柱前衍生-液相色谱-串联质谱法测定红曲中软毛青霉酸含量

伍勋; 施思; 靳祖珑; 沈国芳; 刘宇文

doi:10.13748/j.cnki.issn1007-7693.20242744

柱前衍生-液相色谱-串联质谱法测定红曲中软毛青霉酸含量

Determination of Puberulic Acid Content in Monascus by Pre-column Derivatization-liquid Chromatography with Tandem Mass Spectrometry

摘要

摘要:
目的利用了三甲基硅烷化重氮甲烷对软毛青霉酸进行甲基化衍生，建立红曲中软毛青霉酸高灵敏度检测方法。
方法采用利用超高效液相色谱-三重四极杆质谱对软毛青霉酸2种异构体的甲基化衍生产物进行检测。色谱柱为Phenomenex C₈(2.1 mm×100 mm，2.7 μm)，流动相以10 mmol·L⁻¹甲酸铵为流动相A，50%甲醇水(含有10 mmol·L⁻¹甲酸铵)为流动相B，梯度洗脱(0~0.5 min，90%→10%A；0.5~4 min，10%A)，流速0.3 mL·min⁻¹，柱温40 ℃，进样量10 μL；离子化模式为ESI⁺，进行多反应监测，选择软毛青霉酸甲基化衍生物m/z 255.0→225.0为定量离子，m/z 255.0→194.0作为定性离子。
结果对9批红曲样品进行检测，均未检出软毛青霉酸，合格率100%。
结论所建立的方法经验证，专属性强，可用于红曲中软毛青霉酸成分的检测。

Abstract:
OBJECTIVE To establish a highly sensitive detection method for puberulic acid in monascus by using trimethylsilylated diazomethane for methylation derivation of puberulic acid.
METHODS High performance liquid chromatography triple quadrupole mass spectrometry was used to detect the methylated derivatives of two isomers of puberulic acid. The chromatographic column was Phenomenex C₈(2.1 mm×100 mm, 2.7 μm), and the mobile phase A was 10 mmol·L⁻¹ ammonium formate, mobilephase B was 50% methanol in water(containing 10 mmol·L⁻¹ ammonium formate), gradient elution(0–0.5 min, 90%→10%A; 0.5–4 min, 10%A), flow rate was 0.3 mL·min⁻¹, column temperature was 40 ℃, injection volume was 10 μL. The ionization mode was ESI⁺, and multiple reaction monitoring was carried out. The methylated derivative of puberulic acid m/z 255.0→225.0 was selected as the quantitative ion, and the mass to charge ratio m/z 255.0→194.0 was selected as the qualitative ion.
RESULTS Nine batches of monascus samples were tested, and no puberulic acid was detected, with a pass rate of 100%.
CONCLUSION The established method has been validated and has strong specificity, and can be used for the detection of puberulic acid in monascus.

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