experiment-1

Period: september, 2 nd week～4th week

Preparation:
1. incubators (5, 10, 15, 20, 25, 30℃) &Chambers ( 4 pcs)
2. Freezer & Sample boxes
3. GC, HPLC, HPLC-MS,Colorymeter, HPLC column C 18 and
guard column, filter 0,45µm
4. Camera + Label for taking photo
5. Sample tubes (50 mL pink lid sample tube ×180)
6. Knife, chopping board, tissue, etc…
7. Solvent
• O2, N2, Air, Methanol (HPLC), ACN (HPLC), Formic acid
(HPLC),L-ascorbic acid,tris(2-carboxylethyl)phosphine (TCEP),
metaphosporic acid, H3PO4.
 Experiment Plan
Analytical methods development
 Content analysis(HPLC)
 Vit C (HPLC)

Sampling
Sample : selected vegetables
Each sample approx : 200 g
Replication : 3 ,Sub replication : 2 ,Total sample for each treatment : 1000 g

Experiment
5℃ : Day 0 and day 3
20℃ : Day 0 and day 3
Parameter
Respiration rate, content , color, Vit C, phenolic compounds profile,
antioxidant

Data analysis
 Statistic etc
 After Arrival:
• sample preparation
• Sample selection :
• Weighing the sample weight
Sampling for chemical
analysis

Respiration rate

• Direct analysis on • Sample preparation for

GC experiment day (day 0 indirect analysis
and day 3) • Liquid nitrogen and
• Color freeze dryer
• Amarathin profile • Vit C
day 1 ～day 3 • Phenolic
compounds
profile
• Antioxidant
Experiment plan
Storage treatment

Storage 1 = 5˚C Storage 2 = 20˚C

Plant = 6 Plant = 6 Plant = 6 Plant = 6

ambient 2 % O2 ambient 2 % O2
1 2 3 4 5 6 control

ambient
7 8 9 10 11 12
5 °C 1. Respiration
rate
2 % O2 2. content
13 14 15 16 17 18 3. Color
4. Vitamin C
5. Phenolic
compounds
ambient 6. antioxidant
19 20 21 22 23 24

20 °C

25 26 27 28 29 30
2 % O2
extraction

15 mL
Methanol
80%

1 gram
sample
Homogenized, HPLC Measurement
25 mL
(20000 rpm, 40 s)

Note : extraction process must use

Methanol with HPLC grade
HPLC condition

•Column : C18, 3 µm, 250 mm

•Col Temp : 35°C
•Det : UV = 540 nm and 470 nm
•Mobile Phase:
A : 0,5 % Acetic acid in H2O, B : Methanol
SGP system: Time A B
0 100 0
14 70 30
15 70 30
17 50 50
18 50 50
19 100 0
21 100 0