CN104007040A - Method for detecting collagen content in covering-type collagen modified fiber fabrics - Google Patents

Abstract

The invention discloses a method for detecting the collagen content in covering-type collagen modified fiber fabrics. The method comprises qualitative analysis by adopting a scanning electron microscope, and quantitative analysis by adopting a dissolution method, wherein the qualitative analysis comprises the following steps: carrying out vacuum metal spraying treatment on a tested sample, and then observing the surface morphology of the tested sample by the scanning electron microscope at 1K-10K, and if a fiber membrane exists, the tested sample is determined to be collagen but not denatured collagen; a dissolution method is adopted for carrying out quantitative analysis. The quantitative analysis comprises the following steps: dissolving and removing collagen from the covering-type collagen modified fiber fabrics with known drying mass by an oxidizing agent solution or an alkaline solution at certain temperature, filtering by a sand core crucible, collecting residues, and washing, drying and weighing the residues, carrying out quality correction, calculating the mass percentage of residues accounting for the covering-type collagen modified fiber fabrics, and calculating the content in percentage of collagen by a difference value method.

Description

A kind of method that detects collagen content in cover type collagen modification fabric

Technical field

The present invention relates to a kind of detection method of collagen content, be specifically related to a kind of method that detects collagen content in cover type collagen modification fabric.

Background technology

Collagen is a kind of directly protein from extracting in biosome, and it has retained the triple-helix structure of molecule.Different from the denatured collagen that uncoiling and Fragmentation have occurred, it be medically proved to be can Promote cell's growth protein, there is the skin wound of reparation, antiallergy, pinning moisture, nourish the functions such as skin care and antiultraviolet, in fields such as health care, beauty treatment, medical treatment, be widely used, in recent years, collagen starts to be applied to garment industry, portioned product is put on market, wherein, by the mode of rear arrangement, collagen being covered on general fibre fabric is a most frequently used method.

After surpassing certain temperature, collagen can produce sex change, finally becomes common protein.Although still there is skin-friendly, lost many premium properties of collagen, can not fully demonstrate the value of collagen modification fabric.Collagen and denatured collagen tool in physics and chemistry performance is very different.Wherein, collagen has the advantages that to form tunica fibrosa, have fibre structure clearly, and denatured collagen does not possess this feature on collagem membrane.

At present, in fabric, the content of collagen adopts nitriding to determine, instrument has Dumars azotometer or triumphant formula azotometer, is multiplied by certain coefficient extrapolates protein content according to surveyed nitrogen content.These class methods have the following disadvantages.One, for fabric and the nitrogenous fabric of fiber itself of introducing nitrogenous dyestuff or auxiliary agent in dyeing and finishing, test result is inaccurate; Its two, can not to tell coverture be collagen or uncoiling has occurred and the metaprotein of peptide bond rupture.

Summary of the invention

The object of the invention is to the problem existing for existing method of testing, a kind of method that detects collagen content in cover type collagen modification fabric is provided.

In order to achieve the above object, the present invention adopts technical scheme to comprise the following steps:

(1) sem observation sample microscopic appearance: get fabric 5～10 * 5～10mm, vacuum drying at room temperature is more than 24 hours, with double faced adhesive tape in scanning electron microscope dedicated, vacuum metal spraying is processed more than 20 minutes, with its microscopic appearance of sem observation, enlargement factor 1k-10k, observes fabric surface, if there is tunica fibrosa, can be defined as collagen rather than denatured collagen;

(2) extraction of tested sample and preparation: uniform sampling, make it have representativeness, approximately 1 gram, every part of sample, need dry, and m dries to constant weight ₀, concrete steps are:

Fabric is cut into the cloth waste of 5～10 * 5～10mm; Fabric is placed in measuring cup, carries out drying operation, drying operation carries out in airtight ventilated drying oven, temperature is (105 ± 3) ℃, time is 4h～16h, after oven dry, measuring cup and fabric is taken out and moves into rapidly in exsiccator in baking oven, until completely cooling, be no less than 2 hours cool time, cooling after, from exsiccator, take out measuring cup, weigh rapidly, calculate the fabric dry weight m after drying ₀, the fabric after being dried;

