal, nerolidol oxide and87

coumarin contents (Bouseta, Collins, & Dufour, 1992; Bouseta, Scheirman, & Collin,88

1996; Castro-Vzquez, Daz-Maroto, Gonzalez-Vias, & Prez-Coello 2009; Guyot-89

Declerck, Renson, Bouseta, & Collin, 2002; Radovic, Careri, Mangia, Musci, Gerboles,90

& Ankla, 2001; Shimoda, Wu, & Osajima, 1996). However, little is known about91

Lavandin hybrid honey (Lavandula angustifolia x latifolia) volatile composition.92

Although phenylacetaldehyde and heptanoic acid contents allowed Lavandin honey to93

be distinguished from Lavender honey in a previous study (Guyot-Declerck et al.,94

2002), the Lavender samples used were from L. angustifolia, which is less frequently95

used for honey production. Therefore, the contents in the previous compounds by96

themselves are seemingly inadequate to characterize Lavandin honey.97

98

Based on the foregoing, and on the increasing demand for products with specific99

characteristics and an identifiable geographical origin, the ability to authenticate100

Lavandin honey may be extremely useful for the beekeeping sector in the protected101


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designation of origin (PDO) La Alcarria (central Spain).Therefore, the purposes of this102

study were as follows: (a) to identify volatile components in Lavandin honey103

(Lavandula angustifolia x latifolia) with a view to its authentication and discrimination104

from lavender honey (L. latifolia); (b) to evaluate physicochemical parameters and105

pollen types associated with Lavandin honey; and (c) to elucidate the sensory profile of106

Lavandin honey in order to facilitate its authentication.107

108

2.Material and methods109

2.1. Samples110

All honeys were collected fromLa Alcarria(Spanish north-central area) sheltered111

by the Origin Denominationand had not been industrially processed. Samples belong to112

several beehives, corresponding with the beekeepers cropping areas.113

Ten Lavender honeys(Lavandula latifolia) and ten Lavandin honeys (Lavandula114

angustifolia x latifolia)were analysed just after harvest by professional beekeepers. The115

samples were stored at 4 C until analysis.116

117

2.2.Isolation and analysis of volatile compounds118

The isolation of the volatile compounds was done on the styrene-divinylbenzene119

cartridges (Lichrolut EN, Merck, 0.5 g of phase), previously conditioned with 10 ml of120

dichloromethane, 5 ml of methanol and 10 ml of ethanol/distilled water (10%) at a flow121

rate of 2 mL/minute. Ten grams of each honey were dissolved in 50 mL of distilled122

water and 25 L of 2-pentanol (1g/L) was added as internal standard. Theses solutions123

were passed through cartridges. Then, sugars were eluted with 25 mL of distilled water,124

in order to eliminate artefacts. Volatile compounds were eluted with 30mL of125

dichloromethane at a flow rate of 2 ml/minute. The organic phase collected was126


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concentrated in a Vigreux column and analysed by GC/MS Castro-Vzquez, Daz-127

Maroto, Guchu, & Prez-Coello, 2006.128

An Agilent 6890 N gas chromatograph, coupled to a 5973 Inertmass selective129

detector was used. Two microliters of extracts were injected in splitless mode (0.6 min)130

on a polyethylene glycol capillary column BP-21 (50 m x 0.32 mm x 0.25 m of film131

thickness). The oven temperature was programmed to remain at 60 C for 3 min and132

then increase 2 C/min to 200 C and held for 30 min. Helium was used as carrier gas at133

a flow rate of 0.8 mL/min-1

. Injector and transfer line temperatures were 250 C and134

280 C, respectively. Mass detector conditions were: electronic impact (EI) mode at 70135

eV; scan mass acquisition range: 40-450 amu.136

137

Peak identifications were based on comparison of their mass spectra with the138

corresponding commercial pure standards purchase from Sigma-Aldrich with a139

chemical purity between 85-99 %, and/or with those reported by the NBS75K and140

Wiley A commercial libraries. The quantitative analysis by total ion was performed141

using 2-pentanol such as internal standard. Further, individual response factors, ranged142

between 0-1, were taken into account. Response factors for each compound were143

calculated in water solution with 350 and 448 g/L of glucose and fructose, respectively,144

like a model honey. For compounds which commercial standards was not available, the145

response factors of compounds with similar chemical structures were used. All the146

samples were injected in duplicate.147

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2.3.Melissopalynological analysis152

Pollen analysis was carried out using the method established by the International153

Commission of Bee Botany described by Louveaux et al., (1978). The study of the154

sediment pollen spectrum was performed by microscopic analysis. Ten grams of honey155

was used for concentrating and counting all the pollen and spores particles. At least 300156

grains of pollen from each honey preparation were counted per slide.157

158

2.4. Physicochemical and Colour Analysis159

Physicochemical parameters were analysed following the specifications of the160

International Honey Commission and Spanish legislation (Bogdanov, Lullmann, Martin,161

