制备方法

方法一、酶解-树脂法酶解 取100kg新鲜肠黏膜(总固体5%-7%),加苯酚200ml (0.2%),如气温低时可不加。在搅拌下加入绞碎胰0.5-1kg(0.5%-1%),用 300-400g/L(30%-40%)NaOH溶液调节pH8.5-9,升温至40-45℃,保温2-3h。pH8,再加5kg粗食盐(5%)升温至90℃左右,用6mol/L HCl调节pH 6.5左右,停止搅拌,保温20min,以布袋过滤,得酶解滤液。肠黏膜[胰浆,NaOH]→[pH8.5-9, 40-45℃, 保温2-3h]肠黏膜液[NaCl, 6mol/L HCl]→[pH8; 90℃, pH6.5]滤液吸附、洗涤、洗脱 取上述滤液冷至50℃以下,用6mol/L NaOH调节pH7,加入5kgD-254强碱性阴离子交换树脂,搅拌5h左右,交换完毕,弃去液体,用自来水漂洗树脂至水清,用与树脂等体积的2mol/L NaCl,搅拌洗涤15min,弃去洗涤液,再加2倍树脂体积的1.2mol/L NaCl洗涤2次。用约树脂体积一半的5mol/L NaCl洗脱1h,收集洗脱液,然后用约树脂体积的1/3的3mol/L NaCl洗脱2次,合并洗脱液。滤液[D-254树脂, NaOH]→[pH7, 5h]吸附物[NaCl]→[2mol/L→1.2mol/L, →5mol/L, →3mol/L]洗脱液沉淀、脱水、干燥 将洗脱液用滤纸浆助滤,过滤1次,澄清滤液以体积计加入用活性炭处理过的0.9倍的95%乙醇,冷处沉淀8-12h。虹吸上清液弃去,按100kg黏膜加300ml的比例向沉淀中补加蒸馏水,再加入4倍量95%乙醇,冷处放置6h,收集沉淀,用无水乙醇脱水1次,丙酮脱水2次,置P2O5干燥器中真空干燥,得肝素钠粗品。洗脱液 → 沉淀物[无水乙醇;丙酮]→[P2O5]肝素钠粗品脱色、沉淀、干燥 用20g/L(2%)氯化钠溶液配成100 g/L(10%)的肝素钠溶液,以40g/L(4%)高锰酸钾溶液进行氧化脱色,每亿单位估计加入0.65mol高锰酸钾。先将药液调至pH8,预热80℃,在搅拌下加入高锰酸钾,保温2.5h,以滑石粉作助滤剂,过滤,收集滤液。将滤液调节pH6.4,用0.9倍的95%乙醇置于冷处理沉淀6h以上,收集沉淀,溶于10 g/L(1%)氯化钠溶液中,用量按肝素钠粗品质量收率60%-70%计,配成50 g/L(5%)肝素钠溶液,再用4倍量的95%乙醇冷处放置6h以上,收集沉淀物,用无水乙醇、丙酮脱水,乙醚洗涤1次,置P2O5干燥器中干燥,得肝素钠精品。肠黏膜换算成总固体7%计,收率20000U/kg,最高效价141.6U/mg。肝素钠粗品[20g/L NaCl]→肝素钠溶液[KMnO4]→[pH8, 80℃, 2.5h]滤液 →[pH6.4, 6h]沉淀物[无水乙醇;丙酮;乙醚]→肝素钠精品注射剂的制备 取肝素钠成品溶于一定量注射用水中,封口,灭菌,即得。