jkra The Journal of the Korean Rheumatism Association J Korean Rheum Assoc 1226-8070 2233-4718 The Korean Rheumatism Association 10.4078/jkra.2010.17.4.368 jkra-17-368 Original Article Measurement of Purine Contents in Korean Alcoholic Beverages JunJae-Bum 1 NaYoung-In 2 KimHyun-Jin 3 KimShin-Hee 3 ParkYoo-Sin 3 KangJuseop 3 Department of Internal Medicine, Hanyang University Hospital for Rheumatic Diseases, College of Medicine, Hanyang University, Seoul, Korea Institute of Rheumatism, Hanyang University, College of Medicine, Hanyang University, Seoul, Korea Department of Pharmacology & Clinical Pharmacology Lab, College of Medicine, Hanyang University, Seoul, Korea 122010 23122010 17 4 368 375 02082010 01092010 24092010 Copyright © 2010 The Korean Rheumatism Association 2010 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Objective

Gout is one of the most common forms of inflammatory arthritides among men, which is caused primarily by chronic hyperuricemia. Although pharmacological therapy is the mainstay treatment to manage gout, limiting the consumption of dietary purine is also important. Several epidemiological studies have reported that alcohol consumption is closely related to hyperuricemia and gout. The objective of this study was to determine the purine content in common Korean alcoholic beverages using high performance liquid chromatography (HPLC) to provide a dietary guideline for those with hyperuricemia or gout.

 Methods

Thirty-five alcoholic beverages were analyzed. Blindly labeled samples of each alcoholic beverage were degassed and frozen. The sample preparation prior to HPLC followed the methods of Japanese researchers. HPLC was performed to analyze adenine, guanine, hypoxanthine, and xanthine content in the alcoholic beverages.

 Results

The standard curves were linear for all purines. Purine contents were as follows: beer (42.26~146.39 μmol/L, n=12), medicinal wine (8.2 and 40.41 μmol/L, n=2), rice wine (13.19 μmol/L), Makgeolri (11.71 and 24.72 μmol/L, n=2), red wine (0, 6.03, and 17.9 μmol/L, n=3). No purines were found in fruit wine (n=2), Kaoliang (n=1), white wine (n=1), or distilled alcoholic beverages, such as soju (n=10) or whiskey (n=1).

 Conclusion

Among popular Korean alcoholic beverages, beer contained a considerable amount of purines, whereas distilled alcoholic beverages did not. Patients with either gout or hyperuricemia should avoid alcoholic beverages, especially those containing large amounts of purines.

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Chromatogram of standard purine bases and a sample.

Standard curves for for purine bases. (A) Adenine, (B) Guanine, (C) Hypoxanthine, (D) Xanthine.

High performance liquid chromatography (HPLC) conditions

1. HPLC apparatus (PDA-HPLC)
Pump Binary Pump 126 (Beckman Coulter Co., Fullerton, USA)
Sampler Autoinjector (Beckman Coulter Co., Fullerton, USA)
Column 5mm i.d.×300 mm length (Shodex Asahipak HQ-310, Shoko America, Inc. Japan)
Column oven Photodiode Array detector 168 (Beckman Coulter Co., Fullerton, USA)
Detector Karat 32 (Beckman Coulter Co., Fullerton, USA)
Software VS-6000CFI (Vision Scientific Co., Ltd., Korea)
Centrifuge KMC-1300V (Vision Scientific Co., Ltd., Korea)
Vortex-mixer Sartorius AC2115 (Sartorious Mechatronics, Ltd., Germany)
Balance Model 420 A+ (Thermo Scientific Inc. USA)
pH meter PC-420 (Corning Co., USA)
Stirrer & hot plate SH-3600 (Saehan Ultrasonic Co., Korea)
Sonicator Binary Pump 126 (Beckman Coulter Co., Fullerton, USA)
2. Mobile phase
Sodium phosphate Monobasic Anhydrous 98% (Yakuri Pure Chemicals Co., Ltd., Japan)
Purelab Classic triple distilled water (ELGA, UK)
O-Phosphoric acid 85% (Sigma-Aldrich Co., Ltd., USA)
Perchloric acid 70% (Junsei Chemical Co., Ltd., Japan)
Potassium Hydroxide 95% (Samchun Pure Chemical Co., Ltd., Korea)
Standard Adenine 99% (Sigma-Aldrich Co., Ltd., USA)
Guanine hydrochloride (Sigma-Aldrich Co., Ltd., USA)
hypoxanthine 99% (Sigma-Aldrich Co., Ltd., USA)
Xanthine sodium salt 99% (Sigma-Aldrich Co., Ltd., USA)
3. Experimental conditions
Mobile phase 150 mM Sodium Phosphate buffer (pH : 2.5)
Apparatus Flow Rate: 0.6 mL/min
UV Wavelength: 260 nm
Column oven: 35.0°C
Column: Shodex Asahipak HQ-310

