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Research Article | Open Access

Development and validation of a rapid gas chromatography for simultaneous quantification of major fatty acids in royal jelly

Xinyu Yu, Wenting Wei, Yuqi Wu, Yuanyuan Lu, Fuliang Hu(), Shanshan Li()
College of Animal Sciences, Zhejiang University, Hangzhou 310058, China
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Abstract

Fatty acids are distinctive components of royal jelly (RJ), serving as a crucial indicator for assessing the quality of RJ. This study aimed to establish a rapid and simultaneous quantification method for various fatty acids in RJ utilizing gas chromatography. An optimized two-step extraction process, incorporating ethanol and diethyl ether followed by derivatization with N,O-bis-(trimethylsilyl) trifluoroacetamide, was developed to enhance sensitivity and precision in detecting fatty acids. Validation of the methodology revealed excellent linearity (R2 > 0.999), precision (relative standard deviation (RSD) < 1%), repeatability (RSD < 1%), and recoveries (94.4%–104%). Furthermore, the limits of detection and quantification were found to be low. The results indicated that the method offered reliable and consistent quantification of major fatty acids, thereby improving quality control for RJ and facilitating its applications in food, pharmaceutical, and biochemical research.

Keywords

royal jellyfatty acidsrapid determinationgas chromatography

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Food Science of Animal Products
Volume 3 Issue 1,
March 2025
Article number: 9240099
DOI: 10.26599/FSAP.2025.9240099
Cite this article:
Yu X, Wei W, Wu Y, et al. Development and validation of a rapid gas chromatography for simultaneous quantification of major fatty acids in royal jelly. Food Science of Animal Products, 2025, 3(1): 9240099. https://doi.org/10.26599/FSAP.2025.9240099
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Determination of the linear range for major fatty acids in RJ.

Fatty acidLinear range (μg/mL)Regression equationCorrelation coefficient (R2)
10-HDAA13.4–134y = 0.883 1x + 0.045 90.999 5
3-HDAA4.9–49y = 0.667 8x + 0.008 20.999 5
SEA2.0–20y = 0.770 0x + 0.006 40.999 7
8-Hydroxycaprylic acid8.2–82y = 0.585 4x – 0.041 80.999 3
2-Decene-1,10-dicarboxylic acid8.0–80y = 0.746 4x – 0.009 30.999 8

LODs and LOQs for determination of fatty acids in RJ (µg/mL).

Fatty acidLODLOQ
10-HDAA0.7192.398
3-HDAA0.9443.147
SEA0.8072.690
8-Hydroxycaprylic acid1.0853.615
2-Decene-1,10-dicarboxylic acid0.6952.316

Precision for determination of fatty acids in RJ (n = 6).

ItemSample
123456
Sample quality (g)0.512 340.505 010.496 820.509 800.491 020.508 42
Internal standard quality (g)0.003 250.003 250.003 250.003 250.003 250.003 25
Peak area ratio of 3-HDAA and internal standard0.147 3850.144 6190.141 1190.144 3610.138 7340.146 530
Peak area ratio of 10-HDAA and internal standard1.041 0471.037 6821.019 0531.049 2640.973 9611.033 674
Peak area ratio of SEA and internal standard0.195 4010.191 5780.188 9560.193 0900.182 2390.191 797
Peak area ratio of 8-hydroxycaprylic acid and internal standard0.279 1690.277 7340.269 3200.278 3050.260 0920.274 578
Peak area ratio of 2-decene-1,10-dicarboxylic acid and internal standard0.223 8050.216 7500.217 8130.221 7740.2112460.223 101
3-HDAA content (%)0.0930.0930.0920.0920.0920.094
Mean ± RSD (%)0.093 ± 0.078
10-HDAA content (%)0.6600.6680.6670.6690.6450.661
Mean ± RSD (%)0.662 ± 0.902
SEA content (%)0.1240.1230.1240.1230.1210.123
Mean ± RSD (%)0.123 ± 0.119
8-Hydroxycaprylic acid content (%)0.1770.1790.1760.1770.1720.176
Mean ± RSD (%)0.176 ± 0.226
2-Decene-1,10-dicarboxylic acid content (%)0.1420.1390.1420.1410.1400.143
Mean ± RSD (%)0.141 ± 0.135

Repeatability for determination of fatty acids in RJ (n = 6).

Fatty acidIntra-day precision (%)Inter-day precision (%)
10-HDAA0.9081.308
3-HDAA0.1320.947
SEA0.1910.453
8-Hydroxycaprylic acid0.2760.338
2-Decene-1,10-dicarboxylic acid0.3380.480

Recovery for determination of fatty acids in RJ (%).

Fatty acidLow concentration recoveryMiddle concentration recoveryHigh concentration recoveryAverage spiked recovery
10-HDAA103.0105.0104.0104.0
3-HDAA94.894.593.994.4
SEA95.998.295.696.6
8-Hydroxycaprylic acid102.1102.4102.0101.7
2-Decene-1,10-dicarboxylic acid100.798.399.899.6