(3) quantitative test: adopt different solution and different treatment temperatures, be divided into two kinds of methods, be i.e. method A or method B, its test result equivalence;

Analytical procedure is as follows: method A: adopt oxygenant and NaOH to be combined into lysate, fabric after step (2) is dried is put into 250mL with the Erlenmeyer flask of stopper, fabric by the lysate of preparation after by every gram of oven dry heavily adds 100～120mL to Erlenmeyer flask, by the residue washing existing in solution after the dissolving m that dries to constant weight ₁after, by differential technique, calculate collagen content; Method B: adopt sodium hydroxide solution to dissolve collagen, the residue existing in solution after dissolving is washed to the m that dries to constant weight ₁after, adopt differential technique to calculate collagen content, collagen content is pressed formula (1) and is calculated:

$P=(1-\frac{m_{1}d}{m_0})\×100\%---\left(1\right)$

P is collagen dry mass mark, %;

D is the mass change correction factor of insoluble fiber component.

In said method, described fabric is one or more knitted fabric or the woven in cotton fiber, Modal fibre, viscose rayon, CUP, dacron, polyamide fibre, mekralon, polyacrylonitrile fibre, polyurethane fiber, chlorofibre.

In said method, described in step (3), oxygenant is hypochlorite or hydrogen peroxide.

In said method, when oxygenant is hypochlorite, in the lysate of preparing: hypochlorite solutions is 0.9～1.1mol/L, NaOH mass percent concentration is 0.3%-0.6%, and the fabric after oven dry is isothermal vibration 30～40min dissolving under 20 ℃ ± 2 ℃ conditions in lysate.

In said method, described hypochlorite is sodium hypochlorite or lithium hypochlorite.

In said method, when oxygenant is hydrogen peroxide, prepare in lysate: hydrogen peroxide mass percent concentration is 4～7%, and NaOH mass percent concentration is 2～3%, the fabric after oven dry in lysate under 70 ℃ ± 2 ℃ conditions isothermal vibration 8～12min dissolve.

In said method, in the described method B of step (3), described sodium hydroxide solution mass percent concentration is 4～7%, and the fabric after oven dry is isothermal vibration 8～12min dissolving under 97 ℃ ± 2 ℃ conditions in sodium hydroxide solution.

In said method, in step (3), the value of d is: in method A, take hypochlorite as solution, when described fabric is cotton fiber, viscose rayon or Modal fibre, d is 1.01, and when described fabric is CUP, dacron, polyamide fibre, mekralon, polyacrylonitrile fibre, polyurethane fiber or chlorofibre, d is 1.00; Take hydrogen peroxide as solution, when described fabric is cotton fiber or CUP, d is 1.02, when described fabric is viscose rayon or Modal fibre, d is 1.03, and when being dacron, polyamide fibre, mekralon, polyacrylonitrile fibre, polyurethane fiber or chlorofibre for described fabric, d is 1.00; In method B, when described fabric is cotton fiber, viscose rayon or Modal fibre, d is 1.03, when described fabric is dacron, d is 1.04, and when described fabric is CUP, polyamide fibre, mekralon, polyacrylonitrile fibre, polyurethane fiber or chlorofibre, d is 1.00.

In said method, when described fabric is knitted fabric two or more in cotton fiber, Modal fibre, viscose rayon, CUP, dacron, polyamide fibre, mekralon, polyacrylonitrile fibre, polyurethane fiber, chlorofibre or woven, d value is pressed formula (2) and is calculated:

$d=\frac{m_3}{m_4}---\left(2\right)$

M ₃for the described fabric that does not contain collagen is processed to the fabric dry weight obtaining, the g of unit according to step (2);

M ₄for the fabric obtaining after not processing according to step (2) containing the described fabric of collagen, then through step (3) method, process the residue dry weight obtaining, the g of unit.

The present invention compares with the method for nitriding test collagen modification fabric collagen content, and tool has the following advantages:

(1) the present invention, by electron microscopic observation, can determine that coverture is collagen or collagenous degeneration protein hydrolysate;

(2) method of the present invention can be determined the content of collagen in the multiple conventional fabric that covers collagen, and is not subject to nitrogen element that dyeing and finishing processing belt enters and the impact of the institute of fiber Nitrogen element own;

(3) method of the present invention is simple, easy to operate.

