Liquid chromatography-tandem mass spectrometry analysis of pyrrolizidine alkaloids in Polish honey

Compound	Q1 mass (Da)	Q3 mass (Da)	DP (V)	EP (V)	CE (V)	CXP (V)
Echimidine	398.2	120.1*/220.1/238.1	121	10	33/25/43	14/12/12
Echimidine N-oxide	414.2	254.2*/352.2/220.2	126	10	43/35/55	14/18/11
Echinatine	300.7	138.2*/156.2/120.2	104	10	31/37/39	18/8/18
Echinatine N-oxide	316.3	172.2*/111.2/94.2	110	10	39/51/53	8/14/16
Erucifoline	350.2	120.3*/138.1/220.1	152	10	38/38/45	14/16/12
Erucifoline N-oxide	366.2	118.0*/94.0/120.2	150	10	58/40/40	11/15/15
Europine	330.2	138.2*/254.1/156.3	127	10	30/27/40	16/13/10
Europine N-oxide	346.2	172.1*/256.3/111.2	60	10	42/35/57	21/12/12
Heliosupine	398.0	120.2*/220.2/336.2	101	10	37/29/27	16/12/16
Heliosupine N-oxide	414.0	254.3*/94.2/120.2	122	10	41/61/45	14/12/18
Heliotrine	314.1	138.2*/156.2/94.2	114	10	29/37/45	18/20/16
Heliotrine N-oxide	330.2	172.0*/138.1/94.1	158	10	37/38/54	21/16/12
Indicine	300.4	94.2*/156.3/138.0	130	10	35/38/28	11/15/16
Indicine N-oxide	316.4	172.3*/138.2/94.2	120	10	37/37/53	8/18/18
Intermedine	300.3	94.3*/156.2/138.3	135	10	35/38/27	11/17/12
Intermedine N-oxide	316.2	172.2*/94.0/111.3	130	10	38/51/53	15/11/13
Integerrimine	336.2	120.1*/94.0/308.0	150	10	38/42/36	15/11/16
Integerrimine N-oxide	352.2	118.1*/94.1/136.2	149	10	60/39/42	11/14/12
Jacobine	352.2	120.1*/280.1/155.2	136	10	33/39/39	14/14/8
Jacobine N-oxide	368.2	296.2*/120.3/94.0	137	10	46/34/56	15/15/11
Jaconine	388.1	352.2*/94.1/120.0	66	10	41/49/49	18/12/18
Jaconine N-oxide	404.0	118.1*/94.2	100	10	43/69	16/18
Lasiocarpine	412.2	120.2*/220.3/336.2	81	10	35/27/27	16/12/18
Lasiocarpine N-oxide	428.1	254.2*/120.2/94.2	130	10	41/45/57	12/18/12
Lycopsamine	300.3	94.2*/156.1/138.1	135	10	36/38/29	11/16/16
Lycopsamine N-oxide	316.2	172.3*/94.2/111.2	142	10	56/38/51	11/15/13
Monocrotaline	326.2	120.2*/121.1/237.1	151	10	45/39/33	14/14/12
Retrorsine	352.2	120.2*/138.4/324.0	150	10	39/39/38	7/6/16
Retrorsine N-oxide	368.2	94.1*/120.0/136.0	124	10	62/42/43	11/15/18
Rinderine	300.4	138.2*/156.3/94.2	156	10	31/37/45	18/8/16
Rinderine N-oxide	316.4	172.3*/111.1/94.2	110	10	39/51/53	8/20/14
Seneciphylline	334.2	120.1*/306.3/94.1	146	10	37/35/41	14/16/14
Seneciphylline N-oxide	350.2	94.1*/118.1/136.3	140	10	57/40/42	11/14/15
Senecionine	336.2	120.1*/308.2/138.1	151	10	39/37/39	12/16/16
Senecionine N-oxide	352.1	118.0*/94.3/136.1	131	10	40/61/45	14/11/16
Senceivernine	336.2	308.2*/120.1/138.2	161	10	37/39/41	16/14/6
Senecivernine N-oxide	352.1	94.1*/120.2/136.0	155	10	58/45/43	11/14/14
Senkirkine	366.2	168.2*/150.2/122.2	126	10	39/37/43	8/8/16
Spartioidine	334.2	120.1*/94.2/138.3	142	10	36/43/39	14/11/12
Trichodesmine	354.3	222.4*/120.1/138.1	150	10	38/46/46	11/6/18
Usaramine	352.2	120.0*/94.0/138.0	151	10	38/44/39	19/16/16
Usaramine N-oxide	368.2	94.0*/120.0/136.2	150	10	63/42/43	16/19/17

Q1 – precursor ion; Q3 – product ion; DP – declustering potential; EP – entrance potential; CE – collision energy; CXP – collision cell exit potential;

* – quantifier ion

Table 2.

