Analysis of Organic Acid Standards
Organic acids can be easily analyzed by using a highly versatile reversed phase column with the UV absorbance detection at 210 nm. This is the analysis of highly polar organic acids and thus, LaChrom C18-AQ column which can be used with 100% aqueous eluents was used. The simultaneous analysis of 10 organic acid standards and the analyses of the organic acids in drinks, wine, and vinegar were performed. As components other than organic acids are detected by this analytical system, this system is suitable for samples containing organic acid as a main component and with little contaminants. One of the simple methods to distinguish target components from contaminants is to compare the elution positions with the standards when the column temperature is changed. 100100.0 1 8080.0 3 8 2 4 7 6060.0 6
mAU 5
mAU 4040.0 9 10 2020.0
0.00
0.000 2.002 4.004 6.006 8.008 10.0010 12.0012 14.0014 16.0016 18.0018 20.0200 分 min * Standard Solution Component Conc. (mg/L) Component Conc. (mg/L) 1. Tartaric acid 500 6. Pyroglutamic acid 100 2. Formic acid 1000 7. Citric acid 1000 3. Malic acid 1000 8. Fumaric acid 10 4. Lactic acid 1000 9. Succinic acid 1000 5. Acetic acid 1000 10. Propionic acid 1000
Calibration curves with good linearities were obtained for the concentration ranges shown below. Formic acid, Malic acid, Lacticc acid, Acetic acid, Citric acid, Succinic acid, Propionic acid : 5 - 1000 mg/L Tartaric acid : 2.5 - 500 mg/L, Pyroglutamic acid : 0.5 - 100 mg/L, Fumaric acid : 0.05 - 10 mg/L
SAMPLE 10 μL of Std. Soln. * PRESSURE
PACKING MATERIAL HITACHI LaChrom C18-AQ (5 μm) TEMPERATURE 25°C
COLUMN SIZE 4.6 mm I.D. × 250 mm (P/N : 891-5059) SEPARATION METHOD Partition/Adsorption
ELUENT DETECTOR UV 210 nm
1 mmol/L H SO + 8 mmol/L Na SO (pH 2.8) INSTRUMENTS 2 4 2 4 Chromaster 5110 (Pump), Chromaster 5210 (Autosampler), Chromaster 5310 (Column Oven), FLOW RATE 1.0 mL/min Chromaster 5420 (UV-VIS Detector) KEY WORDS High Performance Liquid Bio/Medical Science/Food/Pharmaceutical, Food, Food Component, Chromatograph (HPLC) Organic Acid, Sports Drink, Supplement Drink, Wine, Vinegar, UV-VIS Spectrometry, Health, Chromaster, LaChrom C18-AQ, Sheet No. LC100029-01 Partition/Adsorption Hitachi High-Technologies Corporation Analyses of Organic Acids in Sports Drink and Supplement Drink
4040.0 4. 乳酸 7. クエン酸
3030.0 7
2020.0 mAU mAU
1010.0 4
0.00
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 0 2 4 6 8 10分 12 14 16 18 20 min [Chromatogram of Sports Drink]
5050.0 3. Malic Acid 6. Pyroglutamic Acid 4040.0 7. Citric Acid
7 3030.0
mAU 2020.0 3
1010.0 6
0.00
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 0 2 4 6 8 10分 12 14 16 18 20 min [Chromatogram of Supplement Drink]
[Sample Preparation Method] High Performance Liquid Dilute to 5 times with purified water and filter through a 0.45 μm filter Chromatograph (HPLC) Sheet No. LC100029-02
Hitachi High-Technologies Corporation Analyses of Organic Acids in Red Wine and White Wine
150150.0 15.015 1. Tartaric Acid [Enlarged Chromatogram] 1 3. Malic Acid 5. Acetic Acid 10.010 6. Pyroglutamic Acid 100100.0 7. Citric Acid mAU 8. Fumaric Acid 5.05 6 7 8 5
mAU 0.00 mAU 5050.0 6.006 8.008 10.0010 12.0120 分
3 5 6 7 8 0.00
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 0 2 4 6 8 10分 12 14 16 18 20 min [Chromatogram of Red Wine]
1. Tartaric Acid 6060.0 3. Malic Acid 1 5. Acetic Acid 6. Pyroglutamic Acid 7. Citric Acid 8. Fumaric Acid 4040.0 9. Succinic Acid mAU mAU
2020.0 3
8 9 5 6 7 0.00
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 0 2 4 6 8 10分 12 14 16 18 20 min [Chromatogram of White Wine]
[Sample Preparation Method] High Performance Liquid Dilute to 5 times with purified water and filter through a 0.45 μm filter Chromatograph (HPLC) Sheet No. LC100029-03
Hitachi High-Technologies Corporation Analyses of Organic Acids in Apple Vinegar and Grain Vinegar
50.0 50 1.01.0 3. Malic Acid [Enlarged Chromatogram] 5. Acetic Acid 5 4040.0 6. Pyroglutamic Acid 8. Fumaric Acid 0.50.5 mAU
30.0 mAU 30 8 6 0.00.0 mAU mAU 2020.0 6.006 8.008 10.0010 12.0120 分min 1010.0
3 6 8 0.00
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 0 2 4 6 8 10分 12 14 16 18 20 min [Chromatogram of Apple Vinegar]
5050.0 2.02 5. Acetic Acid [Enlarged Chromatogram] 6. Pyroglutamic Acid 4040.0 5 1.01
mAU 6 3030.0 mAU
mAU 0.0
mAU 0 2020.0 6.006 8.008 10.0010 12.0120 分min 1010.0
6 0.00
0.000 2.002 4.004 6.006 8.008 10.0010 12.0012 14.0014 16.0016 18.0018 20.0200 分 min [Chromatogram of Grain Vinegar]
[Sample Preparation Method] High Performance Liquid Dilute to 50 times with purified water and filter through a 0.45 μm filter Chromatograph (HPLC) Sheet No. LC100029-04
Hitachi High-Technologies Corporation Comparison of Column Temperature
The elution positions of 10 organic acid standards were compared by changing the column temperature. The results obtained at the temperatures of 25°C and 40°C show that the elution positions of pyroglutamic acid and citric acid become switched, indicating that the elution behavior changes depending on the analysis temperature. 300300.0
40 C 250.0 3 ° 1 7 8 4 2 6 5 200200.0 9 10
mAU 150.0 mAU
100100.0 1 3 8 25°C 2 4 7 6 50.0 5 9 10 0 0.0
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 0 2 4 6 8 10分 12 14 16 18 20 min
1. Tartaric acid 6. Pyroglutamic acid 2. Formic acid 7. Citric acid 3. Malic aicd 8. Fumaric acid 4. Lactic acid 9. Succinic acid 5. Acetic acid 10. Propionic acid
High Performance Liquid Chromatograph (HPLC) Sheet No. LC100029-05
Hitachi High-Technologies Corporation