Analysis of aflatoxins and ochratoxin a in chilli powder using ultrahigh performance liquid chromatography with fluorescence detection and tandem mass spectrometry

Manisha Dhanshetty; Raviraj Shinde; Arnab Goon; Dasharath Oulkar; Christopher T. Elliott; Kaushik Banerjee

doi:10.1007/s12550-022-00460-4

Analysis of aflatoxins and ochratoxin a in chilli powder using ultrahigh performance liquid chromatography with fluorescence detection and tandem mass spectrometry

Manisha Dhanshetty, Raviraj Shinde, Arnab Goon, Dasharath Oulkar, Christopher T. Elliott, Kaushik Banerjee^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

17 Citations (Scopus)

Abstract

Chilli powder, a popular spice, is predominantly contaminated with aflatoxins (AFs) and ochratoxin A (OTA), posing a menace to public health. As no validated method exists for the simultaneous and direct analysis of AFs and OTA in chilli powder, it was imperative to develop one to ensure their effective monitoring and promote trade. In this research, we developed and validated a multi-mycotoxin analysis method that allows the simultaneous determination of AFs (AFB₁, AFB₂, AFG₁ and AFG₂) and OTA in chilli powder with high sensitivity, accuracy and precision. The optimised sample preparation workflow started with the extraction of chilli powder (25 g) with methanol–water (100 mL, 80:20). An aliquot (3 mL) was cleaned on a multi-mycotoxin, immunoaffinity column (AFLAOCHRA PREP^®) and analysed using ultrahigh performance liquid chromatography with fluorescence (UHPLC-FLD) and tandem mass spectrometric (LC–MS/MS) detection in a single chromatographic run. The method performance was evaluated through intra- and inter-laboratory validation (ILV) studies, and also by analysing a certified reference material. A direct analysis using UHPLC-FLD (without derivatisation) provided the limits of quantification (LOQ) of 0.25 and 1 ng/g for AFs and OTA, respectively, while the LOQ for all these mycotoxins in LC–MS/MS was 0.5 ng/g. These LOQs are much lower than the maximum levels (MLs) specified by the European Commission. The recoveries of these analytes at LOQ and higher levels were above 75% (RSD_r < 12%). The ILV study demonstrated satisfactory method-reproducibility (RSD_R < 25%). The analysis of the certified reference material provided accuracies of AFs and OTA in the range of 83–101%. The analysis by UHPLC-FLD and LC–MS/MS provided very similar results. The incurred levels of B1 in market samples were estimated with a precision-RSD of < 6%. Considering its efficiency and alignment with the regulatory requirements, this method can be implemented for the routine analysis of AFs and OTA in chilli powder.

Original language	English
Pages (from-to)	193-203
Number of pages	11
Journal	Mycotoxin Research
Volume	38
Early online date	14 Jul 2022
DOIs	https://doi.org/10.1007/s12550-022-00460-4
Publication status	Published - 2022

Bibliographical note

Funding Information:
This work received partial funding from the Food Safety Standards Authority of India under the National Reference Laboratory (NRL) scheme.

Publisher Copyright:
© 2022, The Author(s) under exclusive licence to Society for Mycotoxin (Research Gesellschaft für Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.

Keywords

Aflatoxins
Chilli powder
LC–MS/MS
Method validation
Ochratoxin A
UHPLC-fluorescence

ASJC Scopus subject areas

Biotechnology
Microbiology
Toxicology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Cite this

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title = "Analysis of aflatoxins and ochratoxin a in chilli powder using ultrahigh performance liquid chromatography with fluorescence detection and tandem mass spectrometry",

abstract = "Chilli powder, a popular spice, is predominantly contaminated with aflatoxins (AFs) and ochratoxin A (OTA), posing a menace to public health. As no validated method exists for the simultaneous and direct analysis of AFs and OTA in chilli powder, it was imperative to develop one to ensure their effective monitoring and promote trade. In this research, we developed and validated a multi-mycotoxin analysis method that allows the simultaneous determination of AFs (AFB1, AFB2, AFG1 and AFG2) and OTA in chilli powder with high sensitivity, accuracy and precision. The optimised sample preparation workflow started with the extraction of chilli powder (25 g) with methanol–water (100 mL, 80:20). An aliquot (3 mL) was cleaned on a multi-mycotoxin, immunoaffinity column (AFLAOCHRA PREP{\textregistered}) and analysed using ultrahigh performance liquid chromatography with fluorescence (UHPLC-FLD) and tandem mass spectrometric (LC–MS/MS) detection in a single chromatographic run. The method performance was evaluated through intra- and inter-laboratory validation (ILV) studies, and also by analysing a certified reference material. A direct analysis using UHPLC-FLD (without derivatisation) provided the limits of quantification (LOQ) of 0.25 and 1 ng/g for AFs and OTA, respectively, while the LOQ for all these mycotoxins in LC–MS/MS was 0.5 ng/g. These LOQs are much lower than the maximum levels (MLs) specified by the European Commission. The recoveries of these analytes at LOQ and higher levels were above 75% (RSDr < 12%). The ILV study demonstrated satisfactory method-reproducibility (RSDR < 25%). The analysis of the certified reference material provided accuracies of AFs and OTA in the range of 83–101%. The analysis by UHPLC-FLD and LC–MS/MS provided very similar results. The incurred levels of B1 in market samples were estimated with a precision-RSD of < 6%. Considering its efficiency and alignment with the regulatory requirements, this method can be implemented for the routine analysis of AFs and OTA in chilli powder.",

keywords = "Aflatoxins, Chilli powder, LC–MS/MS, Method validation, Ochratoxin A, UHPLC-fluorescence",

author = "Manisha Dhanshetty and Raviraj Shinde and Arnab Goon and Dasharath Oulkar and Elliott, {Christopher T.} and Kaushik Banerjee",

note = "Funding Information: This work received partial funding from the Food Safety Standards Authority of India under the National Reference Laboratory (NRL) scheme. Publisher Copyright: {\textcopyright} 2022, The Author(s) under exclusive licence to Society for Mycotoxin (Research Gesellschaft f{\"u}r Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.",

year = "2022",

doi = "10.1007/s12550-022-00460-4",

language = "English",

volume = "38",

pages = "193--203",

journal = "Mycotoxin Research",

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T1 - Analysis of aflatoxins and ochratoxin a in chilli powder using ultrahigh performance liquid chromatography with fluorescence detection and tandem mass spectrometry

