Eel extract rich in branched chain amino acid and application of eel extract in anti-seasickness products

Abstract

The invention relates to the technical field of preparation of bioactive substances, in particular to an eel extract rich in branched chain amino acid and application of the eel extract in anti-seasickness products. According to the method, the small-molecule oligopeptide is obtained through directional composite enzymolysis, degreasing and membrane separation technologies; directional composite enzymolysis is formed by compounding animal protease and neutral protease, whole wheat flour degreasing and green natural degreasing raw materials are adopted, a net structure of beta-glucan and araboxylan adsorbs grease through capillary action and surface tension, and oil-water interfacial tension can be reduced in an enzymatic hydrolysate liquid-phase system; the contact between fat droplets and the fiber surface is promoted, the adsorption efficiency and the bonding strength are enhanced, denaturation separation is synchronously carried out by enzyme deactivation and degreasing, and the production energy consumption is reduced; the obtained branched-chain amino acid-enriched eel oligopeptide, gamma-aminobutyric acid and the like are combined to prepare tabletted candies, so that the motion sickness effect of model seasickness mice can be remarkably improved, and the tabletted candies have an obvious dose-dependent effect, high-efficiency retention of active ingredients and function improvement.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of bioactive substance preparation, and particularly relates to a branched-chain amino acid-rich eel extract and application thereof in anti-seasickness products. BACKGROUND

[0002] Motion sickness is a multisystem syndrome mainly caused by autonomic nervous response induced by abnormal acceleration movement or virtual movement visual stimulation. Motion sickness shows symptoms and signs of multiple systems, mainly including stomach discomfort, nausea, vomiting, cold sweat, salivation, pale face, headache, dizziness and sleep syndrome. In addition to showing high-level nervous function disorders (dizziness, anxiety, fear, headache) and nystagmus, the seasickness patients also have symptoms of nausea, vomiting, diarrhea, pale skin, cold sweat, and changes in heart rate, blood pressure and respiration in severe cases. In addition to the above-mentioned high-level nervous function and changes in internal organs and cardiovascular function, the seasickness patients also show obvious mental fatigue (mental fatigue) and physical fatigue (physical fatigue). However, the existing anti-seasickness drugs such as anticholinergic drugs (M receptor blockers such as hyoscine, anisodamine, etc.) and antihistamine drugs (H1 receptor blockers such as diphenhydramine, etc.) can obviously inhibit the occurrence of seasickness symptoms, but due to the obvious inhibition of mental activity, the mental and physical fatigue phenomenon is more serious. Therefore, how to effectively reduce the incidence of seasickness and reduce the degree of mental and physical fatigue caused by seasickness and promote the rapid recovery of brain function is a difficult problem.

[0003] At present, a large amount of by-products such as fish heads, fish bones and internal organs in the processing of roasted eels are often used as raw materials for fish meal production without further development and utilization. The mineral content in eel head bones and the amino acid content in eel head meat are relatively high, and there is a large space for development and utilization.

[0004] In order to improve the utilization rate of eel processing leftovers, solve the problem of anti-motion sickness and reduce the mental and physical fatigue caused by seasickness, the roasted eel head leftovers are used as raw materials, and the existing biological active peptide extraction and protein delivery system utilization status are combined to prepare branched-chain amino acid-rich eel peptides by using composite enzyme hydrolysis. On the one hand, the high-value utilization of eel processing by-products can improve the bioavailability of eel peptides; on the other hand, the extracted branched-chain amino acid-rich eel peptides can promote the recovery of motion sickness and reduce fatigue. SUMMARY

[0005] The present application aims to provide a branched-chain amino acid-rich eel extract and application thereof in anti-seasickness products to solve the problems in the background.

