KHOA HOC CONG NGHE
XAC DfiVH CHi TH! CHOIXIG OXI HOA CUA MAT OIVG DE TRUY XUAT SAlXi PHAIVI C H I D A | \ I DIA LY MAT OIVG
DAC HA CAO IMGUYEIM D A DOIMG VAIM, HA GIAIMG
Le Quang Trung\ Kun Bich Nguy^t^, Cam Thi Hing^, Nguyen Dinh Anh^, U m Thu Hang^, Vu Thiiy Nga^, Nguyen Thi Tliiiy H6a^, Phgm Mmh Giang^
TOMTAT
San pham chi dan dia ly mat ong bac ha Cao nguyen da Dong Van, tinh Ha Giang (MBH) co gia ban cao nhat tren thi tnrdng trong nuoc hien nay, nen kho tranh khoi gian lan thuong mai bang each tron \hn MBH vdi cac loai mat ong co san lucmg cao va gia ban thap nhu mat keo tai tugng (M0R2) nham tang lgi nhuan.
De giii thuong hieu MBH, trong nghien ciiu nay, ham lupng DL-S-phenyllacUc axit (3-PA) trong mat ong phan tich tiieo phuang phap LC-MS/MS, Fe^* hinh thanh tiieo phep tiiii FRAP va %DPPH da phan ling theo phep thii DPPH dugc phan tich tren cac loai nen mau mat ong, bao gom cac mau MBH nguyen chat, cac miu M0R2 va 4 loai mat pha tron giua M0R2 va MBH tiieo cac ty le 1:9, 1:4, 3:7 va 1:1. Gia tri cua 3-PA, Fe^' va DPPH ciia MBH nguyen chat deu thap hon vdi tiiong ke tin cay (0,0001 <P<0,05) so voi M0R2 va 4 loai nen mAu pha tron giOra MBH vdi M0R2 a cac ty le khac nhau. Gia tri dac thu, do lech chuan va dao dong ve gia tri ciia m6i chat giua cac loai ndn mlu cho thay ham lugng 3-PA va Fe^' co the su dung nhu cac chi till de phan biet MBH nguyen chat voi MBH bi pha trpn voi >20% M0R2, trong khi %DPPH co tiie truy xuat MBH bi pha tron vol >30% M0R2. Cu tfie, MBH nguyen chat co ham lupng 3-PA <1,36 mg/kg.
Fe^"<263,09 mg/kg, va DPPH<16,54%, trong khi MBH trpn >20% M0R2 co ham lugng 3-PA >1,64 mg/kg, Fe^'>308,23 mg/kg va MBH trpn >30%, M0R2 co ham lugng 3-PA >2,10 mg/kg, Fe^'>348,07 mgAg va DPPH>22,00%. Ket qua va phuong phap trong nghien cmi nay la ca sa khoa hpc de ap dung truy xuat ngudn goc nhIm giam thidu gian lan thuong mai cac loai mat ong a nude ta, trong do co san pham chi din dia ly mat ong bac ha Cao nguyen da Dong Van cua tinh Ha Giang.
Ttr khda: Truy xuat nguon goc, DL-S-phenyllactic axit, DPPH, FRAP, m$t ong bac ha Cao nguyen da Dong Van, Ha Giang.
I.MdBAU scavenging effect assay) da va dang duge ap dung nhu nhirng ehi thi phd bidn va hieu qua de truy xuat Gian lan temgmm^trsn thi I r a o n g m ^ ong ^ ^ . ^ ^.^ ^^^^^ ^^^ ^^^ ^^^ ^ . ^ ^ . ^^
dang la van de toan cau (McDonald etal., 2018). Su (jy^p^^^y ^f^_ 2018; Colucci etal.. 2016).
phat trien ciia ky thuat phan tich trong nhirng nam
gin day da dua ra mot sd chi thi hoa hoc dac hifeu d« San phSm chi dan dia ly mat ong bac ha Cao truy xu& ngudn gde nhim giam thifo gian lan "fniyta da Dong Van (goi tat la mat ong bac ha, thuong mai mat ong. Trong sd dd, ham luong cac MBH) chu ydu do cac giong ong mat Apis cerana thu chat chdng oxi hoa nhu axit DL-3 phenyllactic (3-PA) ve va che bien ttr mat hoa cay bac ha {Elsholtzia xac dinh bang phuong phap LC/MS-MS, Fe^* hinh ^pp.) moc tu nhien d cao nguyen. Vao miia khai thac, thanh sau phep thu FRAP (ferric reducing MBH cd gia 400-500 nghin dong/kg, cao nhat d thi anUoxidant power assay) va %DPPH (2,2-diphenyl-l- * " * " « ™ « ^ hien nay, nen de xay ra viec trpn mat picrylhydrazyl) da phan ling do mat ong sau phep ™8. ^d san luong cao va gia ban thap nhu keo toi thu DPPH (1,1 diphenyl-2-piciylhydrazyl -radical- t"™« (M0R2) vao MBH nguyen chat de tang loi nhuan. M0R2 la mat la, khi mdi khai thac cd mau sac tuong ddi gidng vdi MBH d mot sd vimg ciia cao nguyen, khong cd miu dac tnmg nen khong the phan ' Vien An loan Thirc phim, Cong ty CP Chiing nhan va (jjjj j,j„g ^am quan glQa MBH nguyen chat va MBH
" " bi pha trpn ttr 20-30%. 6 nude ta, ham lupng 3^PA trong mat ong, ham luong Fe^* tao thanh sau phep Giam djnh VinaCen
'Trung lam Ki^m nghiem linh Ha Giang, So Y te Ha Giang
^ Sa Khoa hoc va Cong nghe Ha Giang
thu FRAP va %DPPH hinh thanh sau phep thu DPPH da duoe chung minh la eae ehi thi de phan biet MBH vdi M0R2 nguyen chat (Le Quang Trung et al, 2018), nhung chira cd cdng bd nao sii dung eac ehi thi nay de phan biet MBH nguyen chat voi mat ong pha tron giua MBH vdi M0R2.