Accompanying drawing explanation

Fig. 1 is the scanning electron microscope (SEM) photograph of embodiment 1 sample enlargement factor 1k;

Fig. 2 is the scanning electron microscope (SEM) photograph of embodiment 1 sample enlargement factor 10k;

Fig. 3 is the scanning electron microscope (SEM) photograph of embodiment 2 sample enlargement factor 3k;

Fig. 4 is the scanning electron microscope (SEM) photograph of embodiment 2 sample enlargement factor 10k;

Fig. 5 is the scanning electron microscope (SEM) photograph of embodiment 3 sample enlargement factor 1k;

Fig. 6 is the scanning electron microscope (SEM) photograph of embodiment 3 sample enlargement factor 10k;

Fig. 7 is the scanning electron microscope (SEM) photograph of embodiment 4 sample enlargement factor 1k;

Fig. 8 is the scanning electron microscope (SEM) photograph of embodiment 4 sample enlargement factor 10k.

Embodiment

Below in conjunction with embodiment, the present invention is described in further detail, but the scope of protection of present invention is not limited to the scope that embodiment represents.

Embodiment 1

The test of collagen content in collagen modification pure cotton knitting cloth: in known this cotton fabrics, collagen content is 2.75%.

(1) sem observation sample microscopic appearance.Get cloth specimen 5～10 * 5～10mm, vacuum drying at room temperature is more than 24 hours, and with double faced adhesive tape, in scanning electron microscope dedicated, vacuum metal spraying is processed more than 20 minutes, with its microscopic appearance of sem observation, amplifies respectively 1k and 10k., there is tunica fibrosa in observation fabric fibre surface, can be defined as collagen rather than denatured collagen (as depicted in figs. 1 and 2).

(2) extraction of tested sample and preparation: uniform sampling, makes it have representativeness, approximately 1 gram, each sample.As dissolving analytical test sample, cloth specimen is cut into the cloth waste of 5 * 5mm, then dry.All drying operation carries out in airtight ventilated drying oven, and temperature is (105 ± 3) ℃, 5 hours time.After oven dry, measuring cup and sample are taken out and moved in exsiccator, until completely cooling rapidly in baking oven.After cooling, from exsiccator, take out measuring cup (sample is in measuring cup), weigh rapidly, weigh up respectively measuring cup and sample weight, calculation sample dry weight m ₀for 1.3214g.

(3) quantitative test.Analytical procedure is as follows: the 1. preparation of solution.The sodium hydroxide solution that preparation mass percent concentration is 5%; 2. the ready sample of step (2) is put among 250mL tool plug Erlenmeyer flask, the lysate that 1. step is prepared heavily adds 100mL to Erlenmeyer flask by every gram of cloth, concuss 10min at 95-98 ℃ of temperature; 3. then, material in flask is poured into the core crucible of known dry weight, vacuum filtration, then successively with the residue in clear water cleaning, the neutralization of 5g/L acetic acid aqueous solution, clear water continuous wash core crucible, each washing all first keeps gravity discharge opeing 5min, then vacuum draw discharge opeing, by crucible and residue dry, cooling, weigh, calculate residue weight m ₁for 1.2489g.4. getting d is 1.03, and calculating collagen dry mass content in fabric is 2.65%, compares with collagen content 2.75% in known fabric, and error is 3.64%<10%.

Embodiment 2

The test of collagen content in collagen modification cotton/Polyurethane Knitted Fabric: in known this looped fabric, collagen content is 2.20%.

(1) sem observation sample microscopic appearance.Get cloth specimen 5～10 * 5～10mm, vacuum drying at room temperature is more than 24 hours, with double faced adhesive tape in scanning electron microscope dedicated, vacuum metal spraying is processed more than 20 minutes, with its microscopic appearance of sem observation, amplify respectively 3k and 10k and observe fabric fibre surface, have tunica fibrosa, can be defined as collagen rather than denatured collagen (as shown in Figure 3 and Figure 4).