[0006] To solve the above technical problems, the technical scheme adopted by the present application is as follows: a preparation method of branched-chain amino acid-rich eel extract, comprising the following steps: S1, providing eel offal as raw material and performing pretreatment: The eel offal is mixed with water and crushed, and 0.1%-0.3% ginger powder is added during the cooking of the eel offal to remove the fishy smell by boiling, and then the eel offal is filtered and washed, and then put into an enzyme hydrolysis tank, and 0.1%-0.3% ginger powder is added again, and cooked for 1-2h to form a slurry; S2, using a complex enzyme system containing neutral protease and animal protease to hydrolyze the slurry to produce a filtrate; S3, performing solid-liquid separation on the filtrate to obtain a clear enzyme hydrolysis filtrate; S4, filtering the enzyme hydrolysis filtrate through an ultrafiltration membrane with a molecular weight cut-off value of 1000 daltons to collect enzyme hydrolysis liquid with a molecular weight of less than 1000 daltons; S5, mixing the enzyme hydrolysis liquid with soft water at a weight ratio of 1:5 to obtain a dilution liquid, and then filtering the dilution liquid to collect a retentate; S6, performing concentration reaction on the retentate to make the solid content reach 15%-20%; S7, drying the concentrated material to obtain branched-chain amino acid-rich eel peptide powder; the branched-chain amino acid-rich eel extract is branched-chain amino acid-rich eel peptide powder.

[0007] Preferably, in the step S1, the eel offal is selected from at least one of eel head, eel bone and eel offal.

[0008] Preferably, in the step S2, the temperature of the slurry is first kept constant at 55-60℃, and then the pH is adjusted to neutral, and then the complex enzyme is added for enzyme hydrolysis for 4-6h.

[0009] Preferably, in the step S2, the total amount of the complex enzyme added is 0.1%-0.3% of the weight of the raw material slurry, and the mass ratio of the neutral protease to the animal protease is 1:1-2.

[0010] The present application also provides a branched-chain amino acid-rich eel extract prepared by the above method, which is branched-chain amino acid-rich eel peptide powder. The peptide powder has a molecular weight of <10000 Da, high purity, a branched-chain amino acid content of ≥15%, no other raw and auxiliary materials, and good palatability, solubility and flowability.

[0011] The use of the branched-chain amino acid-rich eel extract in the preparation of an anti-seasickness product, the anti-seasickness product being an anti-seasickness tablet candy, wherein the branched-chain amino acid-rich eel peptide powder accounts for 33-46% by weight. The anti-seasickness tablet candy has a single serving size of 10 g, and contains 1.1-3.3 g of the branched-chain amino acid-rich eel peptide powder.

[0012] Preferably, the anti-seasickness tablet candy is composed of the following raw materials by weight percentage: branched-chain amino acid-rich eel peptide powder 33-46%, gamma-aminobutyric acid 2.15-4.15%, ferric pyrophosphate 0.01-0.05%, dried tangerine peel powder 5-10%, ginger powder 0.1-1%, isomalt 20-40%, and lemon mint flavor 0.2-0.8%.

[0013] The application also provides a preparation method of the anti-seasickness tablet candy, comprising the following steps: (a) weighing and mixing: accurately weighing each raw material according to the formula amount, and then putting the weighed raw materials into a three-dimensional mixer, mixing for 20-40 min until uniform in a closed state; (b) boiling granulation: boiling granulating the mixed material, drying, and obtaining granules with a water content of <5%; (c) granulating: passing the granulated granules through an 80-mesh sieve; (d) tabletting: tabletting the granules; (e) packaging: sub-packaging the tabletted tablet candy.

[0014] Preferably, in the step (b), the boiling granulation is performed in a closed container, and the material is granulated after being sucked into a hopper.

[0015] Preferably, in the step (d), the tabletting is processed in a tablet press.

[0016] A mouse motion sickness model is constructed, and the motion sickness mice are modeled by a three-dimensional rotating balance during feeding. The effect of the branched-chain amino acid-rich eel peptide tablet candy on improving motion sickness is evaluated by determining the resting time, total moving distance, crossing line number, and position preference of the mice in the open field experiment.

[0017] After modeling, the motion sickness behavior of the mice is shown in the open field experiment as deviating from the activity habit of the mice themselves, i.e., the normal mice walk along the edge path of the open field, the position preference tends to the corner, and the crossing line number of the body balance moving track is low, while the modeled mice are opposite. Therefore, the above mouse habits can be used as an index for judging the severity of motion sickness of the mice.

