CN112690459B - Hydrolyzed whey protein, composition and clinical application thereof - Google Patents
Hydrolyzed whey protein, composition and clinical application thereof Download PDFInfo
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- CN112690459B CN112690459B CN202011532404.7A CN202011532404A CN112690459B CN 112690459 B CN112690459 B CN 112690459B CN 202011532404 A CN202011532404 A CN 202011532404A CN 112690459 B CN112690459 B CN 112690459B
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- whey protein
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Abstract
The present invention relates to hydrolysed whey protein, compositions and clinical uses thereof. The hydrolyzed whey protein has a degree of hydrolysis of between 10 and 30% and an average molecular weight of between 500 and 3000Da, and wherein the hydrolyzed whey protein has a sodium content ranging from 80 to 250mg/100g, a magnesium content ranging from 60 to 100mg/100g, a phosphorus content ranging from 150 to 300mg/100g, a potassium content ranging from 450 to 530mg/100g, a calcium content ranging from 350 to 800mg/100g, an essential amino acid content ranging from 31 to 36g/100g, and a branched amino acid content ranging from 14 to 17g/100g. The hydrolyzed whey protein of the invention can be used as a formula food protein component for special medical application of patients with renal disease dialysis, and provides nutrition for patients with renal disease dialysis, diabetic renal disease dialysis or patients with chronic renal failure at the end stage.
Description
Technical Field
The invention relates to the field of foods, in particular to a special medical purpose formula food protein component for a patient suffering from renal disease dialysis and a preparation method thereof.
Background
Hemodialysis is a treatment method for prolonging the life of patients with nephrotic uremia, but patients who are continuously subjected to dialysis treatment have protein nutrition deficiency with different degrees, symptoms such as inappetence and dyspepsia appear in the patients during hemodialysis, the concentration of essential amino acids in plasma is remarkably reduced clinically, complications and death rate of the patients with nephrotic uremia are increased, and the factors are also important factors influencing the prognosis of the patients. The fatality rate is increased by 6 percent when the serum albumin is reduced by 1 g/L.
The existing kidney disease nutritional food for patients with kidney disease dialysis is mainly prepared by taking maltodextrin as a main raw material, adding whole proteins such as whey protein, soybean protein and the like, adding fat, vegetable fat and the like as energy supply substances, and adding dietary fiber, a small amount of vitamins and minerals. In clinical practice, as renal dialysis patients are dialyzed for a plurality of times for a long time, intestinal mucosa barriers are damaged, serum albumin is reduced obviously, immune functions are penetrated and absorbed, whole proteins, grease and dietary fibers cannot be well digested and absorbed, and the renal dialysis patients and the gastrointestinal patients are burdened; in addition, some patients have the symptoms of hyperglycemia, hypertension and hyperlipidemia, and the eating of maltodextrin and vegetable oil is unfavorable for the blood sugar and the blood fat of the patients. For example, chinese patent application CN201510528459.3 discloses a nutritional composition comprising the following components in parts by weight: 1-50 parts of protein powder, 1-20 parts of short peptide, 10-70 parts of edible vegetable oil microcapsule powder, 1-30 parts of dietary fiber, 1-30 parts of maltodextrin and 0.1-1 part of B vitamins. Chinese patent application CN201510701888.6 discloses a special dietary food suitable for diabetic nephropathy nutritional mode, which comprises the following raw material components in parts by weight: 2200 to 3500 parts of maltodextrin, 2000 to 3000 parts of powdered oil, 1000 to 2000 parts of potato starch, 400 to 900 parts of whey protein, 400 to 850 parts of inulin, 400 to 800 parts of white granulated sugar, 300 to 700 parts of puffed powder, 300 to 700 parts of whole milk powder, 50 to 150 parts of hydrolyzed whey protein, 40 to 140 parts of composite mineral substance and 5 to 10 parts of composite vitamin.
The intake of high-quality protein, especially low-molecular weight high-quality hydrolyzed protein or short peptide can reduce the risk of malnutrition of a renal disease dialysis patient and delay the progress of renal failure, but at present, a proper renal dialysis nutritional preparation is not available.
Disclosure of Invention
The invention provides a high-quality protein, low-sodium, low-potassium and low-phosphorus hydrolyzed protein component, and the formula meets the nutritional requirements of a patient with dialysis kidney by adjusting the levels of hydrolyzed protein and electrolyte according to the requirements of the patient on nutrients; the molecular weight of the short peptide in the formula is lower than 3000Da, the content of essential amino acid is high, the kidney burden such as metabolic acidosis, calcium-phosphorus metabolic disorder, secondary hyperparathyroidism, renal osteopathia and the like can be reduced, and the immunity of the organism can be improved. The hydrolyzed whey protein is suitable for patients with kidney disease dialysis, has the characteristics of small bitter taste and less whole protein, and increases the compliance and the absorption capacity of the patients.
In another aspect, the present invention provides a hydrolyzed whey protein. The hydrolyzed whey protein may have a degree of hydrolysis between 10 and 30% and an average molecular weight between 500 and 3000, and wherein the hydrolyzed whey protein has a sodium content ranging from 80 to 250mg/100g, a magnesium content ranging from 60 to 100mg/100g, a phosphorus content ranging from 150 to 300mg/100g, a potassium content ranging from 450 to 530mg/100g, a calcium content ranging from 350 to 800mg/100g, an essential amino acid content ranging from 31 to 36g/100g, and a branched amino acid content ranging from 14 to 17g/100g.
In another aspect, the present invention provides a hydrolyzed whey protein. The hydrolyzed whey protein is obtained by a method comprising enzymatic hydrolysis of whey protein with pepsin and trypsin. The method further comprises a desalting step. The desalting step may result in a desalting rate of the hydrolysed whey protein of above 90%, e.g. 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%. For example, the desalting step is carried out as follows: desalting with DA201-C macroporous resin, using deionized water and 40-70% ethanol as desorbent, loading at flow rate of 0.5-2.5mL/min and loading concentration of 10-50mg/mL.