Trong nghien cuu nay, ham lugng eae chat ehdng oxi hda 3-PA, Fe-* va %DPPH duge xae dinh va so sanh giua cac nen mlu MBH nguyen chat, M0R2 nguyen ehat va cac nen mSu mat ong pha tron giiia M0R2 voi MBH nhSm dtra ra cae chi thj de phan biet giua MBH nguyen chat va MBH hi pha tron vdi M0R2 a ty le khac nhau. Ket qua nghien cuu cd th^
ap dung de kiem scat chat lugng nham giam thieu gian lan thuong mai ddi vdi san pham ehi dan dia ly mat ong bac ha Cao nguyen da Dong Van.
2 . NGUYEN UEU VA PHUDNG PHAP 2.1. Nguyen li^u
Miu mat ong nguyen chat (12 mau) bao gdm 9 miu mat ong bac ha (MBH1-MBH9) duoe thu trong miia mat 2017-2018 d 9 trai ong cua eae huyen Quan Ba, Yen Minh, Dong Van va Meo Vac eua Cao Bang 1. Ky hieu ckc n^n mfiu n i | t ong nguyen eh^t v^
c^e ty le pha
nguyen da Dong Van; 3 mdu mat ong keo tai tugng (M0R2.1-M0R2.3) thu b Phu Thg, Hoa Bmh va Nghe An tir thang 6-9 nam 2017 va 1 mhi mat trdn 3 loai keo b eac vimg thu mhi theo ty le 1:1:1 (MDCIO).
Cae mhi mat tron (MIX1-MIX9) giiia mat keo bdn hgp (MKIO) vdi timg mlu MBH nguyen chat duoc chuan bi theo cae ty le ve khdi luong ttr 1:1 (Mcc SO- SO), 3:7 (Mix30-70), 1:4 (Mix2O80) den 1:9 (Mbc 10- 90). Trong dd, M K l pha h-dn giira MKIO vdi MBH Quan Ba thu (MBHl); MIX2 va MIX3 giua MIXIO vdi MBH Yen Minh trai 1 va 2 (MBH2, MBH3);
MIX4, MIX 5, MIX 6 giua MIXIO vdi MBH Dong Van trai 1, 2 va 3 (MBH4, MBH5, MBH6); MIX7, MIX 8, MIX 9 giua MDCIO vdi MBH Meo Vac trai 1, 2 va 3 (MBH7, MBH8, MBH9). Cae mhi mat ong ddng deu ve thuy phan (20-21%) va ham lugng HMF (22- 25mgAg) diroe dung trong Ip thuy tinh day nip kin va bao quan b dieu kien nhiet do <0''C den khi phan tieh. Ky hieu mau MBH, M0R2 nguyen chat, cac mlu MIX va cong thiie ciing nhu ty le pha trdn eac MIX giira MBH va MIXIO duge th^ hien ehi tiet b bang 1.
mat pha trdn gifta mat bsic h i vdi m^t keo tai tuong a khicnhau
STP 1 2 3 4 5 6 7 8 9 10
Mat ong nguyen chat Mat ong
bac ha (MBH) MBHl MBH2 MBH3 MBH4 MBH5 MBH6 MBH7 MBH8 MBH9
Mat ong keo tai tuong
(M0R2) M0R2.1 MOR2.2 MOR2.3
Mat ong pha tron (MIX) Ten MIX
M K l MIX2 M1X3 MIX4 MIX5 MIX6 MIX7 MIX8 M1X9 MKIO
Cong thiic pha MIX MKIO+MBHI MlX10tMBH2 MKIO+MBHS MIX10+MBH4 M1X10+MBH5 MIX10+MBH6 MIX10+MBH7 MIX10+MBH8 MIX10-IMBH9 MOR2.1-1-MOR2.2 +MOR2.3
Mbc SO- SO 1:1 1:1 1:1 1:1 1:1 1:1 1:1 1:1 1:1 1:1:1
Ty le pha tr6n Mix 30-
70 3:7 3:7 3:7 3:7 3:7 3:7 3:7 3:7 3:7 1:1:1
Mix 20- 80 1:4 1:4 1:4 1:4 1:4 1:4 1:4 1:4 1:4 1:1:1
Mix lo- go 1:9 1:9 1:9 1:9 1:9 1:9 1:9 1:9 1:9 1:1:1 Chii thich: MBH: mat bac ha nguyen chat; MBH1-MBH9: MBH a9a-ai ong khac nhau; MOS2: mat keo tai hrang nguyen chat; M0R2.1' M0R2.3: M0R2 a 3 trai ong khac nhau; MIXIO: hSn hop M0R2.1- M0R2.3:
MIX1-MIX9: mat ong tron giua MIXIO vai timg miu MBH a cac tyle khac nhau.
