基于化学衍生和质谱技术的苹婆酸定量方法研究

doi:10.16431/j.cnki.1671-7236.2026.01.024

Abstract

Abstract:

Objective This study aimed to establish an accurate method for determining the sterculic acid content in cottonseed meal and feed to facilitate the rational utilization of cottonseed meal protein resources. Method By optimizing the reaction conditions of 2-pyridinecarboxylic hydrazide with sterculic acid， the extraction process of sterculic acid and mass spectrometry parameters， a liquid chromatography tandem mass spectrometry （LC-MS/MS） method for quantifying sterculic acid using external standards was successfully established. The performance of the method was evaluated by examining its linear range， limit of detection， limit of quantification， recovery， precision （intra-assay and inter-assay） and accuracy， along with application to analyze actual samples. Result After optimization， the optimal reaction mixture composition for the reaction of sterculic acid with 2-pyridinecarboxylic hydrazide was determined as 500 mmol/L EDC， 15 mmol/L HOAt and 40 mmol/L 2-pyridinecarboxylic hydrazide. The optimal reaction temperature and time were established as 20 ℃ and 60 min， respectively. Methanol was found to be the best extracting solution for free sterculic acid， while the best hydrolysis conditions for bound sterculic acid were 50 ℃ for 12 h. The method demonstrated a good linear relationship for sterculic acid over the range of 1 to 1 000 ng/mL， with correlation coefficients （r） all greater than 0.99. The limit of detection and limit of quantification were were determined to be 5 and 12.5 μg/kg， respectively. For free sterculic acid， the recovery ranged from 80.13% to 95.30%， with intra coefficient of variation of 1.01% to 2.21% and inter coefficient of variation of 1.61% to 2.38%. For esterified sterculic acid， the recovery ranged from 85.46% to 105.20%， with intra coefficient of variation of 0.79% to 2.01% and inter coefficient of variation of 3.40% to 6.22%. The optimized method had been successfully applied to quantified sterculic acid in cottonseed oil， cure cottonseed meal， degossypoled cottonseed meal and cottonseed protein samples， with concentrations ranging from 0.09 to 1 812.76 mg/kg. Conclusion The established LC-MS/MS method had the advantages of high sensitivity， high accuracy and high precision. It was suitable for quantifying free and esterified sterculic acid in feed and provides technical support for the appropriate utilization of cotton meal.

Key words: sterculic acid; cottonseed meal; derivatization; quantification

CLC Number:

S816.43

WANG Furong, FENG Xiaohui, FENG Shengnan, ZHANG Zhengjie, WANG Yujiao, MENG Qingshi, ZHANG Chunxiang. Quantification of Sterculic Acid Based on Chemical Derivatization and Mass Spectrometry Technology[J]. China Animal Husbandry & Veterinary Medicine, 2026, 53(1): 266-275.