In one embodiment, the enzymatic treatment is carried out at an enzymatic temperature of 30-45 ℃ and/or at a pH of 2-4.
In one embodiment, the weight ratio of enzyme to whey protein is 1/30 to 1/20 and the whey protein concentration is 5wt% to 6wt%.
In one embodiment, the enzymatic time is 200-260 minutes.
The invention provides a preparation method of hydrolyzed whey protein, which comprises a step of enzymolysis of the whey protein by pepsin and trypsin and a desalting step. Preferably, the enzymatic treatment is carried out at an enzymatic temperature of 30-45 ℃ and a pH of 2-4. Preferably, the ratio of enzyme to whey protein is 1/30-1/20 and the whey protein concentration is 5wt% to 6wt%. Preferably, the enzymolysis time is 200-260 minutes. The ratio of pepsin to trypsin is 1.
In one aspect, the present invention provides a composition for renal dialysis patients comprising hydrolysed whey protein according to the invention.
In one aspect, the present invention provides a specific medical use formula comprising the hydrolysed whey protein of the invention. The hydrolysed whey protein may be the sole protein component or may be used in combination with other protein components. Preferably, the hydrolysed whey protein is the only protein component.
In one embodiment, the composition comprises the following ingredients by weight:
75 to 96 percent of hydrolyzed whey protein,
1.5 to 15 percent of sour agent,
0.1% -15% of sweetening agent.
In one embodiment, the acidulant is selected from the group consisting of lactic acid, citric acid, monosodium fumarate, adipic acid, tartaric acid, phosphoric acid, malic acid, acetic acid, and sodium carbonate. In one embodiment, the sweetener is selected from the group consisting of sucralose, neotame, cyclamate, maltitol, sorbitol, thaumatin, aspartame, steviol glycosides, acesulfame k, isomaltulose, erythritol and xylitol.
In another aspect, the invention provides the use of the composition or hydrolysed whey protein of the invention in the preparation of a special medical use formula for patients with renal disease. For example, the special medical use formula may be used in renal dialysis patients, diabetic renal dialysis patients, or end stage chronic renal failure patients.
In another aspect, the invention provides the use of hydrolysed whey protein as a protein component of a food product for subjects in need of a low electrolyte content food (e.g. low sodium, low calcium, low magnesium, low phosphorus and/or low potassium) and/or subjects that are not compliant to bitter taste. For example, the subject may be a child. The food is a formula food for special medical application. Preferably, the special medical formula is used for patients with renal disease, such as patients with renal disease dialysis, patients with diabetic nephropathy dialysis or patients with end stage chronic renal failure. Hydrolyzed whey protein may also provide the subject with abundant protein, increasing the subject's blood albumin level.
Detailed Description
The present invention is based in part on the inventors' discovery that: the hydrolyzed whey protein of the prior art does not meet the needs of renal dialysis patients for special medical use of formula protein components. The present invention is also based, in part, on: a hydrolyzed whey protein suitable for satisfying such needs can be obtained by treating whey protein with an enzyme under appropriate conditions.
For example, conventional desalination processes subject whey to electrodialysis and/or ion exchange to reduce the mineral content by 80%. The inventor finds that after the conventional hydrolyzed whey protein is measured, the sodium content of the hydrolyzed whey protein in the prior art is 80-805 mg/100g, the magnesium content is 15-120 mg/100g, the phosphorus content is 180-1100 mg/100g, the potassium content is 120-540 mg/100g, and the calcium content is 120-1700 mg/100g. In addition, the amino acid composition of the hydrolyzed whey protein varies from place to place, from manufacturer to manufacturer, and in different specifications. Some products have obviously low content of essential amino acids such as isoleucine, leucine, lysine, methionine, threonine, tryptophan, valine and the like, and have obviously high content of non-essential amino acids such as glutamic acid. The hydrolyzed whey proteins of the prior art are not suitable for use as a protein component in a specific medical use formula for renal dialysis patients. The inventors have found through long-term screening that the hydrolysed whey proteins of the invention differ from the hydrolysed whey proteins of the prior art in specific contents of elements, such as sodium content, magnesium content, phosphorus content, potassium content and/or calcium content, as well as essential and non-essential amino acids content, and are suitable for use as a special medical use formula protein component in renal dialysis patients.
The invention provides a method for producing hydrolyzed whey protein, which comprises providing whey protein; recovering; adjusting the pH value; adding enzyme for hydrolysis; keeping the pH constant; measuring the degree of hydrolysis; inactivating enzymes; ultrafiltration; desalting; freeze drying; and a step of finishing. The preparation method of the hydrolyzed whey protein comprises the steps of carrying out enzymolysis on the whey protein by using pepsin and trypsin and desalting. The invention provides a preparation method of hydrolyzed whey protein, which comprises a step of enzymolysis of the whey protein by pepsin and trypsin and a desalting step. Preferably, the enzymatic treatment is carried out at an enzymatic temperature of 30-45 ℃ and a pH of 2-4. In one embodiment, the temperature is 35 or 40 ℃. In one embodiment, the pH is 2.5, 3, or 3.5. Preferably, the ratio of enzyme to whey protein is 1/30-1/20 and the whey protein concentration is 5wt% to 6wt%. In one embodiment, the ratio of enzyme to whey protein is 1/25. In one embodiment, the whey protein concentration is 5.5wt%. Preferably, the enzymatic time is 200-260 minutes, such as 210, 220, 230, 240, or 250 minutes. In the method, concentrated whey protein is used as a raw material, pepsin and trypsin are selected, multiple factors and multiple levels are arranged by adopting uniform experimental design, the optimal optimization condition for whey protein enzymolysis is sought, the test result is subjected to regression analysis by using DPS (data processing System) data processing software, a regression equation with high fitting degree is obtained, and the optimal hydrolysis condition for whey protein enzymolysis by pepsin and trypsin is determined as follows: the enzymolysis temperature is 42 ℃, the pH value is 2.5, the ratio of enzyme to substrate is 1/25, the substrate concentration is 5.25%, the enzymolysis time is 240min, and the hydrolysis degree is 10-30%. The method adopts an improved ion exchange technology and regulates the pH value of the whey protein, determines the optimal combination condition of ion exchange resin desalination (DA 201-C macroporous resin is used for desalination, deionized water and 40-70% ethanol are used as desorbents, the sampling flow rate is 0.5-2.5mL/min, and the sampling concentration is 10-50 mg/mL), and the desalination rate of the prepared hydrolyzed whey protein reaches 96.5% by comprehensively considering the exchange capacity, the regeneration easiness, the protein adsorption rate, the dry matter adsorption rate and the pH regulation. The electrolyte in the whey protein can be better reduced, the equipment investment is low, and better economic benefit is achieved.
