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Preparation of Neferine Sample

Application Introduction

Neferine is a bisbenzylisoquinoline alkaloid extracted from the green germ of nelumbo nuficera gaertn mature seed. Research shows it has the pharmacological effects, including vascular dilation, antihypertension,

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Introduction

Neferine is a bisbenzylisoquinoline alkaloid extracted from the green germ of nelumbo nuficera gaertn mature seed. Research shows it has the pharmacological effects, including vascular dilation, antihypertension, resistance to arrhythmia, resistance to platelet aggregation, resistance to thrombosis, resistance to oxidation, resistance to organophosphorus pesticide poisoning, resistance to cicatrization, chemosensitization, etc. This experiment establishes a preparative purification method to prepare neferine monomer with extract.

Table 1 Neferine Sample Information

Sample name

English name

Structural formula

Molecular formula

Molecular weight

CAS No.

Neferine

Neferine

C38H44N2O6

624.77

2292-16-2

Experimental

Instrument, reagents and materials

Cheetah HP100 high-pressure preparative instrument

Chromatographic solvent: acetonitrile; water: deionized water

HPLC columns: Innoval ODS-2 (7 µm, 100 Å, 50×250 mm)

Sample pretreatment

The sample is liquid in brownish black (concentration: 2 g/15 mL). Filter the sample with a 0.22 µm membrane, and then perform the sample loading directly.

Preparation conditions

Chromatographic columns: Innoval ODS-2 (7 µm, 100 Å, 50×250 mm);

Mobile phases: A: TFA aqueous solution (concentration: 0.05%), B: acetonitrile;

Detector: UV (wavelength: 254 nm)

Flow rate: 80 mL/min

Sample size: 5 mL

Gradient:

Table 2 Gradient Elution Conditions of Liquid Chromatography

Time/min

Flow rate/(mL/min)

A/%

B/%

0.0

80.0

90.0

10.0

30.0

80.0

55.0

45.0

Detection conditions

Chromatographic columns: Innoval ODS-2 (5 µm, 100 Å, 4.6×250 mm);

Mobile phases: A: TFA aqueous solution (concentration: 0.05%), B: acetonitrile

Detector: 254 nm

Flow rate: 1.0 mL/min

Sample size: 5 µL

Column temperature: 30 °C

Gradient:

Table 3 Gradient Elution Conditions of Liquid Chromatography

Time/min

Flow rate/(mL/min)

A/%

B/%

0.0

1.0

90.0

10.0

30.0

1.0

55.0

45.0

Results and Discussion

In this experiment, neferine sample is prepared with Innoval ODS-2 (7 µm, 100 Å, 50 × 250 mm) packing. As shown in Table 2 and Figure 1, the sample prepared shows a good peak shape after going through the gradient elution in the mobile phases including TFA aqueous solution (concentration: 0.05%) and acetonitrile. Besides, the purity of the product prepared is conforming to the requirement of customer.

Figure 1 Preparation Chromatogram

Figure 2 Detection Chromatogram of Fraction at 12.4min

Conclusion

In this experiment, neferine is prepared with Innoval ODS-2 (7 µm, 100 Å, 50 × 250 mm) chromatographic columns. Results show that Innoval ODS-2 packing has a good effect on this sample in separation, and the purity of product prepared is conforming to the requirement of customer.


  • Innoval ODS-2

    Innoval ODS-2 has excellent mechanical strength, which could be used under pressure up to 6000psi. Its low surface area, provide fast separation speed, also more tolerance ability for dirty sample. Innoval ODS-2 could provide good separation ability for hydrophilic compounds, compatible with 100% aqueous mobile phase.

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