肝素钠精品[注射用水]→注射剂方法二、盐解-树脂法提取 取新鲜猪肠黏膜投入反应锅中,按30g/L(3%)加入氯化钠,NaOH调节pH9,逐步升温至50-55℃,保温2h,继续升温至95℃,维持10min,冷却50℃以下,用30目双层纱布过滤,收集滤液。猪肠粘膜[NaCl; NaOH]→[pH9; 50-55℃]滤液吸附、洗涤、洗脱、沉淀 取冷却50℃以下的滤液,加入714型(强碱性)Cl-型树脂(新树脂用量为2%),搅拌8h后静置过夜。次日虹吸除去上层液,收集树脂,用水冲洗至洗液澄清,滤干,用2倍量1.4mol/L氯化钠搅拌2h,滤干;再用1倍量1.4mol/L氯化钠搅拌2h,滤干;继续改用2倍树脂量的3mol/L氯化钠搅拌洗脱8h,滤干;再用1倍量3mol/L氯化钠搅拌洗脱2h,滤干,合并滤液,加入等量95%乙醇沉淀,过夜,虹吸除去上清液,收集沉淀。丙酮脱水干燥,得粗品。滤液[714型树脂]→吸附物[NaCl]→[1.4mol/L, 3mol/L]洗脱液[95%乙醇,丙酮]→粗品溶解、脱色、沉淀、干燥 粗品溶于15倍10 g/L(1%)氯化钠溶液中,加入6mol/L盐酸调pH1.5左右,过滤至清,随即用5mol/L氢氧化钠调节pH11,按3%加入过氧化氢(浓度为30%)25℃放置。开始不断调整pH至11,第2天再按1%加入过氧化氢,调整pH11,继续放置,共48h。过滤,用6mol/L盐酸调节pH6.5,加入等量的95%乙醇沉淀。过夜。次日虹吸除去上清液,沉淀用丙酮洗涤脱水,干燥,即得肝素钠精品。总收率20000U/kg肠黏膜(按肠黏膜的固体量为5%-7%计算)。粗品[1%NaCl, 6mol/L HCL]→[pH1.5]滤液[NaOH, H2O2]→[pH11, 25℃, 48h]滤液[6mol/L HCl, 95%乙醇]→[pH6.5, 24h]沉淀[丙酮]→肝素钠精品方法三、CTAB*提取法(*十六烷基三甲基溴化胺为长链季铵盐)提取、络合 按猪肠黏膜液(V):Na2SO4(m):硫酸铝(m):CTAB(V)=1:0.15:0.04:0.001的配料比投料。取新鲜猪小肠黏膜投入反应罐中,搅拌下加入硫酸钠,溶解后,用碱液调节pH11-11.5,升温50℃,保温搅拌2h,再加硫酸铝,溶解后,用氢氧化钠调节pH7.5-8,升温至95℃,保温10min,趁热过滤,待滤液冷却至60℃以下时,缓缓加入CTAB和硅藻土,搅拌吸附1h,静置过夜,滤饼用40℃热蒸馏水洗至无色,抽干即得络合物。猪肠黏膜液[Na2SO4,碱液, Al2(SO4)3]→[pH11-11.5, 50℃, 2h; pH7.5-8, 95℃, 10min]提取液[CTAB, 硅藻土]→[60℃以下,1h]滤饼[蒸馏水]→ 络合物解离、沉淀 取络合物溶于适量氯化钠溶液中,以冰醋酸调pH4,升温60℃,保温30min,静置过