Ohe, Russmann, & Vorwohl, 2000; B.O.E., 1986). Water content (moisture) was162

determined using an ABBE refractometer model NAR-1T. Free acidity was analysed by163

potentiometric titration, and pH using pH-meter model Eutech Systems XS PC510.164

Hydroxymethylfurfural content was determined based on the method of White, 1979165

using a Hitachi U-1100 Spectrophotometer. Conductivity was measured with a166

Radiometer CDM-83 conductivimeter.167

168

Color parameters were established in the CIELAB system by reflection method169

(measuring geometry d/8, illuminant D65, range 380 to 780 nm, observer 10) using a170

Hitachi model U-1100 spectrophotometer (CIE 1986). Colour was also determined171

using a Pfund colour grader Lovibond Visual Comparator 2000 Series.172

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2.5.Descriptive Sensory Analysis176

Lavender (L. latifolia) and Lavandin honeys were evaluated according to177

quantitative descriptive analysis (QDA), where each honey was individually evaluated178

using a list with previously defined descriptors. The panel was formed by eight179

assessors, members of Food Technology Department (UCLM) and beekeeping research180

centre (Marchamalo) with aged ranging from 28 to 45 years. Sessions were performed181

in a tasting room with individual chambers that were separated of the area where the182

samples were prepared (ISO 8589, 1998). During the evaluation, room temperature was183

21C and 70% humidity. Three different coded samples were presented in random order184

to each assessor in 40-mL glass vials sealed with a twist-off cap and at room185

temperature. Mineral water was provided for the assessors mouth-rinsing between186

samples. Sessions were carried out between 11:00 and 13:00.187

The screening of assessors was carried out in previous studies to determine188

sensory profiles of unifloral honeys (Castro-Vazquez et al., 2009; Castro-Vzquez,189

Daz-Maroto, de Torres, & Prez-Coello 2010) All of them had prior experience in190

discrimination and descriptive sensory test.191

Despite that, a training of judges was done during six sessions. In the first one,192

aqueous standards solutions of linalool, -decalactone, and benzaldehyde (50 ppb), and193

eugenol, 1,8-cineol, (15ppb) were used to identify floral, fresh fruit, bitter194

almond, spice and balsamic notes. Those levels were chosen based on their195

proximity to their respective odour detection thresholds in air. Each panellist correctly196

described the standards and then knowing their identities.197

To complete the training period, the judges evaluated sixteen lavender (L.198

latifolia) and lavandin honeys during four sessions. In each session, the subjects199

received four honey samples and individually described the flavour and taste sensory200


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attributes with their own terminology. The judges generated a total of ten olfactory and201

eight gustatory descriptors.202

A new session was carried out in order to discuss the terms generated by each203

individual, and, with the supervision of a panel leader, they consensually defined the204

terms that adequately described the aroma and taste sensory similarities and differences205

among the evaluated honeys. Finally, seven olfactory and six flavour attributes were206

considered among the best at describing the sensory characteristics of lavender (L.207

latifolia)and lavandin honeys and the main differences between them. The aim of judge208

training was twofold: reaching a consensus about the meaning of each attribute and209

achieving intensity rating in a reliable way.210

211

The formal evaluation of honeys consisted in ten sessions realized in different212

days. The panellist used a 10 cm non-structured scale, from 0 to 10, to rate the intensity213

of each attribute, previously selected. Scales were delimited at the ends by the terms214

"weak" and "strong. Two replications of all sample assessments were performed.215

216

2.6. Statistical Analysis217

The statistical methods used for data analysis was a Student T-test for independent218

samples, to determine significant differences between volatile composition of honeys,219

and among sensory profiles of honey types. Step-way Discriminant analysis to select the220

aroma compounds most useful in differentiating the two honey sources was performed.221

One way ANOVA was used to check differences among physicochemical values.222

Statistical processing was carried out by using the SPSS 17.0 statistical software223

package for Windows.224

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3. Results and discussion226

3.1.Identification of honey volatile markers227

A total of eighty volatile organic compounds (VOCs) were quantified in Lavandin228

(Lavandula angustifolia x latifolia) and Lavender (Lavandula latifolia) honeys.229

Although most are present in both types of honey, the two honey types can be230

differentiated in terms of their concentrations.231

232

Table 1 shows the volatile fingerprints off the two types of honey as obtained by233

SPE/GCMS analysis. Students t-test for independent samples was used to identify234

significant differences according to monofloral origin. As can be seen, there were235

marked differences between the two types of honey. There were none, however, in236

hexanal or phenylacetaldehyde levels, which, according to Guyot-Declerck et al., can be237

used to differentiate Lavender and Lavandin honeys. This was probably a result of the238

particular honey species investigated by these authors being L. angustifoliarather than239

L. latifolia.240

241

The most interesting finding of this study was that lactones are the volatile242

compounds most markedly contributing to distinguishing Lavandin honey from243

Lavender honey (L. latifolia). Thus, the mean contents in -butyrolactone, pantolactone244

and -nonalactone of Lavandin honey were 57.4% higher than those of Lavender honey.245