Figures/Tables 7

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Table 1

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References 22

[1]	ZINN R A， MONTAÑO M， ALVAREZ E， et al. Feeding value of cottonseed meal for feedlot cattle［J］. Journal of Animal Science， 1997， 75（9）： 2317-2322.
[2]	LIU X， ZOU D， WANG Y， et al. Replacement of fish meal with cottonseed protein concentrate in Chinese mitten crab （Eriocheir sinensis）： Nutrient digestibility， growth performance， free amino acid profile， and expression of genes related to nutrient metabolism［J］. Animal Nutrition， 2024， 12： 447-462.
[3]	PELÁEZ R， PARIENTE A， PÉREZ-SALA Á， et al. Sterculic acid： The mechanisms of action beyond stearoyl-CoA desaturase inhibition and therapeutic opportunities in human diseases［J］. Cells， 2020， 9（1）： 140-159.
[4]	AO Y， CONG Z， BEIYU Z， et al. Effects of dietary cottonseed oil and cottonseed meal supplementation on liver lipid content， fatty acid profile and hepatic function in laying hens［J］. Animals， 2021， 11（1）： 78-90.
[5]	YANG M， LUOYI Z， AO Y， et al. The effects of dietary cottonseed meal and oil supplementation on laying performance and egg quality of laying hens［J］. Food Science & Nutrition， 2019， 7（7）： 2436-2447.
[6]	穆杨. 棉籽油和棉籽粕在鸡“橡皮蛋”形成中的作用及机制研究［D］.武汉：华中农业大学， 2019.
	MU Y. Study on the effect and mechanism of cottonseed oil and cottonseed meal on the formation of “rubber eggs”［D］.Wuhan： Huazhong Agricultural University， 2019.（in Chinese）
[7]	TAO A， WANG J， LUO B， et al. Research progress on cottonseed meal as a protein source in pig nutrition： An updated review［J］. Animal Nutrition， 2024， 25： 220-233.
[8]	ZHU Z B， WEI Y， SHI M. Recent developments of cyclopropene chemistry［J］. Chemical Society Reviews， 2011， 40（11）： 5534-5563.
[9]	BIANCHINI J， RALAIAMANERIVO A， GAYDOU E M. Determination of cyclopropenoic and cyclopropanoic fatty acids in cottonseed and kapok seed oils by gas-liquid chromatography［J］. Analytical Chemistry， 1981， 53（13）： 2194-2201.
[10]	贾怡文，陈小威，孙尚德，等. 气相色谱技术测定棉籽及其饼（粕）中环丙烯脂肪酸含量［J］. 中国油脂， 2022， 47（8）： 121-127.
	JIA Y W， CHEN X W， SUN S D， et al. Determination of cyclopropene fatty acids in cottonseed and its cake （meal） by gas chromatography［J］. China Oils and Fats， 2022，47（8）： 121-127.（in Chinese）
[11]	OBERT J C， HUGHES D， SORENSON W R， et al. A quantitative method for the determination of cyclopropenoid fatty acids in cottonseed， cottonseed meal， and cottonseed oil （Gossypium hirsutum） by high-performance liquid chromatography［J］. Journal of Agricultural and Food Chemistry， 2007， 55（6）： 2062-2067.
[12]	黄龙会，石幸玉，谢海洲，等.基于化学衍生化的质谱技术在脂质分析中的研究进展［J］.分析测试学报，2025，44（1）：73-81.
	HUANG L H， SHI X Y， XIE H Z， et al. Research progress on mass spectrometry techniques based on chemical derivatization for lipid analysis［J］ Journal of Instrumental Analysis， 2025，44（1）： 73-81.（in Chinese）
[13]	LIU M， WEI F， LV X， et al. Rapid determination of aflatoxin B₁ in medicinal and edible food by ultra-performance liquid chromatography-tandem mass spectrometry［J］. Food Chemistry， 2018， 242： 338-344.
[14]	XIANG L， ZHU L， HUANG Y， et al. Application of derivatization in fatty acids and fatty acyls detection： Mass spectrometry-based targeted lipidomics［J］. Small Methods， 2020， 4（8）： 2000160.
[15]	MANTZOURANI C， KOKOTOU M G. Liquid chromatography-mass spectrometry （LC-MS） derivatization-based methods for the determination of fatty acids in biological samples［J］. Molecules， 2022， 27（17）： 5717.
[16]	刘天宇，张成龙，杨瑞琴，等. 衍生化在脂肪酸质谱分析中的应用研究进展［J］. 理化检验-化学分册， 2025，61（3）， 364-372.
	LIU T Y， ZHANG C L， YANG R Q， et al. Research progress on application of derivatization in mass spectrometry analysis of fatty acid［J］. Physical and Chemical Testing-Chemical Volume， 2025，61（3）： 364-372.（in Chinese）
[17]	FENG X H， WANG J， TANG Z H， et al. A strategy for accurately and sensitively quantifying free and esterified fatty acids using liquid chromatography mass spectrometry［J］. Frontiers in Nutrition， 2022， 9： 977076.
[18]	陈喆，云永欢，马飞，等. 基于特征脂肪酸的食用油中棉籽油掺假鉴别技术［J］. 食品安全质量检测学报， 2023， 14（9）： 122-127.
	CHEN Z， YUN Y H， MA F， et al. Adulteration detection of edible oil with cottonseed oil based on characteristic fatty acid［J］. Journal of Food Safety and Quality， 2023， 14 （9）： 122-127.（in Chinese）
[19]	MITCHELL B， ROZEMA B， VENNARD T， et al. Determination of nutritional and cyclopropenoid fatty acids in cottonseed by a single GC analysis［J］. Journal of the American Oil Chemists’ Society， 2015， 92（7）： 947-956.
[20]	CRISCUOLO A， ZELLER M， COOK K， et al. Rational selection of reverse phase columns for high throughput LC-MS lipidomics［J］. Chemistry and Physics of Lipids， 2019， 221： 120-127.
[21]	曹正，李水军，沈敏，等. 液相色谱串联质谱临床检测方法的开发与验证［J］. 检验医学， 2019， 34（3）： 189-196.
	CAO Z， LI S J， SHEN M， et al. Consensus of method development and validation of liquid chromatography-tandem mass spectrometry in clinical laboratories［J］. Laboratory Medicine， 2019， 34（3）， 189-196.（in Chinese）
[22]	YANG A， ZHANG C， ZHANG B， et al. Effects of dietary cottonseed oil and cottonseed meal supplementation on liver lipid content， fatty acid profile and hepatic function in laying hens［J］. Animals （Basel）， 2021， 11（1）： 78.

项目 Items	苹婆酸浓度 Concentration of sterculic acid/（μg/kg）
	12.5		25		50
	批内 Intra-assay	批间 Inter-assay	批内 Intra-assay	批间 Inter-assay	批内 Intra-assay	批间 Inter-assay
游离态苹婆酸的RSD RSD of free sterculic acid	1.57	2.38	1.01	1.94	2.21	1.61
结合态苹婆酸的RSD RSD of esterified sterculic acid	1.77	4.28	0.79	6.22	2.01	3.40

样品 Sample	游离态苹婆酸 Free sterculic acid	结合态苹婆酸 Esterified sterculic acid
棉油 Cottonseed oil	29.72±2.48	1 812.76±475.47
棉粕 Cure cottonseed meal	0.46±0.01	25.96±2.39
脱酚棉粕 Degossypoled cottonseed meal	0.60±0.05	24.13±5.21
棉籽蛋白 Cottonseed protein	0.09±0.02	12.14±1.75

Quantification of Sterculic Acid Based on Chemical Derivatization and Mass Spectrometry Technology

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