Determination of hippuric acid by RP-HPLC using two different analytical columns - a short report

• Autorzy: Pegg R. B. , Rybarczyk A. , Amarowicz R.

Warianty tytułu

PL

Oznaczenie kwasu hipurowego metoda RP-HPLC przy uzyciu dwoch roznych kolumn analitycznych - krotki komunikat

• Języki publikacji: EN

Abstrakty

EN

Reversed-phase HPLC of liberated hippuric acid (HA) from an ACE assay in the presence of ACE inhibitory peptides derived from a crackling hydrolysate was conducted. The efficacies of two different analytical HPLC columns using identical mobile phases with an isocratic system were tested. Chromatograms revealed that the shorter C8 column (4.6 × 150 mm, 5 μm) was just as efficient as the longer C18 column (4.6 × 250 mm, 5 μm) in resolving liberated HA, but achieved this in a much shorter time (i.e., 3.67 cf 12.52 min). The presence of the crackling hydrolysate exhibited ACE-inhibiting activity by retarding the liberation of HA from the substrate hippuryl-L-histydyl-L-leucine (HHL) in a dose-dependent manner, and did not interfere with the chromatography. Hence, a quick reliable analytical methodology is at hand for the in vitro examination of various “bioactive peptide concoctions” for possible use in the development of functional foods.

PL

W pracy oznaczano kwas hipurowy (HA) metodą HPLC w układzie odwróconych faz. Posłużono się metodą stosowaną przy pomiarze aktywności peptydowych inhibitorów ACE, których źródłem w tym przypadku był hydrolizat białek skwarek wieprzowych. Do dwóch kolumn analitycznych zastosowano identyczną fazę ruchomą w układzie izokratycznym. Skuteczność rozdziału kwasu hipurowego przy użyciu krótszej kolumny C8 (4,6 × 150 mm, 5 μm) była podobna do tej, którą otrzymano stosując dłuższą kolumnę C18 (4,6 × 250 mm, 5 μm). Czas retencji dla HA w przypadku kolumny C8 był krótszy (3,67 min) niż przy zastosowaniu kolumny C18 (12,52 min). Użyty w badaniach hydrolizat wykazywał aktywność hamowania ACE poprzez hamowanie uwalniania HA z substratu HHL (hipurylo-L-histydylo-L-leucyna). Zawarte w hydrolizacie peptydy nie interferowały z HA. Zaprezentowana w pracy szybka metoda chromatograficzna może być stosowana w analizie in vitro biologicznie aktywnych peptydów w badaniach z obszaru żywności funkcjonalnej.

Słowa kluczowe

EN

hippuric acid; determination; angiotensin I converting enzyme; hypertension; bioactive peptide; analytical column; functional food

• Wydawca: -
• Czasopismo: Polish Journal of Food and Nutrition Sciences
• Rocznik: 2007
• Tom: 57
• Numer: 4
• Opis fizyczny: p.447-450,fig.,ref.

Twórcy

autor

Pegg R. B.

• The University of Georgia, Athens, GA, USA

autor

Rybarczyk A.

autor

Amarowicz R.