[0018] As can be seen from the above description of the structure of the application, compared with the prior art, the application has the following advantages: 1. The present application obtains small molecule oligopeptide by directional complex enzymolysis, degreasing and membrane separation technology; wherein the directional complex enzymolysis is composed of animal protease and neutral protease in a certain proportion, whole wheat flour is used for degreasing, green and natural degreasing raw materials, the network structure of beta-glucan and arabinoxylan can adsorb oil and fat through capillary action and surface tension, the oil-water interfacial tension can be reduced in the enzymolysis liquid phase system, the contact between fat droplets and fiber surface is promoted, the adsorption efficiency and binding strength are enhanced, and denaturation separation is realized simultaneously with enzyme inactivation and degreasing, thereby reducing the production energy consumption; the obtained branched chain amino acid type eel oligopeptide can significantly improve the motion sickness effect of the molded motion sickness mice, has obvious dose-dependent effect, and the active ingredients are efficiently reserved and the function is improved.

[0019] 2. The branched chain amino acid type eel peptide powder obtained by the present application has a molecular weight of <10000 Da, high purity, a branched chain amino acid content of ≥15%, no other raw and auxiliary materials, good palatability, solubility and flowability, stable shelf life, and wide application range.

[0020] 3. The composition of the present application is reasonably matched, ensures the branched chain amino acid content of eel peptide, has good palatability, and contains no fat; the combination of gamma-aminobutyric acid and a specific formula system is verified by anti-motion animal experiment, and an effective action time is provided, which can significantly improve the balance ability of motion sickness mice and promote the recovery after motion sickness. DETAILED DESCRIPTION

[0021] The accompanying drawings, which form a part of this application, are included to provide a further understanding of the application and are incorporated in and constitute a part of this application. The embodiments illustrated in the drawings are provided to explain the present application and should not be considered as an improper limitation on the present application. In the drawings: Figure 1 Molecular weight distribution of the branched chain amino acid type eel peptide in Preparation Example 3 of the present application; Figure 2 Comparison of branched chain amino acid (BCAA) contents of eel peptides in Preparation Examples 1-3 of the present application; Figure 3 Mouse open field route map in the anti-motion animal experiment of the present application. DETAILED DESCRIPTION

[0022] In order to further explain the technical solutions of the present application, the following specific embodiments are used for detailed description.

[0023] Preparation Example 1 The present application provides a preparation method of branched chain amino acid type eel peptide powder, comprising the following steps: S1, pretreatment: take out the eel head and put it into the thawing pool, soak in water, soak out blood water until no blood water is leached out, mix the treated eel head with water in a mass ratio of 1:2, pass through a meat grinder and then through a colloid mill for further refinement to obtain uniform slurry, first add 0.1% ginger powder to heat and boil to remove fishy smell, filter and wash the material after boiling, then introduce the slurry into an enzyme hydrolysis tank and add 0.1% ginger powder again for continuous boiling for 1h to form the slurry for enzyme hydrolysis; S2, enzyme hydrolysis: first heat the slurry to 55℃, adjust the pH of the slurry to neutral, add the compound enzyme with a mass ratio of 1:1 of neutral protease and animal protease activated in advance, stir for 4h, then measure the solid content every half hour with a solid content meter when the solid content is basically stable, the enzyme hydrolysis is completed, and the temperature is raised to 98℃ for enzyme inactivation to obtain a filtrate; S3, separation: cool the enzyme-inactivated filtrate, and then separate the solid and liquid by a tubular centrifuge with a speed of 4000r / min and a mesh size of 100 microns to obtain a clear enzyme hydrolysis filtrate; S4, membrane filtration: the enzyme hydrolysis filtrate is filtered through an ultrafiltration membrane system with a molecular weight cut-off of 1000 daltons to collect enzyme hydrolysate with a molecular weight of less than 1000 daltons; S5, desalination: mix the enzyme hydrolysate with softened water in a weight ratio of 1:5 to obtain a dilution liquid, and then filter the dilution liquid through a sodium membrane to collect a retentate; S6, concentration: concentrate the retentate by a double-effect vacuum concentration device, with an evaporation pressure of 0.06Mpa, an evaporation temperature of 68℃ for the first effect and an evaporation temperature of 56℃ for the second effect, so that the solid content reaches 15%, and then inject the concentrated retentate into a blending tank for standby; S7, drying: homogenize the concentrated retentate by a homogenizer, and then introduce it into a spray tower for spray drying, with an inlet temperature of 170℃ and an outlet temperature of 95℃, and a vibrating screen set at the powder outlet of the spray tower to screen once every 8 minutes to finally form a branched-chain amino acid-rich eel extract; the branched-chain amino acid-rich eel extract is a branched-chain amino acid-rich eel peptide powder.