Cac hda chat de thu nghiem tren cac loai ndn diing cho HPLC nhu axit formic, methanol, mau mat ong dui^c cung cElp td hang Sigma va acetonitrile (ACN) ... de phan tich ham lupng 3-PA Merck. Cac hda chat chu yeu bao gdm: 1) chat chuan trong mat ong; 2) cac hda chit nhu ethanol, dung DL 3-phenyllatic axit (3-PA) va cac hda chat khac dich mudi Fe^*, dung dich mudi natri, o-
70 NONG NGHIEP VA PHAT TRIEN NONG THON - KY 2 - THANG 11/2018
KHOA HOC CONG NGHE phenanthroUne, ethanol, DPPH (l,l-diphenyl-2-
picrylhydrazyl)... de thu nghiem kha nang ehdng oxi hda tong sd cua mat ong.
2.2. Phuong phlp
Ham lugng 3-PA (mg/kg mat ong) trong eae mlu mat ong dupe xae dinh bang phuong phap UPLC-MS/MS theo Trautvetter v l ddng tac gia (2009) va duge tinh theo cdng thue:
|[(Si*N)/S2l*V)}/M. Trong dd. Si la dien tich peak ciia chat phan tich trong mlu mat ong tren sic ky do;
N - nong do mlu cd them chuan; S^ - dien tich peak tren sic ky do eua chat phan tich trong mlu eo them chuan; V - the tich pha loang mlu; M - khdi lugng mlu.
Kha nang chdng oxi hda tong so cua mat ong dupe danh gia bing phep thu FRAP va DPPH theo Dobre va dong tac gia (2010). Trong phep thu FRAP, ehi so ehdng oxi hda tong duge xae dinh dua vao ham luong Fe^* tao tiianh theo cong thiic: X = [A*V*56* 10^]/[A(l%)*m* 100*284], trong do, X la ham lugng Fe^"^ (mg/kg mat ong); A- D6 h^p thu quang eua mlu thu; A(l%) - Do hap thu quang eua dung dieh mudi Morh 1%; V - Th^ tich chiet mlu (ml); m - Khdi lugng mlu (g); 56 - Khdi lugng phan tu ciia Fe; va 284 - Khdi lugng phan tu cua mudi Morh (NH4)2Fe(S04)2. Trong phep thur DPPH, chi sd chdng oxi hda tdng dugc xac dinh bing %DPPH da
phan ung (K) theo cong thire: X=100 -Xi vol Xj la
%DPPH eon lai ti-ong mlu sau phan ling, duge tinh tiieo cong thuc: X^ {[(A^ + 2,58)*10-^]/((A, + 2,58)*10-^]}*100. Trong do. A, la do h i p thu quang cua dung dich mlu va A„ - do hap thu quang cua dung dieh DPPH. Moi chi tieu cho tung mlu mat duge phan tich vdi 3 lan lap lai. Ham lugng 3-PA trong mat ong, Fe^* va phan tram DPPH sau khi thu nghiem kha nang chdng oxi hoa tong sd cua mdi mlu mat ong dupe tinh gia tri trung binh cua 3 lan lap lai. Sai khac tin cay ve thdng ke khi so sanh gia tri eua tung ehi tieu giua cae nhom n^n mlu su dung Tiesf So sanh va hien thi su dao dong ve gia tri eua timg chi tieu giiia cac nen mlu bang bi^u do duong thing.
S.KFTQUANGMBUCIAI
3.1. Phan tich ham lugng DLr3-Phenyllactic axit (3-PAl ti-ong mat ong
Ket qua phan tich 6 hang 2 vl hinh 1 cho thay, ham lugng 3-PA eua 9 mlu MBH dao dong 0,70-1,36 mg/kg mat ong, thap hon (P<0,001) so vdi him lugng chat nay trong cac mlu M0R2 (3,08-6,54 mg/kg). Ddng thdi, h i m luong 3-PA eiia cac mlu MBH nguyen chat eung thap hon vdi sai sd thdng ke tin eay (0,01<P<0,05) vdi h i m lugng chat nly cua 2 nhom mat tron MIX20-80 vl MIX30-70 lan lugt 11 1,64-2,17 mg/kg va 2,10-2,56 mg/kg mat ong.
Bang 2. Ham lugng 3-PA (mg/kg mat ong) tren clc loai n^n mlu: MBHl-9, MOR2.1-2.3&MIX10 va MIX giOa MDQO va MBH 6 3 tj^ Ife khlc nhau
STP
1 2 3 4 S 6 7 8 9
Trong mat on Mat bac ha (MBH) MBHl
MBH2 MBH3 MBH4 MBHS MBH6 MBH7 MBHS MBH9 W Sd Min Max
0,70 0,73 1,25 0,73 0,71 1,16 1,36 1,22 1,3S 1,02 0,30 0,70 1,36
g nguyen chat Mat keo (M0R2) M0R2.1 MOR2.2 MOE2.3 MKIO
TB Sd Min Max
6,54 5,55 3,08 5,38
5,14 1,46 3,08 6,54
Ten va ty le M K (M0R2-MBH) Ten
MIX M K l MIX2 M K 3 M K 4 M K 5 M K 6 MK7 MK8 MK9 TB Sd Min Max
MKlO-90 (1:9) 1,16 1,19 1,6S 1,27 1,17 1,61 1,75 1,68 1,73 1,47 0,26 1,16 1,75
MIX20-80 (1:4) 1,64 1,67 2,08 1,67 1,66 2,02 2,17 2,06 2,17 1,90 0,24 1,64 2,17
MK30-70 (3:7) 2,10 2,12 2,48 2,12 2,11 2,43 2,56 2,46 2,56 2,33 0,21 2,10 2,56 Ghi ehii: Ky hieu mau: xem bang 1.