-Nonalactone was especially relevant in this respect, as it was identified as a246

component of honey aroma for the first time in this work. This lactone was present at247

significantly higher concentrations in Lavandin honey, which contained it at levels248

above its odour threshold (Table 2). Therefore, -nonalactone can be an effective249


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volatile marker for authenticating Lavandin honey. Figure 1 shows a typical250

chromatogram for Lavandin honey with its peaks clearly identified and numbered.251

One other outstanding compound quantified in Lavandin honey was decanal. This252

compound was recently reported to occur in similar amounts in chestnut and acacia253

honeys (Castro-Vazquez et al.,2010; Plutowska, Chmiel, Dymerski, & Wardencki,254

2011; Radovic et al., 1996). The mean concentration of decanal can also be used to255

authenticate Lavandin honey since, as found in this study, it was twice higher in it than256

in Lavender honey.257

258

Worth special note are the statistical differences in farnesol mean levels between259

honeys fromL. latifoliaand L. angustifolia x latifolia cultivated in the Alcarria region260

(Spain). Thus, the farnesol contents of Lavandin honey were 8 times higher than those261

found in Lavender honey. The fact that this compound was never previously identified262

in honey warrants its use as a floral marker for authenticating Lavandin honey.263

264

One other interesting result was the presence of acetovanillone in substantial265

amounts in Lavandin honey (27 g/kg vsonly 2.7 g/kg in Lavender honey). Such a266

large difference can also be useful with a view to differentiating honeys by botanical267

origin. Acetovanillone has a flavour threshold of 100 ppb (Campo, Ferreira, Escudero,268

Marques, & Cacho, 2005), so it can hardly contribute to Lavandin honey aroma;269

however, the high levels found in Lavandin (L. angustifolia x latifolia) honey can be of270

help for its authentication and differentiation from Lavender honey.271

272

The honeys were also found to contain substantial amounts of 4-273

methoxyacetophenone, a compound involved in the shikimic acid conversion pathway.274

The differences in 4-methoxyacetophenone contents between Lavandin and Lavender275


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(L. latifolia)honeys can also be used to discriminate between the two. The sweet aroma276

of 4-metohoxyacetophenonemay influence the sensory profile of Lavandin honey.277

Dehydrovomifoliol levels in the studied honeys were similar to or greater than278

those in honeys from other blossom sources, particularly eucalyptus eucalyptus (Castro-279

Vzquez et al. 2006; DArcy, Rintoul, Rowland & Blackman, 1997; Guyot,280

Scheirmann, & Collin, 1999; Piasenzotto, Gracco, & Conte, 2003). In any case, such281

levels were significantly higher in Lavandin honey than in Lavender honey.282

283

Finally, several compounds involved in plant metabolism such as linalool,284

hotrienol, camphor,p-cymen-8-ol and 1,8-cineole (isomer I) were all absent or present285

at low levels in Lavandin honey relative to Lavender honey (L. latifolia).286

287

The previous results constitute a breakthrough towards authenticating Lavandin288

honey and can therefore be highly interesting for the beekeeping industry in the Alcarria289

region, which has long sought effective ways to differentiate these two types of290

monofloral honeys.291

292

The chemical data were subjected to stepwise discriminant analysis in order to293

confirm the two groups of honeys on the basis of volatile compounds. Using the data for294

all VOCs and their standard deviations afforded considerable data simplification. Two295

different independent discriminant functions were computed. Table 3 shows the296

individual variables most markedly contributing to discrimination between Lavender (L.297

latifolia) and Lavandin (L. angustifolia x latifolia) honeys, and their coefficients. Cross-298

validation afforded 100% correct assignment of the honey samples.299

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3.2. Palynological profile301

Several pollen types were found in Lavander and Lavandin honeys, although only302

those whose taxa appear in at least 50% of the samples have been represented (Fig. 2).303

In Lavender honeys, the most frequent species associated to Lavandula latifolia304

pollen were Leguminosae, Lonicera, Satureja, Cichorioideae, Asteroideae, Lamiun,305

Plantago and Rosaceae. From a botanical point of view, these species are related to306

wild vegetation.307

308

On the other hand, in Lavandin honeys (L. latifolia), Rubus was the most309

characteristic pollen since it was present in all samples, although Retama,310

Scrophulariaceae, Brassica, Crataegus, Cytisus, Prunus, Rosmarinus officinalis, Salix,311

Trifolium and Thymus pollens were also frequently found. The three first species seem312

to be a reflection of the cropping areas.313

314

The exclusive presence of pollen from Conium and Heliotropium-europaeum in315

Lavandin honeys, and Boraginaceae in Lavender ones (Figure 1) cant be used as316

markers of the type of honey since their presence didnt occur in all the honey samples.317

Despite that, the presence of the cited pollens can be considered an extra tool for honeys318

differentiation.319

320

Although palynologycal data were not enough to differentiate these honey types,321

the joint information of pollen composition with other physico-chemical, volatile and322

sensory markers seem to be a better way to distinguish Lavander (L. latifolia) and323