Accuracy and precision of the purine base standards

1. Adenine
Standard (μg/mL) Adenine (μg/mL) Average SD Precision (% CV) Accuracy (% RE)
1 2 3
20 20.89 22.02 21.99 21.63 0.65 2.98 108.17
10 10.35 10.75 10.57 10.55 0.20 1.89 105.53
8 8.46 8.81 9.29 8.85 0.42 4.69 110.68
5 5.48 5.77 5.86 5.70 0.20 3.51 114.04
2.5 2.78 2.88 2.88 2.84 0.06 2.02 113.76
1 1.19 1.24 1.22 1.21 0.02 1.97 121.49
0.5 0.63 0.63 0.64 0.63 0.01 1.06 126.40
0.25 0.37 0.38 0.36 0.37 0.01 2.31 147.98
2. Guanine
Standard (μg/mL) Guanine (μg/mL) Average SD Precision (% CV) Accuracy (% RE)
1 2 3
20 21.42 22.45 22.51 22.13 0.61 2.76 110.63
10 10.65 11.02 10.86 10.84 0.18 1.68 108.44
8 8.72 9.07 9.21 9.00 0.26 2.84 112.49
5 5.76 6.03 6.47 6.09 0.36 5.93 121.74
2.5 3.03 3.09 3.14 3.09 0.05 1.73 123.44
1 1.34 1.36 1.39 1.36 0.03 2.03 136.27
0.5 0.82 0.87 0.80 0.83 0.04 4.57 166.60
0.25 0.55 0.55 0.54 0.55 0.01 1.14 218.91
3. Hypoxanthine
Standard (μg/mL) Hypoxanthine (μg/mL) Average SD Precision (% CV) Accuracy (% RE)
1 2 3
20 21.63 22.73 22.82 22.39 0.67 2.98 111.97
10 10.82 11.35 10.94 11.04 0.28 2.51 110.37
8 8.78 9.17 9.14 9.03 0.22 2.44 112.87
5 5.78 6.01 6.11 5.97 0.17 2.87 119.34
2.5 3.03 3.14 3.19 3.12 0.08 2.59 124.67
1 1.29 1.38 1.36 1.34 0.04 3.31 134.47
0.5 0.85 0.95 0.85 0.89 0.06 6.61 177.16
0.25 0.53 0.55 0.67 0.58 0.08 12.98 233.50
4. Xanthine
Standard (μg/mL) Xanthine (μg/mL) Average SD Precision (% CV) Accuracy (% RE)
1 2 3
20 23.42 24.56 24.54 24.18 0.65 2.70 120.88
10 11.65 12.22 11.86 11.91 0.29 2.44 119.11
8 9.51 9.87 9.76 9.71 0.18 1.87 121.41
5 6.11 6.81 6.55 6.49 0.35 5.45 129.87
2.5 3.14 3.36 3.54 3.35 0.20 5.89 133.89
1 1.75 1.63 1.61 1.66 0.07 4.32 166.33
0.5 0.91 1.07 0.92 0.97 0.09 9.58 193.22
0.25 0.65 0.59 0.81 0.68 0.11 16.34 273.94

SD, standard deviation; CV, coefficients of variation; RE, relative error

Purine contents (μmol/L) of co mmon Kor ean alcoholic beverages

Category No. Adeni ne Guani ne Hypoxanthine Xanthine Total
Beer Local 1 11.25 26.44 3.09 31.99 72.77
Local 2 11.03 29.53 2.2 38.43 81.19
Local 3 13.32 31.24 6.39 45.09 96.04
Local 4 10.43 12.53 0 19.3 42.26
Local 5 15.84 48.94 3.01 28.03 95.82
Imported 6 19.76 85.67 13.96 27 146.39
Imported 7 10.51 20.74 2.35 10.05 43.65
Imported 8 16.87 30.6 2.28 12.12 61.87
Imported 9 10.29 16.1 2.64 20.28 49.31
Imported 10 29.16 58 4.11 28.32 119.59
Imported 11 16.87 26.33 4.56 25.85 73.61
Imported 12 9.77 10.71 3.09 29.75 53.32
Soju 1 0 0 0 0 0
2 0 0 0 0 0
3 0 0 0 0 0
4 0 0 0 0 0
5 0 0 0 0 0
6 0 0 0 0 0
7 0 0 0 0 0
8 0 0 0 0 0
9 0 0 0 0 0
10 0 0 0 0 0
Medicinal wine 1 0 2.03 6.17 0 8.2
2 3.4 12.9 7.57 16.54 40.41
Makgeolri 1 6.96 13.06 4.7 0 24.72
2 1.85 1.92 2.2 5.74 11.71
Fruit wine 1 0 0 0 0 0
2 0 0 0 0 0
Kaoliang 0 0 0 0 0
Whisky 0 0 0 0 0
Wine Red 1 0 0 0 6.03 6.03
Red 2 0 0 0 0 0
Red 3 2.29 4.58 0 11.03 17.9
White 4 0 0 0 0 0
Rice wine 2.07 2.67 8.45 0 13.19