Validation of High Performance Liquid Chromatography Methods for Determination of Bioactive Sulfur Compounds in Garlic Bulbs

다국어 초록 (Multilingual Abstract)

High performance liquid chromatography (HPLC)methods for determination of organosulfur compounds (OSCs) before their enzymatic decomposition and allicin in garlic were optimized and validated. Four γ-glutamyl peptides, 2 S-alk(en)yl cysteine sulfoxides, and 2 Salk(en)yl cysteine were simply extracted with water,followed by the sensitive and selective determination of all compounds in a single run using reverse phase HPLC.
Allicin was extracted with dichloromethane and diethyl ether and analyzed by normal phase HPLC/UV system.
OSCs and allicin were separated on Hypurity Elite C18 and silica gel column, respectively. These methods exhibited excellent linearity for all of the analytes with good correlation coefficient (r>0.984). Average recoveries were in the range of 81.89-102.38% with good standard deviations. The limits of quantitation (LOQs) of OSCs and allicin ranged from 0.76 to 5.79 and 20.09 μg/mL,respectively. The precision results were expressed as relative standard deviation (RSD), ranged from 0.03 to 3.31% for intra-day and from 1.22 to 14.00% for the interday.
These analytical methods and validation data may be useful for quality evaluation of garlic bulbs.

참고문헌 (Reference)

1 Lawson LD, "r-Glutamyl-S-alkylcysteines in garlic and other Alliums spp.: Precursors of age-dependent trans-1-propenyl thiosulfinates" 54 : 436-444, 1991

2 Diego MD, "Validated liquid chromatographic method for quantitative determination of allicin in garlic powder and tablets" 30 : 2703-2707, 2007

3 Pruthi JS, "Thermal stability of allinase and enzymatic regeneration of flavor odourless garlic powder" 28 : 403-404, 1950

4 Rahman AU, "Studies in Natural Products Chemistry" Elsevier Science 455-467, 2000

5 Iberl B, "Quantitative determination of allicin and alliin from garlic by HPLC" 56 : 320-326, 1990

6 Yang CS, "Mechanisms of inhibition of chemical toxicity and carcinogenesis by diallyl sulfide (DAS) and related compounds from garlic" 131 : 1041S-1045S, 2001

7 Itakura Y, "How to distinguish garlic from the other Allium vegetables" 131 : 963S-967S, 2001

8 Arnault I, "High-performance ion-pair chromatography method for simultaneous analysis of alliin, deoxyalliin, allicin, and dipeptide precursors in garlic products using multiple mass spectrometry and UV detection" 991 : 69-75, 2003

9 Ziegler SJ, "HPLC of S-alk(en)yl-L-cysteine derivatives in garlic including quantitative determination of (+)-S-allyl-L-cysteine sulfoxide (alliin)" 55 : 372-378, 1989

10 Kubec R, "Gas chromatographic determination of S-alk(en)ylcysteine sulfoxides" 862 : 85-94, 1999

11 Mukherjee S, "Freshly crushed garlic is a superior cardioprotective agent than processed garlic" 57 : 7137-7144, 2009

12 Lamm DL, "Enhanced immunocompetence by garlic: Role in bladder cancer and other malignancies" 131 : 1067S-1070S, 2001

13 Banerjee SK, "Effect of garlic on cardiovascular disorders: A review" 1 : 1-14, 2002

14 Ichikawa M, "Determination of seven organosulfur compounds in garlic by highperformance liquid chromatography" 54 : 1535-1540, 2006

15 Kubec R, "Chromatographic methods for determination of S-substituted cysteine derivatives - A comparative study" 1216 : 6957-6963, 2009

16 Yeh YY, "Cholesterol-lowering effect of garlic extracts and organosulfur compounds: Human and animal studies" 131 : 989S-993S, 2001

17 Ichikawa M, "Changes in organosulfur compounds in garlic cloves during storage" 54 : 4849-4854, 2006

18 US DHHS, "Center for Veterinary Medicine (CVM): Guidance for Industry, Bioanalytical Method" Center Evaluation and Research (CDER), Food and Drug Administration, US Department of Health and Human Services 2001

19 Block E, "Allium chemistry: HPLC analysis of thiosulfinates from onion, garlic, wild garlic (Ramsoms), leek, scallion, shallot, elephant (great-headed) garlic, chive, and Chinese chive uniquely high allyl to methyl ratios in some garlic sample" 40 : 2418-2430, 1992

20 Karasaki Y, "A garlic lectin exerted an antitumor activity and induced apoptosis in human tumor cell" 34 : 7-13, 2001