4.2 检测方法4.2.1 效价（羊血浆法） 标准：大于等于150IU

4.2 检测方法
4.2.1 效价（羊血浆法）
标准：大于等于150IU∕mg。
4.2.1.1、 试剂：羊血浆（冰冻保存）、0.9%生理盐水、0.25%CaCl2（用生理盐水配置）、8%柠檬酸钠溶液（用生理盐水配置）、8IU∕mg肝素钠标准溶液。

4.2.1.2 判断8%柠檬酸钠溶液的加入量
取三只试管，编号1、2、3，各加入标品110ul，然后分别加入8%柠檬酸钠溶液10ul、20ul、30ul，再各加入0.25%CaCl2 0.8ml及过滤后的羊血浆1ml，每只试管盖好管盖，倒转3—4次混匀，使内壁湿润，垂直放入37℃水域5min。5min后取出各管，检查各管的凝固程度，取三支试管中凝固最好试管对应的微升数，然后在下面实验中加入对应微升数的8%柠檬酸钠溶液x ul。

4.2.2.1 待检样品溶液的配置：精确称取肝素钠固体待检样品M mg，溶解并用0.9%生理盐水定容至100ml，再取V ml定容至25ml即得待检样品溶液，冷藏保存。
估计M及V的计算公式为：样品估计效价×MV∕100=25×8
估算时V只尽量小于10且取整数，从而计算出M值。

4.2.2.2 检测方法：
将恒温水域调制37℃恒温，取出冰冻羊血浆，融化，并过滤，待用。取2组10只试管，分别加入标样110ul、120ul……200ul，然后每管加入8%柠檬酸钠溶液x ul、0.25%CaCl2 0.8ml 及过滤后的羊血浆1ml，另一组加入待检样品溶液110ul、120ul……200ul，同样每管加入8%柠檬酸钠溶液x ul 、0.25%CaCl2 0.8ml及过滤后的羊血浆1ml。并将每只试管盖好管盖，倒转3—4次混匀，使内壁湿润，垂直放入37℃水域1h。1h后取出各管，检查各管的凝固程度，并记录确定标样及待检样的1∕2凝固点及体积。

4.2.2.4 凝固程度标准：
①完全凝固：溶液完全凝固，倒转试管并猛敲一下时，凝块不从管壁脱落。
②半凝固：溶液完全凝固，但当猛敲一下时，凝块能从管壁脱落。
③未凝固：溶液无凝固或少量凝固现象。
若半凝固则取半凝固点微升数，若相邻两只试管一只未完全凝固，一只为未凝固，则取这两只试管凝固点微升数的中间值。
4.2.2.5 待检样品效价计算公式：
F=标品微升数*预估微升数/样品微升数
4.2.2.6 注意事项：
4.2.2.6.1 实验室操作环境必须控制在30℃以下。

4.2.3吸光度的检测（标准260nm：小于等于0.15；280nm：小于等于0.15）
原理：测量样品的吸光度，以判定样品中蛋白和核酸的浓度大小。
仪器：紫外分光光度计法、玻璃棒、万分之一天平、烧杯等。
溶液的配制：配置0.4%肝素溶液（g/ml)，在260nm、280nm测吸光度。

4.2.4 比旋度的检测（标准：比旋度+45°~+55°）
配置4%的肝素溶液测旋光度，根据下列公式计算出比旋度（α）：

式中： α- 测得的旋光度
C - 溶液的浓度（g/ml)
L - 溶液的长度

結果 (英文) 1:

4.2 potency of detection methods
4.2.1 (sheep plasma method)
standard: 150IU/territory mg or more.
4.2.1.1, reagents: plasma of sheep (frozen), normal saline, 0.9% 0.25%CaCl2 (configured with the saline solution), 8% sodium citrate solution (saline solution configuration), 8IU/territory mg sodium heparin standard solution.

4.2.1.2 judge 8% sodium citrate solution by addition
take three tubes, numbered 1, 2, 3, and joined 110ul, and then joined 10ul, 20ul 8% sodium citrate solution, 30ul, sheep plasma after joining the 0.25%CaCl2 filter 0.8ml and 1ml, each tube cover tube coverReverse 3-4 mix, moist inner wall, vertically in a 37 ° c water 5min. 5min remove the tubes, check the pipe freezing level, solidified the best three test tube test tube slightly, then joined in the experiment below the corresponding micro-litres x UL of 8% sodium citrate solution.

4.2.2.1 configuration of the test sample solution: precise preparation of solid test sample m mg sodium heparin, saline volume dissolved and 0.9% to 100ml, then take the v ml fixed volume to 25ml the sample solution in quarantine, refrigerated.
Estimates of m and v is calculated as: the sample estimated estimated potency xMV/territory 100=25x8
v to less than 10 and rounded when,To calculate m.

4.2.2.2 test method: modulation 37 ° c temperature
thermostatic water, take out the frozen sheep plasma melting and filtering, for later use. Take 2 set of 10 tubes, 110ul, 120ul were added to standard sample ... ... 200ul and then each tube add 8% sodium citrate solution x UL 1ml, 0.25%CaCl2 0.8ml and filtered sheep plasma, another group to join the test sample solution 110ul, 120ul ... ... 200ul, as each tube add 8% sodium citrate solution x UL 1ml, 0.25%CaCl2 0.8ml and filtered sheep plasma. Place each tube cover tube cover, reverse 3-4 mix, inner wall of moist,Vertically in a 37 ° c water 1h. 1H remove the hose, check all pipe freezing levels and records to determine standard freezing point and quarantine-like 1/2 and volume.

