CN104904706A - Formaldehyde-free animal specimen fixed liquid and preparation method thereof - Google Patents
Formaldehyde-free animal specimen fixed liquid and preparation method thereof Download PDFInfo
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Abstract
本发明公开了一种无甲醛动物标本固定液及其制备方法，包括如下成分：杀菌防霉剂，蛋白变性剂，多元醇保湿剂和水，其中若杀菌防霉剂为固态，则其质量浓度为5-20g/L，若杀菌防霉剂为液态，则其体积浓度为1-2％；蛋白变性剂的体积浓度为5％-40％；多元醇保湿剂的体积浓度为2％-10％；余量为水。本发明所公开的动物标本固定液配方不含甲醛，无挥发性，对人体无害；将新鲜动物器官用生理盐水冲洗后，保存在本固定液中，一年后器官无腐败，无霉斑，色泽、形状无明显变化，固定液颜色无明显变化，保存效果良好。The invention discloses a formaldehyde-free animal specimen fixative and a preparation method thereof. 5-20g/L, if the bactericidal antifungal agent is liquid, its volume concentration is 1-2%; the volume concentration of protein denaturant is 5%-40%; the volume concentration of polyol moisturizing agent is 2%-10% %; the balance is water. The animal specimen fixative formula disclosed by the present invention does not contain formaldehyde, is non-volatile, and is harmless to the human body; after fresh animal organs are washed with physiological saline and stored in the fixative, the organs will not be corrupted or moldy after one year , there is no obvious change in color and shape, no obvious change in the color of the fixative, and the preservation effect is good.
Description
技术领域technical field
本发明涉及动物标本固定液，具体的说涉及一种无甲醛动物标本固定液及其制备方法。The invention relates to an animal specimen fixative, in particular to a formaldehyde-free animal specimen fixative and a preparation method thereof.
背景技术Background technique
动物组织在动物死亡后受到细胞内酶(溶酶体酶)的作用而引起各组织器官变软或液化，这种现象叫自溶。各器官中脑、脊髓、胰腺和肾上腺等自溶发生较快，甚至细胞结构完全消失；肌肉、肝、脾、肾等自溶表现为质地变软，结构模糊，最后细胞溶解消失；胃、肠等器官也会因消化液的作用而溶解。另外动物体内的细菌和环境中的细菌在动物死亡后会大量繁殖而引起组织中蛋白质的腐败分解。因此标本的保存液要具有杀菌、防霉和蛋白质变性的作用。Animal tissues are affected by intracellular enzymes (lysosomal enzymes) after animal death, causing various tissues and organs to soften or liquefy. This phenomenon is called autolysis. The autolysis of the midbrain, spinal cord, pancreas, and adrenal gland in various organs occurred relatively quickly, and even the cell structure completely disappeared; Other organs will also dissolve due to the action of digestive juices. In addition, the bacteria in the animal body and the environment will multiply after the death of the animal and cause the decomposition of protein in the tissue. Therefore, the preservation solution of the specimen should have the functions of sterilization, anti-mildew and protein denaturation.
目前，动物标本的保存普遍使用的是甲醛水溶液(福尔马林)。福尔马林的渗透能力强，能与蛋白质中氨基结合，使蛋白质凝固，防腐效果好。但是甲醛的挥发性极强，具有强烈的刺激性气味，能够对人的多种器官造成损害。2004年6月，世界卫生组织国际癌症研究机构(IARC)的153号公告将甲醛由2类致癌物质上升为1类致癌物质。由于甲醛的强挥发性，会刺激接触人员的眼结膜和呼吸道黏膜，导致眼刺痛、流眼泪、头晕，皮肤接触会造成过敏反应、皮肤炎和湿疹，长期接触还可能导致癌症，严重危害接触人员的身体健康；在福尔马林中长期保存的标本还会变硬，褪色。另外由于甲醛的挥发性，在标本保存的过程中，需要定期补充或更换。At present, formaldehyde solution (formalin) is commonly used in the preservation of animal specimens. Formalin has a strong penetrating ability and can combine with amino groups in proteins to coagulate proteins and have a good antiseptic effect. However, formaldehyde is extremely volatile, has a strong pungent smell, and can cause damage to various human organs. In June 2004, the No. 153 Bulletin of the International Agency for Research on Cancer (IARC) of the World Health Organization raised formaldehyde from a Class 2 carcinogen to a Class 1 carcinogen. Due to the strong volatility of formaldehyde, it will irritate the conjunctiva and respiratory mucosa of the contact personnel, causing eye stinging, tearing, and dizziness. Skin contact can cause allergic reactions, dermatitis and eczema. Long-term exposure may also cause cancer, which seriously endangers exposure to The health of the personnel; specimens preserved in formalin for a long time will harden and fade. In addition, due to the volatility of formaldehyde, it needs to be replenished or replaced regularly during the preservation of specimens.