The hydrolyzed whey protein can be produced by the method for producing hydrolyzed whey protein, wherein the hydrolysis degree of the hydrolyzed whey protein is between 10 and 30 percent and the average molecular weight is between 500 and 3000Da, and wherein the hydrolyzed whey protein has a sodium content in the range of 80 to 250mg/100g, a magnesium content in the range of 60 to 100mg/100g, a phosphorus content in the range of 150 to 300mg/100g, a potassium content in the range of 450 to 530mg/100g, a calcium content in the range of 350 to 800mg/100g, an essential amino acid content in the range of 31 to 36g/100g, and a branched amino acid content in the range of 14 to 17g/100g. In one embodiment, the glycine content is from 1.2 to 1.4g/100g. In one embodiment, the alanine content is from 3.6 to 4.1g/100g. In one embodiment, the arginine content is 1.9 to 2.1g/100g. In one embodiment, the proline content is 3.7 to 4.2g/100g. As demonstrated in the examples, the hydrolysed whey proteins of the present invention are suitable for use as a special medical use formula protein component in renal dialysis patients, providing nutrition to renal dialysis patients, diabetic renal dialysis patients or end stage chronic renal failure patients.
The hydrolyzed whey protein may be derived from whey protein, including whey protein isolate, whey protein concentrate, whey powder, desalted whey powder, etc.; more preferably isolated whey protein (higher protein content in isolated whey protein); more preferably low lactose isolated whey protein (also suitable for patients with lactose intolerant renal disease); more preferably, the whey protein isolate is free of lactose and low in salt content (the cost of a further desalting process can be saved, and adverse reactions intolerant to lactose and the like can be reduced).
The hydrolysed whey protein may be provided in the form of a composition. The composition may comprise hydrolysed whey protein. In one embodiment, the composition may comprise 75% to 96% hydrolyzed whey protein, 1.5% to 15% acidulant, and 0.1% to 15% sweetener. The content of hydrolysed whey protein may be 90%, 91%, 92%, 93%, 94% or 95%. The amount of acidulant may be 3%, 4%, 4.8%, 5% or 6%. The sweetener may be present in an amount of 3%, 4%, 4.8%, 5% or 6%. In one embodiment, the composition comprises hydrolyzed whey protein powder, lactic acid, citric acid, sucralose, and neotame. The content of lactic acid may be 2-4%. The citric acid may be present in an amount of 0.1-3%, for example 1%, 2% or 2.5%. The sucralose may be present in an amount of 30mg to 100mg. The neotame content may be 100-200mg.
The souring agent can be one or more of lactic acid, citric acid, monosodium fumarate, adipic acid, tartaric acid, phosphoric acid, malic acid, acetic acid, and sodium carbonate. Preferably, the source of the sour agent is one or more of lactic acid, citric acid, phosphoric acid, malic acid. Preferably, the source of the acidulant is lactic acid and/or citric acid.
The sweetener can be one or more of sucralose, neotame, sodium cyclamate, maltitol, sorbitol, thaumatin, aspartame, stevioside, acesulfame potassium, isomaltulose, erythritol and xylitol. Preferably, the source of the sweetener is one or more of sucralose, neotame, steviol glycosides, erythritol. Preferably, the source of sweetener is sucralose and/or neotame.
The invention proves that the composition with the composition can be used for providing nutrition for nephrotic dialysis patients, diabetic nephrotic dialysis patients or chronic renal failure end-stage patients, and effectively controlling the albumin, blood phosphorus and blood calcium levels of dialysis patients.
Examples
Example 1: preparation of the hydrolysed protein component
Prior art hydrolyzed whey protein (manufacturer 1: fivelyland company; manufacturer 2 and manufacturer 3: ara company; manufacturer 4 and manufacturer 5: hengyuan company)
The inventors purchased different batches of hydrolyzed whey protein and measured the hydrolyzed whey protein (the measurement party was assigned to the institute of nutrient sources, beijing), and obtained the following results.
Table 1: electrolyte content in hydrolyzed whey protein
Sodium content range of hydrolyzed whey protein: 80-805 mg/100g
Magnesium content range of hydrolyzed whey protein: 15-120 mg/100g
Phosphorus content range of hydrolyzed whey protein: 180-1100 mg/100g
Potassium content range of hydrolyzed whey protein: 120-540 mg/100g
Calcium content range of hydrolyzed whey protein: 120-1700 mg/100g.
