BibTex RIS Kaynak Göster

Detection of the Milk Proteins by RP-HPLC (in English)

Yıl 2010, Cilt: 35 Sayı: 1, 1 - 7, 01.02.2010

Öz

In this study the bovine κ-, α- and β-caseins, α-lactalbumin and β-lactoglobulin by HPLC-UV using an RP column was detected by developed a simple and rapid method. Gradient elution was carried out at a 1 ml/min flow rate, and a temperature of 25 oC, using a mixture of two solvents. Solvent A was acetonitrile: water: trifluoroacetic acid (100:900:1) and Solvent B was acetonitrile:water:trifluoroacetic acid (900:100:1). The effluent flow was monitored by a UV detector at 220 nm. Different chromatographic profiles were obtained for bovine milk proteins which was prepared with different methods for RP-HPLC. It was suggested that using of Solvent A and Solvent B during the sample preparation by addition into the sample before injection at given ratio was allowed the separation more clear and effective than the other sample preparation methods. By the current sample preparation technique and elution practice which was occurred in this study, it was achieved detection of bovine milk proteins was being simple, rapid and sensitive provide.

Kaynakça

  • Walstra P, Jennes R. 1984. Dairy Chemistry and Physics, John Willey and Sons Inc., New York, 467 pp.
  • Ng-Kwai-Hang KF, Kroeker EM. 1984. Rapid separation and quantification of major caseins and whey proteins of bovine milk by polyacrylamide gel electrophoresis. J Dairy Sci, 67: 3052-3056.
  • Basch JJ, Douglas FW, Procino LG, Holsinger VH, Farrel HM. 1985. Quantification of caseins and whey proteins of processed milks and whey protein concentrates, application of gel electrophoresis and comparison with Harland-Ashworth procedure. J Dairy Sci, 68: 23-31.
  • Andrews AT, Taylor MD, Owen AJ. 1985. Rapid analysis of bovine milk proteins by fast protein liquid chromatography. J Chrom, 348: 177-185.
  • Dumay E, Cheftel JC. 1989. Heat treatments of a β-lactoglobulin concentrate at slightly alkaline pHs. Effects on the solubility and the chromatographic behaviour of β-lactoglobulin and α-lactalbumin. Sci Aliment, 9: 561-582.
  • Visser S, Slangen CJ, Rollema HS. 1991. Phenotyping of bovine milk proteins by reverse-phase high-performance liquid chromatography. J Chrom, 548: 361-370.
  • de Jong N, Visser S, Olieman C. 1993. Determination of milk proteins by capillary electrophoresis. J Chrom A, 652: 207-213.
  • Leonil J, Molle M, Gaucheron F, Arpino P, Guenot P, Maubois JL. 1995. Analysis of major bovine milk proteins by on-line high-performance liquid chromatography and electrospray ionization mass spectrometry. Lait, 75: 193-210.
  • Ferreira IMPLVO, Cacote H. 2003. Detection and quantification of bovine, ovine and caprine milk percentages in protected denomination of origin cheeses by reversed-phase high-performance liquid chromatography of beta-lactoglobulins. J Chrom A, 1015: 111-118.
  • Veloso ACA, Teixeira N, Peres AM, Mendonca A, Ferreira IMPLVO. 2004. Evaluation of cheese authenticity and proteolysis by HPLC and urea-polyacrylamide gel electrophoresis. Food Chem, 87: 289-295.
  • Bovenhuis H, Verstege AJM. 1989. Improved method for phenotyping milk protein variants by isoelectric focusing using Phast System. Neth Milk Dairy J, 43: 447- 451.
  • Nieuwenhuijse JA, van Boekel MAJS, Walstra P. 1991. On the heat-induced association and dissociation of proteins in concentrated skim milk. Neth Milk Dairy J, 45: 3-22.
  • Groen AF, van der Vegt R, van Boekel MAJS, de Rouw OLAM, Vos H. 1994. Case study on individual animal variation in milk protein composition as estimated by high-pressure liquid chromatography. Neth Milk Dairy J, 48: 201-212.
  • Parris N, White AE, Farrell HM. 1990. Identification of altered proteins in nonfat dry milk powder prepared from heat-treated skim milk. J Agric Food Chem, 38: 824- 829.
  • Bobe G, Beitz DC, Freeman AE, Lindberg GL. 1998. Separation and quantification of bovine milk proteins by reverse-phase high-performance liquid chromatography. J Agric Food Chem, 46: 458-463.
  • Bobe G, Beitz DC, Freeman AE, Lindberg GL. 1998. Sample preparation affects separation of whey proteins by reverse-phase high-performance liquid chromatography. J Agric Food Chem, 46: 1321-1325.
  • Bordin G, Raposo FC, de la Calle B, Rodriguez AR. 2001. Identification and quantification of major bovine milk proteins by liquid chromatography. J Chrom A, 928: 63-76.
  • Veloso ACA, Teixeira N, Ferreira IMPLVO. 2002. Separation and quantification of the major casein fractions by reverse-phase high-performance liquid chromatography and urea-polyacrylamide gel electrophoresis: Detection of milk adulterations. J Chrom A, 967: 209-218.