AU - Dhanshetty, Manisha

AU - Shinde, Raviraj

AU - Goon, Arnab

AU - Oulkar, Dasharath

AU - Elliott, Christopher T.

AU - Banerjee, Kaushik

N1 - Funding Information: This work received partial funding from the Food Safety Standards Authority of India under the National Reference Laboratory (NRL) scheme. Publisher Copyright: © 2022, The Author(s) under exclusive licence to Society for Mycotoxin (Research Gesellschaft für Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.

PY - 2022

Y1 - 2022

N2 - Chilli powder, a popular spice, is predominantly contaminated with aflatoxins (AFs) and ochratoxin A (OTA), posing a menace to public health. As no validated method exists for the simultaneous and direct analysis of AFs and OTA in chilli powder, it was imperative to develop one to ensure their effective monitoring and promote trade. In this research, we developed and validated a multi-mycotoxin analysis method that allows the simultaneous determination of AFs (AFB1, AFB2, AFG1 and AFG2) and OTA in chilli powder with high sensitivity, accuracy and precision. The optimised sample preparation workflow started with the extraction of chilli powder (25 g) with methanol–water (100 mL, 80:20). An aliquot (3 mL) was cleaned on a multi-mycotoxin, immunoaffinity column (AFLAOCHRA PREP®) and analysed using ultrahigh performance liquid chromatography with fluorescence (UHPLC-FLD) and tandem mass spectrometric (LC–MS/MS) detection in a single chromatographic run. The method performance was evaluated through intra- and inter-laboratory validation (ILV) studies, and also by analysing a certified reference material. A direct analysis using UHPLC-FLD (without derivatisation) provided the limits of quantification (LOQ) of 0.25 and 1 ng/g for AFs and OTA, respectively, while the LOQ for all these mycotoxins in LC–MS/MS was 0.5 ng/g. These LOQs are much lower than the maximum levels (MLs) specified by the European Commission. The recoveries of these analytes at LOQ and higher levels were above 75% (RSDr < 12%). The ILV study demonstrated satisfactory method-reproducibility (RSDR < 25%). The analysis of the certified reference material provided accuracies of AFs and OTA in the range of 83–101%. The analysis by UHPLC-FLD and LC–MS/MS provided very similar results. The incurred levels of B1 in market samples were estimated with a precision-RSD of < 6%. Considering its efficiency and alignment with the regulatory requirements, this method can be implemented for the routine analysis of AFs and OTA in chilli powder.

AB - Chilli powder, a popular spice, is predominantly contaminated with aflatoxins (AFs) and ochratoxin A (OTA), posing a menace to public health. As no validated method exists for the simultaneous and direct analysis of AFs and OTA in chilli powder, it was imperative to develop one to ensure their effective monitoring and promote trade. In this research, we developed and validated a multi-mycotoxin analysis method that allows the simultaneous determination of AFs (AFB1, AFB2, AFG1 and AFG2) and OTA in chilli powder with high sensitivity, accuracy and precision. The optimised sample preparation workflow started with the extraction of chilli powder (25 g) with methanol–water (100 mL, 80:20). An aliquot (3 mL) was cleaned on a multi-mycotoxin, immunoaffinity column (AFLAOCHRA PREP®) and analysed using ultrahigh performance liquid chromatography with fluorescence (UHPLC-FLD) and tandem mass spectrometric (LC–MS/MS) detection in a single chromatographic run. The method performance was evaluated through intra- and inter-laboratory validation (ILV) studies, and also by analysing a certified reference material. A direct analysis using UHPLC-FLD (without derivatisation) provided the limits of quantification (LOQ) of 0.25 and 1 ng/g for AFs and OTA, respectively, while the LOQ for all these mycotoxins in LC–MS/MS was 0.5 ng/g. These LOQs are much lower than the maximum levels (MLs) specified by the European Commission. The recoveries of these analytes at LOQ and higher levels were above 75% (RSDr < 12%). The ILV study demonstrated satisfactory method-reproducibility (RSDR < 25%). The analysis of the certified reference material provided accuracies of AFs and OTA in the range of 83–101%. The analysis by UHPLC-FLD and LC–MS/MS provided very similar results. The incurred levels of B1 in market samples were estimated with a precision-RSD of < 6%. Considering its efficiency and alignment with the regulatory requirements, this method can be implemented for the routine analysis of AFs and OTA in chilli powder.

KW - Aflatoxins

KW - Chilli powder

KW - LC–MS/MS

KW - Method validation

KW - Ochratoxin A

KW - UHPLC-fluorescence

U2 - 10.1007/s12550-022-00460-4

DO - 10.1007/s12550-022-00460-4

M3 - Article

C2 - 35834161

AN - SCOPUS:85134325903

SN - 0178-7888

VL - 38

SP - 193

EP - 203

JO - Mycotoxin Research

JF - Mycotoxin Research

ER -

Analysis of aflatoxins and ochratoxin a in chilli powder using ultrahigh performance liquid chromatography with fluorescence detection and tandem mass spectrometry

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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