The Sheep Plasma Method for the Bioassay of Heparin Preparations

Swoap, Orlo F.; Kuizenga, Marvin H.

doi:10.1002/jps.3030381012

Cited by 9 publications

(3 citation statements)

References 6 publications

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“…The centrifuge tube was inverted three times and placed in a 37 • C water bath. The tube was gently inverted every 30 s, and the clotting time was recorded until the blood stopped flowing [49].…”

Section: Assay Of Anticoagulant Activity In Vitromentioning

confidence: 99%

Antithrombotic Activity of Heparinoid G2 and Its Derivatives from the Clam Coelomactra antiquata

et al. 2022

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Clam heparinoid G2 (60.25 kDa) and its depolymerized derivatives DG1 (24.48 kDa) and DG2 (6.75 kDa) prepared from Coelomactra antiquata have been documented to have excellent fibrinolytic and anticoagulant activity. In this study, to further explore the antithrombotic activity of G2, DG1 and DG2, azure A, sheep plasma, and clot lytic rate assays were used to determine their anticoagulant and thrombolytic activity in vitro. The results indicated that the anticoagulant titer of G2 was approximately 70% that of heparin and the thrombolytic activity of DG2 was greater than G2, DG1, and heparin activities. Moreover, in a carrageenan-induced venous thrombosis model, oral administration of G2 and DG1 each at 20 mg/kg and 40 mg/kg for 7 days significantly reduced blacktail thrombus formation, increased tissue-type plasminogen activator, fibrin degradation products, and D-dimer levels, decreased von Willebrand factor and thromboxane B2 levels, and restored phylum and genus abundance changes of intestinal bacteria. DG2 had no antithrombotic effect. At 20 mg/kg, G2, DG1, and heparin had comparable antithrombotic activities, and DG1 at 40 mg/kg had more muscular antithrombotic activity than G2. Thus, DG1 could be an antithrombotic oral agent owing to its more robust antithrombotic activity and lower molecular weight.

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Section: Assay Of Anticoagulant Activity In Vitromentioning

confidence: 99%

Antithrombotic Activity of Heparinoid G2 and Its Derivatives from the Clam Coelomactra antiquata

et al. 2022

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“…The coagulation was triggered by addition of 0.2 mL 1% CaCl2 to 0.5 mL citrated plasma. The absorbance of the solution was read at 470 nm for every 10 s until the absorbance reaches 0.8 (coagulation) (Swoap and Kuzienga, 1949).…”

Section: Anticoagulantmentioning

confidence: 99%

Anti-coagulant Activity of Isolated and Partially Characterized Proteoglycans and Glycosaminoglycans from African Night Crawler (Eudrilus eugeniae Kinberg)

Bercansil

Belgado

2015

KIMIKA

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Proteoglycans and glycosaminoglycans were isolated from African night crawler (Eudrilus eugeniae Kinberg) and partially characterized proteoglycans (3.04 % of lyophilized worm) were liberated from the defatted and depurinated worm samples by dissociative method using 4M urea in acetate buffer. Glycosaminoglycans (12.47% of proteoglycan extract) were extracted using enzymatic hydrolysis of the proteoglycan extract with papain. Gel filtration chromatography using Sepharose CL-4B was used to purify and estimate the molecular weights of the proteoglycan and glycosaminoglycan fractions. Three proteoglycan fractions PGF1, PGF2 and PGF3 with estimated molecular weigths 860 kDa, 181 kDa and 3 kDa, respectively were identified as monitored by the Bradford and modified carbazole assay. Two glycosaminoglycan fractions -GF1 (MW = 860 kDa) and GF2 (MW=140 kDa) were identified using the modified carbazole assay. Infrared spectroscopy of the GF1 and GF2 showed the possible identities of the fractions. GF1 may be a hyaluronic acid and GF2 is possibly chondroitin. Anti-coagulant assay for the extracts and fractions revealed that the glycosaminoglycan isolate has anti-coagulant activity but not the GF1 and GF2 fractions individually.

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“…Previously developed methods for quantitative determination of unfractionated heparin include various enzymatic bioassays (16)(17)(18)(19) and colorimetric methods Fig. 1.…”

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confidence: 99%

Development and validation of a simple and sensitive size-exclusion chromatography method for quantitative determination of heparin in pharmaceuticals

et al. 2015

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Heparin is widely used as an anticoagulant for the treatment and prevention of various thrombotic diseases. However, due to its high anionic charge, heterogeneity in size distribution and high polarity, its analysis is very challenging. In this paper, a novel method based on size-exclusion chromatography (SEC) for quantitative determination of intact heparin in pharmaceuticals is presented. Analyses were performed on a BioSep-SEC-S 2000 column with Larginine solution at pH 6.5 as mobile phase and UV detection at 210 nm. The proposed method was found to be selective, linear (R 2 > 0.997) over the concentration range of 3.1 to 1222 µg mL -1 , with a limit of detection of 1.0 µg mL -1 . Intraday and inter-day precision were below 5.1 % and inaccuracy expressed as bias did not exceed 6.5 %. The reported method is simple, selective, sensitive, and requires no laborious sample preparation, which makes it appropriate for routine quantitative analysis of heparin in pharmaceuticals.

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The Sheep Plasma Method for the Bioassay of Heparin Preparations

Cited by 9 publications

References 6 publications

Antithrombotic Activity of Heparinoid G2 and Its Derivatives from the Clam Coelomactra antiquata

Antithrombotic Activity of Heparinoid G2 and Its Derivatives from the Clam Coelomactra antiquata

Anti-coagulant Activity of Isolated and Partially Characterized Proteoglycans and Glycosaminoglycans from African Night Crawler (Eudrilus eugeniae Kinberg)

Development and validation of a simple and sensitive size-exclusion chromatography method for quantitative determination of heparin in pharmaceuticals

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