UPLC-UV-QTOF-MS/MS method of quantitative detection of triterpenoid saponins in Camellia sinensis (L.) O.Kuntze

A UPLC-UV-QTOF-MS, quantitative detection technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of high detection results, large sample requirements, long time consumption, etc., to achieve accurate quantitative results and sample requirements. Low volume and high sensitivity

Inactive Publication Date: 2019-05-17
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is only suitable for the detection of tea saponin samples with a content of 30%-70%, which requires a large amount of samples, takes a long time, and has large errors, so it is not suitable for the detection of a small amount of samples
Concentrated sulfuric acid (or perchloric acid)-vanillin colorimetric method is a common method for the determination of saponin, but it is easily interfered by other impurities in the sample, which makes the detection result higher; in addition, it cannot accurately quantify the saponin monomer compound

Method used

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  • UPLC-UV-QTOF-MS/MS method of quantitative detection of triterpenoid saponins in Camellia sinensis (L.) O.Kuntze
  • UPLC-UV-QTOF-MS/MS method of quantitative detection of triterpenoid saponins in Camellia sinensis (L.) O.Kuntze
  • UPLC-UV-QTOF-MS/MS method of quantitative detection of triterpenoid saponins in Camellia sinensis (L.) O.Kuntze

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Quantitative detection of saponin in tea tree seed extract

[0035] (1) Sample preparation:

[0036] The tea seeds are dehulled, the seeds are ground and passed through a 20-mesh sieve, 70% methanol is added in a ratio of 1:15, and the mixture is refluxed for 5 hours at 70° C., and then filtered while hot to obtain a crude extract. The crude alcohol extract is concentrated by rotary evaporation, lyophilized into a powder in a vacuum freeze dryer, and stored in a refrigerator below -40°C for later use. The sample is made into a 60% methanol solution (2mg / ml) and passed through a 0.22μm filter membrane to obtain the test solution.

[0037] (2) Sample detection:

[0038] AcquityTM ultra high performance liquid chromatograph (Waters, USA), Triple TOF 5600+ time-of-flight mass spectrometer, ACQUITY UPLC HSS T3 chromatographic column (1.8μm, 150mm×2.1mm i.d., Waters) chromatographic column was used.

[0039] Chromatographic conditions are: mobile phase A (0.1% formic acid / ...

Embodiment 2

[0046] Example 2: Quantitative detection of the total saponin fraction of tea tree seeds

[0047] (1) Sample preparation:

[0048] The tea seeds are dehulled, the seeds are crushed, passed through a 20-mesh sieve, 70% methanol is added in a ratio of 1:15, and extracted at 70°C for 5 hours under reflux, and then filtered while hot to obtain a crude extract. The crude alcohol extract is rotated Concentrated by evaporation (45-60°C). After concentration, the extract is extracted with n-hexane, ethyl acetate and n-butanol three times in sequence. The n-butanol phase is collected and concentrated under reduced pressure to obtain the extract.

[0049] Use D-101 macroporous resin for purification. After the sample is adsorbed, it is eluted with distilled water, 30% ethanol, 50% ethanol, and 70% ethanol successively. The elution volume of each elution solvent is 2 column volumes, the flow rate is 3ml / min, and finally the layer is washed with 95% ethanol. Analyze the column for next use. ...

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Abstract

The invention belongs to the technical field of natural product detection, and relates to a quantitative detection method of UPLC-PDA-MS/MS of triterpenoid saponins in Camellia sinensis (L.) O.Kuntze.The UPLC-UV-QTOF-MS/MS method of quantitative determination of the triterpenoid saponins in the Camellia sinensis (L.) O.Kuntze comprises the following steps that (1) samples to be detected are pretreated; (2) any one or more saponin compounds known in the Camellia sinensis (L.) O.Kuntze are taken, and a standard product solution is prepared; and (3) the structure of high performance liquid chromatography/ultraviolet/mass spectrometry combination equipment (UPLC-PDA-QTOF-MS/MS) of a test solution and a standardized product solution is identified by tandem mass spectrometry, and an external standard method is used for quantitative detection under a PDA detector. According to the UPLC-UV-QTF-MS/MS method of quantitative detection of the triterpenoid saponins in the Camellia sinensis (L.) O.Kuntze, quantitative results are more accurate; the sensitivity is high; and the sample demand amount is less.

Description

Technical field [0001] The invention belongs to the technical field of natural product detection, and specifically relates to a UPLC-PDA-MS / MS quantitative detection method for triterpenoid saponins in tea seed (Camellia sinensis (L.) O. Kuntze). Background technique [0002] Camellia sinensis (L.) O.Kuntze has a variety of biological activities and is considered to be another important resource of tea trees after tea. In particular, the triterpene saponin not only has good surface activity and decontamination ability, but also has biological pharmacology such as antibacterial and anti-inflammatory, anti-tumor, improving gastrointestinal damage caused by alcohol, inhibiting gastric emptying, and promoting the absorption of trace elements by plants. Features. At present, more than 50 kinds of triterpene saponins have been isolated from tea seed species in China, Japan, India, Sri Lanka and other countries, but they have similar structures and similar polarities, which brings grea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 李博屠幼英吴学进吴嘉璠刘亚文
Owner ZHEJIANG UNIV
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