Hinh 1. Dao dOng v^ ham lirgrng J-PA gifla cac loai nto miu: M0R2 nguySn chit & MKIO, MK30-70, MK2(>80, MKIOOO wk MBH nguyta ch& (Kyhieu
man: xem bang 1).
Gia tn dac thii, dp lech chuan va dao dong ve gia tri cua 3-PA tren cac mSu MBH (trung binh, TB: 1,02
± 0,30 mg/kg), M0R2 (TB: 5,14 ± 1,46 mg/kg), MIX20-80 (1,90±0,24 mgAg) va MIX30-70 (2,33+0,21 mg/kg) cho thay su khac biet ve ham luong 3-PA giOa 4 16 mSu (Bang 2 va Hinh 1). Ham luong 3-PA cua cac miu trpn MKlO-90 tir 1,16-1,75 mg/kg, dii cao hon cua 9 miu MBH nguyen chat (0,70-1,36 mg/kg) voi tin cay vd thdng ke (P<0,004), nhung do lech chuan va dao dong ve gia tri 3-PA ciia cac miu MBH nguyen chat (TB: 1,02±0,30 mg/kg) va MKlO- 90 a B : 1,47±0,26 mg/kg) cho thay mpt sd miu MKlO-90 cd ham lupng 3-PA trong khoang gia tri chat nay cua MBH nguyen chat Nhu vay, ket qua phan tich ham lupng 3-PA theo LC-MS/SM tren 4 loai ndn miu mat ong khac nhau trong nghien cuu nay cho thay ham luong 3-PA cd thd sir dung nhu chi Bang 3. Ham luong Fe^ (mg/kg mat ong) tao th^nh
MOR2.1-2.3&MK10vaMKl-9 giiia
thi de phan biet MBH nguyen chit vdi MBH bi pha tron tir 20% mat keo trd len.
3.2. Phan tich ham lupng Fe^ hinh th^nh sau phdp thu FRAP
Ham luong Fe^* tao thanh sau phep thu FRAP tren cac loai ndn miu mat MBH, mat keo va 3 nhdm mat trpn MKlO-90, MK20-80, MK30-70 duoc the hien chi tiet d biing 3 va hinh 2. Ham lupng Fe^* tao thanh tir 9 miu MBH (59,74-263,09 mg/kg mat ong), thap hon dang kd ve mat thdng ke (P<0,0002) khi so voi mat keo (994,34-1.053,65 mg/kg). Tuong tu, ham lupng Fe^^ tao thanh tir MBH cung thap hon (0,0001<P<0,001) so voi ciia MIX20-80 va MK3O-70, lan luot tir 308,23 - 460,46 mg/kg va 348,07 - 493,82 mg/kg mat ong. Do lech chuan va dao dong ve ham lupng Fe^' tao thanh do cac miu MBH (TB: 148,63 + 59,92 mg/kg), mat keo (TB: 1.019,92 ± 24,87 mg/kg), MK2O-80 (375,89 ± 44,69 mg/kg) va MK30-70 (412,55 + 42,44 mg/kg) cho thay su khac biet tin cay ve ham lupng Fe^* dupe tao thanh giira 4 loai ndn miu. Trong khi dd, ham luong Fe^^ tir 9 miu MKlO-90 (180,7 - 365,70 mg/kg), dii cao hon cua cac miu MBH (59,74 - 263,09 mg/kg mat ong) voi tin cay ve thdng ke (P<0,006), nhung dp lech chuan va dao dong ve gia tri Fe^* ttr cac mau MBH (PB: 148,63 ± 59,92 mg/kg) va MKlO-90 (TB: 260,37 ± 52,78 mg/kg) cho thay ham lupng Fe^' tao thanh ttr mot sd miu MKlO-90 nim trong khoang gia tri cua chat nay tao thanh tir cac miu MBH. Ket qua phan tich cho thay ham luong Fe^^ tao thanh sau phep thir FRAP giira cac loai nen miu mat ong co the sir dung nhu chi thi dd phan biet MBH vdi MBH bi pha trpn tir 20%MOR2n-dlen.
sau phep thu FRAP cua cac loai ndn miu: MBHl-9, MKIO vdi MBH or 3 ty 18 khAc nhau
STT
1 2 3 4 5 6 7 8
Trong mat ong nguyen chat Mat bac ha (MBH)
MBHl MBH2 MBH3 MBH4 MBH5 MBH6 MBH7 MBHS
263,09 174,66 203,03 121,92 137,36 143,94 59,74 92,29
Mat keo (M0R2) M0R2.1 MOR2.2 MOR2.3 MKIO
1.019,67 1.012,01 994,34 1.053,65
Ten va ty le MK (KEO-MBH) Ten
M K M K l MIX2 MK3 MK4 MIX5 MK6 MK7 MK8
MKlO-90 (1:9)
365,70 281,14 301,74 238,88 249,87 259,13 1S0,71 215,15
MIX20-80 (1:4)
450,46 396,78 415,41 357,33 368,30 367,53 308,23 333,73
MK30-70 (3:7)
493,82 432,48 448,96 395,41 402,93 412,15 348,07 374,06
72 NONG NGHIEP VA PHATTRIEN NONG THON - KY 2 - THANG 11/2018
KHOA HOC CONG NGHE 9 MBH9
TB Sd Min Max
141,66 148,63 59,92 59,74 263,09
TB Sd Min Max
1.019,92 24,87 994,34 1.053,65
MK9 TB Sd Min Max
251,05 260,37 52.78 180,71 365,70
375,27 375,89 44,69 308,23 460,46
405,09 412,55 42,44 348,07 493,82 Ghi ehu: Ky hieu m&u: xem bang 1.