[0024] Preparation Example 2 The present application provides a preparation method of a branched-chain amino acid-rich eel peptide powder, comprising the following steps: S1, pretreatment: take out the eel head and put it into the thawing pool, soak in water, soak out blood water until no blood water is leached out, mix the treated eel head with water in a mass ratio of 1:2.5, pass through a meat grinder and then through a colloid mill for further refinement to obtain uniform slurry, first add 0.2% ginger powder to heat and boil to remove fishy smell, filter and wash the material after boiling, then introduce the slurry into an enzyme hydrolysis tank and add 0.2% ginger powder again for continuous boiling for 1.5h to form the slurry for enzyme hydrolysis; S2, enzymolysis: first, the slurry is constant temperature to 58℃, adjust the pH of the slurry to neutral, add the mass ratio of 1:1.5 of the compound enzyme of the neutral protease and animal protease activated in advance, stir for 5h, measure the solid content every half hour with the solid content meter after 5h, when the solid content is basically stable, the enzyme hydrolysis is completed, and the temperature is raised to 99℃ for enzyme inactivation to obtain the filtrate; S3, separation: the enzyme inactivated filtrate is cooled, and the cooled material is separated by a tubular centrifuge to obtain a clear enzymatic hydrolysate; S4, membrane filtration: the enzymatic hydrolysate is filtered through an ultrafiltration membrane system with a molecular weight cut-off of 1000 daltons to collect an enzymatic hydrolysate with a molecular weight of less than 1000 daltons; S5, desalination: the enzymatic hydrolysate is mixed with softened water at a weight ratio of 1:5 to obtain a dilution liquid, and the dilution liquid is filtered through a sodium membrane to collect a retentate; S6, concentration: the retentate is concentrated by a double-effect vacuum concentration device, the evaporation pressure is 0.06Mpa, the one-effect temperature of evaporation is set to 69℃, and the two-effect temperature is set to 58℃, so that the solid content reaches 18%, and the concentrated retentate is injected into a blending tank for standby; S7, drying: the concentrated retentate is homogenized by a homogenizer and then introduced into a spray tower for spray drying, the inlet temperature of the spray tower for spray drying is 180℃, the outlet temperature is 100℃, and the powder outlet of the spray tower is provided with a vibrating screen which is screened every 10 minutes, so as to finally form a branched-chain amino acid-rich eel extract; the branched-chain amino acid-rich eel extract is a branched-chain amino acid-rich eel peptide powder.

[0025] Preparation Example 3 The present application provides a preparation method of a branched-chain amino acid-rich eel peptide powder, comprising the following steps: S1, pretreatment: the eel head is taken out and placed in a thawing pool for thawing, soaked in water, and soaked to remove blood water until no blood water is leached out, the treated eel head is mixed with water at a mass ratio of 1:3, first passed through a meat grinder and then further refined by a colloid mill to obtain a uniform slurry, 0.3% ginger powder is added for the first time to raise the temperature and boil to remove odor, the material is filtered and washed after boiling, and the slurry is introduced into an enzymolysis tank and 0.3% ginger powder is added again for continuous boiling for 2h to form a slurry for enzymolysis; S2, enzymolysis: first, the slurry is constant temperature to 58℃, adjust the pH of the slurry to neutral, add the mass ratio of 1:1.5 of the compound enzyme of the neutral protease and animal protease activated in advance, stir for 5h, measure the solid content every half hour with the solid content meter after 5h, when the solid content is basically stable, the enzyme hydrolysis is completed, and the temperature is raised to 99℃ for enzyme inactivation to obtain the filtrate; S3. Separation: The filtrate after enzyme inactivation is cooled and then separated into solid and liquid components by a tubular centrifuge. The centrifuge is set to 4000 r / min to remove insoluble residues larger than 100 mesh, resulting in a clear enzymatic hydrolysis filtrate. S4. Membrane filtration: The enzymatic hydrolysate is passed through an ultrafiltration membrane system with a molecular weight cutoff of 1000 Daltons to collect the enzymatic hydrolysate with a molecular weight of less than 1000 Daltons. S5. Desalination: The enzymatic hydrolysate and softened water are mixed at a weight ratio of 1:5 to obtain a diluted solution. The diluted solution is then filtered through a sodium membrane, and the retentate is collected. S6. Concentration: The retentate is concentrated using a double-effect vacuum concentration device. The evaporation pressure is 0.06 MPa, the first-effect temperature is set to 70°C and the second-effect temperature is set to 60°C, so that the solid content reaches 20%. The concentrated retentate is then injected into a mixing tank for later use. S7. Drying: The concentrated retentate is homogenized by a homogenizer and then introduced into a spray drying tower for spray drying. The inlet temperature of the spray drying tower is 190℃ and the outlet temperature is 105℃. A vibrating screen is installed at the powder outlet of the spray tower, and the screen is sieved once every 12 minutes to finally form a branched-chain amino acid-rich eel extract. This branched-chain amino acid-rich eel extract is a branched-chain amino acid-rich eel peptide powder.

