David Wiles

The identification of salicylates as normal constituents of serum: a link between diet and health?

AIM: To examine sera for the presence of salicylic acid and 2,3- and 2,5-dihydroxybenzoic acids (2,3- and 2,5-DHBA), in individuals not taking salicylate drugs. METHODS: Extracts of acidified serum samples were analysed by high performance liquid chromatography with electro-chemical detection. The chromatographic conditions were altered, and the retention times of the unknown compounds compared against authentic salicylic acid, 2,3-DHBA, and 2,5-DHBA. Serum samples (some spiked with salicylic acid) were incubated with salicylate hydroxylase and analyses undertaken. An extract of acidified serum was derivatised using N-methyl-N-trimethylsilyltrifluoroacetamide and the salicylic acid derivative identified by gas chromatography-mass spectrometry. RESULTS: Salicylic acid, 2,3-DHBA, and 2,5-DHBA were identified as being normal constituents of serum. CONCLUSIONS: Salicylic acid, 2,3-DHBA, and 2,5-DHBA possess anti-inflammatory properties. The finding that these compounds are present as normal constituents of serum, possibly arising from diet, raises important questions as to their role in the promotion of health.

Identification and determination of salicylic acid and salicyluric acid in urine of people not taking salicylate drugs

Background Salicylic acid (SA) is present in the serum of people who have not taken salicylate drugs. Now we have examined the urine of these subjects and found that it contains SA and salicyluric acid (SU). We have established the identities of these phenolic acids and determined their concentrations. Methods and Results The acidic hydrophobic compounds of urine were separated using high-performance liquid chromatography (HPLC) and were detected and quantified electrochemically. Two approaches were used to establish the identity of SA and SU. First, the retention times (Rt) of the substances extracted and those of SA and SU were compared under two sets of chromatographic conditions; the Rt of the compounds suspected to be SA and SU and those of the authentic substances were very similar under both sets of conditions. Second, the unknown substances, isolated by HPLC, were treated with acetyl chloride in methanol and compared with the methyl esters of SA and SU by using gas chromatography-mass spectrometry; the unknown compounds after esterification had very similar mass spectra and gas chromatographic Rt to those of methyl salicylate and methyl salicylurate. The median (n = 10) urinary concentration of SA was 0·56 μmol/L (range 0·07-0·89 μmol/L) and that of SU was 3·20 μmol/L (range 1·32-6·54 μmol/L). SA and its major urinary metabolite, SU, were found in the urine of all of the 10 people examined.

Clinical and Biochemical Determinants of Plasma Lipid Peroxide Levels in Type 2 Diabetes

The enhanced risk and increased severity of atheroma in diabetes is well recognized but, as yet, incompletely explained. A cross-sectional study of vascular disease risk factors in a group of type 2 diabetic patients from South West Scotland has revealed an association between glycaemic control, assessed by HbA1 level and plasma lipid peroxides measured by a specific high-performance liquid chromatography method. Duration of diabetes appeared to be a subsidiary contributor to lipid peroxidation. We suggest this evidence supports the importance of glycaemic control in modulating glyco-oxidative mechanisms probably crucial to production of diabetic complications. Atherosclerosis prevention in diabetes may hinge on exemplary simultaneous control of both hyperglycaemia and hyperlipidaemia.