JPS602039B2 - Carbohydrate analysis method and device - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method and apparatus for analyzing carbohydrates from pasture, feed crops, etc., and more particularly to an analytical method and apparatus for analyzing carbohydrates without using multiple types of analytical instruments. It is.

Feed analysis of grasses, forage crops, etc. has become increasingly important in recent years, and emphasis has been placed on non-structural carbohydrates and structural carbohydrates among the analysis items, as well as the measurement of dry matter digestibility using in vitro methods. . However, various analyzes related to these carbohydrates require boiling extraction, filter washing, decomposition treatment with enzymes, etc., and the transfer of analytical samples from one container to another many times, making the analytical operations complicated and time-consuming. It takes time. Various methods have been developed to simplify such analysis operations, but at present all of them require the use of multiple types of expensive equipment. Therefore, the present invention provides carbohydrates (non-structural carbohydrates - TNC, Neutra
l De to r ntFiber-NDF, AcidDe
In addition, the dry matter digestibility by the invitm method (
By making it possible to perform heating, washing, enzyme treatment, etc. of the specimen in one container for analysis of IVDMD, the analysis time can be significantly shortened by eliminating the transfer operations described above. The purpose of the present invention is to provide an analysis method and an analysis device that can simplify and speed up the operational steps for fractionating and quantifying carbohydrates in pasture and feed crops at low cost without using expensive equipment. Hereinafter, the present invention will be explained with reference to the drawings and attached tables by a series of analytical examples regarding the quantification of carbohydrates (TNC, NDF), the degradation rate of cell membrane constituents (CWC), and the quantification of IVDMD.

Conventionally, the quantitative determination of carbohydrates is generally carried out using a number of glass instruments as shown in the attached table, and through many steps from a to m as shown in FIG. 1.

To briefly explain this process, starch ratio is carried out in a, decomposition (shaking) with amylase is carried out in b, sample is transferred in c, washing is carried out in b, and in c and d, INC Perform colorimetric determination. After that, the sample was transferred from d to e, heated and extracted with solvent (N, D solution) in f, and g
The sample was transferred with
C measurement) and ashing (NDF measurement). Attached Table: Types of instruments used during analysis Q King: ○ indicates required instruments, single mark indicates unnecessary.

In addition, the sample was transferred in i, digested with cellulase (shaking) in j, further transferred in k, transient washing was performed in 1, and dry weighed in m to determine the CWC decomposition rate and IVDMD.
Calculate. In this way, in the conventional method, in addition to using a lot of expensive equipment, there are many transfer operations such as c, e, g, i, and k, which are not only complicated and time-consuming, but also have problems with analytical accuracy. This was causing a decline in
On the other hand, the glass sieve (container) with a seal used in the present invention has two closing parts at both ends, as shown in FIG.
, 3, a bottle-shaped tube body 1 made of heat-resistant glass;
A glass filter (G2 or G3) is provided inside the tube body 1 so as to be partitioned in the vertical direction, and a heat-resistant, acid-resistant, and heat-resistant It is composed of caps 5 and 6 having alkalinity. This glass transfer vessel with a sealed stopper is used for the process and analysis shown in FIG. 1A.

First, the strainer is weighed A in advance, the sample for analysis is weighed B and placed in the strainer, and the cap 6 is attached to the opening □ part 3, and the lees are gelatinized by steam heating (large-scale processing is possible). Further, an amylase solution is added, a cap 5 is attached to the entrance part 2, and the container is shaken as a sealed container to decompose the sediment in the sample. After the disassembly is completed, the caps 3 and 5 are removed, and the entrance section 3 is placed on a filtration table, or a rubber tube is attached to the closing section 3 and suction is carried out to perform filtration and cleaning. A fixed amount of non-structural carbohydrates (TNC) transferred to the transferred liquid is quantified by color development after a certain number of customers have passed. The residual liquid in the sieve is heated with steam after attaching the cap 6 to the entrance part 3 and adding Neutral Detergent liquid, and attaching the cap 5 to the closing part 2.
(capable of mass processing). After cooling, remove the caps 5 and 6, and clean the same machine as before while using the transfer device.
After drying the sample containing CWC, structural carbohydrates, etc. remaining in the sieve, the sample is weighed (C) together with the sieve, and then ashed and weighed (D) together with the sieve. CWC=C33×loo(%) NDF=(C-A)×rA2×100(%) Also, C
When determining the decomposition rate (digestion rate) of WC and DM, N
After attaching the cap 6 to the opening 3 of the filter containing the DF extraction residue and adding the cellulase solution into the filter,
Attach the cap 5 to the opening 2 and disassemble it by shaking.

