超高效液相色谱法分析丝瓜酚类物质组分及其含量

建立超高效液相色谱快速测定丝瓜中酚类物质组分及其含量的方法,为丝瓜酶促褐变的后续研究提供技术支持。液相色谱柱为ACQUITY UPLC BEH C18(2.1 mm×100 mm,1.7μm),流动相为甲醇(A)/0.3%乙酸水溶液(B),柱温32℃,进样量2μL,流速为0.25 m L·min~(-1)。采用梯度洗脱,洗脱程序0~0.5 min,95%B;0.5~17.5 min,95%~67.5%B;17.5~18 min,67.5%~95%B。检测波长选取283 nm。14种酚类化合物标准品在18 min内完全分离,样品中检测出13种酚酸。线性范围在0.05~4mg·L-1,检出限(S/N=3)为0.004~0.049 mg·L~(-1),决定系数均大于0.999,精密度、重复性、稳定性相对标准偏差均小于5%。平均回收率在95.70%~106.89%,相对标准偏差在0.32%~4.71%。龙胆酸为丝瓜样品中主要酚酸,其次是绿原酸、多巴胺、L–酪氨酸,再次是焦性没食子酸、原儿茶酸、香草酸、儿茶酚、表儿茶酸、对羟基苯甲酸、丁香酸、4–甲基儿茶酚、对香豆酸。

Determination of Polyphenols in Luffa by Ultra Performance Liquid Chromatography

This study based on ultra performance liquid chromatograph（UPLC）for rapid determination method of polyphenols and its content in the sponge gourd to supports luffa enzymatic browning of follow-up studies to provide technical. The liquid-chromatographic separation was performed on Agilent ZORBAX Eclipse Plus C18（2.1 mm × 100 mm，1.7 μm-Micron）column kept at 32 ℃，using methanol/acetic acid/water mixture as the mobile phase with a flow rate of 0.25 mL · min^-1 through gradient elution：0–0.5 min，95% B；0.5–17.5 min，95%–67.5% B；17.5–18 min，67.5%–95% B. The detection wavelength was set at 283 nm. Results showed that 14 polyphenols standard was completely separated within 18 min where detecting thirteen poly samples. The determination coefficients were higher than 0.999 and linear range was from 0.05 to 4 mg · L^-1. The limits of quantification range from 0.004 to 0.049 mg · mL^-1. Recoveries achieved from the luffa ranged from 95.70% to 106.89%（RSDs，0.32%–4.71%）. The precision，reproducibility and stability were satisfactory with the relative standard deviations（RSDs）less than 5%. Gentisic acid as the main polyphenols，followed by chlorogenic acid，dopamine，L-tyrosine，finally the pyrobitumengallic acid，protocatechuic acid，vanillic acid，catechin，epicatechin，p-hydroxy benzoic acid and clove acid as well as the 4-methyl catechol，coumaric acid.