Validation results of recovery, repeatability and reproducibility

Compound	Recovery (1 μg kg⁻¹ spike) (%)	Recovery (10 μg kg⁻¹ spike) (%)	Recovery (100 μg kg⁻¹ spike) (%)	Repeat ability (1 μg kg⁻¹ spike) (CV, %)	Repeat ability (10 μg kg⁻¹ spike) (CV, %)	Repeat ability (100 μg kg⁻¹ spike) (CV, %)	Reproducibility (1 μg kg⁻¹ spike) (CV, %)	Reproducibility (10 μg kg⁻¹ spike) (CV, %)	Reproducibility (100 μg kg⁻¹ spike) (CV, %)
Echimidine	106.7	110.0	99.0	3.6	2.0	5.2	14.4	2.9	4.9
Echimidine N-oxide	105.2	107.9	97.1	5.4	3.1	1.2	19.0	8.6	7.5
Erucifoline	103.1	117.6	104.1	2.0	1.4	2.0	19.2	5.4	7.2
Erucifoline N-oxide	111.2	109.7	97.8	3.8	5.6	2.7	13.7	16.3	9.1
Europine	109.8	108.6	93.9	3.2	2.3	5.7	10.7	3.9	6.0
Europine N-oxide	103.1	110.2	91.3	5.5	6.0	2.9	18.0	11.8	10.5
Heliosupine	118.1	112.3	97.2	2.8	1.8	2.3	14.9	2.9	5.1
Heliosupine N-oxide	105.5	108.2	95.9	3.1	3.2	4.8	13.9	6.4	5.2
Heliotrine	105.4	111.4	98.9	4.2	2.4	5.6	17.8	4.2	5.4
Heliotrine N-oxide	106.6	107.9	94.8	4.1	1.8	5.5	16.2	7.4	6.7
Intermedine	106.5	110.6	98.8	3.6	2.7	4.5	14.2	4.9	4.0
Intermedine N-oxide	110.3	111.1	93.1	4.1	2.1	4.8	10.5	7.3	7.0
Integerrimine	101.0	116.0	103.8	4.7	3.9	3.3	19.9	4.3	6.3
Integerrimine N-oxide	107.8	107.3	95.4	3.9	4.0	2.9	13.5	9.7	4.7
Jacobine	102.9	118.2	103.6	5.1	2.7	2.9	19.6	6.6	8.9
Jacobine N-oxide	106.1	107.4	92.9	8.7	5.4	6.1	13.3	8.9	6.4
Jaconine	95.8	118.3	109.4	5.6	4.4	4.3	19.3	7.0	8.7
Jaconine N-oxide	101.6	109.7	99.5	4.0	11.6	3.5	19.8	11.2	6.3
Lasiocarpine	105.7	112.8	90.4	7.3	4.4	2.5	18.4	6.8	5.8
Lasiocarpine N-oxide	107.2	111.2	94.2	2.8	1.9	4.1	16.8	6.6	7.2
Lycopsamine	103.3	116.6	99.8	5.0	4.5	3.0	19.8	4.1	4.4
Lycopsamine N-oxide	107.3	109.5	92.2	8.7	3.9	3.4	13.6	13.5	7.6
Monocrotaline	98.3	116.4	105.5	8.3	2.6	3.2	18.5	3.2	6.6
Retrorsine	103.6	110.1	96.4	4.2	3.7	3.3	14.9	3.2	4.2
Retrorsine N-oxide	105.6	106.6	92.8	5.9	3.8	2.5	16.8	9.8	7.7
Rinderine	105.5	114.7	101.8	2.4	2.8	2.0	17.1	4.3	4.2
Rinderine N-oxide	107.9	110.6	93.6	6.2	3.3	5.1	14.9	6.3	6.2
Seneciphylline	104.6	114.3	97.8	4.1	3.2	3.3	19.1	5.3	6.0
Seneciphylline N-oxide	105.1	107.9	96.1	1.6	2.8	2.5	18.7	11.2	7.7
Senecionine	102.8	112.9	100.8	3.6	1.7	1.9	19.4	4.0	6.5
Senecionine N-oxide	111.1	106.4	93.0	2.9	1.7	0.8	12.5	10.5	6.9
Senceivernine	103.8	114.6	99.6	6.5	2.6	2.5	13.4	4.5	6.4
Senecivernine N-oxide	106.5	106.9	95.4	5.6	6.2	4.1	15.1	10.6	8.1
Senkirkine	100.6	110.9	100.5	7.4	4.6	3.7	13.4	6.2	6.3
Spartioidine	106.1	117.1	100.4	5.0	4.1	2.6	20.0	4.9	6.8
Trichodesmine	100.2	115.0	107.6	10.3	1.6	2.5	17.1	3.5	6.4
Usaramine	105.6	114.8	98.9	4.4	4.3	3.0	17.8	5.4	6.3
Usaramine N-oxide	108.1	106.0	94.6	11.1	3.0	3.5	16.0	8.6	6.8

