Assay method of metabolite of bergenin in animal body
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A technology for petracenin and metabolites, which is applied in the field of determination of metabolites of petracenin in vivo, can solve the problems of limited content determination, imperfect metabolism research, incomplete understanding and mastery of drugs, etc., and achieve strong specificity, The effect of high sensitivity
Inactive Publication Date: 2013-06-05
SHANDONG UNIV
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However, most of them are limited to the content determination of the original drug in natural plants or tablets; limited to the detection technology and the low concentration in the body, the study of its pharmacokinetics in vivo has always been difficult at home and abroad. In recent years, there have been very few reports in the literature. The in vivo determination method of the prototype drug after drug administration; and the sensitive and accurate method for determining its metabolites in vivo after administration has not been reported at home and abroad. comprehensive
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[0027] Preparation of test solution:
[0028] The standard product of petracenin was provided by Shanghai Yusen New Drug Development Co., Ltd., the raw material drug of petracenin was provided by Shaanxi Senfu High-Tech Industrial Co., Ltd., acetonitrile was purchased from J.T.BAKER Company in the United States, and experimental water was purchased from Hangzhou Wahaha Co., Ltd., 10% hydrated Chloral was purchased from Qilu Hospital of Shandong University. Eighteen Wistar rats (180-220 g) were purchased from the Animal Center of Shandong University, and fasted for 12 hours before the experiment.
[0029] Eighteen rats were randomly divided into 3 groups, and were given 250 mg / kg petracenin crude drug suspension by intragastric administration.
[0030] Rats in the first group were fed in metabolic cages, and urine and feces were collected 12 hours before and 12 hours after administration.
[0031] The second group was used to collect bile. After being anesthetized with 10% ch...
Embodiment 1
[0035] Determination method of petracenin metabolites in animals:
[0036] (1) Take 300 μL of administered plasma, urine, bile, and unadministered blank, add 2 mL of acetonitrile respectively, vortex and mix for 2 minutes, centrifuge at 10800 rpm for 5 minutes, take the supernatant, dry it with nitrogen in a water bath at 40°C, and make 100 μL Reconstitute with acetonitrile, and take 10 μL for injection.
[0037] Weigh 500 mg of feces, add 4 mL of acetonitrile, vortex for 3 min, and then sonicate the suspension for 1 h, centrifuge at 5000 rpm for 10 min, and separate to obtain the supernatant (I); add 4 mL of acetonitrile to the remaining lower layer, and repeat the above steps to obtain the supernatant (II). The supernatants (I) and (II) were mixed, dried in a water bath with nitrogen at 40°C, reconstituted in 200 μL of acetonitrile, and 10 μL was taken for injection.
[0038] (2) Inject 10 μL of each test solution (including blank sample and drug administration sample) prep...
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Abstract
The invention relates to an assay method of metabolite of bergenin in an animal body. The method includes the following steps: adding acetonitrile into dosed blood plasma, dosed urine, dosed bile and dosed excrement sample to be assayed and non-dosed blank blood plasma, non-dosed urine, non-dosed bile and non-dosed excrement, mixing and spinning, centrifuging, clear-taking, drying and redissolving the samples to obtain liquid to be assayed, analyzing the liquid to be assayed through a high performance liquid chromatograph/ quadrupole rod-time of flight/ tandem mass spectrum (HPLC/Q-TOF/MS) method to obtain a total ions chromatogram of the liquid to be assayed, conducting contrast identification to the total ions chromatogram of a blank sample and the total ions chromatogram of a dosed sample, extracting ions and secondary fragments from the total ions chromatogram of the blank sample and the total ions chromatogram of the dosed sample, analyzing the ions and the secondary fragments, and obtaining that the metabolite of the bergenin in the animal body is a glucuronic acid combined product, a sulfuric combined product, hydrolysate and a methyl combined product. According to the assay method of the metabolite of the bergenin in the animal body, a high-sensitivity, strong-specificity, convenient and feasible HPLC/ Q-TOF MS/ MS method is first established and the metabolite of the bergenin in the animal body is successfully assayed through the HPLC/ Q-TOF MS/ MS method.
Description
technical field [0001] The invention relates to a method for determining metabolites, in particular to a method for determining metabolites of petracenin in organisms. Background technique [0002] Bergenin, the molecular formula is C 14 h 16 o 9 , the molecular weight is 328.27, and the structural formula is as follows: [0003] [0004] Also known as rock cabbage lactone, low tea, saxifrage, rock cabbage, bergamot, ghost lamp, Bergenit, Vakrint, AmlisieacidB. , Cuscutin, Pehaphorin, white loose needle crystal or crystalline powder, discoloration when exposed to light and heat, soluble in methanol, less soluble in water or ethanol. "Chinese Pharmacopoeia" is included as an antitussive and expectorant drug for the treatment of chronic bronchitis. At present, the preparations of petracenin include unilateral and compound preparations. The single prescription is an ordinary tablet, and chlorpheniramine maleate is added to the compound preparation. The main dosage form...
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