Analytical methods for screening and determination of conventional drugs adulterated in herbal products

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Date

2023

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Sokoine University of Agriculture

Abstract

Herbal products are popular worldwide. Their popularity is threatened by untrustworthy manufacturers who add conventional drugs. The addition of conventional drugs increases the risk of developing antimicrobial resistance and herb-drug interactions. To safeguard the users and enhance the safety of herbal products, analytical methods for screening and determining conventional drugs adulterated in herbal products are required. Therefore, this study was carried out to develop analytical methods and apply them in screening and determination of antibiotic, antimalarial, pain killer and erectile dysfunction adulterants in herbal products. Thin layer chromatography methods for screening twelve conventional drugs in herbal products were developed and applied. The analytes were extracted from herbal products using a solvent mixture of acetonitrile:methanol:acetic acid:water (4:4:1:1, v/v). The mobile phase consisting of dichloromethane:ethyl acetate:methanol (75:15:10, v/v) separated well trimethoprim, sildenafil, paracetamol and sulfamethoxazole. Pyrimethamine, metronidazole and sulfadoxine were well separated by dichloromethane:ethyl acetate:methanol (77.5:12.5:10, v/v). In addition, acetyl salicylic acid, ibuprofen, diclofenac, quinine and lumefantrine were well separated by ethyl acetate:methanol:30% ammonia (75:22.5:2.5, v/v). Chromatographic separations were highly reproducible and more than 10 samples were analysed in one run. The developed methods were used to screen 229 herbal products. Consequently, 24.0% of the samples contained one adulterant while 21.4% contained at least two adulterants. A high performance liquid chromatography–tandem mass spectrometry method was developed and used for screening and determining six conventional antibiotics (amoxicillin, ampicillin, metronidazole, trimethoprim, sulfamethoxazole, v and ciprofloxacin) in herbal products. The developed method had linear (r2 ≥ 0.996) calibration curves over the range of 0.005–2.5 µg mL–1 for all compounds except metronidazole, whose range was 0.005–1 µg mL–1 . The limit of detection ranged from 0.012 to 0.046 µg mL–1 while the limit of quantification ranged from 0.066 to 0.153 µg mL–1 . Accuracy, expressed as recovery of spiked herbal products ranged from 45% to 114%. The precision expressed as relative standard deviation at two concentration levels ranged from 1.6% to 15.9%. The matrix effect, expressed as matrix factor ranged from 0.79 to 0.92. The developed method was used to analyse 78 herbal products purchased from Njombe, Morogoro, Manyara, Arusha, Mwanza and Dar es Salaam in Tanzania. Metronidazole was detected in eight samples with the highest concentration of 1.38 µg g–1 . Another high performance liquid chromatography–tandem mass spectrometry method was developed and used to screen and determine eleven conventional antimalarials (chloroquine, quinine, sulfadoxine, pyrimethamine, mefloquine, lumefantrine, amodiaquine, artemisinin, dihydroartemisinin, artesunate and artemether) in herbal products. The developed method had linear (r2 ≥ 0.991) calibration curves over the range of 0.001–0.3 µg mL–1 for all compounds. The limit of detection ranged from 0.002 to 0.02 g mL–1 while the limit of quantification ranged from 0.006 to 0.08 g mL–1 . Accuracy, expressed as recovery of spiked herbal products ranged from 52% to 128%. The precision, expressed as percent relative standard deviation at two concentration levels, ranged from 1.0% to 13.8%. The matrix effect, expressed as the matrix factor ranged from 0.77 to 0.97. The developed method was used to analyse 50 herbal product samples from Njombe, Morogoro, Manyara, Arusha, Mwanza and Dar es Salaam in Tanzania. Ten of the herbal products were found to contain amodiaquine, sulfadoxine, pyrimethamine, mefloquine, dihydroartemisinin, artemether and lumefantrine. Herbal products are popular worldwide. Their popularity is threatened by untrustworthy manufacturers who add conventional drugs. The addition of conventional drugs increases the risk of developing antimicrobial resistance and herb-drug interactions. To safeguard the users and enhance the safety of herbal products, analytical methods for screening and determining conventional drugs adulterated in herbal products are required. Therefore, this study was carried out to develop analytical methods and apply them in screening and determination of antibiotic, antimalarial, pain killer and erectile dysfunction adulterants in herbal products. Thin layer chromatography methods for screening twelve conventional drugs in herbal products were developed and applied. The analytes were extracted from herbal products using a solvent mixture of acetonitrile:methanol:acetic acid:water (4:4:1:1, v/v). The mobile phase consisting of dichloromethane:ethyl acetate:methanol (75:15:10, v/v) separated well trimethoprim, sildenafil, paracetamol and sulfamethoxazole. Pyrimethamine, metronidazole and sulfadoxine were well separated by dichloromethane:ethyl acetate:methanol (77.5:12.5:10, v/v). In addition, acetyl salicylic acid, ibuprofen, diclofenac, quinine and lumefantrine were well separated by ethyl acetate:methanol:30% ammonia (75:22.5:2.5, v/v). Chromatographic separations were highly reproducible and more than 10 samples were analysed in one run. The developed methods were used to screen 229 herbal products. Consequently, 24.0% of the samples contained one adulterant while 21.4% contained at least two adulterants. A high performance liquid chromatography–tandem mass spectrometry method was developed and used for screening and determining six conventional antibiotics (amoxicillin, ampicillin, metronidazole, trimethoprim, sulfamethoxazole, v and ciprofloxacin) in herbal products. The developed method had linear (r2 ≥ 0.996) calibration curves over the range of 0.005–2.5 µg mL–1 for all compounds except metronidazole, whose range was 0.005–1 µg mL–1 . The limit of detection ranged from 0.012 to 0.046 µg mL–1 while the limit of quantification ranged from 0.066 to 0.153 µg mL–1 . Accuracy, expressed as recovery of spiked herbal products ranged from 45% to 114%. The precision expressed as relative standard deviation at two concentration levels ranged from 1.6% to 15.9%. The matrix effect, expressed as matrix factor ranged from 0.79 to 0.92. The developed method was used to analyse 78 herbal products purchased from Njombe, Morogoro, Manyara, Arusha, Mwanza and Dar es Salaam in Tanzania. Metronidazole was detected in eight samples with the highest concentration of 1.38 µg g–1 . Another high performance liquid chromatography–tandem mass spectrometry method was developed and used to screen and determine eleven conventional antimalarials (chloroquine, quinine, sulfadoxine, pyrimethamine, mefloquine, lumefantrine, amodiaquine, artemisinin, dihydroartemisinin, artesunate and artemether) in herbal products. The developed method had linear (r2 ≥ 0.991) calibration curves over the range of 0.001–0.3 µg mL–1 for all compounds. The limit of detection ranged from 0.002 to 0.02 g mL–1 while the limit of quantification ranged from 0.006 to 0.08 g mL–1 . Accuracy, expressed as recovery of spiked herbal products ranged from 52% to 128%. The precision, expressed as percent relative standard deviation at two concentration levels, ranged from 1.0% to 13.8%. The matrix effect, expressed as the matrix factor ranged from 0.77 to 0.97. The developed method was used to analyse 50 herbal product samples from Njombe, Morogoro, Manyara, Arusha, Mwanza and Dar es Salaam in Tanzania. Ten of the herbal products were found to contain amodiaquine, sulfadoxine, pyrimethamine, mefloquine, dihydroartemisinin, artemether and lumefantrine.

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Keywords

Analytical methods, Screening determination, Conventional drugs adulterated, Herbal products

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