Süt Proteinlerinin RP-HPLC ile Saptanması (İngilizce)

Yıl 2010, Cilt: 35 Sayı: 1, 1 - 7, 01.02.2010

Öz

Bu çalışmada, κ-, α- and β-kazein ile α-laktalbumin and β-laktoglobulin, bir RP kolonunun kullanıldığı HPLC-UV ile basit ve hızlı bir yöntem geliştirilerek saptanmıştır. Gradient elüsyonu, 1 ml/dk akış hızında ve 25 oC sıcaklıkta iki çözücü karışımı kullanılarak yürütülmüştür. A çözücüsü asetonitril: su:trifloroasetik asit (100:900:1) ve B çözücüsü asetonitril: su: trifloroasetik asit (100:900:1) karışımlarından oluşmaktadır. Akış, bir UV dedektörü kullanılarak 220 nm’de kaydedilmiştir. Farklı yöntemler kullanılarak RP-HPLC analizi için hazırlanan örneklerde, süt proteinlerine ait farklı kromatografik profiller elde edilmiştir. Diğer örnek hazırlama yöntemlerine kıyasla, enjeksiyondan önce belirtilen oranlarda A ve B çözücülerinin kullanıldığı örnek hazırlama yöntemi ile separasyonun daha keskin ve efektif olduğu ortaya konulmuştur. Bu çalışmada ortaya konulan örnek hazırlama tekniği ve elusyon pratiği ile, süt proteinlerinin analizi basit, hızlı ve duyarlı bir şekilde gerçekleştirilmiştir.

Kaynakça

  • Walstra P, Jennes R. 1984. Dairy Chemistry and Physics, John Willey and Sons Inc., New York, 467 pp.
  • Ng-Kwai-Hang KF, Kroeker EM. 1984. Rapid separation and quantification of major caseins and whey proteins of bovine milk by polyacrylamide gel electrophoresis. J Dairy Sci, 67: 3052-3056.
  • Basch JJ, Douglas FW, Procino LG, Holsinger VH, Farrel HM. 1985. Quantification of caseins and whey proteins of processed milks and whey protein concentrates, application of gel electrophoresis and comparison with Harland-Ashworth procedure. J Dairy Sci, 68: 23-31.
  • Andrews AT, Taylor MD, Owen AJ. 1985. Rapid analysis of bovine milk proteins by fast protein liquid chromatography. J Chrom, 348: 177-185.
  • Dumay E, Cheftel JC. 1989. Heat treatments of a β-lactoglobulin concentrate at slightly alkaline pHs. Effects on the solubility and the chromatographic behaviour of β-lactoglobulin and α-lactalbumin. Sci Aliment, 9: 561-582.
  • Visser S, Slangen CJ, Rollema HS. 1991. Phenotyping of bovine milk proteins by reverse-phase high-performance liquid chromatography. J Chrom, 548: 361-370.
  • de Jong N, Visser S, Olieman C. 1993. Determination of milk proteins by capillary electrophoresis. J Chrom A, 652: 207-213.
  • Leonil J, Molle M, Gaucheron F, Arpino P, Guenot P, Maubois JL. 1995. Analysis of major bovine milk proteins by on-line high-performance liquid chromatography and electrospray ionization mass spectrometry. Lait, 75: 193-210.
  • Ferreira IMPLVO, Cacote H. 2003. Detection and quantification of bovine, ovine and caprine milk percentages in protected denomination of origin cheeses by reversed-phase high-performance liquid chromatography of beta-lactoglobulins. J Chrom A, 1015: 111-118.
  • Veloso ACA, Teixeira N, Peres AM, Mendonca A, Ferreira IMPLVO. 2004. Evaluation of cheese authenticity and proteolysis by HPLC and urea-polyacrylamide gel electrophoresis. Food Chem, 87: 289-295.
  • Bovenhuis H, Verstege AJM. 1989. Improved method for phenotyping milk protein variants by isoelectric focusing using Phast System. Neth Milk Dairy J, 43: 447- 451.
  • Nieuwenhuijse JA, van Boekel MAJS, Walstra P. 1991. On the heat-induced association and dissociation of proteins in concentrated skim milk. Neth Milk Dairy J, 45: 3-22.
  • Groen AF, van der Vegt R, van Boekel MAJS, de Rouw OLAM, Vos H. 1994. Case study on individual animal variation in milk protein composition as estimated by high-pressure liquid chromatography. Neth Milk Dairy J, 48: 201-212.
  • Parris N, White AE, Farrell HM. 1990. Identification of altered proteins in nonfat dry milk powder prepared from heat-treated skim milk. J Agric Food Chem, 38: 824- 829.
  • Bobe G, Beitz DC, Freeman AE, Lindberg GL. 1998. Separation and quantification of bovine milk proteins by reverse-phase high-performance liquid chromatography. J Agric Food Chem, 46: 458-463.
  • Bobe G, Beitz DC, Freeman AE, Lindberg GL. 1998. Sample preparation affects separation of whey proteins by reverse-phase high-performance liquid chromatography. J Agric Food Chem, 46: 1321-1325.
  • Bordin G, Raposo FC, de la Calle B, Rodriguez AR. 2001. Identification and quantification of major bovine milk proteins by liquid chromatography. J Chrom A, 928: 63-76.
  • Veloso ACA, Teixeira N, Ferreira IMPLVO. 2002. Separation and quantification of the major casein fractions by reverse-phase high-performance liquid chromatography and urea-polyacrylamide gel electrophoresis: Detection of milk adulterations. J Chrom A, 967: 209-218.
Toplam 18 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Bölüm Makaleler
Yazarlar

