Dr Peter Brooks amp Daniel Meloncelli USC Honey Research Lab pbrooksuscedu au 61 7 5430 2828 Some Basic Facts Compounds collected from Nectar and Honey Dew by the Bees are responsible for the properties of honey ID: 549906
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Slide1
Chromatographic Fingerprints of Honeys
Dr Peter Brooks &
Daniel MeloncelliUSC Honey Research Labpbrooks@usc.edu .au61 7 5430 2828Slide2
Some Basic Facts
Compounds collected from Nectar and Honey Dew by the Bees
are responsible for the properties of honey, eg. Taste and Aroma.Compounds unique to one Nectar or Honey Dew, are marker compounds of the honey source.
There is no such thing as 100%
monofloral
Honey.
Question?
Can chromatography be used to authenticate honey type?Slide3
Phenolic fingerprints by HPLC
Authenticating Floral Source by RP-HPLC
New Zealand
L.
scoparium
honey
Tasmanian
L.
scoparium
honey0.8 min: Kojic acid4.3 min Leptosperin8.3 min: 4-methoxyphenyllactic acid10.3 min: Methyl syringate
Lep
KA
MSR
Lep
MSR
4MPL
KASlide4
Phenolic
fingerprints by HPLC
NZ
L.
scoparium
honey
NZ
K.
ericoides
honey
Leptosperin
Methyl syringate
Methyl syringate
KA
KA
4MPLSlide5
Phenolic
fingerprints by HPLC
NZ
Pohutukawa
honey
NZ Rata honeySlide6
Phenolic
fingerprints by HPLC
NZ Clover
honey
NZ Kamahi honeySlide7
Phenolic
fingerprints by HPLC
NZ Rewarewa
honey
NZ Blue Borage honeySlide8
Phenolic
fingerprints by HPLC
NZ Thyme
honey
NZ
Tarawi
honeySlide9
Phenolic
fingerprints by HPLC
NZ Honey Dew
honey
NZ
Ulmo
Chile honeySlide10
Phenolic fingerprints by HPLC
L.
laevigatum
honey
(Tas.)
L. whitei
honey
(NSW)
Methyl syringate
Lep
Lep
KA
KA
Methyl
syringateSlide11
Identifying “Mislabelled” honeys
Reverse-Phase High Performance Liquid Chromatography (RP-HPLC-PDA)
L. polygalifolium
honey
“Fake”
L.
polygalifolium
honey
Leptosperin
Methyl syringateSlide12
DHA, MGO and HMF
NZ
L. scoparium honey
NZ
K.
ericoides
honey
DHA
(4623 ppm)
DHA
(103 ppm)
HMF
(5 ppm)
HMF
(9 ppm)
MGO
(547 ppm)
MGO
(25 ppm)Slide13
Marker Compounds in Honey
)
Honey sample
Mean (SE) concentration of compounds (µg/g, ppm)
DHA
HMF
MGO
KJA
LEP
3PA
2MB
4MPL
MSR
L. scoparium (TAS)
1169 (83)
4 (1)
237 (31)
125 (17)
1051 (105)
906 (187)
14 (14)
623 (71)
97 (12)
L. scoparium (NZ)
2665 (398)
10 (3)
519 (91)
252 (24)
681 (127)
1199 (165)
70 (18)
41 (41)
87 (18)
L. polygalifolium
1475 (314)
20 (4)
543 (91)
197 (28)
399 (80)
665 (86)
183 (33)
