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Method for separating and testing campesterol, stigmasterol and beta-sitosterol by using high performance liquid chromatography method

A high-performance liquid chromatography and campesterol technology, which is applied in the field of separation of phytosterols, achieves the effects of shortening retention time, high sample recovery and high precision

Pending Publication Date: 2018-03-09
JIANGNAN UNIV
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  • Application Information

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Problems solved by technology

[0003] Since campesterol and stigmasterol only have structural differences at the C-22 and C-24 positions, the polar neutralization effect between the two will cause campesterol and stigmasterol to co-elute during chromatographic separation At present, most of the researches at home and abroad focus on the simultaneous separation and determination of one or two sterols, but no simultaneous separation and determination of the three sterols

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  • Method for separating and testing campesterol, stigmasterol and beta-sitosterol by using high performance liquid chromatography method
  • Method for separating and testing campesterol, stigmasterol and beta-sitosterol by using high performance liquid chromatography method
  • Method for separating and testing campesterol, stigmasterol and beta-sitosterol by using high performance liquid chromatography method

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Embodiment 1

[0024] Preparation of standard solution:

[0025] Mixed standard stock solutions containing campesterol, stigmasterol and β-sitosterol were prepared with methanol, dried by nitrogen blowing apparatus, and then prepared with acetonitrile containing campesterol 2.5, 5, 10, 20, 40, 50 μg / mL, soybean Sterols and β-sitosterol 5, 10, 20, 40, 80, 100 μg / mL standard series solutions.

[0026] The high performance liquid chromatography conditions are:

[0027] Column: Water C18 column (4.6mm×250mm, 5μm)

[0028] Detector: Waters2489 UV-Vis detector

[0029] Detection wavelength: 208nm

[0030] Column temperature: 35°C

[0031] Mobile phase: acetonitrile:water=98:2(v / v)

[0032] Mobile phase speed: 1.0mL / min

[0033] Injection volume is 20μL

[0034] Determination of recovery and precision:

[0035] Homogeneous soy sterol samples were taken and divided into 3 groups, and three mixed standard series solutions with different concentration levels were added (camesterol: 10, 20, 4...

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Abstract

The invention discloses a method for separating and testing campesterol, stigmasterol and beta-sitosterol by using a high performance liquid chromatography method. The method comprises the following steps: preparing a standard solution, namely preparing a mixed standard standby solution with campesterol, stigmasterol and beta-sitosterol from solar solvents, treating the mixed standard standby solution, and further respectively preparing mixed standard standby solutions with campesterol, stigmasterol and beta-sitosterol from solar solvents. By adopting the method, three sterol monomers are subjected to base line separation, the retention time is greatly shortened, an optimal spectrum condition of 1.0mL / minute is finally confirmed, the detection wavelength is 208nm, the column temperature is35 DEG C, the method is relatively high in both precision and sample feeding recycling rate, and the detection requirement that contents of three sterol monomers are accurately and quantitatively detected is met.

Description

technical field [0001] The invention belongs to the technical field of separation of phytosterols, in particular to a method for separating and measuring campesterol, stigmasterol and β-sitosterol by high-performance liquid chromatography. Background technique [0002] Phytosterol is a natural active compound with cyclopentane perhydrophenanthrene as the basic skeleton, a hydroxyl group is attached to the C-3 position, and a steroid compound with 8 to 10 carbon atoms forming a side chain is attached to the C-17 position. It belongs to the unsaponifiable matter in lipids, among which campesterol (Campesterol), stigmasterol (Stigmasterol) and β-sitosterol (Sitosterol) are the sterol monomers with the highest content in phytosterols, and their structures are also shown in figure 1 . Phytosterols have physiological functions such as reducing cholesterol and cardiovascular disease risk, anti-cancer, anti-inflammation, and anti-oxidation. They are receiving more and more attentio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/34
CPCG01N30/06G01N30/34G01N2030/065
Inventor 常明冯文欢王兴国金青哲刘睿杰吴正章张鹏
Owner JIANGNAN UNIV
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