Zerrin Yüksel Bu kişi benim

Yaşar Kemal Erdem Bu kişi benim

Yayımlanma Tarihi 1 Şubat 2010
Yayımlandığı Sayı Yıl 2010 Cilt: 35 Sayı: 1

Kaynak Göster

APA Yüksel, Z. ., & Erdem, Y. K. . (2010). Süt Proteinlerinin RP-HPLC ile Saptanması (İngilizce). Gıda, 35(1), 1-7.
AMA Yüksel Z, Erdem YK. Süt Proteinlerinin RP-HPLC ile Saptanması (İngilizce). GIDA. Şubat 2010;35(1):1-7.
Chicago Yüksel, Zerrin, ve Yaşar Kemal Erdem. “Süt Proteinlerinin RP-HPLC Ile Saptanması (İngilizce)”. Gıda 35, sy. 1 (Şubat 2010): 1-7.
EndNote Yüksel Z, Erdem YK (01 Şubat 2010) Süt Proteinlerinin RP-HPLC ile Saptanması (İngilizce). Gıda 35 1 1–7.
IEEE Z. . Yüksel ve Y. K. . Erdem, “Süt Proteinlerinin RP-HPLC ile Saptanması (İngilizce)”, GIDA, c. 35, sy. 1, ss. 1–7, 2010.
ISNAD Yüksel, Zerrin - Erdem, Yaşar Kemal. “Süt Proteinlerinin RP-HPLC Ile Saptanması (İngilizce)”. Gıda 35/1 (Şubat 2010), 1-7.
JAMA Yüksel Z, Erdem YK. Süt Proteinlerinin RP-HPLC ile Saptanması (İngilizce). GIDA. 2010;35:1–7.
MLA Yüksel, Zerrin ve Yaşar Kemal Erdem. “Süt Proteinlerinin RP-HPLC Ile Saptanması (İngilizce)”. Gıda, c. 35, sy. 1, 2010, ss. 1-7.
Vancouver Yüksel Z, Erdem YK. Süt Proteinlerinin RP-HPLC ile Saptanması (İngilizce). GIDA. 2010;35(1):1-7.

by-nc.png

GIDA Dergisi Creative Commons Atıf-Gayri Ticari 4.0 (CC BY-NC 4.0) Uluslararası Lisansı ile lisanslanmıştır. 

GIDA / The Journal of FOOD is licensed under a Creative Commons Attribution-Non Commercial 4.0 International (CC BY-NC 4.0).

https://creativecommons.org/licenses/by-nc/4.0/