High performance liquid chromatography method for measuring content of hypocrellin A

A high-performance liquid chromatography, rhododendron A technology, which is applied in measurement devices, color/spectral property measurement, instruments, etc., can solve the problems of low silanol activity, unsatisfactory degree of symmetry, unsuitable detection wavelength, etc.

Active Publication Date: 2010-05-19
YUNNAN PHYTOPHARML
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0013] (1) The selected mobile phase methanol-water=83:17, or acetonitrile-KH 2 PO 4 (0.02mmol / L)=70:30, all can not well separate HA in the sample, the HA peak in the chromatogram is inclined, and the symmetry does not reach " Chinese Pharmacopoeia " 2005 edition one appendix VI D for HPLC analysis tailing factor at 0.95 Requirements between ~1.05
[0014] (2) The fluorescence detector adopted in the above method ① is an uncommonly used instrument, and its applicable area is narrow.
Method ① The patented technology only uses Zorbax Extend C18 column with double end-capping to ensure very low silanol activity, which has a narrow application range, and the price of this type of chromatographic column is generally high, which increases the analysis cost
[0015] (3) The above-mentioned methods ② and ③ adopt the detection of ultraviolet absorption at 300nm, while the ultraviolet-visible absorption curve of HA (as attached figure 2 Shown) is not an absorption peak at 300nm, and is not suitable for use as a detection wavelength for quantitative analysis

Method used

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  • High performance liquid chromatography method for measuring content of hypocrellin A
  • High performance liquid chromatography method for measuring content of hypocrellin A
  • High performance liquid chromatography method for measuring content of hypocrellin A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1: the content determination of hypocrellin A in bamboo red fungus medical material

[0067] Take the bamboo red fungus medicinal material, crush it, pass through a 60-mesh sieve, weigh 1g precisely, put it in a 100ml round bottom flask, add 50ml of acetone accurately, weigh, extract in a water bath for 1 hour, let it cool, and make up for the lost weight with acetone , filtered to obtain the test solution. Take an appropriate amount of hypocrellin A, accurately weigh it, and dissolve it in anhydrous methanol to prepare a reference solution containing 30 μg hypocrellin A per 1 ml.

[0068] According to the method determined by the research of the present invention, the reference substance solution and the test solution are accurately drawn, injected into a liquid chromatograph, and measured to obtain final product. Wherein, the chromatographic column uses carbon octadecyl bonded silica gel as a filler, the detection wavelength is 462nm, and the mobile phase ...

Embodiment 2

[0071] Embodiment 2: the content determination of hypocrellin A in bamboo red fungus medical material

[0072] Take bamboo red fungus medicinal material (produced in Ninglang County, batch number 20071201), pulverize, pass through a 60-mesh sieve, accurately weigh 1g, put it in a 100ml round bottom flask, add 50ml of acetone accurately, weigh, extract in a water bath for 1 hour under reflux, and put Cool, make up the lost weight with acetone, and filter to obtain the test solution. Take an appropriate amount of hypocrellin A, accurately weigh it, and dissolve it in anhydrous methanol to prepare a reference solution containing 30 μg hypocrellin A per 1 ml.

[0073] Using a chromatographic column with carbon octadecyl bonded silica gel as a filler, a detection wavelength of 462nm, and using the mixed solvents in Table 4 as mobile phases, the content analysis of hypocrellin A was carried out. Precisely draw the reference substance solution and the test solution, inject it into t...

Embodiment 3

[0076] Embodiment 3: Determination of content of hypocrellin A in Zhuhuang medical material

[0077] The method is the same as that in Example 1, and the medicinal material measured is Bamboo Yellow. The results of determination of the content of hypocrellin A in Zhuhuang medicinal materials from different origins are shown in Table 5.

[0078] Table 5 Determination results of hypocrellin A content in Zhuhuang medicinal materials from different origins

[0079]

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Abstract

The invention discloses a high performance liquid chromatography (HPLA) method for measuring content of hypocrellin A. The conditions of the liquid chromatography are as follows: octadecylsilane chemically bonded silica is used as a filler for a chromatographic column, and a mobile phase is a mixed solvent consisting of methanol and dilute phosphoric acid (or dilute formic acid, dilute acetic acid and dilute sulfuric acid), wherein the volume ratio of the methanol and the dilute phosphoric acid (or dilute formic acid, dilute acetic acid and dilute sulfuric acid) is 80:20-70:30, and the detection wavelength is 462 nm. The invention overcomes the defects of traditional analysis method and can be directly applied to the quantitative analysis of hypocrellin A in traditional Chinese medicine and preparation including the hypocrellin A. The method has the technical characteristics of good separation effect, precise measurement, sensitivity, strong specificity, fast measurement and the like. The absorption peak of the hypocrellin A in the medicine and preparation including the hypocrellin A can fully flow out the chromatographic column within 30 minutes, and the analysis work is finished.

Description

technical field [0001] The invention relates to the technical field of drug analysis, in particular to a high-performance liquid chromatography (HPLC) method for determining the content of hypocrellin A (Hypocrellin A). Background technique [0002] Photodynamic therapy (Photodynamic Therapy, PDT) is a new type of clinical treatment technology. Its development began in the 1970s for clinical diagnosis and treatment of tumors. For the treatment of port wine stains, microvascular diseases, macular degeneration, etc. Since the photosensitive drug Photofrin was first approved in Canada for photodynamic therapy of bladder cancer and esophageal cancer in 1993, photodynamic therapy has been approved in many countries, and many patients have benefited from photodynamic therapy. [0003] As a new type of therapy, PDT is to introduce a photosensitizer into the body and distribute it among tissues, then irradiate the target tissue with light that matches the absorption spectrum of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/33G01N30/02
Inventor 郑立雄张慧颖李俊饶高雄
Owner YUNNAN PHYTOPHARML
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