Lavandin honeys.324

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3.3. Physicochemical and colour parameters326

The physico-chemical parameters of Lavender (Lavandula latifolia) and Lavandin327

(Lavandula angustifolia x latifolia) honeys are shown in Table 4. The mean value,328

standard deviation and maximum and minimum values were shown.329

Water content of these honeys was below 20%, not exceeding the maximum limit330

established by the European Honey Directive., 2001. The same occurs with HMF331

content, which only reflects honey freshness, regardless its botanical origin. The results332

obtained for the studied honeys were below the legal limit established as 40 mg/kg.333

334

After the application of the One-Way ANOVA test to the studied parameters, only335

electrical conductivity and moisture showed significantly statistical differences (p


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3.4. Sensory Differentiation between Lavender and Lavandin Honeys351

352

Lavender (L. latifolia)and Lavandin honeys were compared in sensory terms for353

the first time in this study. A comprehensive sensory description was obtained from354

their aroma and flavour profiles. Figure 3 shows a spiderweb diagram displaying the355

average intensity scores for the different attributes.356

357

The data set was subjected to Students t-test in order to identify significant358

differences between the two types of honey. All sensory impressions except aftertaste359

intensity and quality exhibited significant differences at thep< 0.05 level.360

361

The most relevant differences noted by the judges in Lavandin honey were much362

less strong (p > 0.05) in balsamic, fresh, citric, lavender and floral aromas363

and tastes, in comparison with Lavender honey. The lower concentrations of hotrienol364

and linalool in Lavandin honey (Table 1) account for the significantly lower scores365

given to lavender and floral flavours in Lavandin samples. On the other hand, the366

decreased levels of p-cymen-8-ol and 1,8-cineole which confer fresh, citric and367

balsamic aromas (Amoore and & Venstrom, 1966; http://www.flavornet.org) in368

Lavandin honey may also be associated to its characteristic sensory profile.369

Caramel and jam/peach compote were the specific attributes receiving the370

highest scores in Lavandin honey relative to Lavender honey (L. latifolia). The371

increased caramel scores for Lavandin honey are consistent with its also increased372

amounts of 4-methoxyacetophenone and, mainly, lactones, which are largely associated373

to this attribute (http://www.flavornet.org). In fact, the mean sensory scores confirm that374


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lactone levels largely account for and influence the typical caramel notes of Lavandin375

honey.376

377

Moreover, the presence of -nonalactone, which occurs naturally in peaches and378

apricots(Cullere, Escudero, Cacho, & Ferreira, 2004; Sevenants & Jennings, 1996), is379

consistent with the jam/peach compote aroma and flavour notes best reflecting the380

distinguishing features identified by the assessors in Lavandin floral honey.381

382

The higher intensities of caramel and jam/peach compote notes appear to be383

an unequivocal support for

Lavandin honeys authentication from a sensory point of384

view.385

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4. Conclusions400

The results of this study reveal that the volatile composition and sensory profile of401

Lavandin honey (L. angustifolia x latifolia), and its palynological characteristics last402

one with smaller contribution, are specific enough to allow its floral source to be403

identified and certified. Also, the previous factors afford discriminating between honeys404

from Lavandin and Lavender (L. latifolia), which is a breakthrough for the honey405

research and beekeeping sectors.406

SPE/GC-MS determinations revealed marked differences in volatile profile407

between the two monofloral types of honey. The increased levels of -nonalactone,408

farnesol and acetovanillone, which were for the first time identified in honey here, are409

proposed as Lavandin floral markers. These compounds, together with 1-heptanol,410

decanal, 4-methoxyacetophenone and dehydrovomifoliol, allow Lavandin honey to be411

accurately discriminated from Lavender honey (L. latifolia) in terms of volatile412

composition.413

Although pollen profiles are inadequate to differentiate Lavender (L. latifolia) and414

Lavandin honey, the joint use of palynological data and physicochemical markers may415

be useful to distinguish them.416

Lavandin (L. angustifolia x latifolia) honey and Lavender (L. latifolia) honey are417

also distinguishable in sensory terms. Thus, quince compote and caramel418

sensations, which are closely associated to the volatile composition of Lavandin honey,419

are its key sensory descriptors.420

The proposed volatile and sensory markers provide a powerful authentication tool421

for Lavandin (L. angustifolia x latifolia) honey and a self-obvious marketing advantage422

for the beekeeping sector since it can enable its marketing as monofloral honey with a423

characteristic profile different from that of Lavender honey (L. latifolia).424


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AUTHOR INFORMATION425

Corresponding Author426

427

Phone:+34-926-295300, Fax: +34-926-295318.428

E-mail: [email protected]

430

431

Funding Sources432

The authors wish to thank the professional beekeeper for supplying the samples. This433

study was supported by the Junta de Comunidades de Castilla-La Mancha (05-299/IA-434

47, PAI09-0018-9267), INIA-FEDER funds (RTA2007-00072-C03 and Support435

through Researcher Contract INIA-CCAA).436

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TABLE CAPTIONS

Table 1. Minimum, maximum, and average concentrations (g/Kg) of volatile

compounds determined in Lavender and Lavandin honeys fromLa Alcarriaarea.

