Quantitative determination of total lipid hydroperoxides by a flow injection analysis system

Abstract

A flow injection analysis (FIA) system coupled with a fluorescence detection system using diphenyl-1-pyrenylphosphine (DPPP) was developed as a highly sensitive and reproducible quantitative method of total lipid hydroperoxide analysis. Fluorescence analysis of DPPP oxide generated by the reaction of lipid hydroperoxides with DPPP enabled a quantitative determination of the total amount of lipid hydroperoxides. Use of 1-myristoyl-2-(12-((7-nitro-2-1,3-benzoxadiazol-4-yl)amino) dodecanoyl)-sn-glycero-3-phosphocholine as the internal standard improved the sensitivity and reproducibility of the analysis. Several commercially available edible oils, including soybean oil, rape-seed oil, olive oil, corn oil, canola oil, safflower oil, mixed vegetable oils, cod liver oil, and sardine oil were analyzed by the FIA system for the quantitative determination of total lipid hydroperoxides. The minimal amounts of sample oils required were 50 microg of soybean oil (PV = 2.71 meq/kg) and 3 mg of sardine oil (PV = 0.38 meq/kg) for a single injection. Thus, sensitivity was sufficient for the detection of a small amount and/or low concentration of hydroperoxides in common edible oils. The recovery of sample oils for the FIA system ranged between 87.2+/-2.6% and 102+/-5.1% when PV ranged between 0.38 and 58.8 meq/kg. The CV in the analyses of soybean oil (PV = 3.25 meq/kg), cod liver oil (PV = 6.71 meq/kg), rapeseed oil (PV = 12.3 meq/kg), and sardine oil (PV = 63.8 meq/kg) were 4.31, 5.66, 8.27, and 11.2%, respectively, demonstrating sufficient reproducibility of the FIA system for the determination of lipid hydroperoxides. The squared correlation (r2) between the FIA system and the official AOCS iodometric titration method in a linear regression analysis was estimated at 0.9976 within the range of 0.35-77.8 meq/kg of PV (n = 42). Thus, the FIA system provided satisfactory detection limits, recovery, and reproducibility. The FIA system was further applied to evaluate changes in the total amounts of lipid hydroperoxides in fish muscle stored on ice.

Determination of lipid and protein hydroperoxides using the fluorescent probe diphenyl-1-pyrenylphosphine

“…This concentration is lower than that of 1 lEq/L reported in a batch method (Meguro, Akasaka, & Ohrui, 1990) but also from those using flow injection analysis system and HPLC methods (Kemmo, Soupas, Lampi, & Piironen, 2005;Sohn et al, 2005).…”

“…The fluorimetric determination of lipid hydroperoxides by means of the fluorescent probe diphenyl-1-pyrenylphosphine (DPPP) has been reported to be accurate and very sensitive in foodstuffs and biological materials using flow injection and HPLC post-column methods (Akasaka & Ohrui, 2000;Akasaka, Takamura, Ohrui, Meguro, & Hashimoto, 1996;Sohn, Taki, Ushio, & Ohshima, 2005). According to these latter authors, the phosphine moiety of the DPPP reacts with various HP producing a high intensity fluorescent DPPP oxide which is normally measured at 380 nm (excitation at 352 nm).…”

Control of lipid oxidation and meat color deterioration in skipjack tuna muscle during ice storage

“…Total lipid hydroperoxides were analyzed by a flow injection analysis (FIA) system equipped with a fluorescent detection system using DPPP as a fluorescent reagent, as previously described [16]. Briefly, a 1 mL aliquot of NBDlabeled PC (4.82 nmol/mL in methanol) was added to a 5 g portion of minced muscle as an internal standard, and TL were extracted and purified according to the Bligh and Dyer procedure [15].…”

Bioactive Properties of Japanese Fermented Fish Paste, Fish Miso, Using Koji Inoculated With Aspergillusoryzae

“…The degree of linoleic acid oxidation was measured at 48-h intervals by monitoring the total hydrogen peroxide content as well as oxygen absorption. The total amount of hydroperoxide was measured using DPPP as a fluorescent reagent and a flow-injection analysis system equipped with a fluorescence detection system [31]. The oxygen uptake was measured by gas chromatography using the method described by Ohshima et al [32].…”