The caps 5 and 6 are removed again and filtered and washed.
Dry the residue with a sieve and weigh it. CWC decomposition rate = (
C-ri-go-AZxloo(%)C--AlvD plate=B
-Hair Xloo (%) As described above, in the present invention, as shown in FIG. As shown in the attached table, the glass strainer with a seal according to the invention can be used as a funnel for the secondary method, a glass strainer (or paper), a tall beaker, and a decomposition bottle for enzymatic decomposition.

In FIG. 1, the analysis steps indicated by a to m in the conventional method are indicated by uppercase letters A to M in the present invention indicated by A. As is clear from the above description, according to the carbohydrate analysis method and apparatus of the present invention, by using a glass filter with a sealed stopper, in particular,
■Boiling, extraction, filtration, washing, drying, ashing, and even enzymatic decomposition processing can be performed in the same container.

Therefore, there is no need to transfer samples for analysis, making it possible to shorten analysis time and quickly analyze a large amount of samples. ■The number of various instruments used for powder analysis is reduced, there is no need to use expensive equipment, and facility costs are reduced.

■Analysis costs are economical as only a small amount of reagent is required.

■It has versatility and can be applied not only to the analysis of carbohydrates in grasses, feed crops, etc., but also to various analytical and decomposition processes including the two steps of extraction and filtration.

■Analysis accuracy is improved because there is less wear and tear due to sample transfer.

It is possible to achieve the following effects.

[Brief explanation of the drawing]

FIG. 1 is a process diagram showing a comparison of analysis steps of the present invention (A) and the secondary method opening, and FIG. 2 is a vertical sectional side view showing the glass transition device with a seal of the present invention. 1... Pipe body, 2, 3... Sekiguchi part, 4... Glass filter, 5, 6... Cap. Takashi Z-zu Cedar Diagram

Claims (1)

[Claims] 1. In a method for analyzing carbohydrates in pasture, feed crops, etc., a sample is placed in a container that has a filter inside and removable caps at the top and bottom openings. After removing the cap and gelatinizing the starch by heat treatment, an enzyme solution is added, the cap is placed in the upper opening, the container is sealed, and the starch in the sample is decomposed. The cap of the container is removed, filtered and washed, and the non-structural carbohydrates transferred to the filtrate are quantified, while the residue inside the container is removed by attaching the cap at the bottom opening, adding the solvent, and then closing the cap at the top opening. The sample is heated in the attached closed container, and after cooling, the caps at the top and bottom openings are removed, and the sample containing CWC and structural carbohydrates remaining in the container is dried and removed together with the container. By weighing the sample, ashing it, and weighing the entire container, the various steps of heating, filtering, washing, drying, weighing, and ashing the specimen can be carried out without the specimen being removed from the container even once. Carbohydrate analysis method. 2. In a method for analyzing carbohydrates in grasses, feed crops, etc., the sample is heated in a container with a filter inside and removable caps at the top and bottom openings, with the caps at the top openings removed. After gelatinizing the starch, an enzyme solution is added, a cap is attached to the top opening, and the container is sealed to decompose the starch in the sample.Furthermore, the caps from the top and bottom openings are removed and filtered and washed. While the non-structural carbohydrates transferred to the filtrate are quantified, the residue in the container is heated in a closed container with a cap attached to the bottom opening and a solvent added, followed by a cap attached to the top opening. After cooling, remove the caps at the top and bottom openings, perform filtration and washing as described above, dry the sample containing CWC and structural carbohydrates remaining in the container, weigh the container together, and then remove the solvent extraction residue. Attach a cap to the bottom opening of the container containing the enzyme solution, add the enzyme solution into the container, then attach the cap to the top opening to disassemble the container, remove the caps from the top and bottom openings again, filter, After washing, the residue is dried and weighed in the container, and the various steps such as heating, filtration, drying, weighing, and ashing of the sample are performed without the detection object being ever removed from the container. A method for analyzing carbohydrates. 3. In an apparatus for analyzing carbohydrates of pasture, forage crops, etc., the container used for analysis includes a tube made of heat-resistant glass with open upper and lower ends, a glass filter that partitions the inside of this tube in the radial direction, and the above-mentioned. 1. A carbohydrate analysis device comprising a heat-resistant, acid-resistant, and alkali-resistant cap that can be sealed or removed at openings at the upper and lower ends.