Table 2. Aroma descriptors and Olfactive perception threshold (ppb) of some

compounds that statistically differentiate Lavender and Lavandin honeys

Table 3. Standard Canonical Discriminant Functions coefficients of Lavender

(Lavandula latifolia) and Lavandin (Lavandula angustifolia x latifolia) honeys

cultivated in La Alcarria, according to their volatile composition.

Table 4. Physicochemical and colour parameters in the honey types studied, including

the results of the One-Way ANOVA test applied to the mean values.


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ACCEPTED MANUSCRIPT

Table 1. Minimum, maximum, and average concentrations (g/Kg) of volatile

compounds determined in Lavender and Lavandin honeys fromLa Alcarriaarea.

Lavender honeys(L.latifolia)

Lavandin honeys(L. angustifolia x latifolia)

n COMPOUNDS RI Min Max Mean SD Min Max Mean SD1 Hexanal 1044 15.12 26.04 20.18 4.43 6.88 51.74 27.41 15.22

2 3-Penten-2-ol 1136 21.15 102.14 46.46 27.51 19.85 177.74 73.54 50.99

3 Heptanal 1168 1.60 7.52 4.01 2.00 1.94 7.47 4.03 1.67

4 3-Methyl-1-butanol 1214 2.89 24.25 11.82 6.85 6.22 25.84 14.05 6.21

5

2-Methyl-3-(2H)

dihydrofuranone*1245 1.68 14.29 4.39 3.90 tr 14.51 8.98 4.26

6 3-Methyl-3-buten-1-ol 1252 9.37 46.95 23.03 12.37 11.18 47.96 29.22 13.55

7 3-Methoxy-1-butanol* 1260 tr 15.86 4.40 5.92 tr 19.91 7.78 7.72

8 3-Hydroxy-2-butanone 1289 7.26 34.32 13.88 8.04 3.01 56.55 17.55 18.20

9 2-Hexanol 1293 1.74 56.35 18.77 19.76 2.13 72.39 28.02 25.43

10 2-Methyl-2-buten-1-ol 1313 8.10 36.77 16.16 8.41 9.35 54.20 29.17 16.1711 1-Hexanol 1352 20.93 118.54 65.31 37.43 2.25 325.91 131.65 112.03

12 1-Hydroxy-2-butanone 1363 0.48 13.31 3.96 3.77 tr 9.29 3.76 3.39

13 Nonanal 1388 9.68 67.38 26.10 19.41 7.40 33.26 21.66 8.39

14 2-Butoxyethanol 1391 tr 28.52 9.71 9.26 tr 16.16 8.89 6.39

15 Ethyl octanoate 1430 tr 10.38 3.10 3.56 tr 8.32 2.72 2.81

16 (Z)-Linalool oxide 1438 10.10 52.40 20.44 13.22 4.20 27.40 11.93 7.58

17 1-Heptanol 1445 2.31 10.14 6.99a 3.12 9.25 34.85 21.63

b 11.80

18 Acetic acid 1450 3.52 20.99 8.32 5.33 2.46 14.97 8.22 3.94

19 (E)-Linalool oxide 1459 3.30 31.95 8.70 9.20 2.30 38.18 9.74 11.10

20 Furfural 1470 1.48 20.82 8.16 6.94 3.37 22.77 12.49 6.09

21 2-Ethyl-1-hexanol 1478 3.83 31.21 11.57 8.55 4.07 17.94 10.22 5.04

22 -Campholene aldehyde* 1483 1.40 5.74 2.40 1.33 1.48 4.48 2.97 1.06

23 Decanal 1489 2.90 13.28 8.17a 4.19 12.57 29.51 17.61

b 5.19

24 1-(2-Furanyl)ethanone 1508 2.05 23.90 5.91 6.92 2.43 5.82 3.81 0.94

25 Camphor 1517 tr 1.51 0.85a 0.58 n.d. n.d. n.d.

b

26 Benzaldehyde 1532 7.30 50.51 21.95 12.86 9.62 45.88 26.05 9.47

27 Propanoic acid 1544 3.39 12.07 6.37 2.63 1.12 12.46 6.23 3.70

28 Linalool 1553 11.01 28.23 17.54a

5.80 0.87 15.15 7.05b

4.33

29 Octanol 1564 1.91 10.90 4.74 3.19 3.67 10.61 6.49 2.48

30 2-Methylpropanoic acid 1576 3.83 26.02 9.05 6.75 5.82 21.82 11.19 5.32

31 Hotrienol * 1616 27.96 219.87 113.31

a

66.61 7.28 111.56 53.57

b

34.9632 -Valerolactone 1621 3.55 18.32 7.69

4.29 2.19 28.15 7.63

7.61

33 Butanoic acid 1627 4.84 133.18 31.58 39.43 10.50 197.41 74.66 78.19

34 Ethyl decanoate 1632 n.d. 436.45 59.47 143.04 n.d. 9.11 2.57 3.52

35 -Butyrolactone 1646 2.36 12.55 9.11a 3.65 13.57 36.04 19.29

b 7.28

36 Phenylacetaldehyde 1661 19.53 142.58 85.74 48.68 51.26 243.72 107.68 78.82

37 1-Nonanol 1670 2.79 32.69 7.79 9.43 n.d. 12.09 3.45 4.53


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Table 1. (Continued)

