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Merck

HPLC Analysis of Caseins on BIOshell A400 Protein C4

HPLC Analysis of Caseins on BIOshell A400 Protein C4 application for HPLC

Materials

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BIOshell A400 Protein C4, 3.4 ฮผm HPLC Column

3.4 μm particle size, L ร— I.D. 10 cm ร— 2.1 mm

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ฮฒ-Casein from bovine milk

BioUltra, ≥98% (PAGE)

ฮบ-Casein from bovine milk

≥70% (PAGE), lyophilized powder

ฮฑ-Casein from bovine milk

โ‰ฅ70% ฮฑs-casein basis (electrophoresis), lyophilized powder

CONDITIONS

column

BIOshell A400 Protein C4, 10 cm x 2.1 mm I. D., 3.4 μm particles (66825-U)

mobile phase

[A] 80:20 Water (0.1% trifluoroacetic acid): Acetonitrile (0.1% trifluoroacetic acid); [B] 20:80 Water (0.1% trifluoroacetic acid): Acetonitrile (0.1% trifluoroacetic acid)

gradient

20 to 53% B in 5 min

flow rate

0.3 mL/min

pressure

1150 psi (79 bar)

column temp.

50 ยฐC

detector

UV, 215 nm

injection

10 μL

sample

Three component casein mix, 200 μg/mL, 0.1% trifluoroacetic acid

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๋ถ„์„ ๋ฉ”๋ชจ

Caseins are a group of phosphoproteins found predominantly in mammalian milk. Each type of mammalian milk has a different profile of caseins (alpha, beta, etc.) based on the mammal′s diet and where it is located. Therefore, this profile can be used to authenticate where certain dairy products (i.e. cheeses) came from. A RP-HPLC method for the separation of three caseins (alpha, beta, and kappa-casein) was developed on the BIOshell A400 Protein C4 column.

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Due to the heterogeneity of the standards, band broadening and extra peaks resulted in the chromatogram.
Two isoforms of beta-casein exist in mammalian milk: A1-beta-casein and A2-beta-casein; therefore, two peaks are identified for beta-casein.

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