乙醇(酒精)是医学上常用的消毒剂。用于固定时浓度以80-95％为好，具有蛋白凝固、硬化、脱水等作用。能够避免组织器官的自溶和微生物引起的器官腐败，但其渗透力较弱，用高浓度酒精固定时，会使组织变硬，收缩较大。Ethanol (alcohol) is a commonly used disinfectant in medicine. When used for fixation, the concentration is preferably 80-95%, and it has the functions of protein coagulation, hardening, and dehydration. It can avoid autolysis of tissues and organs and organ corruption caused by microorganisms, but its penetration is weak. When fixed with high-concentration alcohol, the tissue will harden and shrink greatly.
专利CN 200910003987.1公开了一种含双乙酸盐、硝酸盐、多元醇的尸体保存液。专利CN 1233391 A公开了一种含有钠盐或钾盐防腐剂、多羟基类保色剂和水组成的动物标本保存液，分别以双乙酸盐和氯化钠、苯甲酸钠、氯化钾、亚硝酸钾等作为防腐剂，以乙二醇、丙三醇等多元醇作为保色剂。这两种保存液均缺少蛋白变性剂，不能很好的保存大体标本和内脏标本。专利CN 102175496 B公开了无甲醛组织标本固定液，主要包括乙醇71％-79％、2-4％的冰乙酸、4-6％的多元醇、4-6％的表面活性剂。此固定液的主要成分是乙醇，其渗透力较差，对大的器官固定效果较差。Patent CN 200910003987.1 discloses a corpse preservation solution containing diacetate, nitrate and polyol. Patent CN 1233391 A discloses a kind of animal specimen preservation solution containing sodium salt or potassium salt preservative, polyhydroxy color retaining agent and water. Potassium nitrite and the like are used as preservatives, and polyalcohols such as ethylene glycol and glycerin are used as color retention agents. Both of these two preservation solutions lack protein denaturants and cannot preserve gross specimens and visceral specimens well. Patent CN 102175496 B discloses a formaldehyde-free tissue specimen fixative, which mainly includes 71%-79% ethanol, 2-4% glacial acetic acid, 4-6% polyol, and 4-6% surfactant. The main component of this fixative is ethanol, which has poor penetration and poor fixation effect on large organs.
发明内容Contents of the invention
本发明所要解决的技术问题，就是提供一种不含甲醛和挥发性试剂的动物标本固定液及其制备方法。The technical problem to be solved by the present invention is to provide a formaldehyde-free and volatile reagent-free animal specimen fixative and a preparation method thereof.
为了解决上述技术问题，本发明采用如下技术方案：In order to solve the above technical problems, the present invention adopts the following technical solutions:
一种无甲醛动物标本固定液，其改进之处在于，包括如下成分：杀菌防霉剂，蛋白变性剂，多元醇保湿剂和水，其中若杀菌防霉剂为固态，则其质量浓度为5-20g/L，若杀菌防霉剂为液态，则其体积浓度为1-2％；蛋白变性剂的体积浓度为5％-40％；多元醇保湿剂的体积浓度为2％-10％；余量为水。A formaldehyde-free animal specimen fixative, which is improved in that it includes the following components: a bactericidal and antifungal agent, a protein denaturant, a polyol moisturizer and water, wherein if the bactericidal and antifungal agent is solid, its mass concentration is 5 -20g/L, if the bactericidal antifungal agent is liquid, its volume concentration is 1-2%; the volume concentration of protein denaturant is 5%-40%; the volume concentration of polyol moisturizer is 2%-10%; The balance is water.