Raw material production process data of hydrolyzed whey protein of the present application
The hydrolyzed whey protein is prepared by the following preparation process: whey protein-recovering-adjusting pH-adding enzyme for hydrolysis-keeping pH constant-measuring degree of hydrolysis-enzyme inactivating-ultra-filtering-desalting-freeze drying-finished product
Concentrated whey protein is used as a raw material, pepsin and trypsin are selected, multi-factor and multi-level are arranged by adopting uniform experimental design, the optimal optimization conditions (weak bitter taste, moderate hydrolysis degree and low whole protein content) for enzymolysis of the whey protein are sought, the regression analysis is carried out on the test results by utilizing DPS data processing software, the regression equation with higher fitting degree is obtained, and the optimal hydrolysis conditions for enzymolysis of the whey protein by pepsin and trypsin are determined as follows: the enzymolysis temperature is 42 ℃, the pH value is 2.5, the ratio of enzyme to substrate is 1/25, the substrate concentration is 5.25%, the enzymolysis time is 240min, and the hydrolysis degree is 10-30%. Degree of hydrolysis of hydrolyzed whey protein: 10-30% and average molecular weight less than 3000Da.
Hydrolysate bitterness Change test
The enzymolysis temperature is 42 ℃, the pH value is 2.5, the ratio of enzyme to substrate is 1/25, the substrate concentration is 5.25 percent, the enzymolysis time is 240min, and the hydrolysis degree is 10-30 percent; the following results were obtained with one of the terms as a variable. Bitterness assessment was performed using a quinine sulfate standard solution. A50ml 5% ethanol solution of 0.1g quinine sulfate was used as a standard solution. The bitterness is grade 5 standard, and the standard solution concentration is C (3 x 10) -6 mole/L) is non-bitter; 2C slightly bitter; 4C medium bitter taste; strong 8C bitterness; 10C is very bitter; 12C is strongly bitter.
Table 2: bitterness testing for different treatment conditions
Method for measuring whole protein content, hydrolysis degree and Gly-Leu-Phe
The molecular weight of whole proteins and short peptides can be determined by ultracentrifugation-sedimentation rate method (AUC-SV) or SDS-polyacrylamide gel electrophoresis. Both are commonly used assay methods.
The degree of hydrolysis of proteins and short peptides is expressed as: the peptide bonds that are hydrolytically cleaved from a protein molecule account for the proportion of total peptide bonds in the protein molecule. The currently commonly used measurement methods are: OPA method, TNBS method, TCA method, pH-Stat method, formaldehyde fixation method, ninhydrin colorimetric method.
The sequence determination method of the short peptide fragment such as Gly-Leu-Phe and the like comprises the following steps: a method for measuring a peptide fragment sequence by a mass spectrometer, a method for identifying the peptide fragment by a liquid chromatography-mass spectrometry (LC-MS) method, and a method for identifying a liquid chromatography/capillary electrophoresis tandem flight time mass spectrometry.
Table 3-1: degree of hydrolysis index for different treatment conditions
Tables 3-2: whole protein content of different treatment conditions
The effective amino acid sequence of the short peptide fragment in the hydrolyzed whey protein in the prior art is not clear, and only few reports exist, for example, the hydrolyzed whey protein contains Gly-Leu-Phe (0.5%) short peptide fragment, and animal experiments show that the hydrolyzed whey protein has the function of activating phagocytosis and can improve the adhesion force to aged erythrocytes. The prior art reports that the content of Leu-Glu is not more than 2%, val-Thr-Iso is not more than 1%, phe-Ser-Tyr is not more than 1%, and Tyr-Gly content is not reported yet.
In this example, the hydrolyzed whey protein obtained by hydrolysis with physiologically-derived pepsin and trypsin contains about 6% of Leu-Glu fragments, about 3% of Val-Thr-Iso fragments, about 2% of Phe-Ser-Tyr fragments, and about 1% of Tyr-Gly fragments. The fragments are more compliant with the physiological absorption of human bodies, and the literature supports that the fragments can promote the synthesis of immune protein, induce the proliferation and differentiation of B lymphocytes and regulate the effects of T lymphocytes. Is suitable for patients with nephropathy to improve immunity, relieve negative nitrogen balance, and maintain homeostasis.
Degree of hydrolysis of hydrolyzed whey protein: the hydrolysis degree is required to be between 10 and 30 percent, and the average molecular weight is less than 3000Da. Enzyme preparation for hydrolysis of whey protein: pepsin and trypsin. The desalting process comprises the following steps: desalting with DA201-C macroporous resin, using deionized water and 40-70% ethanol as desorbent, loading at flow rate of 0.5-2.5mL/min and loading concentration of 10-50mg/mL, and determining desalting rate after desalting. Removing the raw materials and adding sodium, potassium, calcium, phosphorus, magnesium and the like when adjusting the pH value, and ensuring that the low electrolyte in the raw materials is hydrolyzed. The hydrolyzed whey protein was measured (the measurement side was assigned to the institute of nutritional sources, beijing, inc.), and the following results were obtained.
Sodium content range of hydrolyzed whey protein: 80-250 mg/100g
Magnesium content range of hydrolyzed whey protein: 60-100 mg/100g
Phosphorus content range of hydrolyzed whey protein: 150-300 mg/100g
Potassium content range of hydrolyzed whey protein: 450-530 mg/100g
Calcium content range of hydrolyzed whey protein: 350-800 mg/100g.
In the selection of the hydrolyzed whey protein material, the material with low electrolytes is selected as much as possible in principle, but in practice, the electrolytes in the material are somewhat too high and too low, so that the application risks, and a reasonable range needs to be comprehensively considered to reduce the risks as much as possible.
The present invention further performs the side chain of amino acid content on the hydrolyzed whey protein of the present invention and the purchased hydrolyzed whey protein, and the results are shown in table 1.
Table 4: comparison of amino acid content of the hydrolyzed whey protein of the present invention with that of commercially available hydrolyzed whey protein
Based on the above assay results, the inventors considered that the hydrolyzed whey protein was more suitable than commercially available hydrolyzed whey protein for use as a protein component of a formula for special medical use for renal disease dialysis patients.
The inventors also compared the amino acid content of the hydrolysed whey protein of the invention with soy oligopeptides, beef protein peptides or isolated whey protein.
