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透明质酸酶 来源于绵羊睾丸

  • Accession #: P12964
  • 生物活性: ≥300 IU/mg dry weight
  • 内毒素水平: <0.2 EU/IU
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货号 (SKU) 包装规格 是否现货 价格 数量
H755162-100mg
100mg 现货 Stock Image
H755162-500mg
500mg 现货 Stock Image
H755162-1g
1g 现货 Stock Image
H755162-5g
5g 期货 Stock Image

基本描述

产品名称 透明质酸酶 来源于绵羊睾丸
别名 醌还原酶 | 玻璃酸酶
英文别名 Hyaluronate 4-glycanohydrolase | Hyaluronoglucosaminidase | HYAL2 | oHyal2
规格或纯度 生物活性, ActiBioPure™, 天然, 通过内毒素测试, 高性能, EnzymoPure™, ≥300 IU/mg dry weight 
生化机理 透明质酸酶是一种糖苷内切酶,可分解细胞外基质的成分透明质酸(HA)。它用于改善药物扩散,并逆转HA填料注射剂(filler injection)的作用。透明质酸酶可溶解皮下结节,并可修正过量的注射填料。它可以消除胞浆内精子注射过程中形成的卵冠丘复合体。透明质酸酶可抑制多种物质溢出以后导致的组织损伤。它用于减少水肿治疗玻璃体积血。
产品介绍

从哺乳动物睾丸(例如羊睾丸)中提取的酶,能够水解透明质酸类型的粘多糖。它可能包含一个合适的稳定器。效力:每毫克(干燥物质)透明质酸酶活性最低300 IU。

生产

产生透明质酸酶的动物必须符合适合人类食用的动物的健康要求。 

特征

外观:白色或黄白色,无定形粉末。

溶解度:可溶于水,几乎不溶于丙酮和无水乙醇。

鉴别

一种含有相当于100 IU的溶液透明质酸酶在1 mL的9g/L的氯化钠溶液中,将10 g/L的透明质酸钠BRP溶液在20℃下解聚,粘度显著降低。将透明质酸酶在100℃加热30 min会破坏这种作用。 

检查

溶液外观:溶液应澄清. 在水中溶解0.10 g,用相同的溶剂稀释至10 mL。

pH:4.5 to 7.5.

将30 mg溶解在无二氧化碳水中,用相同的溶剂稀释至10 mL。

干燥失重:最大浓度为5.0%,在60℃下,在不超过670Pa的压力下干燥2小时,测定的浓度为0.500克。

细菌内毒素:<0.2 EU/IU.

分析

透明质酸酶的活性是通过比较它水解透明质酸钠BRP的速率与使用国际标准获得的速率,或使用国际单位校准的参考制剂,使用斜率比测定。

基质溶液:在25 mL锥形瓶中加入0.10 g透明质酸钠BRP,在4℃下缓慢加入20.0 mL水。加入速率必须足够慢,以使基质颗粒膨胀(约5 min)。保持在4摄氏度,搅拌至少12小时。4℃储存,4天内使用。

对于测试溶液和参考溶液,准备溶液,并在0℃至4C下进行稀释。

检测溶液:将适当量的物质溶解在透明质酸酶稀释剂中,以获得每毫升含有0.6±0.3 IU的透明质酸酶的溶液。

对照溶液:溶解适当数量的 透明质酸酶BRP在透明质酸酶稀释液中,得到每毫升含0.6 IU透明质酸酶的溶液。

在反应容器中,混合1.50 mL的pH为6.4 的磷酸盐缓冲溶液和1.0 mL的底物溶液,并在37±0.1℃下平衡。在时间t:=0(第一计时计)添加0.50 mL含有E毫克检测酶的测试溶液,混合,使用合适的粘度计保持在37±0.1℃并记录流出时间t,使用第二计时计(0.1秒间隔),多次在约20 min(读第一计时计)。以下粘度计:微粘度计(DIN51 562,第2部分),毛细管型MII,粘度计常数约0.1 mm²/s²。

使用0.50 mL含透明质酸酶BRP的参考溶液重复该步骤。根据以下表达式计算粘度比:

K = 粘度计常数为mm²/s²(在粘度计上表示);

t2 =溶液的流出时间(以秒为单位);

0.6915 = 缓冲溶液在37℃时mm²/s的运动粘度。

由于酶反应在流出时间测量期间继续,实际反应时间等于t,+t,/2,流出时间(t,/2)的一半,被加到测量开始的时间t中。图(In r)-l作为反应时间的函数,(t.+ t/2),以秒为单位。得到了一个线性关系。计算待检测物质的坡度(b)和参比制剂(b.)。 根据表达式计算每毫克国际单位的比活性:

A = 透明质酸酶BRP的比活性,国际单位每毫克。

至少执行三次完整的程序,并计算出待检查物质的平均活性。

贮存

密闭容器,温度 2℃ 到 8℃。如果该物质是无菌的,则该容器也是无菌的和防被篡改的。


Enzymes extracted from mammalian testes (e.g., sheep testes) can hydrolyze mucopolysaccharides of the hyaluronic acid type. They may contain a suitable stabilizer. Potency: Minimum 300 IU of hyaluronidase activity per milligram (of dry substance).