进一步的，所述的杀菌防霉剂为壳聚糖、季铵盐类表面活性剂、山梨酸钾、Na2EDTA中的一种或几种。Further, the bactericidal antifungal agent is one or more of chitosan, quaternary ammonium salt surfactant, potassium sorbate, and Na 2 EDTA.
进一步的，所述的壳聚糖为壳聚糖乳酸盐，所述的季铵盐类表面活性剂为十二烷基二甲基苄基溴化铵、十二烷基三甲基氯化铵、二癸基二甲基溴化铵中的一种或几种。Further, the chitosan is chitosan lactate, and the quaternary ammonium salt surfactant is dodecyl dimethyl benzyl ammonium bromide, dodecyl trimethyl chloride One or more of ammonium and didecyldimethylammonium bromide.
进一步的，所述的蛋白变性剂为乙醇、乙酸、丙酮中的一种或几种，其中乙醇的体积浓度为5％-40％、乙酸的体积浓度为0-2％、丙酮的体积浓度为0-2％。Further, the protein denaturant is one or more of ethanol, acetic acid, and acetone, wherein the volume concentration of ethanol is 5%-40%, the volume concentration of acetic acid is 0-2%, and the volume concentration of acetone is 0-2%.
进一步的，所述的多元醇保湿剂为乙二醇、二乙二醇、丙二醇、丙三醇中的一种或几种。Further, the polyhydric alcohol moisturizing agent is one or more of ethylene glycol, diethylene glycol, propylene glycol, and glycerol.
一种无甲醛动物标本固定液的制备方法，其改进之处在于，包括如下步骤：A kind of preparation method of formaldehyde-free animal specimen fixative, its improvement is, comprises the following steps:
(1)称取壳聚糖乳酸盐5-20g或山梨酸钾5-20g，溶解于水中，加入1-3g的Na2EDTA，搅拌均匀；(1) Weigh 5-20g of chitosan lactate or 5-20g of potassium sorbate, dissolve in water, add 1-3g of Na 2 EDTA, and stir evenly;
(2)分别加入1-2％体积浓度的季铵盐表面活性剂、5％-40％体积浓度的95％乙醇、0-2％体积浓度的乙酸、0-2％体积浓度的丙酮、2％-10％体积浓度的二乙二醇等试剂，搅拌均匀；(2) add respectively the acetic acid of the quaternary ammonium salt surfactant of 1-2% volume concentration, the 95% ethanol of 5%-40% volume concentration, the acetic acid of 0-2% volume concentration, the acetone of 0-2% volume concentration, 2 %-10% volume concentration of diethylene glycol and other reagents, stir evenly;
(3)加水定容至1000ml。(3) Add water to make up to 1000ml.
本发明的有益效果是：The beneficial effects of the present invention are:
(1)本发明所公开的动物标本固定液配方不含甲醛，无挥发性的试剂。将新鲜动物器官用生理盐水冲洗后，保存在本固定液中，一年后器官无腐败，无霉斑，色泽、形状无明显变化，固定液颜色无明显变化，保存效果良好。(1) The animal specimen fixative formulation disclosed in the present invention does not contain formaldehyde and has no volatile reagents. Fresh animal organs are washed with physiological saline and stored in this fixative solution. After one year, the organs have no corruption, no mildew spots, no obvious changes in color and shape, and no obvious changes in the color of the fixative solution. The preservation effect is good.
(2)本发明所公开的动物标本固定液可以用于动物解剖学器官的保存和动物组织学组织切片制作过程中的固定，其不含甲醛，所以无挥发性气味，对人体无害，学生也不会因为刺激性而不愿意靠前，教学效果良好。(2) The animal specimen fixative disclosed by the present invention can be used for the preservation of animal anatomical organs and the fixing in the process of making animal histological tissue sections. It does not contain formaldehyde, so it has no volatile odor and is harmless to the human body. They will not be reluctant to move forward because of stimulation, and the teaching effect is good.