Table 5: amino acid content comparison of hydrolyzed whey protein and Soybean oligopeptide
Table 6: amino acid content comparison of hydrolyzed whey protein and beef protein peptide
Based on the above assay results, the inventors believe that the hydrolyzed whey protein is more suitable than soy oligopeptide, beef protein peptide or isolated whey protein for use as a protein component of a special medical use formula for renal disease dialysis patients.
Table 7: comparison of amino acid content of hydrolyzed whey protein with other whey protein sources
From the above measurement results, the inventors considered that the hydrolyzed whey protein of the present invention has a higher amino acid content and a lower electrolyte content than hydrolyzed whey protein isolate, hydrolyzed whey protein concentrate, hydrolyzed whey powder, hydrolyzed desalted whey powder, and the like, which are derived from other different sources.
Bitterness testing of compositions
The addition amount of each component in each 100g of the product is as follows:
hydrolyzed whey protein powder: 95g
Lactic acid: 2.4g
And (3) citric acid: 2.4g
Sucralose: 80mg of
Neotame: 120mg.
Table 8: taste testing of compositions
The inventors have found that a composition comprising specific amounts of hydrolyzed whey protein powder, lactic acid, citric acid, sucralose and neotame can eliminate the bitter taste of the hydrolyzed whey protein powder.
Example 2: clinical efficacy observation of experimental product contrast casein peptide intervention treatment renal dialysis patients
To further verify the applicability of the hydrolysed whey proteins of the present invention clinically for renal dialysis patients, the inventors prepared the following experimental products for patient use.
Experimental products
The addition amount of each component in each 100g of the product is as follows:
hydrolyzed whey protein powder: 95g
Lactic acid: 2.4g
And (3) citric acid: 2.4g
Sucralose: 80mg of
Neotame: 120mg.
Control product
The hydrolyzed whey protein powder of the experimental product was replaced with casein peptide.
Experimental methods
The albumin determination method comprises the following steps: the measurement was carried out using an Albumin (ALB) measurement kit (bromocresol green method).
Serum phosphorus, serum calcium, serum sodium, serum potassium, serum magnesium: measurement with a Biochemical Analyzer
A biochemical analyzer: the biochemical analyzer is mainly used for measuring various chemical components in human serum, and the detection items are used for measuring items such as liver function, kidney function, myocardial enzyme, blood sugar, blood fat and the like besides potassium, sodium, calcium, magnesium, phosphorus and chloride ions, and is a necessary instrument for clinical examination departments of hospitals. Blood ions can be measured 6: calcium (ca), phosphorus (p), magnesium (mg), potassium (k), sodium (na), chlorine (cl).
Blood amino acid: the amino acid content was measured by an amino acid analyzer for clinical analysis. The expected application of the product is as follows: the metabolic condition of human body is judged by analyzing the content of amino acid in physiological body fluid, and various amino acid metabolic diseases are diagnosed.
Renal blood flow: measurement by the p-Aminomaleic acid method
Renal blood flow is mainly measured using specific substances such as the clearance test for aminohippuric acid.
1. Test the morning on empty stomach and lying still, 07.
2.07.
3. Sterile p-aminohippuric acid solution (200 g/L) was immediately injected intravenously at 0.4ml/kg body weight over 10min, and then a maintenance infusion was performed by adding 10ml of p-aminohippuric acid solution (200 g/L) to 500ml of physiological saline.
4.08. The bladder was flushed with 20ml of saline and the bladder was emptied by injecting 20ml of air. The washing liquid and the urine are combined and mixed evenly, and 10ml is taken to determine the content of the p-amino hippuric acid.
5.09, repeatedly drawing blood for the 1 st time, and reserving a urine specimen; the blood drawing and urine specimen collection are repeated for the 2 nd time at 20 min, and the operation method refers to the step 4.
6. Measuring the concentration of p-amino hippuric acid, firstly, sequentially adding distilled water, p-amino hippuric acid standard solution (0.04 mg/ml) and diluted urine (4 ml) into a blank tube, a standard tube and a measuring tube respectively; then, 0.8ml of hydrochloric acid (2 mol/L), 0.4ml of sodium nitrite solution (1 g/L) and 0.4ml of ammonium sulfamate solution (5 g/L) are added into each tube in sequence; finally, 0.4ml of color developing agent is added into each tube. Adding the reagent each time, mixing, and standing for 3min. The color development is stable for 10-15 min, the blank tube is used for zero calibration at 540nm, and the absorbance value of each tube is read.
7. Renal blood flow was measured.
All the above measurements of the index were made by the hospital. The following examples all used the same method and were determined by the hospital side.
Malnutrition and calcium-phosphorus metabolic disorders are common complications of patients with Maintenance Hemodialysis (MHD), and are one of the main causes of cardiovascular and cerebrovascular diseases, secondary hyperparathyroidism and renal bone diseases. Hemodialysis replaces the function of kidney, so that toxin generated by human body metabolism is discharged out of the body, and the dialysis influences the nutritional status and calcium and phosphorus metabolism of MHD patients and is closely related to the survival prognosis of the patients.
Malnutrition of patients in renal dialysis is mainly protein energy malnutrition, and is mainly caused by accumulation of uremic toxins, metabolic disorders, amino acid loss during dialysis, insufficient dialysis, and slight inflammatory reaction. Phosphorus plays an important role in mineral metabolism, 70-80% of phosphorus taken by the body from food is metabolized by the kidney, and hemodialysis patients often have kidney injury of different degrees. Therefore, the decrease in the phosphorus excretion function of these patients is likely to lead to the occurrence of hyperphosphatemia if the decrease is not controlled. Hyperphosphatemia can continue to cause metabolic disorder complications such as hyperparathyroidism and hypocalcemia in patients, thereby affecting the quality of life and survival rate of the patients. On the other hand, protein is also an indispensable nutrient for human body, and for dialysis patients, the more protein is taken in daily, the more phosphorus and calcium are taken in by protein intake, the higher the blood phosphorus and calcium concentration of patients is, and the high protein intake is an important cause of hyperphosphatemia and hypercalcemia. However, too little protein intake is a risk factor that leads to increased mortality in dialysis patients. Therefore, theoretically, the blood phosphorus level of a patient can only be improved if the patient has sufficient protein intake while controlling the intake of phosphorus and calcium.