Lavender honeys

(L.latifolia)Lavandin honeys

(L. angustifolia x latifolia)

COMPOUNDS RI Min Max Mean SD Min Max Mean SD

38 Furfuryl alcohol 1679 10.03 27.61 21.71 12.48 2.33 44.20 17.83 13.90

39 3-Methylbutanoic acid 1682 7.91 71.92 20.71 19.70 11.16 41.96 25.86 10.98

40 Ketoisophorone 1686 2.34 12.75 6.17 3.77 1.68 9.45 4.61 2.48

41 -Terpineol 1691 0.45 10.19 2.95 2.88 n.d. 2.87 1.13 1.01

42 Epoxylinalool (isomer I) * 1732 2.15 26.09 13.27 10.92 4.02 86.99 28.62 26.49

43 Epoxylinalool (isomer II) * 1745 0.73 15.02 5.63a 5.51 2.55 57.12 17.07b 19.01

44 Myrtenol * 1796 0.39 15.96 2.96 4.96 n.d. 14.16 3.37 4.21

45 Hexanoic acid 1857 15.13 97.25 52.42 29.02 32.27 126.50 81.08 31.15

46 p-Cymen-8-ol * 1878 0.85 12.40 4.79a 4.29 n.d. 4.29 0.73b 1.53

47 2-Hydroxycineole * 1882 0.75 12.71 4.52 3.86 n.d. 5.23 2.16 1.84

48 Mequinol *

1891 n.d. 11.37 4.20 3.14 1.54 26.51 6.92 7.28

49 Benzylalcohol 1903 11.47 186.83 51.78 53.79 12.01 143.74 58.55 44.31

50 Nonadecane 1921 n.d. n.d. n.d. n.d. 16.88 4.65 6.68

51 1,8-Cineole (isomer I) 1929 tr 18.16 10.27a

3.92 n.d. n.d. n.d.b

52 2-Phenylethanol 1940 36.68 374.94 107.83 104.65 19.29 268.57 79.12 71.46

53

2,6-Dimethyl-3,7-octadien-2,6-

diol * 1968189.46 1382.46 526.33 381.17 53.63 758.65 390.36 245.57

54 Heptanoic acid 1972 4.54 21.07 10.48 6.13 2.76 27.20 16.61 9.10

55 (E)-3-Hexenoic acid 1981 n.d. 116.10 26.29 35.83 4.24 49.81 19.81 15.41

56 6-Methyl-2-methoxypyrazine 1996 6.08 375.54 59.31 119.14 4.45 48.53 12.03 12.95

57 Methylfurancarboxylate 2027 17.43 443.10 77.00 138.01 17.45 54.58 31.15 10.74

58 Pantolactone 2031 11.98 33.92 24.57a

7.52 34.87 62.40 50.07b

8.81

59 Methylfurancarboxylate 2036 tr tr tr n.d. 38.23 10.16 14.52

60 -Nonalactone

2041 7.23 16.30 12.33a

3.20 30.06 63.28 38.97b

10.51

61 Octanoic acid 2051 20.22 52.30 29.13 11.34 16.00 68.45 36.48 16.70

62 p-Cresol 2068 1.95 9.88 4.51 2.31 1.75 5.64 3.30 1.37

63 p-Mentha-1(7),2-dien-8-ol * 2071 n.d. 4.92 1.34 1.77 n.d. 9.47 2.94 3.67

64 Nonanoic acid 2154 9.87 170.87 45.29 54.32 10.51 53.21 33.19 14.00

65 Eugenol 2144 4.07 44.67 12.63 12.28 2.57 12.46 8.49 3.43

66 4-Methoxyacetophenone* 2168 17.05 36.25 27.57a 5.99 40.20 109.30 64.85b 25.07

67 8-Nonen-2-one 2183 tr 19.22 8.66 5.95 7.22 34.73 16.48 12.67

68 1,8-Cineole (isomer II) 2228 18.22 35.66 25.04 5.76 16.29 40.28 25.17 7.94

69 1-Hydroxylinalool* 2269 20.23 148.90 57.00 41.26 17.05 106.33 58.64 32.80

70 trans-Sobrerol* 2276 11.02 102.30 40.79 41.34 4.02 139.52 42.03 46.35

71 Triethylenglycol 2283 127.02 2489.44 634.44 730.32 150.82 1653.92 643.02 483.03

72 Hexadecanoic acid > 2280 n.d. 1120.68 263.08 345.09 62.46 337.43 175.23 104.39

73 Benzoic acid 36.25 481.18 128.91 138.10 42.75 477.24 192.18 160.59

74 Coumarin 23.82 355.02 142.14 103.59 n.d. 230.06 100.35 79.72

75 Pentacosane n.d. 110.60 12.29 36.87 n.d. 174.68 23.85 56.66


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Table 1. (Continued)