10S3,6S 994.34
493,82
* 1 — ^
•^—\'.Z.
_ 348.07 '^mji ^ ,
59.7V
MIX3a-70 MIX20-80 MIX10-9D
MBH 1 3 3 4 5 6 7 8 9
S6 liFong mlu
ffinh 2. Dao ddng v^ ham lugmg F e ^ t^o tiianh sau phep thu FRAP giOa cac loai n^n mau: M0R2 &
MKIO, MIX3a-70. MIX2(m), MIXlO-90 va MBH nguyen ch^t
{Kyhieu miu: xem phan nguydn Ueu va bang 1).
3.3. Phan tich p h ^ tram DPPH da phan ting sau phep thu DPPH
Bang 4 va hmh 3 trinh bay k^t qua phan tich va so sanh kha nang chong oxi hoa tong so dua vao phin tram DPPH da phan ung sau phep thu DPPH cua cac loai n^n mhx MBH, M0R2, mat tron MIX20-
80, MIX30-70 va MIX50-50. %DPPH da phan ting cua MBH (10,63 -16,54%), thap hon (P<0,02) so voi mat keo tai tuong (26,20 - 28,55%). D6ng thai, %DPPH da phan ung do cac man MBH cung thap hon (0,001<P<0,02) so voi cac mhi MIX30-70 va MDCSO- 50, lan luot 18,03 - 21,32% va 22,00 - 25,22. Do lech chuan va dao dong ve %DPPH tao thanh do cac man MBH (JB: 13,59±1,85%), M0R2 a B : 27,54+ 0,99%), mat MIX30-70 (19,17+1,09%) va MIX50-50 (24,22+1,12%) cho thay su dac thu va khac biet ve
%DPPH duoc tao thanh giiia cac loai nen mSu. Trong khi, %DPPH cua MIX20-80 (14,83 - 18,75%), du cao hon (P<0,002) so voi gia tri nay cua cac mhx MBH (10,63 -16,54%), nhung do lech chuan va dao d6ng ve
%DPPH cua cac mlu MBH nguyen chat (TB:
13,59+1,85%) va MIX20-80 a B : 16,80+1,23%) cho th£ty mot so mhx MIX20-80 co %DPPH frong khoang
%DPPH cua cac mhi MBH. Ket qua phan tich cho thay %DPPH da phan tmg sau phep thu DPPH voi cac loai nen mhi mat ong chi co th^ su dung nhu chi thi de phan biet mat ong bac ha nguyen chat voi MBH bi pha tron hr 30% mat keo h a len.
Bang 4. Phan tram DPPH da phan ling sau phd 2.3&MIX10v!iMK giiia MIXl-9 STT
1 2 3 4 5 6 7 8 9
Trong mat ong nguyen chat Mat bac ha (MBH)
MBHl MBH2 MBH3 MBH4 MBH5 MBH6 MBH7 MBH8 MBH9 TB Sd
16,54 12,25 14,90 11,89 12,68 10,63 13,96 14,80 14,67 13,59 1,85
Mat keo (M0R2) M0R2.1 MOR2.2 MOR2.3 MKIO
TB Sd
27,87 26,20 28,55 27,56
27,54 0,99
p thu DPPH Ciia cic loai n^n miu: MBHl-9, M0R2.1- gitia MKIO v i MBH 6t 3 ty 1* khic nhau
Ten va ty le M K (KEO-MBH) Ten
M K M K l MK2 M K 3 MK4 M K 5 MK6 MK7 MIX8 MK9 TB Sd
MK20-80 (1:4)
18,75 16,20 18,19 15,71 17,02 14,83 16,20 17,49 16,83 16,80 1,23
MK30-70 (3:7)
20,39 18,03 21,32 18,31 19,35 18,45 18,31 19,33 19,06 19,17 1,09
MK50-50 (1:1)
25,16 24,51 24,62 23,18 23,33 22,00 24,99 25,22 24,95 24,22 1,12
Min Max
10,63 Mm 16,54 Max
26,20 Min 28,55 Max
14,83 18,75
18,03 21,32
22,00 25,22 Ghi ehu: Ky hidu miu: xem phan nguyen Ueu va bang 1.