[0026] According to GB5009.6-2016 "National Food Safety Standard - Determination of Fat in Food - Soxhlet Extraction Method", the fat content and branched-chain amino acid content of three batches of eel peptide powder produced by different processes and eel peptide powder produced by defatting whole wheat flour were tested.

[0027] Preparation Example 4 An anti-vertigo compressed candy is composed of the following ingredients in weight percentage: 33%-46% of the above-mentioned branched-chain amino acid-rich eel peptide powder, 2.15%-4.15% of γ-aminobutyric acid, 0.01%-0.05% of ferric pyrophosphate, 5%-10% of dried tangerine peel powder, 0.1%-1% of ginger powder, 20%-40% of isomaltitol, and 0.2%-0.8% of lemon mint flavoring.

[0028] The preparation method of the above-mentioned anti-vertigo compressed candy includes the following steps: (1) Weighing and mixing: Weigh each raw material accurately according to the formula dosage, put the weighed raw materials into the three-dimensional mixer, seal it, and mix for 20-40 minutes until uniform; (2) Fluidized bed granulation: Turn on the fluidized bed granulation equipment, suck the mixture into the sealed container of the hopper, granulate, dry, and obtain granules with a moisture content of <5%; (3) Granulation: The granulated particles are passed through an 80-mesh sieve to obtain relatively uniform particles; (4) tabletting: the environment humidity is controlled at 35%, and the granules are put into a tablet press to be tabletted; (5) packaging: the tabletted candies are packed into the packaging boxes, and the packaging is completed.

[0029] The anti-dizziness tabletted candies of branched-chain amino acid-enriched eel peptides prepared above are applied to experimental animals, and the experimental process and results are described as follows.

[0030] Experimental animals SPF Kunming healthy male mice provided by the Experimental Animal Center of Shandong University are selected, and the body weight is 18-22 g. The experimental environment temperature is 22℃±2℃, and the relative humidity is 50%±10%. The mice are randomly divided into a control group, a model group, a low-dose test sample group, a medium-dose test sample group, and a high-dose test sample group, 8 mice in each group. The sample is weighed to prepare a high-dose sample solution and a medium-dose sample solution (gavage according to 0.02 mL / g body weight). The control group and the model group are given distilled water, and gavage is performed once a day. Each group is fed with SPF special feed. Distilled water is drunk. After 4 weeks of intervention, the mice are placed in a LABGIC disc mixer L-RMO-80Pro dizziness modeling instrument, and appropriate dizziness stimulation parameters such as rotation speed, time, frequency, and acceleration are set. The dizziness stimulation is repeated for several consecutive days to establish a stable dizziness model. The mice are observed in the open field test (OFT) for still time, total moving distance, crossing times, and position preference, and the dizziness anxiety and exploration behavior are evaluated.

[0031] 2. Test grouping Table 1 Gavage dose and grouping 3. Experimental results 3.1 Gavage 30 min in advance Table 2 Open field of mice in each group gavaged 30 min in advance 3.2 Gavage 120 min in advance Table 3 Open field of mice in each group gavaged 120 min in advance 3.3 Gavage for 7 days Table 4 Open field of mice in each group gavaged for 7 days According to the above experimental results, the medium-dose and high-dose test samples have obvious improvement effect on the dizziness modeling mice, and the improvement effect of the medium-dose sample compounded with γ-aminobutyric acid is better than that of the high-dose group; and the effect of the test sample gavaged 30 min in advance is the best, and the long-term gavage for 7 days can significantly improve the dizziness response of the mice.