Lavender honeys

(L.latifolia)Lavandin honeys

(L. angustifolia x latifolia)

COMPOUNDS RI Min Max Mean SD Min Max Mean SD

76 Hydroxymethylfurfural > 2280 n.d. 3713.08 618.59 1181.38 34.34 246.69 149.29 78.21

77 Farnesol *

n.d. 8.22

3.05a

3.25 9.33 59.01 24.83b 16.77

78 Benzenacetic acid 71.71 214.20 126.27 44.92 63.69 332.41 154.51 78.42

79 Acetovanillone

n.d. 6.21

2.76a

2.81 10.02 73.22 27.09b 19.60

80 Dehydrovomifoliol* 21.30 66.01

40.00a 16.75 72.55 145.28 113.23

b 29.60

RI: Retention indices based on polyethylene glycol capillary column BP-21 (50 m x

0.32 mm x 0.25 mm of film thickness

*Compounds tentatively identified using Wiley and NBS75KLibraries

Tr:Traces; n.d.: not detected compoundsa,b: Different letters in the same row indicate statistical differences (P


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Table 2. Aroma descriptors and Olfactive perception threshold (ppb) of some

compounds that statistically differentiate Lavender and Lavandin honeys

Compounds IdentificationAroma descriptors

a

Olfactiveperception

threshold

(ppb)

Lavenderhoneys

Mean Con.(ppb)

LavandinhoneysMean Con.

(ppb)

Decanal AB soap, orange peel, tallow(1)

3(5)

8.17 17.61

Camphor AB Camphor(1)

unknown 0.85 n.d.

Linalool AB floral, fruity(1)

25(6)

17.54 7.05

Hotrienol B floral, fresh, fruity(1)

110(7)

113.31 53.57

p-Cymen-8-ol AB Musty(2)

unknown 4.79 0.73

1,8-Cineole AB eucalyptus, fresh, balsamic(3)

12(8)

10.27 n.d.

Pantolactone AB cotton candy(1)

unknown 24.53 50.07

-Nonalactone AB coconut, candy, peach

(4)

30

(4)

12.33 38.974-Methoxyacetophenone B sweet, anisic, coumarin, fruity

(1) unknown 27.57 64.85

Farnesol B flower, oil(1)

20(9)

3.05 24.83

The identification of the volatile compounds is indicated by the following:

A: mass spectrum and retention time consistent with those of an authentic standard;

B structural proposals are given on the basis of mass spectral data (NBS75K and Wiley A )

The reference from which the olfactive perception threshold has been taken is taken is given in

parentheses: (1) http://www.flavornet.org; (2) Quian &Wang, 2005; (3) Faria, Boido, Carrau,

Versini, & Dellacassa, 2005; (4)Ferreira, Lopez, & Cacho, 2000; (5) Ahmed, Dennison,

Dougherty, & Shaw 1978; (6) Aznar, Lpez, Cacho, & Ferreira, 2003; (7) Riberau-Gayon,

Glories, Maujean, &Dubourdieu, 2000; (8) Amoore and & Venstrom, 1966; (9) Ohloff, 1978.


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Table 3. Standard Canonical Discriminant Functions coefficients of Lavender

(Lavandula latifolia) and Lavandin (Lavandula angustifolia x latifolia) honeys

cultivated in La Alcarria, according to their volatile composition.

Coefficientsattached toFunction 1

Coefficientsattached toFunction 2

Linalool 1.853 -3.714

Hexanoic acid -0.135 0.388

p-Cymen-8-ol 1.156 -2.667

-Nonalactone -0.453 1.604

p-Mentha-1(7),2-dien-8-ol 0.661 -3.486

Dehydrovomifoliol -0.058 0.555

(Constant) -12.563 -59.795


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Table 4. Physicochemical and colour parameters in the honey types studied, including

the results of the One-Way ANOVA test applied to the mean values.

