Method for rapidly detecting anticoagulant activity of poecilobdella manillensis

Abstract

The invention provides a method for rapidly detecting anticoagulant activity of poecilobdella manillensis, which comprises the following steps: step 1, taking living poecilobdella manillensis, draining, weighing and recording; step 2, adding water which is 1.5 times of the weight of the poecilobdella manillensis according to the weighed weight; and step 3, carrying out homogenization treatment on the poecilobdella manillensis living body added with water by using high-speed stirring equipment. And 4, sucking 1g of poecilobdella manillensis slurry by using a dropper, adding 5ml of 0.9% sodium chloride solution, uniformly shaking, extracting for 30 minutes, filtering by using filter paper to obtain filtrate, and titrating. 5, according to the titration detection result, the obtained U value is substituted into a deduced formula y = 10.287 x-13.211, the value x is the U value obtained in the step 4, and the result obtained after calculation is the expected activity of the freeze-dried powder prepared from the poecilobdella manillensis raw material. According to the method, the activity of the sample slurry is directly detected by adding a certain proportion of water into the living body sample for homogenization, so that the process is simplified, and the time is saved.

Description

technical field [0001] The invention belongs to the technical field of content detection of pharmaceutical raw materials, in particular to a method for rapidly detecting the anticoagulant activity of leech. Background technique [0002] Poecilobdella manillensis (Poecilobdella manillensis), also known as Mani leech, commonly known as Phnom Penh leech, is an animal of the genus Poeciloba Block, stroke hemiplegia, fall injury, and the treatment of cerebral thrombosis, cerebral hemorrhage, hyperviscosity, hyperlipidemia, hypertension, pulmonary heart disease, etc., are commonly used characteristic ethnic Chinese medicinal materials. [0003] At present, leech is included in the "Yunnan Provincial Standard for Chinese Herbal Medicine" (YNZYC-0357-2013) and "Yunnan Provincial Standard for Chinese Herbal Medicine" (YPBZ-0199-2013) , There are not a few companies engaged in the production of Philippine leeches freeze-dried powder, and one of the important indicators to evaluate wh...

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Problems solved by technology

[0012] 1. The cost of laboratory freeze-drying equipment is expensive, generally between 50,000 and 200,000, and general institutions do not have the conditions

[0013] 2. The parameters and performance of the laboratory freeze-dryer and the large-scale production freeze-dryer are also different, so unless a lot of time is spent in the early stage to conduct methodological investigation and verification with large-scale production equipment, can the freeze-dried powder made by the laboratory be good? , its stability and reproducibility still have a risk of bias

[0014] 3. Not counting the time required for sending live samples and processing live samples into a form suitable for freeze-drying, only the complete freeze-drying process needs to be processed in freeze-drying equipment for more than 30 hours, and the preparation process for the entire test takes a lot of time

[0017] 1. The anticoagulant active substance of Hirudo phenanthae has biological activity. The temperature of boiling water will cause some effective substances to be inactivated in high temperature environment, and the freeze-drying process is to lose water through low temperature, so that the species that cannot tolerate high temperature can be dehydrated at low temperature. Effective dehydration, so the test sample prepared by high temperature cannot reflect the real activity index of the raw material after being processed into freeze-dried powder

[0018] 2. It takes at least 18-40 hours for drying after scalding, and the sample processing time is also very long

[0019] 3. At present, there is no research report on the anticoagulant activity of the same batch of raw materials of Hirudo phenatus, the two specifications of its medicinal materials and freeze-dried powder decoction pieces have a certain stable quantitative relationship, so the test results of the scalded and dried medicinal materials are not The activity prediction and evaluation of lyophilized powder has limited reference value

[0021] Finally, because it was purchased by a pharmaceutical factory, the anticoagulant activity of Hirudo philippines is not recognized by other detection methods other than titration, that is, the results of detection by other methods do not meet the standards, and the cost of detection that farmers need to pay for detection is relatively high

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