Method for detecting activity of poecilobdella manillensis medicinal material

Abstract

The invention provides an improved poecilobdella manillensis medicinal material activity detection method, which comprises the following steps: taking 0.5 g of poecilobdella manillensis medicinal material, precisely weighing, precisely adding 15 ml of 0.9% sodium chloride solution, fully stirring, leaching for 30 minutes, shaking and filtering in real time, precisely measuring 100 microliters of subsequent filtrate, three times, respectively putting into three test tubes, respectively marking the test tubes as A, B and C, and respectively marking the test tubes as B, C and D; the method comprises the following steps: respectively adding 200 [mu] l of a tris (hydroxymethyl) aminomethane hydrochloric acid buffer solution containing 0.5% of fibrinogen, uniformly shaking, placing 3 parts of tubes to be tested in a water bath, carrying out warm soaking at 37 + / -0.5 DEG C for 5 min, and dropwise adding a thrombin solution containing 40 units per 1 ml. According to the method, the anticoagulant activity of the poecilobdella manillensis medicinal material is rapidly and accurately detected by increasing the dosage multiples of the parallel sample and the thrombin solution within the capacity range of a laboratory technician, and a set of relatively complete operation judgment process is designed.

Description

technical field [0001] The invention belongs to the technical field of detection of traditional Chinese medicinal materials, and in particular relates to a method for detecting the activity of medicinal 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 Mass, 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, the Yunnan provincial standard for the determination of anticoagulant activity is formulated with reference to the thrombin titration method reported by Markwarde in 1970. The principle is that the binding ratio of the effective active substance of leech to thrombin is 1:1, and one unit of activity is equal to one unit of neut...

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Problems solved by technology

[0018] 2. Every titration needs to be shaken to make it uniform. Frequent shaking will cause some fibrin to form a coagulated shape that is difficult to identify with the naked eye and be shaken away, so that the fibrinogen will become thinner, and the test results will be enlarged and even caused. The sample is difficult to solidify and cannot reach the end point

[0021] However, titrating 3 coagulations cannot accurately determine whether it is qualified, because the standard stipulates that it is 60U, then titrating 3 coagulations shows that the sample activity is less than 72U and greater than 48U, then this result is judged as unqualified for 48-60U or qualified for 60-72U Controversial and unclear standards;

[0022] Only when the titration is 4 times or more solidified, that is, the sample is greater than 96U, can it be judged as qualified, so there is a large uncertainty in the judgment results corresponding to the operation method in the standard

[0023] 4. The reproducibility of the experimental results is poor

Due to the large span of the two titration activities, it is often different from the previous titration times in the later re-examination, resulting in a large relative deviation of the results

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