Determination of Phosphatidylcholine Hydroperoxide in Dry Cured Meat Products by Normal-Phase High Performance Liquid Chromatography

doi:10.7506/spkx1002-6630-201614027

Abstract

Abstract:

A normal-phase high-performance liquid chromatographic (HPLC) method was established to determine
phosphatidylcholine monohydroperoxide (PC-OOH) in dry cured meat products. The PC-OOH in dry cured meat samples
was extracted with chloroform/methanol (2:1, V/V), followed by determination by normal-phase HPLC connected to a
photo-diode array detector (HPLC-PAD) with synthetic C16:0/18:2 PC-OOH as the standard. The chromatographic conditions
were established using a Lichrosorb Si60 column (250 mm × 4.0 mm, 5 μm) with a mobile phase consisting of water and
n-hexane/isopropanol (3:2, V/V) in gradient elution at a flow rate of 1.0 mL/min. The detection wavelength was set at
234 nm. Results indicated that the linearity ranged from 20 to 600 nmol/mL (R2 = 0.999 9), and the limits of detection (LOD)
and limits of quantification (LOQ) were 5.8 nmol/mL and 19.4 nmol/mL, respectively. The recoveries from spiked samples
at concentration levels of 32–240 nmol/g were in the range of 79.4%–91.9%. The contents of PC-OOH in farm-made sausage
A, B and Chinese ham samples were 92.4, 106.4 and 102.5 nmol/g, respectively, while PC-OOH in smoked meat was not
detected. This method proved to be of high maneuverability, excellent sensitivity and high accuracy, and could be used for
quantitative analysis of PC-OOH in dry cured meat samples.

Key words: dry cured meat, 1-palmitoyl-2-linoleoyl-phosphatidylcholine, hydroperoxide, normal-phase HPLC

CLC Number:

TS207.3

REN Shuang, SONG Hui, GENG Zhiming, WANG Daoying, ZHANG Muhan, SUN Chong, XU Weimin. Determination of Phosphatidylcholine Hydroperoxide in Dry Cured Meat Products by Normal-Phase High Performance Liquid Chromatography[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201614027.

References

[1] DOBARGANES M C, VELASCO J. Anal y s i s o f l ipid

hydroperoxides[J]. European Journal of Lipid Science and

Technology, 2002, 104(7): 420-428. DOI:10.1002/1438-

9312(200207)104:7<420::AID-EJLT420>3.0.CO;2-N.

[2] SHAHIDI F, ZHONG Y. Lipid oxidation and improving the oxidative

stability[J]. Chemical Society Reviews, 2010, 39(11): 4067-4079.

DOI:10.1039/b922183m.

[3] COHN J S. Oxidized fat in the diet, postprandial lipaemia and

cardiovascular disease[J]. Current Opinion in Lipidology, 2002, 13(1):

19-24. DOI:10.1097/00041433-200202000-00004.

[4] CORSINOVI L, BIASI F, POLI G, et al. Dietary lipids and their

oxidized products in Alzheimer’s disease[J]. Molecular Nutrition

and Food Research, 2011, 55(Suppl 2): 161-172. DOI:10.1002/

mnfr.201100208.

[5] AYAKO K, TOMOHIRO S, TAKAAKI A, et al. Dietary lipid

peroxidation products and DNA damage in colon carcinogenesis[J].

European Journal of Lipid Science and Technology, 2002, 104:

439-447. DOI:10.1002/1438-9312(200207)104:7<439::AIDEJLT439>

3.0.CO;2-K.

[6] RIZVI S I, MAURYA P K. Markers of oxidative stress in erythrocytes

during aging in humans[J]. Annals of the New York Academy of

Sciences, 2007, 1100(1): 373-382. DOI:10.1196/annals.1395.041.

[7] HUI S, CHIBA H, JIN S, et al. Analyses for phosphatidylcholine

hydroperoxides by LC/MS[J]. Journal of Chromatography B

Analytical Technologies in the Biomedical and Life Sciences, 2010,

878(20): 1677-1682. DOI:10.1016/j.jchromb.2010.04.011.

[8] HOPPER T H. Official and tentative methods of AOCS[J]. Journal of

Oil and Fat Industries, 1951, 28: 4-26. DOI:10.1007/BF02648658.

[9] ASAKAWA T, MATSUSHITA S. Colorimetric determination of

peroxide value with potassium iodide-silica gel reagent[J]. Journal

of the American Oil Chemists’ Society, 1978, 55(8): 619-620.

DOI:10.1007/BF02678958.

[10] MIHARA M, UCHIYAMA M. Determination of malonaldehyde

precursor in tissues by thiobarbituric acid test[J]. Analytical

Biochemistry, 1978, 86(1): 271-278. DOI:10.1016/0003-

2697(78)90342-1.

