- Email: [email protected]
Determination of phosphoethanolamine in animal tissues by gas chromatography with flame photometric detection
283
Journal of Chromatography, 494 (1989) 283-288 BLomedzcalApplrcatzons Elsevler Science Pubhshers B V . Amsterdam - Prmted m The Netherlands
CHROMBIO
4879
Note Determination of phosphoethanolamine in animal tissues by gas chromatography with flame photometric detection
HIROYUKI KATAOKA, NORIHISA SAKIYAMA, MITSUYO MAEDA and MASAMI MAKITA* Faculty of Pharmaceutrcal Sczences, Okayama UnrversLty, Tsushtma, Okayama 700 (Japan) (First recewed April 17th, 1989, revwed manuscript recewed May 30th, 1989)
Phosphoethanolamme (PEA) 1s known to be present m most ammal tlssues, but Its blologlcal importance 1sunknown, apart from its role as a possible intermediary m the metabohsm of phospholipld [ 11 However, it 1s reported that urmary excretion of PEA 1s slgmficantly increased m patients with hypophosphatasla [ 1 ] and metabohc born disease [ 21, and the PEA content of the brain 1s slgmflcantly decreased m the patients with Alzhelmer’s disease and Huntmgton’s disease [ 31 In addition, the influence of PEA on the growth of mammary carcinoma m rats [ 4,5] deserves attention The determination of PEA m bloloacal material has been carried out by colorlmetrlc [ 1,2] and fluorometrlc [6] assay methods. However, these methods lack sensltlvlty and usually require time-consummg preliminary clean-up of the sample Chromatographlc procedures utlhzmg an ammo acid analyser [ 7,8] or high-performance hquld chromatography [ 3,9-111 have also been used for the assay, but when they were applied directly to the blologlcal sample there appears to be difficulty m resolving PEA from interfering components [ 12-
141
Recently, we developed a convement and reliable method for the determlnation of ammoalkyl phosphates by gas chromatography (GC) with flame Ionlzatlon detection (FID) in which these compounds were analysed as then Nlsobutoxycarbonyl (N-isoBOC) methyl ester derlvatlves [ 151 This paper describes a sensitive and selective method for the determmatlon of PEA m blo0378-4347/89/$03
50
0 1989 Elsevler Science Publishers B V
288
96 l-100.1%, and the relative standard deviations were 0.2-3.2% (n= 3)) mdicatmg that this method is accurate and precise. The PEA contents of several animal tissues (Table II) shows that PEA is widely distributed. spleen, pancreas and brain contam considerable amounts whereas muscle, heart and hver do not. Similar results were reported by Hayase et al. [lo]. In conclusion, these experiments have conclusively demonstrated that PEA can be accurately and precisely determined by GC-FPD as its N-1soBOC methyl ester derivative This method is selective and sensitive, and complex biological materials can be analysed without any mterference from other substances We beheve that this method provides a useful tool m biochemical and biomedical research where PEA assay is required. REFERENCES
5 6 7 8 9 10 11 12 13 14 15 16 17 18
K Rasmussen, Dan Med Bull , 15 (1968) 1 A A Llcata, N Radfar, F C Bartter and E Bou, Am J Med ,64 (1978) 133 D W Elhson, M F Beal and J B Martin, Bram Res ,417 (1987) 389 T Kano-Sueoka, D M Cohen, i! Yamalzuml, S Nlshlmura, M Marl and H Fupkl, Proc Nat1 Acad Scl U S A, 76 (1979) 5741 T Kano-Sueoka and J E Ernck, Exp Cell Res ,136 (1981) 137 G Gundlach, A Kablsch, R. Voegeh, H Pralle and S Hartenfels, Anal Blochem ,142 (1984) 511 M V Sande and K V Camp, Invest Urol ,16 (1979) 445 J R Eastman and D Blxler, Chn Chem ,26 (1980) 1757 Y Hayase, M. Murakaml, I Kashlwakura and Y Takag, Yakugaku Zasshl, 106 (1986) 615 Y Hayase, M Murakaml and Y Takagl, Yakugaku Zasshl, 106 (1986) 694 D W Elhson, M F Beal and J B Martin, J Neuroscl Methods, 19 (1987) 305 K H Tachlkl, H C Hendrle, J Kellams and M H Apnson, Clm Chum Acta, 75 (1977) 455 B R Larsen, D S Gross0 and S Y Chang, J Chromatogr Scl , 18 (1980) 223 H F Erbersdobler, H G Greuhch and E Trautwem, J Chromatogr ,254 (1983) 332 H Kataoka, N Saklyama and M Maklta, J Chromatogr ,436 (1988) 67 J H Welsburger and W C Schneider, J Org Chem ,26 (1961) 1658 H Schlenk and J L Gellerman, Anal Chem (32 (1960) 1412 E C Horning, W J A VandenHeuvel and B G Creech, Methods Blochem Anal , 11 (1963) 69