(2) extraction of tested sample and preparation: uniform sampling, makes it have representativeness, approximately 1 gram, each sample.As dissolving analytical test sample, cloth specimen is cut into the cloth waste of 5 * 5mm, then dry.All drying operation carries out in airtight ventilated drying oven, and temperature is (105 ± 3) ℃, and the time is 5 hours.After oven dry, measuring cup and sample are taken out and moved in exsiccator, until completely cooling rapidly in baking oven.After cooling, from exsiccator, take out measuring cup, weigh rapidly, weigh up respectively measuring cup and sample weight, calculation sample dry weight m ₀for 1.2923g.

(3) quantitative test.Analytical procedure is as follows: the 1. preparation of solution.Being formulated as hydrogen peroxide mass concentration is 5%, the lysate that NaOH mass concentration is 2%; 2. the ready sample of step (2) is put among 250mL tool plug Erlenmeyer flask, the lysate that 1. step is prepared heavily adds 100mL to Erlenmeyer flask by every gram of cloth; The Erlenmeyer flask that lysate and cloth specimen are housed is placed in 70 ℃ ± 2 ℃ waters bath with thermostatic control, concuss 10min; 3. then residue in flask is poured into the core crucible of known dry weight, vacuum filtration, then successively with the residue in clear water cleaning, the neutralization of 5g/L acetic acid aqueous solution, clear water continuous wash core crucible, each washing all first keeps gravity discharge opeing 5min, then vacuum draw discharge opeing, by crucible and residue dry, cooling, weigh, calculate residue weight m ₁for 1.2409g.4. getting d is 1.02, and calculating collagen dry mass content in fabric is 2.06%, compares with collagen content 2.20% in known fabric, and error is 6.36%<10%.

Embodiment 3

The test of collagen content in collagen modification cotton/viscose glue (50/50): in known this cotton/binding textiles, collagen content is 2.10%.

(1) sem observation sample microscopic appearance.Get cloth specimen 5～10 * 5～10mm, vacuum drying at room temperature is more than 24 hours, and with double faced adhesive tape, in scanning electron microscope dedicated, vacuum metal spraying is processed more than 20 minutes, with its microscopic appearance of sem observation, amplifies respectively 1k and 10k., there is tunica fibrosa in observation fabric fibre surface, can be defined as collagen rather than denatured collagen (as shown in Figure 5 and Figure 6).

(2) extraction of tested sample and preparation: uniform sampling, makes it have representativeness, approximately 1 gram, each sample.As dissolving analytical test sample, cloth specimen is cut into the cloth waste of 5 * 5mm, then dry.All drying operation carries out in airtight ventilated drying oven, and temperature is (105 ± 3) ℃, and the time is 5 hours.After oven dry, measuring cup and sample are taken out and moved in exsiccator, until completely cooling rapidly in baking oven.After cooling, from exsiccator, take out measuring cup, weigh rapidly, weigh up respectively measuring cup and sample weight, calculation sample dry weight m ₀for 1.2648g.

(3) quantitative test.Analytical procedure is as follows: the 1. preparation of solution.The liquor natrii hypochloritis of preparation 1mol/L also adds NaOH, and the mass concentration that makes NaOH is 0.5%; 2. the ready sample of step (2) is put among 250mL tool plug Erlenmeyer flask, the lysate that 1. step is prepared heavily adds 100mL to Erlenmeyer flask by every gram of cloth; The Erlenmeyer flask that lysate and cloth specimen are housed is placed in 20 ℃ ± 2 ℃ waters bath with thermostatic control, concuss 40min; 3. then residue in flask is poured into the core crucible of known dry weight, vacuum filtration, then successively with the residue in clear water cleaning, the neutralization of 5g/L acetic acid aqueous solution, clear water continuous wash core crucible, each washing all first keeps gravity discharge opeing 5min, then vacuum draw discharge opeing, by crucible and residue dry, cooling, weigh, calculate the heavy m of residue ₁for 1.2277g.4. getting d is 1.01, and calculating collagen dry mass content in fabric is 1.96%, compares with collagen content 2.10% in known fabric, and error is 6.67%<10%.

