(CHT Ceramic
Type I, 40 μm) was purchased from Bio-Rad (Munich, Germany).
Formic acid (LC–MS grade), potassium chloride, 85% o-phosphoric
acid, ammonium hydrogen carbonate, 4-chloro-α-cyanocinnamic
acid, and 2,5-dihydroxybenzoic acid were obtained from Sigma-Aldrich
(Taufkirchen, Germany). Acetonitrile (LC–MS grade) was purchased
from Honeywell Fluka (Seelze, Germany) and 1,4-dithiothreitol was
purchased from Carl Roth (Karlsruhe, Germany). Tris(hydroxymethyl)aminomethane
hydrochloride and ammonia (25%) were purchased from Acros Organics
(Geel, Belgium) and urea was purchased from Merck Millipore (Darmstadt,
Germany). Alkaline phosphatase (1500 U, grade I) from calf intestine
was purchased from Roche Diagnostics (Mannheim, Germany). Purified
water was taken from a Milli-Q system (Merck Chemicals, Darmstadt,
Germany).
Three different kefir batches of the same brand were
purchased from a local supermarket. The kefir was industrially produced
from pasteurized and homogenized low-fat organic cow milk (1.5%) using
a complex culture of yeast and lactic acid bacteria.