The purpose is to research the influence of the hydrolyzed protein component (high protein, low phosphorus and low calcium) on the blood protein, the blood phosphorus and the blood calcium of dialysis patients and provide scientific guidance basis for the diet of the dialysis patients.
[ method ] 100 patients with maintenance hemodialysis were randomly divided into a pretreatment group and a control group, 50 patients in each group. The two groups are subjected to diet education, and the high-quality protein in the edible protein accounts for 50-60% of the total protein, the lipid intake is not more than 30% of the total heat, and fruits and vegetables are eaten more so as to ensure that enough vitamins are ingested; reducing intake of food with high potassium content such as herba Spinaciae, radix Dauci Sativae, herba Zosterae Marinae, etc.; the daily water intake of the patient is limited, and the excessive load on the liver is avoided. The intervention group is based on the conventional diet propaganda and is given with a hydrolyzed protein component (high protein, low phosphorus and low calcium) at 25 g/day; the control group was administered casein peptide (high protein, high phosphorus and high calcium) at 25 g/day based on the conventional diet instruction. Protein, phosphorus and calcium levels in the blood of both groups of patients were determined 1 month and 3 months after the intervention.
[ results ]
After 1 month and 3 months of intervention, the albumin level of the intervention group is obviously higher than that of the control group, and the blood phosphorus and blood calcium levels are lower than that of the control group (P < 0.05).
Table 9: two groups of patients intervene in the change of blood protein, blood phosphorus and blood calcium
* Compared with the group before intervention, P is less than 0.05
[ conclusion ] compared with the casein peptide component (high protein, high phosphorus and high calcium), the hydrolyzed protein component (high protein, low phosphorus and low calcium) can effectively control the albumin, blood phosphorus and blood calcium levels of dialysis patients.
Example 3: clinical curative effect observation of experimental product on diabetic nephropathy patient in comparison with soybean oligopeptide intervention treatment
Experimental products
The addition amount of each component in each 100g of the product is as follows:
hydrolyzed whey protein powder: 95g
Lactic acid: 2.4g
Citric acid: 2.4g
Sucralose: 80mg of
Neotame: 120mg.
Control product
The hydrolyzed whey protein powder of the experimental product is replaced by soybean oligopeptide.
Diabetic nephropathy is a common chronic complication of diabetes, and is one of the main factors affecting the life quality and the death rate of diabetic patients. With the rising incidence of diabetes and the increasing aging population, diabetic nephropathy has developed into the main primary disease of end-stage nephropathy. Currently, the medical costs for dialysis for end-stage renal disease worldwide are $ 1.1 trillion, with diabetic nephropathy accounting for 30%. The prevention and delay of progression of diabetic nephropathy has been the focus of chronic disease management and clinical treatment, and protein supply for diabetic nephropathy has become a major problem for current clinical treatment and patient confusion. The research aims at nutritional intervention of milk-derived hydrolyzed whey protein and plant-derived soybean oligopeptide on diabetic nephropathy patients, and compares nutritional treatment effects of the milk-derived hydrolyzed whey protein and the plant-derived soybean oligopeptide.
The serum sodium concentration before dialysis of dialysis patients is inversely related to mortality, and hyponatremia is also related to hypoproteinemia, weight loss, and weight gain during dialysis. Malnutrition is a more common complication of hemodialysis with a frequency of 23% to 76%. Malnutrition, which is associated with increased morbidity and mortality in maintenance hemodialysis patients, is an important factor affecting the prognosis and quality of life of patients. At the same time, both too high and too low sodium levels are detrimental to the patient's nutritional status during dialysis. On the one hand, strict control of low-salt diet and reduced dialysis-interval fluid intake; on the other hand, dysregulation of blood sodium levels is still disadvantageous for patients in malnutrition and inflammation states and causes iatrogenic inter-dialysis weight gain due to increased gradients in serum sodium and dialysate sodium concentrations, and pre-dialysis blood sodium levels below dialysate sodium concentrations can stimulate patient inter-dialysis craving, increase water intake, and lead to inter-dialysis weight gain.
[ purpose ] to research the influence of the hydrolyzed protein component (high essential amino acid, low sodium and low phosphorus) on the blood protein, blood sodium and blood phosphorus of a dialysis patient with diabetic nephropathy and provide scientific guidance basis for the diet of the dialysis patient.
[ method ] 200 dialysis patients with diabetic nephropathy were randomly divided into a pretreatment group and a control group, each of which was 100 patients. And (3) inclusion standard: receive regular hemodialysis treatment >3 months, 3 times a week for 4h each time. Simultaneously excluding: those who had transfused or had bleeding in the last 3 months; those suffering from other hematological disorders besides renal anemia; it has tumor, severe cardiopulmonary insufficiency, severe liver dysfunction, and chronic infectious wasting diseases such as tuberculosis. The two groups are subjected to diet education, and the intervention group gives a protein hydrolysate component (high essential amino acid, low sodium and low phosphorus) 25 g/day on the basis of conventional diet education; the control group was administered 25 g/day of soy oligopeptide module (low essential amino acids, high sodium and high phosphorus) based on the conventional dietary instruction. Protein, sodium and phosphorus levels in the blood of both groups of patients were determined 6 months after the intervention.