Table 3.

Validation results of linearity, matrix effect, uncertainty and limit of detection (LOD)

Compound	Linearity (R²)	Matrix effect (%)	Uncertainty (%)	LOD (μg kg⁻¹)
Echimidine	0.993	106.4	32.5	0.01
Echimidine N-oxide	0.995	97.0	40.5	0.01
Erucifoline	0.990	109.6	40.0	0.03
Erucifoline N-oxide	0.994	113.8	38.7	0.04
Europine	0.995	106.9	29.5	0.01
Europine N-oxide	0.990	127.3	37.5	0.02
Heliosupine	0.997	97.8	33.9	0.01
Heliosupine N-oxide	0.996	105.7	30.7	0.03
Heliotrine	0.995	110.7	38.3	0.01
Heliotrine N-oxide	0.994	99.5	35.9	0.01
Intermedine	0.994	107.5	32.0	0.01
Intermedine N-oxide	0.992	94.7	29.8	0.02
Integerrimine	0.996	102.8	40.1	0.02
Integerrimine N-oxide	0.997	96.9	31.7	0.06
Jacobine	0.997	104.8	40.7	0.08
Jacobine N-oxide	0.997	93.9	29.9	0.01
Jaconine	0.996	104.9	38.4	0.02
Jaconine N-oxide	0.994	110.1	40.9	0.02
Lasiocarpine	0.996	105.0	39.5	0.01
Lasiocarpine N-oxide	0.992	102.3	37.4	0.01
Lycopsamine	0.994	109.2	39.9	0.01
Lycopsamine N-oxide	0.990	119.8	31.6	0.01
Monocrotaline	0.996	108.2	38.3	0.01
Retrorsine	0.995	99.0	31.5	0.03
Retrorsine N-oxide	0.990	96.6	36.4	0.03
Rinderine	0.996	101.4	36.9	0.01
Rinderine N-oxide	0.993	100.9	34.5	0.02
Seneciphylline	0.995	102.0	38.5	0.01
Seneciphylline N-oxide	0.994	99.7	39.8	0.01
Senecionine	0.995	104.0	39.2	0.03
Senecionine N-oxide	0.995	97.8	34.0	0.04
Senceivernine	0.994	100.0	30.7	0.03
Senecivernine N-oxide	0.994	97.1	33.7	0.03
Senkirkine	0.997	98.2	27.5	0.01
Spartioidine	0.994	101.3	40.2	0.02
Trichodesmine	0.998	114.9	35.1	0.01
Usaramine	0.996	99.3	38.4	0.06
Usaramine N-oxide	0.995	104.9	36.6	0.07

Table 4.

Concentrations of open-chain pyrrolizidine alkaloids (PAs) in μg/kg in positive samples of honey

Voiv.	Echimidine	Heliosupine	Echimidine N-oxide	Heliosupine N-oxide	Intermedine	Lycopsamine/indicine	Rinderine / echinatine	Intermedine N-oxide	Lycopsamine N-oxide/indicine N-oxide	Rinderine N-oxide / echinatine N-oxide
LU
LU
LU		< LOQ				< LOQ				< LOQ
LU	9.1				< LOQ
LU
LU
LU
LU
LU					1.3	1.2	12.5			2.8
LU						< LOQ				< LOQ
LU									< LOQ	< LOQ
LU
LU							< LOQ
DS
MP
MP							< LOQ
MP							< LOQ
MP						< LOQ	< LOQ
OP
OP						< LOQ
OP
OP										< LOQ
ZP
ZP		< LOQ
ZP						< LOQ		< LOQ	< LOQ	< LOQ
WM						< LOQ				< LOQ
WP					< LOQ	< LOQ	< LOQ
WP
ŁD							< LOQ
SL										< LOQ
SL									< LOQ
LS						< LOQ
LS
PL						< LOQ
MZ					< LOQ	< LOQ
MZ	2.1		< LOQ		< LOQ	< LOQ	< LOQ	< LOQ	< LOQ
MZ	4.2	13.2			< LOQ	< LOQ	< LOQ
MZ	3.6	7.5	7.1	18.9	< LOQ	1.3	< LOQ	6.0	2.5
PL	< LOQ