tn35 1= 20 .S"15 I Q-10 Q
^ 5 0
^ S » ^
14,8a--*-
1 2 3 4 5 6 7 8 9
3 6 luwng m i u
M0R2 1-234 MixlD -Mix50-50
MIX30-70 MIX20-80
Hinh 3. Dao dOng v4 p h t o tram DPPH da phan ting sau ph6p thii DPPH giiia c ^ loai n^n mau: M0R2 nguyta chat & MKIO, MIX50-50, MIX30-70, MK20- 80 v i MBH nguyta c h i t (Ky hieu miu: xem bang 1)
4.THA01UiN
4.1. DLra-Phenyllactic axit, F e ^ v i %DPPH la c i c chi thi d^ phan bi6t MBH nguyta chat vji MBH bi trpn M0R2 6 cic ty 16 khac nhau
Cong bo gan day (Le Quang Trtmg et al., 2018) da cho thay ham luong DL-3-Phenyllachc axit (3-PA), Fe^* va %DPPH co the su dung nhu cac chi thi de phan biet mat bac ha nguyfen chat (MBH) voi mat keo (M0R2). Trong nghita cuu nay, cac gia tri v^
ham luong cua 3-PA (5,14 + 1,46 mg/kg), Fe^*
(1.019,92 ± 24,87 mg/kg) va %DPPH (27,54 ± 0,99%) cua M0R2 deu cao hon voi khac biet dang ke (P<0,0002 <P<0,02) so vol ciia MBH nguyta chat (3- PA 1,02 ± 0,30 mgAg; Fe^*: 148,63 ± 59,92; %DPPH:
13,59 ± 1,85% (Bang 2, 3 va 4). Dieu do mot lan nita chung minh gia tri su dung ciia cac chi thi nay. Do gia tri cua 3-PA, Fe^* va %DPPH trong MBH thap, nen khi pha h-on M0R2 vao MBH thi gia tri cua 3 chi tita trta se tang Ita theo ty le pha, luong M0R2 tron vao MBH cang cao thi gia tn cua 3 chi tieu cang tang. Day la co so de phan biet MBH bi tron mat keo voi MBH nguyta chat Mat keo duoc pha vao MBH voi 4 ty le khac nhau vj khoi lucmg (Bang 1), g6m ty le 1:9 (MKlO-90); 1:4 (MIX20-80); 3:7 (MK30-70) va 1:1 (MK50-50) c6 gia tri cua 3 chi tieu tang din va cao hon so voi MBH nguyta chat. Lay ham luong trtmg binh ola Fe''* sau phep thii FRAP lam vi du, co the thay ham luong Fe^* a i a MKlO-90, MK20-80 va MK30-70 tang tit 260,37 d t a 412,55 mg/kg, trong
khi do ham luong Fe^* c i a MBH nguyta chat chi co 148,63 mg/kg mat ong (Bang 3). Nhu vay, nha djc hita v^ gia tri, ham luong 3-PA, Fe^* va %DPPH con la cac chi thi de phan biet MBH nguyta chat voi MBH bi tron M0R2 o cac ty le khac nhau. T r t a thuc te, miic dac hita cua mdi chi thi de truy xuat nguon goc mat ong khong giong nhau va duoc xac dinh dtra vao do lech chuan va dao dong ve gia tri ciia timg chi thi khi so sanh giiia mat ong nguyta chat va mat bi pha tron (i cac ty le khac nhau (McDonald et al., 2018; Dzugan etal., 2018). Trong nghita cuu nay, do lech chuan va dao dong v^ ham luong 3-PA va Fe^
cho phep phan biet MBH nguyta chat voi MBH hi pha tron >20% M0R2; trong khi %DPPH chi cho phep phat hita MBH pha b-pn voi >30% M0R2 (Hinh 1 d t a hinh 3). Dua vao do lech chuan va dao dong ve gia tn, CO the ap dung gia tri dac thii cua cac chi thi 3-PA, Fe^* va %DPPH de phan biet MBH nguyen chat va MBH bi pha tron nhu sau: 1) MBH nguyta chat CO h t a i luong 3-PA <1,36 mg/kg, re^*<263,09 mg/kg va %DPPH<16,54%; 2) MBH tron >20% mat keo: 3-PA
>1,64 mg/kg, Fe^*>308,23 mg/kg; 3) MBH tron >30%
mat keo: 3-PA >2,10 mg/kg, Fe"*>348,07 mg/kg va
%DPPH>22,00%.
Nhita cong trinh nghita ciiu chiing minh rang gia di ciia cac chi thi hoa hoc trong mat ong quy^t dinh tinh dac hieu va do chinh xac de phan biet mat ong, trong dd gia tri thap ciia chat chi thi trong mat ong eta truy xuat CO im viet hon chi thi co gia tri cao (Colucci etal, 2016; Ciulu etal., 2016). Theo cac tac gia nay khi chat chi thi trong mat ong can truy xuat CO ham lugng cang cao thi cang d l bi lam gia bang each bd sung chat chi thi tach chiet tir cac nguon khac vao mat ong re tita hoac mat pha tron d^ dat ham luong cua chat chi thi trong mat ong can truy xuat. Nguoc lai, n t a mat ong can truy xuat co gia tri ciia chat chi thi thap, khong the tach chiet giam bdt ham luong chat chi thi ra khoi mat ong gia thap cd ham luong chat chi thi cao. Nhu vay, trong nghien ciiu nay, ham luong cac chat 3-PA, Fe^* va %DPPH ciia MBH thap hon vdi thdng k6 tin cay so vdi mat keo se la cac chi thi hieu qua de phan biet MBH nguyta chat vdi M0R2 gia thap cung nhu MBH hi pha trdn vdi M0R2 d ty le >20%.