具体实施方式Detailed ways
实施例1：Example 1:
将壳聚糖5g、十二烷基三甲基氯化铵10ml、95％乙醇100ml、乙酸20ml、二乙二醇10ml加水至1000ml，即得到无甲醛标本固定液。Add water to 5 g of chitosan, 10 ml of dodecyltrimethylammonium chloride, 100 ml of 95% ethanol, 20 ml of acetic acid, and 10 ml of diethylene glycol to 1000 ml to obtain a formaldehyde-free specimen fixative.
实施例2：Example 2:
将壳聚糖5g、十二烷基三甲基氯化铵50ml、95％乙醇100ml、乙酸20ml、二乙二醇10ml、丙三醇10ml加水至1000ml，即得到无甲醛标本固定液。Add 5 g of chitosan, 50 ml of dodecyltrimethylammonium chloride, 100 ml of 95% ethanol, 20 ml of acetic acid, 10 ml of diethylene glycol, and 10 ml of glycerin to 1000 ml to obtain a formaldehyde-free specimen fixative.
实施例3：Example 3:
将壳聚糖5g、二癸基二甲基溴化铵25ml，95％乙醇100ml、三氯乙酸(5％)20ml、丙三醇10ml加水至1000ml，即得到无甲醛标本固定液。Add 5 g of chitosan, 25 ml of didecyldimethylammonium bromide, 100 ml of 95% ethanol, 20 ml of trichloroacetic acid (5%), and 10 ml of glycerin to 1000 ml to obtain a formaldehyde-free specimen fixative.
实施例4：Example 4:
将壳聚糖5g、二癸基二甲基溴化铵25ml，95％乙醇100ml、乙酸20ml、丙三醇10ml加水至1000ml，即得到无甲醛标本固定液。Add 5g of chitosan, 25ml of didecyldimethylammonium bromide, 100ml of 95% ethanol, 20ml of acetic acid, and 10ml of glycerin to 1000ml of water to obtain a formaldehyde-free specimen fixative.
实施例5：Example 5:
将壳聚糖5g、二癸基二甲基溴化铵25ml，95％乙醇200ml、乙酸20ml、丙三醇10ml加水至1000ml，即得到无甲醛标本固定液。Add 5g of chitosan, 25ml of didecyldimethylammonium bromide, 200ml of 95% ethanol, 20ml of acetic acid, and 10ml of glycerol to 1000ml of water to obtain a formaldehyde-free specimen fixative.
实施例6：Embodiment 6:
将山梨酸钾10g、十二烷基三甲基氯化铵50ml、Na2EDTA 10g、95％乙醇100ml、丙三醇10ml加水至1000ml，即得到无甲醛标本固定液。Add 10 g of potassium sorbate, 50 ml of dodecyltrimethylammonium chloride, 10 g of Na 2 EDTA, 100 ml of 95% ethanol, and 10 ml of glycerol to 1000 ml to obtain a formaldehyde-free specimen fixative.
实施例7：Embodiment 7:
将山梨酸钾10g、十二烷基二甲基苄基溴化铵50ml、Na2EDTA 10g、95％乙醇100ml、丙三醇10ml加水至1000ml，即得到无甲醛标本固定液。Add 10 g of potassium sorbate, 50 ml of dodecyl dimethyl benzyl ammonium bromide, 10 g of Na 2 EDTA, 100 ml of 95% ethanol, and 10 ml of glycerol to 1000 ml to obtain a formaldehyde-free specimen fixative.
保存效果：Save the effect:
采用实施例1的无甲醛标本固定液保存兔子的肝脏组织。2013年6月16日保存，一年后观察保存效果。将兔子新鲜肝脏组织用生理盐水冲洗后，保存在本固定液中，一年后肝脏无腐败，无霉斑，色泽、形状无明显变化，固定液颜色无明显变化，保存效果良好。而70％酒精保存的广口瓶中溶液颜色变黄。The liver tissue of the rabbit was preserved using the formaldehyde-free specimen fixative of Example 1. It was preserved on June 16, 2013, and the preservation effect was observed one year later. Fresh rabbit liver tissue was washed with normal saline and stored in this fixative solution. One year later, the liver had no corruption, no mildew spots, no obvious changes in color and shape, no obvious change in the color of the fixative solution, and the preservation effect was good. And the color of the solution in the jar preserved by 70% alcohol turns yellow.
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