Embodiment 4: collagen modification cotton/wash the test of collagen content in (70/30) woven fabric: in known this woven fabric, collagen content is 2.20%.

(1) sem observation sample microscopic appearance.Get cloth specimen 5～10 * 5～10mm, vacuum drying at room temperature is more than 24 hours, and with double faced adhesive tape, in scanning electron microscope dedicated, vacuum metal spraying is processed more than 20 minutes, with its microscopic appearance of sem observation, and difference enlargement factor 1k and 10k., there is tunica fibrosa in observation fabric fibre surface, can be defined as collagen rather than denatured collagen (as shown in Figure 7 and Figure 8).

(2) extraction of tested sample and preparation: uniform sampling, makes it have representativeness, approximately 1 gram, each sample.As dissolving analytical test sample, cloth specimen is cut into the cloth waste of 5 * 5mm, then dry.All drying operation carries out in airtight ventilated drying oven, and temperature is (105 ± 3) ℃, and the time is 5 hours.After oven dry, measuring cup and sample are taken out and moved in exsiccator, until completely cooling rapidly in baking oven.After cooling, from exsiccator, take out measuring cup, weigh rapidly, weigh up respectively measuring cup and sample weight, calculation sample dry weight m ₀for 1.3015g.

(3) quantitative test.Analytical procedure is as follows: the 1. preparation of solution.100 grams of lithium hypochlorites that contain 35% effective chlorine are dissolved in the water of 700ml, add sodium hydroxide solution, finally add water to 1L, the mass concentration that makes NaOH in whole solution is 0.5%; 2. the ready sample of step (2) is put among 250mL tool plug Erlenmeyer flask, the lysate that 1. step is prepared heavily adds 100mL to Erlenmeyer flask by every gram of cloth; The Erlenmeyer flask that lysate and cloth specimen are housed is placed in 20 ℃ ± 2 ℃ waters bath with thermostatic control, concuss 40min; 3. then residue in flask is poured into the core crucible of known dry weight, vacuum filtration, then successively with the residue in clear water cleaning, the neutralization of 5g/L acetic acid aqueous solution, clear water continuous wash core crucible, each washing all first keeps gravity discharge opeing 5min, then vacuum draw discharge opeing, by crucible and residue dry, cooling, weigh, calculating residue weight is 1.2632g.4. getting cotton d is 1.01, and terylene d is 1.00, and calculating collagen content in fabric is 2.26%, compares with collagen content 2.20% in known fabric, and error is 2.73%<10%.

Claims (9)

1. a method that detects collagen content in cover type collagen modification fabric, is characterized in that, comprises following method:

(1) sem observation sample microscopic appearance: get fabric 5～10 * 5～10mm, vacuum drying at room temperature is more than 24 hours, with double faced adhesive tape in scanning electron microscope dedicated, vacuum metal spraying is processed more than 20 minutes, with its microscopic appearance of sem observation, enlargement factor 1k-10k, observes fabric surface, if there is tunica fibrosa, can be defined as collagen rather than denatured collagen;

(2) extraction of tested sample and preparation: uniform sampling, make it have representativeness, approximately 1 gram, every part of sample, need dry, and m dries to constant weight ₀, concrete steps are:

Fabric is cut into the cloth waste of 5～10 * 5～10mm; Fabric is placed in measuring cup, carries out drying operation, drying operation carries out in airtight ventilated drying oven, temperature is (105 ± 3) ℃, time is 4h～16h, after oven dry, measuring cup and fabric is taken out and moves into rapidly in exsiccator in baking oven, until completely cooling, be no less than 2 hours cool time, cooling after, from exsiccator, take out measuring cup, weigh rapidly, calculate the fabric dry weight m after drying ₀, the fabric after being dried;

(3) quantitative test: adopt different solution and different treatment temperatures, be divided into two kinds of methods, be i.e. method A or method B, its test result equivalence;

Analytical procedure is as follows: method A: adopt oxygenant and NaOH to be combined into lysate, fabric after step (2) is dried is put into 250mL with the Erlenmeyer flask of stopper, fabric by the lysate of preparation after by every gram of oven dry heavily adds 100～120mL to Erlenmeyer flask, by the residue washing existing in solution after the dissolving m that dries to constant weight ₁after, by differential technique, calculate collagen content; Method B: adopt sodium hydroxide solution to dissolve collagen, the residue existing in solution after dissolving is washed to the m that dries to constant weight ₁after, adopt differential technique to calculate collagen content, collagen content is pressed formula (1) and is calculated:

$P=(1-\frac{m_{1}d}{m_0})\&times;100\%---\left(1\right)$

P is collagen dry mass mark, %;

D is the mass change correction factor of insoluble fiber component.