[ results ]
After 6 months of intervention, the albumin in the intervention group is obviously higher than that in the control group, the blood sodium level is obviously higher than that in the control group, and the blood phosphorus level is obviously lower than that in the control group (P < 0.05).
Table 10: two groups of patients intervened in the change of blood protein, blood sodium and blood phosphorus
* Compared with the group before intervention, P is less than 0.05
[ conclusion ] compared with soybean oligopeptide component (low essential amino acid, high sodium and high phosphorus), the hydrolyzed protein component (high essential amino acid, low sodium and low phosphorus) can effectively control the protein nutrition, blood sodium and blood phosphorus level of dialysis patients.
Example 4: experimental product for observing clinical curative effect of experimental product on treatment of renal dialysis patients by beef protein peptide interventionExperimental products
The addition amount of each component in each 100g of the product is as follows:
hydrolyzed whey protein powder: 95g
Lactic acid: 2.4g
Citric acid: 2.4g
Sucralose: 80mg of
Neotame: 120mg.
Control product
The hydrolyzed whey protein powder of the experimental product is replaced by beef protein peptide.
Maintenance hemodialysis is an effective alternative for end-stage renal patients, but as dialysis time is prolonged, even if sufficient dialysis is performed, a plurality of complications still occur in part of patients, wherein malnutrition is quite common and is a main reason for high hospitalization rate and high fatality rate of patients. In patients with kidney disease, varying degrees of hypoproteinemia and electrolyte metabolism disorders are prevalent. Hyperkalemia is common in renal dialysis patients. The intake of potassium-rich food can easily cause the increase of the blood potassium level of a patient with renal dialysis, so that the restriction of the intake of potassium ions in the food is an important link for preventing premenstrual hyperkalemia. Magnesium is a basic cofactor for more than 300 enzymatic reactions involved in energy metabolism in the body, and proper magnesium ion concentration is very important for maintaining normal electrolyte concentration in the body of dialysis patients. In addition, magnesium deficiency is often accompanied by potassium deficiency, magnesium is not only important for maintaining normal intracellular potassium concentrations, but also for the supplementation and restoration of intracellular potassium ions, sufficient magnesium ions are required, and elevated serum magnesium exacerbates the toxic effects of hyperkalemia.
[ purpose ] to research the influence of the protein hydrolysate component (high branched chain amino acid, low phosphorus, low potassium and low magnesium) on blood protein, blood phosphorus, blood potassium and blood magnesium of a renal dialysis patient, and provide scientific guidance basis for diet of the renal dialysis patient.
[ method ] 80 renal dialysis patients were randomly divided into a pretreatment group and a control group, 40 in each of the two groups. Inclusion criteria were: the patient is diagnosed as the terminal stage of chronic renal failure, and the age and the nature are not limited; dialysis treatments were performed 3 times a week for 4 hours each. Exclusion criteria: antibiotic-treated infectious diseases; patients with liver disease, malignant tumor, acute cerebrovascular accident, hyperlipidemia, and hormone or immunosuppressant; patients with recent acute vascular disease, surgical treatment, and trauma. The two groups are subjected to diet education, and the intervention group is given a protein hydrolysate component (high branched chain amino acid, low phosphorus, low potassium and low magnesium) at 25 g/day on the basis of conventional diet education; the control group is administered with beef protein peptide (low branched chain amino acid, high phosphorus, high potassium and high magnesium) 25 g/day based on conventional diet instruction. Protein, phosphorus, potassium and magnesium levels in the blood of both groups of patients were determined 3 months after the intervention.
[ results ]
After 3 months of intervention, the albumin in the intervention group is obviously higher than that in the control group, and the levels of the blood phosphorus, the blood potassium and the blood magnesium are obviously lower than that in the control group (P is less than 0.05).
Table 11: changes in blood protein, blood sodium and blood phosphorus before and after intervention in two groups of patients
* Compared with the group before intervention, P is less than 0.05
The hydrolyzed protein component (highly branched chain amino acids low in phosphorus, low in potassium, and low in magnesium) provides better control of protein nutrition, blood phosphorus, blood potassium, and blood magnesium levels in dialysis patients than does beef protein peptide (low branched chain amino acids high in phosphorus, high in potassium, and high in magnesium).
Example 5: clinical curative effect observation of experimental product comparing isolated whey protein and hydrolyzed whey protein of other manufacturers for interventional therapy of renal dialysis patients
Experimental products
The addition amount of each component in each 100g of the product is as follows:
hydrolyzed whey protein powder: 95g
Lactic acid: 2.4g
Citric acid: 2.4g
Sucralose: 80mg of
Neotame: 120mg.
Control product
Control group 1: the hydrolyzed whey protein powder of the experimental product was replaced with whey protein isolate.
Control group 2: hydrolyzed whey protein powder prepared by Hengyuan company and replacing the experimental product with hydrolyzed whey protein
The end stage nephropathy is the end stage of various chronic kidney diseases, the clinical common treatment method is hemodialysis, the hemodialysis can inhibit the development of nephropathy of patients to a certain extent, the life of the patients is maintained, and the clinical value is high. However, long-term dialysis treatment can cause a large amount of nutrients such as proteins and trace elements to flow out of the body, which causes malnutrition of patients, influences the treatment effect in the later period and reduces the life quality of the patients.
Among dialysis patients with renal diseases, patients with metabolic disorders of high calcium and high phosphorus account for about 10%. Although long-term dialysis can maintain life, the disorder of calcium and phosphorus hormones in a body is difficult to correct, and a series of complications such as renal bone diseases and secondary hyperparathyroidism are inevitable finally. High calcium and high phosphorus are converted into osteogenic cells through vascular smooth muscle cells, so that calcification is caused to generate cardiovascular complications, coronary heart disease is caused, and the death rate is extremely high. In addition, complications can exacerbate hyperphosphatemia and active vitamin D deficiency, leading to a vicious circle. Therefore, the balance of calcium and phosphorus in blood is maintained, and the control of the calcium and phosphorus at normal concentration is the guarantee of successful treatment.
