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Method for pretreatment of blood sample containing aristolochic acid and lactam ingredients thereof

A technology of sample pretreatment and lactam, which is applied in the field of plasma or serum sample pretreatment and extraction treatment with ethyl acetate, which can solve the problems that the pretreatment method has not been reported yet.

Inactive Publication Date: 2005-11-02
PEKING UNIV
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Problems solved by technology

At present, there are few analytical reports in this area, and the pretreatment methods for human plasma or serum samples containing aristolochic acid and its lactam components that are suitable for high performance liquid chromatography (HPLC) determination have not yet been reported.

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  • Method for pretreatment of blood sample containing aristolochic acid and lactam ingredients thereof
  • Method for pretreatment of blood sample containing aristolochic acid and lactam ingredients thereof

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Embodiment Construction

[0071] Since the situation of aristolochic acid and its lactam components in the patient's plasma or serum is unknown, it cannot be guaranteed that aristolochic acid and its lactam components exist in human plasma or serum samples at the same time, so we use Guan Mutong water decoction ( Concentrations are respectively 6g / mL, 4g / mL) and fed SD male rats to obtain drug-containing plasma, then adopt the method of the present invention to carry out pretreatment and HPLC detection to the plasma of administration, simultaneously carry out the method of the present invention and protein precipitation method commonly used Compare. Implementing the concrete steps of the present invention comprises:

[0072] 1. Chromatographic conditions

[0073] Chromatographic column: ZORBAX SB-C18 (4.6mm×250mm, 5μm)

[0074] Mobile phase: A: B = 1% acetic acid / 30mmol / L triethylamine: acetonitrile (35% B → 100% B)

[0075] Column temperature: 15°C

[0076] Injection volume: 20μL

[0077] Detecti...

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Abstract

The present invention relates to aristolochic acid and processing method for its blood plasma or blood serum of lactam, which is characterized in that exacts to the sample aforementioned by acetic acid ethyl ester after acidification by hydrochloric acid. Adds hydrochloric acid to sample of blood plasma and serum (make sure the concentration is 3.0í‡10-5~ 1.0í‡10-4molíñmL-1)and eddy mix; lay for 30~60min at room temperature. Use acetic acid ethyl ester to exact for 1~2 times with 3~5ml per time, collect supernatant after eddy mixing. Use water-bath and nitrogen for exacted supernatant under 60 Deg to dry; add 150uL carbinol for leavings to dissolve; filter, take 20uL filtrate to check by HPLC. The blood plasma or blood serum aforementioned in the invention comprises human, rat, mouse, rabbit, etc. Rate of extraction and recovery for aristolochic acid and its lactam in sample of blood plasma or blood serum is more than 95%.

Description

technical field [0001] The present invention relates to the pretreatment method of blood plasma or blood serum biological sample, more specifically relate to the blood plasma containing aristolochic acid (aristocholic acid I, II) and its lactam components (aristocholactam I, II) Or the plasma or serum sample pretreatment method that uses ethyl acetate to extract after the serum sample is acidified. Background technique [0002] Aristolochic acids are the main active ingredients of Aristolochia plants, and they are also the direct cause of kidney damage caused by taking such Chinese herbal medicines and their prescriptions. Aristolochic lactams are derivatives of aristolochic acids, which are widely distributed in clinically used Chinese medicinal materials. It has been reported in the literature that aristolochic lactam I has a damaging effect on renal tubular epithelial cells (Li Biao et al., Injury effect of aristolactam I on renal tubular epithelial cells. Chinese Journa...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
Inventor 高小丽王璇蔡少青
Owner PEKING UNIV
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