果聚糖[HK法]检测试剂盒 Fructan HK Assa

产品货号：K-FRUCHK

品牌：Megazyme

产品品名：果聚糖[HK法]检测试剂盒

英文品名：Fructan HK Assay Kit

规格型号：50 assays per kit

A modification of AOAC Method 999.03 and AACC Method 32.32. For the specific measurement of all fructo-oligosaccharides (reducing and non-reducing) and of fructan polysaccharide.

INTRODUCTION:

Fructans are defined as any compound where one or more fructosylfructose
linkage constitutes a majority of the linkages (Lewis, 1993).
This refers to polymeric material as well as oligomers as small as the
disaccharide, inulobiose. Material included in this definition may or may
not contain D-glucosyl substituents. The terms oligomer and polymer
are used by fructan researchers to distinguish between materials which
can be specifically characterised and those which cannot (Lewis, 1993).
Fructans are widely distributed in the plant kingdom. They are present
in monocotyledons, dicotyledons and in green algae. Fructans differ in
molecular structure and in molecular weight. They may be classified
into three main types: the inulin group, the levan group and the
branched group. The inulin group consists of material that has mostly
or exclusively the (2→1) fructosyl-fructose linkage. Levan is material
which contains mostly or exclusively the (2→6) fructosyl-fructose
linkage. The branched group has both (2→1) and (2→6) fructosylfructose
linkages in significant amounts (e.g. graminan from Gramineae).
Several procedures have been described for the measurement of
fructan in plant material and food products. It is generally accepted
that these are best measured after hydrolysis to D-fructose and
D-glucose. This introduces the problem of independently removing,
or measuring, sucrose, D-fructose and D-glucose. Pontis (1966)
has reported the removal of sucrose, D-glucose and D-fructose by
hydrolysing sucrose with a crystalline yeast invertase. However, yeast
invertase also hydrolyses lower degree of polymerisation (DP) fructooligosaccharides
(FOS) (McCleary and Blakeney, 1999).
An alternative approach (Quemener et al., 1993) involves the use of
capillary gas chromatography (CGC) or HPLC to analyse extracts
of samples either untreated, or treated with amyloglucosidase or
amyloglucosidase plus inulinase (fructanase). By measuring sucrose,
D-fructose and D-glucose in the various samples, and with appropriate
calculations, it is possible to get an estimate of free D-glucose and
D-fructose, sucrose, starch and fructan.
The currently described method (McCleary and Blakeney, 1999;
McCleary and Rossiter, 2004) is specific for fructans, including
those from chicory, dahlia, jerusalem artichoke, onion, wheat stems
and leaves and agave. Of these, the highly branched (2→1),(2→6)
fructan from agave (Agavaceae) is the most resistant, but even this is
completely hydrolysed under the described assay conditions. Because
this method does not include a borohydride reduction step (like
K-FRUC), partially hydrolysed inulins (e.g. Raftilose P-95?) are not
underestimated. However, the method is not suitable for use on
samples that contain high levels of D-glucose, D-fructose, sucrose or
maltose as these contribute to the sample blank absorbance. This will
lead to large errors if the fructan content is low.

KITS:

Kits suitable for performing 50 determinations of fructan are available
from Megazyme. The kits contain the full assay method plus:
Bottle 1: Buffer (25 mL, pH 7.6) plus sodium azide (0.02%
w/v) as a preservative.
Stable for > 2 years at 4°C.
Bottle 2: NADP+ plus ATP.
Stable for > 5 years at -20°C.
Bottle 3: Sucrase / maltase, lyophilised powder plus BSA.
Stable for > 5 years at -20°C.
Bottle 4: Fructanase (recombinant, affinity purified exo-inulinase
and endo-inulinase, lyophilised powder.
Stable for > 5 years at -20°C.
Bottle 5: Hexokinase plus glucose-6-phosphate dehydrogenase
and PGI suspension, 2.25 mL.
Stable for > 4 years at 4°C.
Bottle 6: D-Fructose standard solution (0.5 mg/mL) in 0.2%
(w/v) benzoic acid.
Stable for > 4 years at room temperature.
Bottle 7: Fructan control flour. Dahlia fructan freeze-dried in
the presence of α-cellulose.
Stable for > 5 years stored dry at room temperature.

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