Stabilitas Vitamin E dari PFAD (Palm Fatty Acid Distillate) yang Diinkorporasi pada Galaktomanan Kolang-Kaling
Lampiran 1. Kromatogram Hasil Analisis Komponen Senyawa PFAD dengan alat GC
Lampiran 2. Kromatogram Analisis Fraksi yang Tidak Tersabunkan dengan Pelarut Dietil eter Menggunakan Alat GC. Sampel 1 a.
b. Sampel 2
Lampiran 3. Kromatogram Hasil Analisis Fraksi yang Tidak Tersabunkan dengan
Pelarut Petroleum Benzena Menggunakan Alat GCa. Sampel 1
b. Sampel 2
Lampiran 4. Kromatogram Hasil Analisis Fraksi yang Tidak Tersabunkan dengan Pelarut n-heksana Menggunakan Alat GC.
a. Sampel 1
b. Sampel 2
Lampir
0.380 % ksi
5.16
0.85
46.82
5.28
28.80
8
4
2
00
00 pada Frak HPLC
86
50
28
29
E Vitamin E akan Alat H Result
7193288.2 926728.2 6460042.5 203274.8 1189203.0
T T3 T3 T3 T3 Komponen rsabunkan CONC. (m Are Vit
n Senyawa
Mengguna
mg/ml): eaTotal matogram g Tidak Ter
ran 5. Krom yang α-T α-T β-T γ-T δ-T
86.90
Lampir ran 6. Krom matogram Hasil Anal lisis Kandu ungan Vitam min E pada a Fase n-hek ksana deng gan menggu unakan HP PLC Sampe l I
Result R CONC. (mg/m ml): 0.3800 Area %
56 653275.46
22.63 α-T
7 708584.05
4.04 α-T3
47 762932.48
34.52 β-T3
140465.96
0.58 γ-T3
7 775692.04
3.36 δ-T3
Total Vit E
65.14
a. Sampel II
Result CONC . (mg/ml): 0.38 800
A Area % %
5776585
5.08
23.12 α-T
722316
6.33
4.12 α-T3
4806855
5.73
34.84 β-T3
141012
2.38
0.59 γ-T3
689758
8.92
2.99 δ-T3
Tota al Vit E
V
65.66
Lampiran 7. Penentuan Panjang gelombang maksimum Vitamin E dengan alat Spektrofotometer Uv-Vis panjang gelombang maksimum Vitamin E
- 0.45
- 0.35
.
. .
0.12% = 0.0785%
gr Galaktomanan = .
.
0.12% = 0.1117%
gr Galaktomanan = .
0.12% = 0.1167.%
gr Galaktomanan
No Sampel
I II
III Rata-rata
1 Blanko N-Hexan 0.0525 0.0529 0.0526 0.0526
2 Standard Tokoperol A 3.4280 3.3955 3.4122 3.4119
=
0.12% = 0.0475%
2 Standard Tokoperol B 3.4280 3.4280 3.4211 3.4257 3 0.45 gr Galaktomanan 3.2653 3.3688 3.4414 3.3583 4 0.35 gr Galaktomanan 1.3545 1.3579 1.3562 1.3562 5 0.25 gr Galaktomanan 2.2415 2.2467 2.2415 2.2432 6 0.15 gr galaktomanan 3.1542 3.1984 3.2223 3.1916 7 0.05 gr Galaktomanan 3.3338 3.5486 3.4151 3.4325
Sampel = γ
Lampiran 8. Adsorbsi Tokoperol dengan Galaktomanan + Tween 20
Standard
Tocoperol 80 % 3%
dari standard 80% = 3 % = 2.4 %
5 ml dari 2.4 % = V1. N1 = V2 . N2
= 5 mL x 2.4 % = 100 mL . N2 N2 = 0.12% N
γ
.
0.12%
gr Galaktomanan = .
.
0.12% = 0.1181%
gr Galaktomanan = .