Production

Animals used for producing hyaluronidase must meet the health requirements for animals intended for human consumption.

Characteristics

1.Appearance: White or yellowish-white, amorphous powder.

2.Solubility: Soluble in water, almost insoluble in acetone and absolute ethanol.

Identification

A solution containing 100 IU of hyaluronidase in 1 mL of 9 g/L sodium chloride solution depolymerizes a 10 g/L sodium hyaluronate BRP solution at 20°C, resulting in a significant decrease in viscosity. Heating the hyaluronidase at 100°C for 30 minutes destroys this effect.

Tests

1.Appearance of Solution: The solution should be clear. Dissolve 0.10 g in water and dilute to 10 mL with the same solvent.

2.pH: 4.5 to 7.5. Dissolve 30 mg in carbon dioxide-free water and dilute to 10 mL with the same solvent.

3.Loss on Drying: Maximum 5.0%. Determine by drying 0.500 g at 60°C under a pressure not exceeding 670 Pa for 2 hours.

4.Bacterial Endotoxins: ≤ 0.2 EU/IU.

Assay

The activity of hyaluronidase is determined using a slope-ratio assay, by comparing the rate at which it hydrolyzes sodium hyaluronate BRP with the rate obtained using the International Standard or a reference preparation calibrated in International Units.

Substrate Solution

In a 25 mL conical flask, add 0.10 g of sodium hyaluronate BRP, then slowly add 20.0 mL of water at 4°C. The addition rate must be slow enough to allow the substrate particles to swell (approximately 5 minutes). Maintain at 4°C and stir for at least 12 hours. Store at 4°C and use within 4 days.
For both the test solution and the reference solution, prepare the solutions and perform dilutions at 0°C to 4°C.
1.Test Solution: Dissolve an appropriate amount of the substance in hyaluronidase diluent to obtain a solution containing 0.6 ± 0.3 IU of hyaluronidase per mL.

2.Reference Solution: Dissolve an appropriate amount of hyaluronidase BRP in hyaluronidase diluent to obtain a solution containing 0.6 IU of hyaluronidase per mL.
In a reaction vessel, mix 1.50 mL of phosphate buffer solution (pH 6.4) and 1.0 mL of the substrate solution, and equilibrate at 37 ± 0.1°C. At time t₀ = 0 (using the first timer), add 0.50 mL of the test solution containing E milligrams of the enzyme to be tested, mix well. Maintain the mixture at 37 ± 0.1°C using a suitable viscometer, record the flow time t using a second timer (with 0.1-second intervals), and perform multiple measurements over approximately 20 minutes (monitoring with the first timer). Use the following viscometer: microviscometer (DIN 51 562, Part 2), capillary type MII, with a viscometer constant of approximately 0.1 mm²/s².
Repeat the above procedure using 0.50 mL of the reference solution containing hyaluronidase BRP. Calculate the viscosity ratio using the following expression:

K = Viscometer constant (in mm²/s², indicated on the viscometer);

t₂ = Flow time of the solution (in seconds);

0.6915 = Kinematic viscosity of the buffer solution at 37°C (in mm²/s).
Since the enzymatic reaction continues during the flow time measurement, the actual reaction time is equal to t₀ + t/2 (i.e., half of the flow time (t/2) is added to the initial measurement time t₀). Plot (ln η)⁻¹ as a function of the reaction time (t₀ + t/2) (in seconds); a linear relationship should be obtained. Calculate the slope (b) of the substance to be tested and the slope (bᵣ) of the reference preparation. Determine the specific activity in International Units per milligram using the following expression:

A = Specific activity of hyaluronidase BRP (in International Units per milligram).
Perform at least three complete sets of the procedure and calculate the average activity of the substance to be tested.

Storage

Store in a tightly closed container at a temperature of 2°C to 8°C. If the substance is sterile, the container should also be sterile and tamper-proof.

生物活性 ≥300 IU/mg dry weight
内毒素水平 <0.2 EU/IU
种属 绵羊(Sheep)
Accession # P12964
来源 天然

储存与运输

物理形态 固体
浓度 ≥300 IU/mg dry weight 
复溶 Reconstitute in water.
储存温度 2-8°C储存
运输条件 冰袋运输
稳定性与储存 长期储存2-8℃(2年);收货后建议分装。
CAS编号和信息 37326-33-3
酶学委员会编号 3.2.1.35
单位定义 透明质酸酶的活性是通过比较它水解透明质酸钠BRP的速率与使用国际标准获得的速率,或使用国际单位校准的参考制剂,使用斜率比测定。
分子类型

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批号(Lot Number) 证书类型 货号
ZJ25F0928023 分析证书 H755162
ZJ25F0928022 分析证书 H755162
ZJ25F0928021 分析证书 H755162

技术文档和文章

溶液计算器