[0032] The above merely describes preferred embodiments of the present application, and is not used to limit the present application, any modification, equivalent replacement and improvement within the spirit and principle of the present application should be included in the protection scope of the present application.

Claims

1. A method for preparing an eel extract rich in branched-chain amino acids, characterized in that, Includes the following steps: S1. Provide eel offal as raw material and perform pre-processing: Mix and crush eel offal with water. When steaming the eel offal, add 0.1%-0.3% ginger powder and heat to boiling to remove fishy smell. Then filter and wash the eel offal, put it into an enzymatic hydrolysis tank, add 0.1%-0.3% ginger powder again, and steam for 1-2 hours to form a slurry. S2. The slurry is enzymatically hydrolyzed using a complex enzyme system containing neutral protease and animal protease to produce filtrate; S3. Perform solid-liquid separation on the filtrate to obtain a clear enzymatic hydrolysis filtrate; S4. Filter the enzymatic hydrolysate through an ultrafiltration membrane with a molecular weight cutoff of 1000 Daltons and collect the enzymatic hydrolysate with a molecular weight of less than 1000 Daltons. S5. Mix the enzymatic hydrolysate and softened water at a weight ratio of 1:5 to obtain a diluted solution, then filter the diluted solution and collect the retentate. S6. Concentrate the retentate to achieve a solids content of 15%-20%; S7. The concentrated material is dried to obtain a branched-chain amino acid-rich eel extract; this branched-chain amino acid-rich eel extract is a branched-chain amino acid-rich eel peptide powder.

2. The method for preparing a branched-chain amino acid-rich eel extract according to claim 1, characterized in that: In step S1, the eel offal is selected from at least one of eel head, eel bone, and eel viscera.

3. The method for preparing a branched-chain amino acid-rich eel extract according to claim 1, characterized in that: In step S2, the temperature of the slurry is first kept constant at 55-60℃ and its pH is adjusted to neutral. Then, the compound enzyme is added and enzymatic hydrolysis is carried out for 4-6 hours.

4. The method for preparing a branched-chain amino acid-rich eel extract according to claim 1, characterized in that: In step S2, the total amount of compound enzyme added is 0.1%-0.3% of the weight of the raw material slurry, and the mass ratio of the neutral protease to the animal protease is 1:1-2.

5. The branched-chain amino acid-rich eel extract prepared according to any one of claims 1-4, wherein the branched-chain amino acid-rich eel extract is a branched-chain amino acid-rich eel peptide powder.

6. The application of the branched-chain amino acid-rich eel extract according to claim 5 in the preparation of anti-seasickness products, characterized in that: The anti-seasickness product is an anti-seasickness compressed candy, and the weight percentage of the branched-chain amino acid-rich eel peptide powder in the anti-seasickness product is 33%-46%.

7. An anti-vertigo compressed candy, characterized in that, It is composed of the following raw materials in weight percentage: 33%-46% of the branched-chain amino acid-rich eel peptide powder obtained in claim 5, 2.15%-4.15% of γ-aminobutyric acid, 0.01%-0.05% of ferric pyrophosphate, 5%-10% of dried tangerine peel powder, 0.1%-1% of ginger powder, 20%-40% of isomaltitol, and 0.2%-0.8% of lemon mint flavoring.

8. A method for preparing an anti-vertigo compressed candy as described in claim 6 or 7, characterized in that, Includes the following steps: (a) Weighing and mixing: Weigh each raw material accurately according to the formula dosage, put the weighed raw materials into the three-dimensional mixer, seal it, and mix for 20-40 minutes until uniform; (b) Fluidized bed granulation: The mixture is subjected to fluidized bed granulation and dried to obtain granules with a moisture content of <5%; (c) Granulation: The granulated particles are passed through an 80-mesh sieve; (d) Tableting: The granules are compressed into tablets; (e) Packaging: The compressed candies are repackaged.

9. The method for preparing the anti-vertigo compressed candy according to claim 8, characterized in that: In step (b), fluidized bed granulation is carried out in a closed container, where the material is drawn into a hopper and granulated.

10. The method for preparing the anti-vertigo compressed candy according to claim 8, characterized in that: In step (d), the tableting process is performed in a tableting machine.

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