4.2.2.4 freezing level standard:
① fully solidified: solution completely solidified, inverted test tube and, bang, and clots from the wall falls off.
② half solidified: solution completely solidified, but when bang, and clots from the wall falls off.
③ fresh: solidification phenomena of no coagulation or a small amount of the solution.
Semi-solidified the number edged up taking half-freezing point, adjacent to two tubes is not completely solidified, one for fresh, then take both in vitro freezing point slightly middle value.
4.2.2.5 calculation formula of sample titer in quarantine:
F= edged * estimates rose slightly edged up/sample:
4.2.2.6 notes:
4.2.2.6.1 laboratory operations must be controlled at 30 ° c or less.

4.2.3 detection of absorbance (260nm: 0.15;280nm or less: less than or equal to 0.15)
principle: measure the absorbance of the sample in order to determine the concentrations of proteins and nucleic acids in a sample size.
Instruments: ultraviolet photometric methods, glass rods, one out of 10,000 scales, beakers, etc.
Solution preparation: configuration 0.4% heparin solution (g/ml), and measuring absorbance at 260nm and 280nm.

4.2.4 specific optical rotation of inspection (standard:Specific optical rotation of 45 °-55 °)
4% heparin solution configured log rotation, according to the following formula to calculate the specific optical rotation (α):

-: rotation measured by α-
c concentration-solution (g/ml)
l-length of the solution

結果 (英文) 3:

4.2 detection methods of
4.2.1 titer (sheep plasma method)
standard: greater than or equal to 150IU / mg.
4.2.1.1, reagent: sheep plasma (frozen), 0.9%, 0.25%CaCl2 physiological saline (saline configuration), 8% sodium citrate solution (saline configuration), 8IU / Mg heparin sodium standard solution.

4.2.1.2 8% sodium citrate solution added
three test tubes, number 1, 2, 3, added to the standard 110ul, then adding 8% sodium citrate solution 10ul, 20ul, 30ul, and then join the sheep plasma 1ml 0.25%CaCl2 0.8ml and filtered, each tube cover tube cover,Version 3 - 4 times in the inner wall of mixing, wetting, vertically in 37 ℃ water 5min. The tube was removed after 5min, check the solidification degree of each tube, slightly take number three tubes in solidification best corresponding, then add the corresponding L number below the experiment in 8% sodium citrate solution x ul. The

4.2.2.1 sample solution: weigh accurately heparin sodium solid sample M mg, dissolved with 0.9% saline volume to 100ml, then V ml sample solution is to 25ml, cold preservation.
estimate calculation formula of M and V: the sample estimate titer * MV / 100=25 × 8
when estimating V only to less than 10 and the integer,In order to calculate the value of M.

4.2.2.2 detection methods:
thermostatic water modulation constant temperature of 37 ℃, remove the frozen sheep plasma, melting, and filtering, stand-by. The 2 group of 10 rats in vitro, were added to the standard 110ul, 120ul...... 200ul, then each tube sheep plasma 1ml in 8% sodium citrate solution of X UL, 0.25%CaCl2 0.8ml and filtered, another group with the sample solution 110ul, 120ul...... 200ul, every pipe sheep plasma 1ml in 8% sodium citrate solution of X UL, 0.25%CaCl2 0.8ml and filtered. And each test tube cover tube cover, inverted 3 - 4 times mixing, the wall wetting,Vertically in 37 ℃ water 1h. The tube was removed after 1h, check the solidification degree of each tube, and record the standard sample and sample determined to be 1 / 2 freezing point and volume.

4.2.2.4 solidification degree standard:
1: solution completely solidified completely solidified, inverted test tube and banging, clot does not fall off from the wall.
2 semi solidification: solution completely solidified, but when banging, clots can fall off from the wall.
the uncured: solution without solidification or a small amount of solidification phenomenon.
if half solidification is semi solidification point up number, if the adjacent two in a not completely solidified, one is not solidified, the middle and the two in the solidification point of microliter number value.
4.2.The calculation formula of 2.5 sample:
F= titer standard increased number * prediction microliter number / sample increased number
4.2.2.6 note:
4.2.2.6.1 laboratory operating environment must be controlled at 30 ℃. Detection of

4.2.3 absorbance (standard 260nm: less than or equal to 0.15; 280nm: less than or equal to 0.15)
principle: the absorbance measurement sample size determination, the concentration of protein and nucleic acid in the sample.
instrument: UV spectrophotometry, glass rod, 1/10000 balance, the beaker. Preparation of
solution: 0.4% heparin solution (g/ml), in 260nm, 280nm measured absorbance. The detection of

4.2.4 specific rotation (standard:The specific rotation of 45 ° ~ 55 °)
heparin solution configuration 4% measuring rotation, according to the following formula to calculate the specific rotation (alpha):

type: alpha measuring concentration of polarimetry
C - solution (g/ml)
L - solution length.