- 0.25
- 0.15
- 0.05
No Sampel Konsentrasi Tocoperol % Adsorbsi
1 Staandard Tocoperol
- 0.12 %
2 0.1181% 0.19 %
0.45 gr galaktomanan
3 0.0475% 7.25 %
0.35 gr galaktomanan
4 0.0785% 4.15 %
0.25 gr galaktomanan
5 0.1117% 0.83 %
0.15 gr galaktomanan
6 0.1167% 0.33 %
0.05 gr galaktomanan
No Sampel
I II
III Rata-rata
1 Blanko N-Hexan 0.0525 0.0529 0.0526 0.0526
2 Standard Tokoperol A 3.4280 3.3955 3.4122 3.4119
2 Standard Tokoperol B 3.4280 3.4280 3.4211 3.4257 3 0.45 gr Galaktomanan 3.2933 3.3316 3.3668 3.3305 4 0.35 gr galaktomanan 1.8989 1.9012 1.8997 1.8999 5 0.25 gr Galaktomanan 2.2726 2.2617 2.2590 2.2644 6 0.15 gr Galaktomanan 3.2476 3.3445 3.2223 3.2714 7 0.05 gr Galaktomanan 3.4152 3.4152 3.3963 3.4089
Lamiran 9. Adsorbsi Tokoperol dengan Galaktomanan tanpa tween 20
Standard Tocoperol 80 %
3% dari standard 80% = 3 % = 2.4 %
5 ml dari 2.4 % = V1. N1 = V2 . N2
= 5 mL x 2.4 % = 100 mL . N2 N2 = 0.12%
γ N 0.12%
Sampel =
γ .
0.45 gr Galaktomanan = 0.12% = 0.1171% .
. 0.35 0.12%
gr Galaktomanan = = 0.0665%
- .
. 0.25 0.12%
gr Galaktomanan = = 0.0793%
- .
. 0.15 0.12%
gr Galaktomanan = = 0.1145%
- .
.
0.05 0.12%
=
gr Galaktomanan = 0.1194%
- .
No Sampel Konsentrasi Tocoperol % Adsorbsi
1
- Staandard Tocoperol 0.12 %
2 0.1171% 0.29 %
0.45 gr galaktomanan
3 0.0665% 5.35 %
0.35 gr galaktomanan
4 0.0793% 4.07 %
0.25 gr galaktomanan
5 0.1145% 0.55 %
0.15 gr galaktomanan
6 0.1194% 0.06 %
0.05 gr galaktomanan
Lampiran 10. Hasil uji stabilitas oksidasi sampel A dengan alat Rancimat
Lampiran 11. Hasil uji stabilitas oksidasi sampel B dengan alat Rancimat
Lampiran 12. Hasil uji stabilitas oksidasi sampel C dengan alat Rancimat
TGL : 02-2-2014
2
Lampirran 13. Hassil Analisis Galaktom manan kolan ng-kaling d dengan alatt FT-IR
Lampir ran 14. Has Den sil analisis ngan vitam galaktoma min E mengg
anan kolang
gunakan al
g-kaling ya lat FT-IR ang telah diiinkorpora siLampir a.
a.
Perbesa ran 15. Has den Hasil uji S Hasil uji S vitamin E aran 1000 k sil Analisis ngan alat SE EM Galak EM Galak kali Inkorpora EM tomana Ko tomana Ko
asi Vitamin
olang-kalin
olang-kalin
n E pada ga ng ng setelah d alaktomana diinkorpor an kolang-k rasi dengan kaling nPerbesaran 1500 kali
Lampiran 16. Prosedur Analisis TG menggunakan Alat Kromatografi Gas (AOCS Ce5- 86)
1. Definition The content of a group of triglycerides having the same carbon number is a quantity expressed as a percentage relative to the total triglycerides content of the sample, separated according to the present procedure.
2. Principle Separation of the triglyceride groups having the same carbon number by direct Gas Liquid Chromatography (GLC) of a fat or oil solution under temperature programmed conditions.
Identification by reference to a standard triglycerides solution. Content determination by peak
areas ratio.3. Scope Applicable to vegetable oils, especially for palm oils, palm kernel oils, coconut oils and derivatives.
4. Reagents 4.1. n-Hexane, analytical chromatography grade quality, purity 98% min.
4.2. Gases:
a. Carrier gas - hydrogen, ultra high purity grade, minimum purity 99.95% mol, dried and containing max of 10 mg O2/kg. b . Make up gas1 - nitrogen, ultra high purity grade.
c. Detector gases - hydrogen, ultra high purity grade, and compress air, ultra high purity, hydrocarbon free, less than 2 ppm hydrocarbon equivalent to CH4.