Journal of Chromatography, 494 (1989) 283-288 BLomedzcalApplrcatzons Elsevler Science Pubhshers B V . Amsterdam - Prmted m The Netherlands
CHROMBIO
4879
Note Determination of phosphoethanolamine in animal tissues by gas chromatography with flame photometric detection
HIROYUKI KATAOKA, NORIHISA SAKIYAMA, MITSUYO MAEDA and MASAMI MAKITA* Faculty of Pharmaceutrcal Sczences, Okayama UnrversLty, Tsushtma, Okayama 700 (Japan) (First recewed April 17th, 1989, revwed manuscript recewed May 30th, 1989)
Phosphoethanolamme (PEA) 1s known to be present m most ammal tlssues, but Its blologlcal importance 1sunknown, apart from its role as a possible intermediary m the metabohsm of phospholipld [ 11 However, it 1s reported that urmary excretion of PEA 1s slgmficantly increased m patients with hypophosphatasla [ 1 ] and metabohc born disease [ 21, and the PEA content of the brain 1s slgmflcantly decreased m the patients with Alzhelmer’s disease and Huntmgton’s disease [ 31 In addition, the influence of PEA on the growth of mammary carcinoma m rats [ 4,5] deserves attention The determination of PEA m bloloacal material has been carried out by colorlmetrlc [ 1,2] and fluorometrlc [6] assay methods. However, these methods lack sensltlvlty and usually require time-consummg preliminary clean-up of the sample Chromatographlc procedures utlhzmg an ammo acid analyser [ 7,8] or high-performance hquld chromatography [ 3,9-111 have also been used for the assay, but when they were applied directly to the blologlcal sample there appears to be difficulty m resolving PEA from interfering components [ 12-
141
Recently, we developed a convement and reliable method for the determlnation of ammoalkyl phosphates by gas chromatography (GC) with flame Ionlzatlon detection (FID) in which these compounds were analysed as then Nlsobutoxycarbonyl (N-isoBOC) methyl ester derlvatlves [ 151 This paper describes a sensitive and selective method for the determmatlon of PEA m blo0378-4347/89/$03
50
0 1989 Elsevler Science Publishers B V
288
96 l-100.1%, and the relative standard deviations were 0.2-3.2% (n= 3)) mdicatmg that this method is accurate and precise. The PEA contents of several animal tissues (Table II) shows that PEA is widely distributed. spleen, pancreas and brain contam considerable amounts whereas muscle, heart and hver do not. Similar results were reported by Hayase et al. [lo]. In conclusion, these experiments have conclusively demonstrated that PEA can be accurately and precisely determined by GC-FPD as its N-1soBOC methyl ester derivative This method is selective and sensitive, and complex biological materials can be analysed without any mterference from other substances We beheve that this method provides a useful tool m biochemical and biomedical research where PEA assay is required. REFERENCES
5 6 7 8 9 10 11 12 13 14 15 16 17 18
K Rasmussen, Dan Med Bull , 15 (1968) 1 A A Llcata, N Radfar, F C Bartter and E Bou, Am J Med ,64 (1978) 133 D W Elhson, M F Beal and J B Martin, Bram Res ,417 (1987) 389 T Kano-Sueoka, D M Cohen, i! Yamalzuml, S Nlshlmura, M Marl and H Fupkl, Proc Nat1 Acad Scl U S A, 76 (1979) 5741 T Kano-Sueoka and J E Ernck, Exp Cell Res ,136 (1981) 137 G Gundlach, A Kablsch, R. Voegeh, H Pralle and S Hartenfels, Anal Blochem ,142 (1984) 511 M V Sande and K V Camp, Invest Urol ,16 (1979) 445 J R Eastman and D Blxler, Chn Chem ,26 (1980) 1757 Y Hayase, M. Murakaml, I Kashlwakura and Y Takag, Yakugaku Zasshl, 106 (1986) 615 Y Hayase, M Murakaml and Y Takagl, Yakugaku Zasshl, 106 (1986) 694 D W Elhson, M F Beal and J B Martin, J Neuroscl Methods, 19 (1987) 305 K H Tachlkl, H C Hendrle, J Kellams and M H Apnson, Clm Chum Acta, 75 (1977) 455 B R Larsen, D S Gross0 and S Y Chang, J Chromatogr Scl , 18 (1980) 223 H F Erbersdobler, H G Greuhch and E Trautwem, J Chromatogr ,254 (1983) 332 H Kataoka, N Saklyama and M Maklta, J Chromatogr ,436 (1988) 67 J H Welsburger and W C Schneider, J Org Chem ,26 (1961) 1658 H Schlenk and J L Gellerman, Anal Chem (32 (1960) 1412 E C Horning, W J A VandenHeuvel and B G Creech, Methods Blochem Anal , 11 (1963) 69