[11] FURUKAWA I, KUROOKA S, ARISUE K, et al. Assays of serum

lipase by the “BALB-DTNB method” mechanized for use with discrete

and continuous-flow analyzers[J]. Clinical Chemistry, 1982(1): 110-

113. DOI:10.1105/tpc.109.065557.

[12] MAWATARI S, MURAKAMI K. Analysis of membrane phospholipid

peroxidation by isocratic high-performance liquid chromatography

with ultraviolet detection[J]. Analytical Biochemistry, 1998, 264(1):

118-123. DOI:10.1006/abio.1998.2830.

[13] OHSHIMA T, HOPIA A, GEMEN J B, et al. Determination

of hydroperoxides and structures by high-performance liquid

chromatography with post-column detection with diphenyl-1-

pyrenylphosphine[J]. Lipids, 1996, 31(10): 1091-1096. DOI:10.1007/

BF02522467.

[14] MILNE G L, PORTER N A. Separation and identification of

phospholipid peroxidation products[J]. Lipids, 2001, 36(11): 1265-

1275. DOI:10.1007/s11745-001-0841-2.

[15] WANG D, XU W, XU X, et al. Determination of intramuscular

phospholipid classes and molecular species in Gaoyou duck[J].

Food Chemistry, 2009, 112(1): 150-155. DOI:10.1016/

j.foodchem.2008.05.052.

[16] 章建浩, 黄明, 赵改名, 等. 干腌火腿工艺过程蛋白质与脂质分

解氧化及其对风味的贡献[J]. 食品工业科技, 2003(12): 103-105.

DOI:10.3969/j.issn.1002-0306.2003.12.052.

[17] 杨红菊, 乔发东, 马长伟, 等. 脂肪氧化和美拉德反应与肉品风

味质量的关系[J]. 肉类研究, 2004, 18(1): 25-28. DOI:10.3969/

j.issn.1001-8123.2004.01.012.

[18] WANG X H, HIDEKI U, TOSHIAKI O. Distributions of

hydroperoxide positional isomers generated by oxidation of

1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine in liposomes

and in methanol solution[J]. Lipids, 2003, 38(1): 65-72. DOI:10.1007/

s11745-003-1032-x.

[19] HUI S P, MURAI T, YOSHIMURA T, et al. Improved HPLC assay

for lipid peroxides in human plasma using the internal standard of

hydroperoxide[J]. Lipids, 2005, 40(5): 515-522. DOI:10.1007/s11745-

005-1412-2.

[20] AKASAKA K, OHRUI H, MEGURO H, et al. Determination of

triacylglycerol and cholesterol ester hydroperoxides in human plasma

by high-performance liquid chromatography with fluorometric

postcolumn detection[J]. Journal of Chromatography A, 1993, 617(2):

205-211. DOI:10.1016/0378-4347(93)80489-Q.

[21] KAZUAKI A, IKUKO S, HIROSHI O, et al. A simple fluorometry

of hydroperoxides in oils and foods[J]. Bioscience Biotechnology and

Biochemistry, 1992, 56(4): 605-607. DOI:10.1271/bbb.56.605.

[22] FOLCH J, LEES M, SLOANE STANLEY G H. A simple method for

the isolation and purification of total lipides from animal tissues[J].

Journal of Biological Chemistry, 1957, 226(1): 497-509. DOI:10.1016/

j.foodchem.2015.07.102.

[23] GARCIA REGUEIRO J A, GIBERT J, DIAZ I. Determination

of neutral lipids from subcutaneous fat of cured ham by capillary

gas chromatography and liquid chromatography[J]. Journal of

Chromatography A, 1994, 667: 225-233. DOI:10.1016/0021-

9673(94)89071-4.

[24] ARAI H, SUZUKI T, TAKAMA K, et al. Reactivity of mammalian

15-lipoxygenase with phospholipids in large unilamellar liposomes[J].

Biochemical and Biophysical Research Communications, 1996,

228(3): 675-682. DOI:10.1006/bbrc.1996.1716.

[25] MILNE G L, SEAL J R, HAVRILLA C M, et al. Identification and

analysis of products formed from phospholipids in the free radical

oxidation of human low density lipoproteins[J]. Journal of Lipid

Research, 2005, 46(2): 307-319. DOI:10.1194/jlr.M400311-JLR200.

[26] HUI S, YOSHIMURA T, MURAI T, et al. Determination of

regioisomeric hydroperoxides of fatty acid cholesterol esters produced

by photosensitized peroxidation using HPLC[J]. Analytical Sciences,

2000, 16(10): 1023-1028. DOI:10.2116/analsci.16.1023.

[27] PAN X, USHIO H, OHSHIMA T. Photo-oxidation of lipids

impregnated on the surface of dried seaweed (Porphyra yezoensis

Ueda). hydroperoxide distribution[J]. Journal of the American Oil

Chemists’ Society, 2004, 81(8): 765-771. DOI:10.1007/s11746-004-

0976-6.

[28] BOCHKOV V N, OSKOLKOVA O V, BIRUKOV K G, et al.

Generation and biological activities of oxidized phospholipids[J].

Antioxidants and Redox Signaling, 2010, 12(8): 1009-1059.

DOI:10.1089/ars.2009.2597.