Preparative HPLC

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Preparative HPLC

Preparative HPLC is a method that uses high pressure and large flow conveying system to separate sample in high resolution, large inner diameter and high load. It also has been the most popular, most effective and fastest-developing method in relevant technology in the chemical research of natural products for more than ten years. The products obtained by this method are far superior to those obtained by other traditional separation methods in terms of purity, recovery rate and separation efficiency. According to the preparation scale, there are 3 types, namely, semi-preparative HPLC, Gram-grade preparative UPLC and industrial preparative HPLC.

BOC Sciences has a wealth of experience in the preparative HPLC. We often use it for separation and purification of the active ingredient of natural products, such as alkaloid, flavonoid, terpenoid, steroid, and lactone.

Alkaloid

Alkaloids are a kind of nitrogen-containing organic compounds in vivo except for protein, peptide, amino acid and vitamin B. Alkaloids have complex structure and most of them have physiological activity, such as antitumor activity. For example, we use preparative HPLC to purify taxol and choose the column D956 resin as filler, the acetone-water (40:60~58:42) as the mobile phase. The purity can reach 99% and the recovery rate reaches more than 80%.

Flavonoids are composed of two benzene rings connected with each other by three carbons, which is widely distributed in various medicinal plants. At present, many flavonoid compounds with biological activity have been found out, and their effects mainly include cardiovascular protection, anti-tumor, anti-oxidation, antivirus, etc. For example, we use column chromatography to obtain the total flavonoids extract of Rob Twist, and combine with preparative HPLC for purification. The chromatographic condition includes reversed-phase column, gradient elution with the ethyl nitrile dissolved in 0.1% gradient elution and water, in which the ethyl nitrile increases from 5% to 50% in 180 mins. By optimizing the flow rate, sample intake and elution gradient, nine flavonoids can be separated from Rob Twist and the purity is more than 90%.

Terpenoids are natural compounds, which are connected by isoprene or isopentane in various ways, and have physiological activities such as eliminating phlegm, relieving cough and relieving pain. For example, we use HPLC and reversed-phase chromatographic column to separate nine high-purity sesquiterpenoids from matteuccia struthiopteris.

Steroids are a kind of natural compounds based on the cyclic-amyl hydrogenation of Phenanthrene as mother nucleus, which have important physiological and pharmacological functions in sustaining life, regulating sexual function, treating skin diseases and controlling fertility. For example, in order to study the chemical constituents of Ganoderma lucidum, we use semi-preparative chromatographic column to separate and purify it, and obtain three steroid compounds, such as ganoderic acid B9.

Lactones usually have the function of resisting allergy, resisting shock and regulating heart function. The preparative HPLC can be used to purify such compounds, usually by reversed-phase chromatography, which chooses methanol-aqueous solution or acetonitrile-water as a mobile phase, and often can get good separation effect, high purity products in a short time. For example, we use preparative HPLC to separate the nigakilactone in the brucea, the purity of them has reached more than 98%, and can be used as reference substances.

BOC Sciences can use preparative HPLC to separate and purify various types of compounds including alkaloid, flavonoid, terpenoid, steroid and lactone. We also use multiple techniques (NMR, LCMS, GCMS, X-ray) to analyze compounds after the chiral synthesis is completed. And our turnaround time is fast. Welcome to contact us for more information.

Reference

  1. Wu X, Deng Q, Xu ZY, Cen JR, Xu J, Application of Preparative High Performance Liquid Chromatography in Separation of Natural Product. Analytical Chemistry Chemometrics. 2017(1):113-122.
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