[ purpose ] to research the influence of the hydrolyzed protein component (low molecular weight, low phosphorus and low calcium) on the blood protein, the blood phosphorus and the blood calcium of a dialysis patient and provide scientific guidance basis for the diet of the dialysis patient.
300 patients with maintenance hemodialysis were randomly divided into a priming group, a control group 1 and a control group 2, 150 patients in each of the three groups. Both groups were subjected to dietary instruction and nutritional intervention. The method comprises the following steps: the patience explains the reasons and adverse consequences of malnutrition which is easy to occur in long-term hemodialysis to patients and family members and has important significance in establishing scientific and reasonable dietary habits, so that the patients and the family members can be aware of the importance of nutrition supply, actively accept and cooperate in the nutrition nursing process, and improve the effect; according to the specific conditions of the patient, an individual nutritional recipe is formulated, the daily nutritional intake condition of the patient is recorded, and the adjustment is carried out according to the requirement; eating more fresh fruits and vegetables, and supplementing various trace elements and vitamins; the water intake is set according to the daily urine output condition of a patient, spicy, greasy and irritant food is avoided, and the principle of less eating and more eating is followed; strict low-phosphorus diet and proper high-protein diet are forbidden to feed foods rich in phosphorus and calcium, such as milk, eggs, dried small shrimps and the like. The intervention group is based on the conventional diet propaganda and is given with a hydrolyzed protein component (low molecular weight, low phosphorus and low calcium) at 25 g/day; the control group 1 was administered with isolated whey protein (high-molecular, high-phosphorus, high-calcium) at 25 g/day based on the conventional diet instruction; the control group 2 was prepared by administering hydrolyzed whey protein (high-molecular, high-phosphorus, high-calcium) of other manufacturers at 25 g/day based on the conventional diet instruction. Protein, phosphorus and calcium levels in the blood of both groups of patients were determined 9 months after the intervention.
[ results ]
After 9 months of intervention, the blood protein level of the intervention group is higher than that of the control group 1 and the control group 2, and the blood phosphorus level and the blood calcium level are lower than that of the control group 1 and the control group 2 (P < 0.05).
Table 12: three groups of patients intervened in the change of blood protein, blood phosphorus and blood calcium
* Compared with the group before intervention, P is less than 0.05
Compared with the isolated whey protein (high-molecular high-phosphorus high-calcium) and the hydrolyzed whey protein (low-molecular high-phosphorus high-calcium) of other manufacturers, the hydrolyzed protein component (low-molecular low-phosphorus low-calcium) can increase the protein level of a dialysis patient and maintain stable blood phosphorus and blood calcium levels.
Claims (12)
1. A hydrolyzed whey protein, wherein the hydrolysis degree of the hydrolyzed whey protein is 10% -30%, the average molecular weight is 500-3000 Da, the sodium content range of the hydrolyzed whey protein is 80-250mg/100 g, the magnesium content range is 60-100mg/100 g, the phosphorus content range is 150-300mg/100 g, the potassium content range is 450-530mg/100 g, the calcium content range is 350-800mg/100 g, the essential amino acid content is 31-36 g/100g, and the branched amino acid content is 14-17g/100 g, and the hydrolyzed whey protein is obtained by a preparation method of the hydrolyzed whey protein, wherein the preparation method comprises a step of carrying out enzymolysis on the whey protein by pepsin and trypsin, and a desalting step, so that the desalting rate of the hydrolyzed whey protein reaches over 90%; performing enzymolysis at 30-45 deg.C and pH of 2-4; the ratio of enzyme to whey protein is 1/30-1/20 and the whey protein concentration is 5wt% -6wt%; the enzymolysis time is 200-260 minutes.
2. The preparation method of the hydrolyzed whey protein comprises the steps of carrying out enzymolysis on the whey protein by pepsin and trypsin and desalting, so that the desalting rate of the hydrolyzed whey protein reaches over 90 percent; performing enzymolysis at 30-45 deg.C and pH of 2-4; the ratio of enzyme to whey protein is 1/30-1/20 and the whey protein concentration is 5wt% -6wt%; the enzymolysis time is 200-260 minutes.
3. A composition for renal dialysis patients comprising a hydrolyzed whey protein according to claim 1.
4. The composition according to claim 3, comprising the following ingredients by weight:
75 to 96 percent of hydrolyzed whey protein,
1.5 to 15 percent of sour agent,
0.1% -15% of sweetening agent.
5. The composition according to claim 4, wherein the acidulant is selected from the group consisting of lactic acid, citric acid, monosodium fumarate, adipic acid, tartaric acid, phosphoric acid, malic acid, acetic acid and sodium carbonate and/or the sweetener is selected from the group consisting of sucralose, neotame, cyclamate, maltitol, sorbitol, thaumatin, aspartame, steviol glycosides, acesulfame potassium, isomaltulose, erythritol and xylitol.
6. A special medical use formula comprising hydrolyzed whey protein according to claim 1 as protein component.
7. Use of a composition according to any one of claims 3 to 5 in the preparation of a special medical use formula for patients with renal disease.
8. The use of claim 7, wherein the renal disease patient is a renal disease dialysis patient, a diabetic nephropathy dialysis patient, or an end stage chronic renal failure patient.
9. Use of hydrolysed whey protein according to claim 1 as a protein component of a food for subjects in need of low electrolyte content food and/or subjects not compliant to bitter taste, which use does not involve treatment or prevention of disease.
10. The use according to claim 9, wherein the food product is a special medical use formula.
11. The use according to claim 10, wherein the special medical use formula is for patients with renal disease.
12. The use of claim 11, wherein the renal disease patient is a renal disease dialysis patient, a diabetic nephropathy dialysis patient, or an end stage chronic renal failure patient.
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