NONG NGHIEP VA PHAT TRIEN NONG THON - KY 2 - THANG 11/2018
KHOA HOC C 6 N G N G H E
4.2. Uu di^m va nhuoc di^m cua cac chi thi 3- PA,Fe^va96DPPH
DL 3-phenyllactic axit (3-PA) trong mat ong, Fe^*
tao thanh do mat ong sau ph6p thu FRAP va %DPPH da phan ung do mat ong la cac chat hoa hpc hen quan den kha nang chdng oxi hoa cua mat ong. Gia tri dac thii cua 3 chat nay da va dang duoc ap dung rong rai va hieu qua nhu cac chi thi d^ truy xuat nguon goc mat ong, phan bifet mat ong nguyfen chat hoac bi pha trfln (McDonald et al, 2018; Colucci et al., 2016). Trong mat ong, 3-PA ben vimg theo thoi gian va it bien doi theo nhiSt do (McDonald et al, 2018), trong khi Fe^^ va %DPPH dugc phan tich hang cac phep thtr FRAP va DPPH voi ky thuat don gian, thoi gian phan tich nhanh va chi phi thfip (Dzugan et al., 2018). Tuy nhien, k^t qua sau phep thu FRAP va DPPH nhi^u khi khong on dinh do thao tac thi nghiem (Colucci et al., 2016; Dzugan etal, 2018).
Trong khi ham luong 3-PA phan tich bang phtrong phap LC-MS/MS co do chinh xac cao, on dinh v^ k^t qua giua cac lan va cac phong thii nghiem (McDonald etal., 2018), nhung ton thoi gian, chi phi cao do thiet hi va hoa chat chay LC-MS/MS. Ngoai ra, 3 chi thi trong nghien cuu nay moi phan hifit dupe MBH bi pha tron voi >20% mat khac. D^ xac dinh duoc ty le pha tr6n <20% can phai ap dung chi thi d6ng vi ben ^^C/'^C phan tich bang phuong phap 513C-EA/LC-IRMS (Emem, Raezke, 2008). Chi thi va phuang phap nay doi hoi phai co ca sa dir lieu
^K/^K cua MBH, phong thii nghiem va thiet hi hi&n dai, boa chat dat tien. Vl the, de quan ly thuong xuyen ch^t luong MBH, chi can ap dung cac chi thi ve %DPPH sii diing phep thu DPPH va ham luang Fe^* voi phep thii FRAP a cac phong thu nghiem lien quan cua dia phuang. Khi co tranh chap hoac yeu cau cua khach hang dac bi6t la khach hang nhap khau can phai str diing chi thi 3-PA phan tich bang phuong phap LC-MS/MS a cac phong thtr nghiem co du nang luc trong nuoc hoac qudc td. Khi co yfeu cau phan biet MBH nguyen chat voi MBH hi pha h-pn voi
<20% mat khac, phai ap dung chi thi ^K/^K phan tich b cac phdng thu nghiem co du thiet bi va hoa chat phan tich chi tifiu nay.
5.KFTLUAN
Kdt qua phan tich tinh dac thu ve gia tri cua 3 chi tieu H6n quan ddn kha nang chdng oxi hoa cua mat ong h-^n cac nhom nen mau mat ong khai thac
mua mat 2017-2018, bao gom MBH nguydn chat, mat keo tai tupng, mat pha trpn giua mat keo va MBH 6" 4 ty le 1:9, 1:4, 3:7 va 1:1 cho thay ham lupng DL-3- phenyllactic axit (3-PA) trong mat ong va Fe^"" tao thanh sau phep thii FRAP la 2 chi thj phan hiet MBH nguyfen chat voi MBH bi pha trpn voi mat keo a ty Id
>20%, h-ong khi %DPPH da phan ling sau phep thu DPPH CO the xac dinh duoc MBH pha trpn voi mat keo a ty le tir 30% tra len. Gia tri cua cac chi thi dd phan bi6t mat ong bao gdm: 1) MBH nguyen chat co ham lupng 3-PA <1,36 mg/kg, Fe2*<263,09 mg/kg va
%DPPH<16,54%; 2) MBH trpn >20% mat keo: 3-PA
>1,64 mg/kg, Fe2*>308,23 mgAg; 3) MBH trpn >30%
mat keo: 3-PA >2,10 mg/kg, Fe^348,07 mg/kg va
%DPPH>22,00%. Phuang phap va ket qua nghien cuu la cff scr ap dung nham xay dung ca sa du hfiu de truy xuat ngudn goc MBH va cac loai mat ong khac a nuocta.
L d l CAM ON
Day la ket qua nghidn cuu thudc dS tai khoa hpe va cdng nghe cap tinh ciia Hi Giang vd "Nghien cuu phan tieh bd sung chi tieu chat luong san pham chi din dia ly cho mat ong bae ha eao nguydn di Dong Van, Ha Giang". Ma sd: DTKH.HG-02/17 Cac tie gia xin cam on Trung tam Kiem nghiem tinh Hi Giang thudc Sd Y te Ha Giang, Vien Hoa sinh Bidn thudc Vien Han lam Khoa hpc vi Cdng nghd Viet Nam da giiip ddky thuat thu nghiem, ky thuat phan tieh, xu ly sd lieu vi tao dieu kien thuc hidn thinh cdng nghidn cuu niy.
TAI LIEU THAM KHAO
1. Ciulu M, Spano N, Maria Pile MI, Gavino Sanna G (2016). Recent Advances in the Analysis of Phenolic Compounds in Unifloral Honeys. Molecules 21 (451); doi:10.3390/molecules21040451.