Bibliografia

• 1. Arihara K., Nakashima Y., Mukai T., Ishikawa S., Itoh M., Peptide inhibitors for angiotensin I-converting enzyme from enzymatic hydrolysates of porcine skeletal muscle proteins. Meat Sci., 2001, 57, 319–324.
• 2. Cheung H-S., Wang F-L., Ondetti M.A., Sabo E.F., Cushman D.W., Binding of peptide substrates and inhibitors of angiotensin- converting enzyme. J. Biol. Chem., 1980, 255, 401–407.
• 3. Curtis J.M., Dennis D., Waddell D.S., MacGillivray T., Ewart H.S., Determination of angiotensin-converting enzyme inhibitory peptide Leu-Lys-Pro-Asn-Met (LKPNM) in bonito muscle hydrolysates by LC-MS/MS. J. Agric. Food Chem., 2002, 55, 3919–3925.
• 4. Cushman D.W., Cheung H-S., Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung. Biochem. Pharmacol., 1971, 20, 1637–1648.
• 5. FitzGerald R.J., Murray B.A., Walsh D.J., Hypotensive peptides from milk proteins. J. Nutr., 2004, 134, 980S–988S.
• 6. Flaczyk E., Amarowicz R., Korczak J., Antioxidant activity of protein hydrolyzates from by-products of the food industry. J. Food Lipids, 2003, 10, 129–140.
• 7. Hata Y., Yamamoto M., Ohni M., Nakajima K., Nakamura Y., Takano T., A placebo-controlled study of the effect of sour milk on blood pressure in hypertensive subjects. Am. J. Clin. Nutr., 1996, 64, 767–771.
• 8. Hernández-Ledesma B., Recio I., Ramos M., Amigo L., Preparation of ovine and caprine b-lactoglobulin hydrolysates with ACE-inhibitory activity. Identification of active peptides from caprine b-lactoglobulin hydrolysed with thermolysin. Int. Dairy J., 2002, 12, 805–812.
• 9. Kannel W.B., Blood pressure as a cardiovascular risk factor. JAMA, 1996, 275, 1571–1576.
• 10. Kim Y.K., Chung B.H., A novel angiotensin-I-converting enzyme inhibitory peptide from human as1-casein. Biotechnol. Lett., 1999, 21, 575–578.
• 11. Kim S-K., Byun H-G., Park P-J., Shahidi F., Angiotensin I converting enzyme inhibitory peptides purified from bovine skin gelatin hydrolysate. J. Agric. Food Chem., 2001, 49, 2992–2997.
• 12. Li G-H., Le G-W., Shi Y-H., Shrestha S., Angiotensin I-converting enzyme inhibitory peptides derived from food proteins and their physiological and pharmacological effects. Nutr. Res., 2004, 24, 469–486.
• 13. Mehanna A.S., Dowling M., Liquid chromatographic determination of hippuric acid for the evaluation of ethacrynic acid as angiotensin converting enzyme inhibitor. J. Pharm. Biomed. Anal., 1999, 19, 967–973.
• 14. Meng Q.C., Oparil, S., Purification and assay methods for angiotensin-converting enzyme. J. Chromatogr. A, 1996, 743, 105–122.
• 15. Nakashima Y., Arihara K., Sasaki A., Mio H., Ishikawa S., Itoh M., Antihypertensive activities of peptides derived from porcine skeletal muscle myosin in spontaneously hypertensive rats. J. Food Sci., 2002, 67, 434–437.
• 16. Nicholls M.G., Richards, A.M., Agarwal, M., The importance of the renin-angiotensin system in cardiovascular disease. J. Human Hypertension, 1998, 12, 295–299.
• 17. Ondetti M.A., Cushman, D.W., Enzymes of the rennin-angiotensin system and their inhibitors. Ann. Rev. Biochem., 1982, 51, 283–308.
• 18. Pihlanto-Leppälä A., Koskinen P., Piilola K., Tupasela T., Korhonen H., Angiotensin I-converting enzyme inhibitory properties of whey protein digests: concentration and characterization of active peptides. J. Dairy Res., 2000, 67, 53–64.
• 19. Seppo L., Jauhiainen T., Poussa T., Korpela R., A fermented milk high in bioactive peptides has a blood pressure-lowering effect in hypertensive subjects. Am. J. Clin. Nutr., 2003, 77, 326–330.
• 20. Tauzin J., Miclo L., Gaillard J.-L., Angiotensin-I-converting enzyme inhibitory peptides from tryptic hydrolysate of bovine as2-casein. FEBS Lett., 2002, 531, 369–374.
• 21. Wanasundara P.K.J.P.D., Ross A.R.S., Amarowicz R., Ambrose S.J., Pegg R.B., Shand P.J., Peptides with angiotensin I-converting enzyme (ACE) inhibitory activity from defibrinated, hydrolyzed bovine plasma. J. Agric. Food Chem., 2002, 50 6981– 6988.
• 22. Wu J., Aluko R.E., Nakai S., Structural requirements of angiotensin I-converting enzyme inhibitory peptides: Quantitative structure–activity relationship study of di- and tripeptides. J. Agric. Food Chem., 2006, 54, 732–738.
• 23. Yoshii H., Tachi N., Ohba R., Sakamura O., Takeyama H., Itani T., Antihypertensive effect of ACE inhibitory oligopeptides from chicken egg yolks. Comp. Biochem. Physiol. Part C, 2001, 128C, 27–33.