5 (4)
97 (15)
L. whitei
2823 (623)
13 (2)
518 (88)
156 (20)
74 (18)
223 (35)
76 (20)
4 (4)
148 (51)
L. liversidgei
824 (165)
35 (16)
451 (89)
213 (47)
175 (29)
306 (55)
48 (37)
13 (4)
61 (11)
L. laevigatum
78 (42)
68 (61)
40 (19)
107 (57)
20 (20)
1509 (622)
117 (71)
nd
20 (20)
Kunzea ericoides
64 (19)
5 (1)
6 (3)
85 (29)
15 (4)
1265 (192)
nd
1128 (182)
222 (30)
Pohutukawa
36 (13)
30 (14)
2 (2)
38 (38)
nd
nd
nd
7 (7)
3 (3)
Rata
40 (6)
17 (4)
4 (4)
54 (10)
nd
nd
nd
20 (11)
nd
Kamahi
12 (6)
5 (1)
nd
11 (11)
nd
nd
nd
9 (9)
nd
Clover
43 (21)
11 (5)
2 (2)
25 (18)
9 (5)
nd
nd
nd
4 (2)
Rewarewa
80 (28)
20 (7)
10 (4)
197 (44)
48 (12)
104 (36)
nd
37 (18)
21 (7)
Blue Borage
*13-15
*10-32
nd
*70-108
*7-13
nd
nd
*0-59
*2-9
Thyme
*3-12
*35-36
nd
nd
*0-4
nd
nd
nd
nd
Tawari
*4-9
*2-7
nd
nd
nd
nd
nd
nd
ndSlide14
Can Manuka purity be fingerprinted
?Question ?
If a mature high purity Manuka fromNew Zealand is approx. DHA 2600ppm; MGO 520 ppm; and Leptosperin 680 ppm,
Then is a 50% Manuka approx. DHA > 1200 ppm; MGO > 250ppm;
Leptosperin
> 300 ppm ?
And is a 20% Manuka; DHA > 500 ppm; MGO > 100 ppm;
Leptosperin
> 120 ppm ?Slide15
GC-MS of Volatiles in L.
scoparium and K. ericoidesSlide16
GC-MS of Volatiles in L.
scoparium and K. ericoides
Compound in trace
Mean
SD
range
Mean
SD
range
Dihydroxyacetone (DHA),
bisTMS
ether
162.3
89.9
21.6-297.5
1.3
1.3
0-3.5
4-Methoxy-benzaldehyde
0.0
0.0
0
1.5
0.9
0.3-3.2
2-Methoxyacetophenone
6.6
1.4
3.7-8.2
0.0
0.0
0
Benzene acetic acid, TMS ester
0.0
0.0
0
2.6
1.1
1.3-4.4
2-Methyl-4-propylphenol
0.0
0.0
0
11.6
13.3
2.2-41.3
2-Methoxybenzoic acid TMS (STD)
33.1
27.5
6.1-90.0
0.0
0.0
0
Mandelic
acid TMS ether TMS ester
0.0
0.0
0
3.1
0.7
1.7-3.6
4-Methoxy-benzoic acid, TMS ester
0.0
0.0
0
7.0
3.9
2.0-13.5
3-(3,4-Dimethoxyphenyl)-2-propenoic acid, methyl ester
0.0
0.0
0
6.2
3.5
1.5-10.4
4-Methoxy-benzeneacetic acid, TMS ester
0.0
0.0
0
5.4
2.9
3.1-10.0
Myrtenoic
acid, TMS ester
0.0
0.0
0
32.6
6.4
24.2-40.8
3,5-Dimethoxy-benzoic acid, methyl ester
2.7
2.1
0.9-7.4
0.0
0.0
0
3-Phenyl-3-TMSoxypropanoic acid TMS ester
7.0
6.3
2.4-20.3
0.0
0.0
0
4-Methoxymandelic acid, TMS ether TMS ester
0.0
0.0
0
31.1
11.4
16.4-42.8
3,4-Dimethoxy-benzoic acid, TMS ester 3.33.60.8-11.50.00.00o-Methoxymandelic acid TMS 18.510.611.0-38.80.00.003,4-Dimethoxymandelic acid, di-TMS 27.917.411.6-64.20.00.00Caffeic acid (TMS) 0.00.002.32.60.4-8.2
L.
s
coparium
K.
ericoides