2. Colucci G, De Vito V, Varricchio E, De Cunzo F, Coccia E, Paolucci M, Di Stasio M, Boscaino F, Viola C and Volpe MG (2016).
Identification of Traceability Markers in Italian Unifloral Honeys of different Botanical Origin. J Nutr Food Sci. 6 (1). DOI: 10.4172/2155-9600.1000462.
3. Dobre I, Gadei G, Pahacu L, Elisei AM, Segal R (2010). The antioxidant activity of selected Romanian honey. Food Technology 34 (2): 68-73.
4. Dzugan,M, Tomczyk M, Sowa P, Grahek- Lejko D (2018). Antioxidant Activity as Biomarker of
Honey Variety. Molecules, 23 (2069); bac ha Cao nguyen da Dong Van. tinh Ha Giang. Tap doi:10.3390/molecules23082069. chi Ndng nghiep va PTNT. 13 (340): 47-53.
5. Elflein L, Raezke KP (2008). Improved 7. McDonald CM, Keeling SE, Brewer MF, detection of honey adulteration by measunng Hathaway SC (2018). Using chemical and DNA differences between 13C/12C stable carbon isotope marker analysis to authenticate a higtvahte food, ratios of protein and sugar compounds with a manuka honey, npj Science of Food (9). DOt combination of elemental analyzer • isotope ratio 10.1038/s41538-018«llM.
mass spectrometry and liquid chromatography 8. Trautvetter S, KoeHing-Speer I, %eer K isotope ratto mass spechrometry (813C-EA/LC- (2009). Confirmation of phenolic acids and flavonoids IRMS). Apidologie, Springer Verlag, 2008, 39 (5); in honeys by UPLC-MS. Apidologie 40:140-150.
574-587. 9. Tuberose, C. L G., Bifulco, E., Caboni, P., 6. Le Quang Trung, XguySn Diic Td, Nguyfn Sarais, G., Cottiglia, F., Flons, I. Lumichrome and Tho Khiem, Vu Thi lien, Le 1114 Nhu Thiiy, Kim Phenyllactic Acid as Chemical Markers of Thisde Bich Nguyet Cam TM H i i g , NguySn Thi Thiiy Hoa, (Galactites tomentosa Moench) Honey. J. Agiic.
Pham Minh Giang (2018). Nghien a m kha nang Food Chem. 2011,59,364-369.
chong oxi hda cua san pham chi din dia ly mat ong
DETERMINING ANTIOXIDANT MARKERS OF HONEYTO AUTHENTICATE ELSHOLTZIA HONEY - A GEOGRAPHICAL INDICATION PRODUCT OF DONG VAN KARST PLATEAU,
HA GIANG PROVINCE
Le Quang Trung, Kim Bich Nguyet, Cam Tlii Hang, Nguyen Dinh Anh, Lam Tliu Hang, Vu Tliuy Nga, Nguyen TTli Tliuy Hoa, Pham Minh (jiang .Nummary
Elsholtzia honey, a geographical indicaUon product of Dong Van Karst Plateau, Hagiang province (MBH), has recently the highest price in the domestic market and is therefore at risk of tiauding to such honey of high productivity and low price as Acacia honey (M0R2) in order to get higber benefits. For protection of die MBH brand, in this study, DL-3phenyllactic acid (3-P-A) amount determined with LC-MS/MS mediod, Fe*- amount formed after FRAP assay and DPPH percentage reacted after DPPH assay in diferent types of honey sample were analyzed. Honey samples included pure MBH, pure M0R2 and 4 other types of mixture between M0R2 and MBH in weight ratios of 1: 9,1: 4, 3: 7 and 1: 1. The values of J-PA, Fe'', and DPPH in pure MBH were found significantly lower (0.0001 <P <0.05) iban those in M0R2 and in all 4 types of honey mixtures between MBH and M0R2 with different ratios. Analysis results of specificity, standard deviation and ranges of marker values indicated that S^PA and Fe*- amount between different honey types could be used as markers to discriminate the pure MBH and MBH mixed with >20% M0R2. while % DPPH can be used to audienticate pure MBH from MBH mixed wltii >30% .M0R2. In details of mariiers, values of 3-PA.
Fe*^ and DPPH in pure MBH were found to be <.36 mg/kg, ^63.09 mgAg, and <16.54't. respectively, while pure MBH nuxed widi >20* M0S2 has amount of 3-PA >1.64mg/kg and Fe'-^SOSaSmg/kg, and pure MBH mixed with >30% M0R2 were tested widi amount of 3-PA >2.10mgAg, Fe-">348.07mg/kg, va DPPH>18.0396. Results and used mefliods in dus study is a scientific basis to be applied for honey authentication in order to minimize the fraud in Vietnamese honey, including the Elsholtzia honey.
Keywords: Authentication, DLSithenyllactic acid, DPPH, FRAP, Elsholtzia honey, Dong Van Karst Plateau, Ha Giang province.
NgiiM phan biSm GS.TS. Nguyta Cong Khin Ngay nhan bai: 10/9/2018
Ngiy fhOng qua phan bita: 11/10/2018 NgSy duyM dang: 18/10/2018
76 NONG NGHIEP VA PHAT TRIEN NONG THON - KY 2 - THANG 11/2018