用途简介

该品是制抗凝药肝素剂的原料。

海关代码：

3001901000

WGK_Germany：

2
德国有关水污染物质的分类清单

安全说明：

24/25

RTECS号：

MI0850000

CHEMICAL IDENTIFICATION

RTECS NUMBER :

MI0700000

CHEMICAL NAME :

Heparin

CAS REGISTRY NUMBER :

9005-49-6

LAST UPDATED :

199706

HEALTH HAZARD DATA

ACUTE TOXICITY DATA

TYPE OF TEST :

TDLo - Lowest published toxic dose

ROUTE OF EXPOSURE :

Subcutaneous

SPECIES OBSERVED :

Human - man

DOSE/DURATION :

1857 units/kg/13D-I

TOXIC EFFECTS :

Cardiac - other changes Nutritional and Gross Metabolic - changes in potassium

TYPE OF TEST :

TDLo - Lowest published toxic dose

ROUTE OF EXPOSURE :

Subcutaneous

SPECIES OBSERVED :

Human - woman

DOSE/DURATION :

1000 units/kg/5D-I

TOXIC EFFECTS :

Nutritional and Gross Metabolic - changes in potassium

TYPE OF TEST :

TDLo - Lowest published toxic dose

ROUTE OF EXPOSURE :

Intravenous

SPECIES OBSERVED :

Human - man

DOSE/DURATION :

3536 units/kg/13D-I

TOXIC EFFECTS :

Nutritional and Gross Metabolic - changes in potassium

TYPE OF TEST :

LD50 - Lethal dose, 50 percent kill

ROUTE OF EXPOSURE :

Oral

SPECIES OBSERVED :

Rodent - rat

DOSE/DURATION :

1950 mg/kg

TOXIC EFFECTS :

Details of toxic effects not reported other than lethal dose value

TYPE OF TEST :

LDLo - Lowest published lethal dose

ROUTE OF EXPOSURE :

Intraperitoneal

SPECIES OBSERVED :

Rodent - rat

DOSE/DURATION :

420 mg/kg

TOXIC EFFECTS :

Details of toxic effects not reported other than lethal dose value

TYPE OF TEST :

LD50 - Lethal dose, 50 percent kill

ROUTE OF EXPOSURE :

Intraperitoneal

SPECIES OBSERVED :

Rodent - mouse

DOSE/DURATION :

1900 mg/kg

TOXIC EFFECTS :

Behavioral - altered sleep time (including change in righting reflex) Behavioral - somnolence (general depressed activity) Behavioral - muscle weakness

TYPE OF TEST :

LD50 - Lethal dose, 50 percent kill

ROUTE OF EXPOSURE :

Subcutaneous

SPECIES OBSERVED :

Rodent - mouse

DOSE/DURATION :

500 mg/kg

TOXIC EFFECTS :

Behavioral - somnolence (general depressed activity) Behavioral - muscle weakness

TYPE OF TEST :

LD50 - Lethal dose, 50 percent kill

ROUTE OF EXPOSURE :

Intravenous

SPECIES OBSERVED :

Rodent - mouse

DOSE/DURATION :

500 mg/kg

TOXIC EFFECTS :

Behavioral - somnolence (general depressed activity)

TYPE OF TEST :

TDLo - Lowest published toxic dose

ROUTE OF EXPOSURE :

Intravenous

DOSE :

2857 units/kg

SEX/DURATION :

male 5 day(s) pre-mating

TOXIC EFFECTS :

Reproductive - Paternal Effects - other effects on male

MUTATION DATA

TYPE OF TEST :

Unscheduled DNA synthesis

TEST SYSTEM :

Rodent - guinea pig Bone marrow

DOSE/DURATION :

5 mg/L

REFERENCE :

FOBLAN Folia Biologica (Prague). (Academic Press Inc. Ltd., 24-28 Oval Rd., London NW1 7DX, UK) V.1- 1955- Volume(issue)/page/year: 16,196,1970 *** REVIEWS *** TOXICOLOGY REVIEW AEMBAP Advances in Experimental Medicine and Biology. (Plenum Pub. Corp., 233 Spring St., New York, NY 10013) V.1- 1967- Volume(issue)/page/year: 52,375,1975 TOXICOLOGY REVIEW JPMSAE Journal of Pharmaceutical Sciences. (American Pharmaceutical Assoc., 2215 Constitution Ave., NW, Washington, DC 20037) V.50- 1961- Volume(issue)/page/year: 62,517,1973 *** NIOSH STANDARDS DEVELOPMENT AND SURVEILLANCE DATA *** NIOSH OCCUPATIONAL EXPOSURE SURVEY DATA : NOHS - National Occupational Hazard Survey (1974) NOHS Hazard Code - 84488 No. of Facilities: 374 (estimated) No. of Industries: 3 No. of Occupations: 8 No. of Employees: 4525 (estimated) NOES - National Occupational Exposure Survey (1983) NOES Hazard Code - 84488 No. of Facilities: 430 (estimated) No. of Industries: 4 No. of Occupations: 14 No. of Employees: 27273 (estimated) No. of Female Employees: 23149 (estimated)