2. a kind of method that detects collagen content in cover type collagen modification fabric according to claim 1, is characterized in that: described fabric is one or more knitted fabric or the woven in cotton fiber, Modal fibre, viscose rayon, CUP, dacron, polyamide fibre, mekralon, polyacrylonitrile fibre, polyurethane fiber, chlorofibre.

3. a kind of method that detects collagen content in cover type collagen modification fabric according to claim 1, is characterized in that: described in step (3), oxygenant is hypochlorite or hydrogen peroxide.

4. a kind of method that detects collagen content in cover type collagen modification fabric according to claim 3, it is characterized in that: when oxygenant is hypochlorite, in the lysate of preparing: hypochlorite solutions is 0.9～1.1mol/L, NaOH mass percent concentration is 0.3%-0.6%, and the fabric after oven dry is isothermal vibration 30～40min dissolving under 20 ℃ ± 2 ℃ conditions in lysate.

5. a kind of method that detects collagen content in cover type collagen modification fabric according to claim 4, is characterized in that: described hypochlorite is sodium hypochlorite or lithium hypochlorite.

6. a kind of method that detects collagen content in cover type collagen modification fabric according to claim 3, it is characterized in that: when oxygenant is hydrogen peroxide, prepare in lysate: hydrogen peroxide mass percent concentration is 4～7%, NaOH mass percent concentration is 2～3%, and the fabric after oven dry is isothermal vibration 8～12min dissolving under 70 ℃ ± 2 ℃ conditions in lysate.

7. a kind of method that detects collagen content in cover type collagen modification fabric according to claim 1, it is characterized in that: in the described method B of step (3), described sodium hydroxide solution mass percent concentration is 4～7%, and the fabric after oven dry is isothermal vibration 8～12min dissolving under 97 ℃ ± 2 ℃ conditions in sodium hydroxide solution.

8. a kind of method that detects collagen content in cover type collagen modification fabric according to claim 2, it is characterized in that: in step (3), the value of d is: in method A, take hypochlorite as solution, when described fabric is cotton fiber, viscose rayon or Modal fibre, d is 1.01, and when described fabric is CUP, dacron, polyamide fibre, mekralon, polyacrylonitrile fibre, polyurethane fiber or chlorofibre, d is 1.00; Take hydrogen peroxide as solution, when described fabric is cotton fiber or CUP, d is 1.02, when described fabric is viscose rayon or Modal fibre, d is 1.03, and when described fabric is dacron, polyamide fibre, mekralon, polyacrylonitrile fibre, polyurethane fiber or chlorofibre, d is 1.00; In method B, when described fabric is cotton fiber, viscose rayon or Modal fibre, d is 1.03, when described fabric is dacron, d is 1.04, and when described fabric is CUP, polyamide fibre, mekralon, polyacrylonitrile fibre, polyurethane fiber or chlorofibre, d is 1.00.

9. a kind of method that detects collagen content in cover type collagen modification fabric according to claim 2, it is characterized in that: when described fabric is knitted fabric two or more in cotton fiber, Modal fibre, viscose rayon, CUP, dacron, polyamide fibre, mekralon, polyacrylonitrile fibre, polyurethane fiber, chlorofibre or woven, d value is pressed formula (2) and calculated:

$d=\frac{m_3}{m_4}---\left(2\right)$

M ₃for the described fabric that does not contain collagen is processed to the fabric dry weight obtaining, the g of unit according to step (2);

M ₄for the fabric obtaining after not processing according to step (2) containing the described fabric of collagen, then through step (3) method, process the residue dry weight obtaining, the g of unit.