[i] Voiv.– voivodeship; LU – Lubelskie; DS – Dolnośląskie; MP – Małopolskie; OP – Opolskie; ZP – Zachodnio-Pomorskie; WM – Warmińsko-Mazurskie; WP – Wielkopolskie; ŁD – Łódzkie; SL – Śląskie; LS – Lubuskie; PL – Podlaskie; MZ – Mazowieckie; PK – Podkarpackie; LOQ – limit of quantification

Table 5.

Concentrations of macrocyclic pyrrolizidine alkaloids (PAs) in μg/kg in positive samples of honey and the total PA content in μg/kg representing the sum of open-chain and macrocyclic PAs

Voivodeship	Seneciphylline	Spartioidine	Seneciphylline N-oxide	Senecionine	Senecivernine	Total PA content
LU		< LOQ				0.0
LU		< LOQ				0.0
LU					1.0	1.0
LU	1.7			1.0	1.0	12.8
LU		< LOQ				0.0
LU		< LOQ				0.0
LU		< LOQ				0.0
LU	< LOQ					0.0
LU	< LOQ					17.8
LU	< LOQ	< LOQ				0.0
LU		< LOQ				0.0
LU		< LOQ				0.0
LU		< LOQ				0.0
DS		< LOQ				0.0
MP		< LOQ				0.0
MP	< LOQ					0.0
MP	21.3		2.3			23.6
MP	< LOQ	< LOQ				0.0
OP	< LOQ					0.0
OP	< LOQ	< LOQ				0.0
OP		< LOQ				0.0
OP		< LOQ				0.0
ZP		< LOQ				0.0
ZP		< LOQ				0.0
ZP		< LOQ				0.0
WM	< LOQ	1.1				1.1
WP		< LOQ				0.0
WP	< LOQ	< LOQ				0.0
ŁD		< LOQ				0.0
SL		< LOQ				0.0
SL						0.0
LS	< LOQ	1.8			1.1	2.9
LS		< LOQ				0.0
PL					< LOQ	0.0
MZ		1.5				1.5
MZ						2.1
MZ	< LOQ	< LOQ	< LOQ	< LOQ		17.4
MZ						46.9
PL						0.0

[i] LU – Lubelskie; DS – Dolnośląskie; MP – Małopolskie; OP – Opolskie; ZP – Zachodnio-Pomorskie; WM – Warmińsko-Mazurskie; WP – Wielkopolskie; ŁD – Łódzkie; SL – Śląskie; LS – Lubuskie; PL – Podlaskie; MZ – Mazowieckie; PK – Podkarpackie; LOQ – limit of quantification

Extracted ion chromatogram of: 1 – intermedine N-oxide; 2 – lycopsamine N-oxide/indicine N-oxide; 3 – rinderine N-oxide/echinatine N-oxide; 4 – intermedine; 5 – lycopsamine/indicine; 6 – rinderine/echinatine; 7 – integerrimine N-oxide; 8 – senecionine N-oxide; 9 – senecivernine N-oxide; 10 – integerrimine; 11 – senecionine; and 12 – senecivernine

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Articles in this issue

DOI: https://doi.org/10.2478/jvetres-2026-0046 | Journal eISSN: 2450-8608 (formerly 2300-3235)

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Language: English

Page range: 463 - 474

Submitted on: Mar 2, 2026

Accepted on: Aug 21, 2026

Published on: Aug 27, 2026

Published by: National Veterinary Research Institute in Pulawy

In partnership with: Paradigm Publishing Services

Keywords:

food contaminants,

honey,

LC-MS/MS,

pyrrolizidine alkaloids,

toxic plant secondary metabolites

Related subjects:

Life sciences,

Molecular biology,

Microbiology and virology,

Life sciences, other,

Medicine,

Veterinary medicine

© 2026 Ewelina Kowalczyk, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution 4.0 License.

Volume 70 (2026): Issue 3 (September 2026)

Liquid chromatography-tandem mass spectrometry analysis of pyrrolizidine alkaloids in Polish honey

Figures & Tables

Table 1.

Table 2.

Table 3.

Table 4.

Table 5.

Fig. 1.

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