5. Apparatus
5.1 Gas Chromatograph with facilities: a. Column Oven, capable of temperature programming up to at least 360oC.
b. Sample Inlet System, capable on capillary split injection using Pneumatic Split/ Splitless (PSS) Injection.
c. Flame Ionization Detector (FID), capable to be maintained at least 25oC above maximum column temperature.
5.2 Capillary column, type Quadrex 007-65HT.
5.3 Autosampler equipped with syringe (maximum 10 μL), graduated in 0.1μL.
5.4 Transferpette® (capacity 5 – 50 μL).
5.5 Sample vials 2mL.
6. Procedure
6.1 Sample preparation
a. Before test portions are taken from samples, the samples should be mixed thoroughly.
Warm th he samples a as necessary y so that it i s completel ly liquefied. .
b. Pipette 5
5 ample into a a vial. Dilut te with 1.5m ml n-hexane .
μl - 7μl2 sa c. Shake th e vials for 1 1 minute to make sure a all the samp ple is dissolv lved well.
d. Inject fro om those via als into gas chromatog raphy.
6.2 Chr romatograph hic specifica ation Set up the g gas chromat tograph wit th the tempe erature and column as d described in n Table
1. Smal ll changes in n the progra am may be r required du ue to the con ndition of th he column a and properti ies of the sa amples bein ng analyzed. .
7. Expr ression of R Results
a. Determin nation of the e triglycerid des group co omposition is carried o out by ident ifying each peak usin ng the graph hic edit func ction.
b. Determin ne the corre ected peak a areas of each h group of t triglycerides s using the
correction .factors d determined b by interpola ation obtain ed from the e standard tr riglycerides
Table 1 Pro oposed optim mum GLC c conditions f for triglycer rides identif fication and d
qua antification of specified d sample pr roduct.
8. Calib bration
Purpose e : T To ensure th he accuracy of the gas c chromatogra aphy instrum ment is with hin the
s specify Prod duct analysi is.Frequen ncy : W When a spec ify time per riod is elaps sed and ever ry change c column. Apparat tus : G Gas-liquid ch hromatograp ph with faci ilities for ca apillary colu umn, PSS sp plit/splitless s injection a and FID (Fl lame Ioniza tion Detecto or) Transfer rpette
S Sample vials s 2 mL (cap pacity 5 - 50 nce (standa ard) sample μL) Refere Reagen nt : n- -hexane (Li chrosolv gr rade).
Lampiran 17. Prosedur analisis vitamin E menggunakan alat HPLC (AOCS Ce8-89)
1. Objective To determine the content of tocopherols and tocotrienols.
2. Scope Applicable to palm oil phytonutrients / vitamin e sample.
3. Reagents
3.1. Acetic Acid Glassial 3.2. HPLC mobile phase – n-Hexane:Isopropanol:Acetic Acid (1000:5:1, v/v). 3.3. n-Hexane for Liquid Chromatography
3.4. Methanol for Liquid Chromatography
4. Apparatus 4.1. Analytical Balance.
4.2. Interchangeable Hypodermic Syringe
4.3. PTFE Membrane Filter 0.2 μm
4.4. Volumetric Flask 100 mL
5. Method’s Parameters
5.1. Weigh accurately the sample and standard sample (Adjust sample weight based on vitamin E concentration. See Notes 8.1) into a 100 mL volumetric flask. Add a quantity of n-Hexane (Reagents 4.3), make up to volume and swirling to dissolve the sample.
5.2. Inject 10 μL of the test solution and standard solution onto the column HPLC by mobile
phase : n- Hexane:Isopropanol:Acetic Acid (1000:5:1, v/v ). and identify the tocopherols
(and tocotrienols) present by reference to the chromatograms obtained from standards.5.3. Carry out two determinations (each consisting of duplicate injections of the prepared test.
solutions) in rapid succession, using a fresh test portion for each determination.
6. Calculation
First step we have to do is to calibrate the standart value with standart reference based on this
formula : Vol injection : (a)μL
Sample Weight : (b) gr Concentration : (c) mg/mL: (d) ng/ μL Sample Weight per injection : (e) ng
7. Expr ress of resu ult Express s the result i in 2 decima al points.
8. Note es
8.1. Alw ways filterin ng every rea agent that w we use as a s solvent or a s a mixture for HPLC mobile
ph hase, with 0. .2 Filter μm PTFE8.2. Alw ways test sta andard sam mple minimu um once a w week to mak ke sure the r results of ou ur test still con nstant and v valid.