Lavender honeys(L.latifolia)

Lavandin honeys(L. angustifolia x latifolia)

Mean SD(min - max)

Mean SD(min - max)

Moisture (%)* 16.62 1.60 15.43 0.67(14.53 - 19.40) (14.45 - 16.25)

HMF (mg/kg) 4.60 4.97 2.28 2.69(0.15 - 16.02) (0.15 - 9.28)

0.22 0.07 0.29 0.05Electrical conductivity (mS/cm)* (0.13 - 0.32) (0.21 - 0.39)

pH 3.88 0.25 4.02 0.13(3.64 - 4.46) (3.91 - 4.26)

Free acidity (meq/kg) 19.34 4.17 20.73 3.25(14.40 - 25.80) (15.25 - 25.00)

Pfund colour (mm of Pfund) 40.06 12.91 45.41 15.48(24.21 61.48) (19.22 67.84)

C.I.E. L a*b* colour

X10 0.42 0.03 0.43 0.04(0.38 - 0.47) (0.37 - 0.48)

Y10 0.43 0.02 0.44 0.04(0.40 - 0.47) (0.39 - 0.54)

Z10 0.15 0.05 0.14 0.06(0.06 - 0.22) (0.06 - 0.25)

2.08 3.37 4.59 3.48a*10(chromaticity + red / green) (-2.03 - 7.28) (-0.17 - 12.22)

52.75 14.30 54.33 14.95b*10(chromaticity + yellow / blue) (36.83 - 80.53) (27.90 - 70.34)

C*ab (chroma) 52.86 14.40 54.58 15.09(36.88 - 80.85) (27.90 - 70.53)

hab.10 (hue angle) 88.45 3.29 85.70 3.16(84.83 - 93.15) (79.02 - 90.32)

L*10 (lightness) 80.93 4.99 77.39 8.49(74.70 - 88.96) (56.94 - 87.84)

*Significant difference between groupsp< 0.05SD: standard deviation.


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1

FIGURE CAPTIONS

Figure 1. Typical chromatogram obtained from Lavandin honey samples by GC-MS

Figure 2. Comparison between percentage of pollen of the more abundant botanical

species that occur in lavender and Lavandin honeys

Figure 3. Descriptive Sensory Analysis of Lavandin and Lavender honeys. Mean

scores of eight judges (two replicates) are shown.


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ACCEPTED MANUSCRIPT

2

Fig. 1.Typical chromatogram obtained from Spanish Lavandin honey samples (Lavandula angustifolia x latifolia

2: 3-Penten-2-ol6: 3-Methyl-3-buten-1-ol

8: 3-Hydroxy-2-butanone9: 2-Hexanol

10: 2-Methyl-2-buten-1-ol

11: 1-Hexanol13: Nonanal

17: 1-Heptanol

21: 2-Ethyl-1-hexanol

23: Decanal

26: Benzaldehyde28: Linalool31: Hotrienol

35: -Butyrolactone

38: Furfuryl alcohol

42: Epoxylinalool (isomer I)

43: Epoxylinalool (isomer II)45: Hexanoic acid

49: Benzylalcohol

52: 2-Phenylethanol53: 2,6-Dimethyl-3,7-octadien-2,6-diol58: Pantolactone

60: -Nonalactone

61: Octanoic acid

65: Eugenol

66: 4-Methoxyacetophenone68: 1,8-Cineole (isomer II)

IS

2

69108

11

1317

21 2623 28

31

35 38 4243

49

45 52

53

5860 61

65

6669

71

70

68 74

73

77


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3

Fig. 2. Comparison between percentage of pollen of the more abundant botanical

species that occur in Lavender and Lavandin honeys

0

50

100

Lavan

du

lalatifolia

Me

dik

.

Legum

inosae

Lon

icera

Cichorio

ideae

Sa

ture

jaspp.

Ru

bus

Re

tamaspp.

Scrop

hu

lariaceae

Con

ium

As

tero

idae

Borag

inaceae

Brass

icasp.

Cra

taegus

t.

Cruc

iferae

Cy

tisus

t.

Ec

hium

spp.

He

lio

trop

ium

europaeum

Lam

ium

Lo

tus

t.

Plan

tagospp.

Prunus

t.

Rosaceae

Rosmarinuso

fficina

lis

L.

Sa

lixspp.

Tri

folium

t.

Thymus

Lavender honeys (L.latifolia) Lavandin honeys (L. angustifolia-L.latifolia)


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4

Fig. 3. Descriptive Sensory Analysis of Lavandin and Lavender honeys. Mean scores

of eight judges (two replicates) are shown.

Lavender honey (Lavandula latifolia) Lavandin honey (L. angustifolia-L. latifolia)

Odor Evaluation

0,0

1,0

2,0

3,0

4,0

5,0

6,0

7,0

Balsamic

Fresh

Lavander

CitricJam/Peach compote

Floral

Caramel

Flavor evaluation

0,0

2,0

4,0

6,0

8,0

10,0

Sweet taste

Citric taste

Caramel taste

Balsamic taste

Jam/Peach compote taste

Floral taste


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Highlights

- Lavandin honey (Lavandula angustifolia x latifolia) had been characterized for the first

time.

- Lavandin monofloral honeys showed botanical markers that allow typify this floral

origin.

- -Nonalactone, farnesol and acetovanillone are proposed as volatile markers for

Lavandin honey authentication.

- The sensory profile of Lavandin honey was characterized by intensities of caramel and

peach compote notes.

- Results provide a complete differentiation between Lavender and Lavandin honeys.

aroma in honey article in press.pdf

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