Browsing by Author "Mwankuna Christopher Johnson"

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A HPLC-MS/MS method for screening of selected antibiotic adulterants in herbal drugs
(The Royal Society of Chemistry, 2022) Mwankuna Christopher Johnson; Uwamaliya Grâce Ange; Mariki Eliapenda Elisante; Mabiki Faith; Malebo Hamisi M.; Mdegela Robinson; Styrishaveb Bjarne
The use of herbal products adulterated with conventional drugs increases the risk of developing microbial resistance and causes herb-to-drug interaction, leading to severe clinical consequences. The complex nature of herbal products has been a challenge for the unambiguous identification of adulterants. The improved analytical selectivity and sensitivity of hyphenated techniques such as high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) enable the confirmatory screening of adulterants in herbal products. Simultaneous screening of adulterants is necessary and efficient because it has been established that more than one chemical adulterant may be present in one herbal product. An HPLC-MS/MS method for the simultaneous detection and quantification of amoxicillin, ampicillin, metronidazole, ciprofloxacin, sulfamethoxazole, and trimethoprim in powdered herbal drugs was developed. Deuterated metronidazole-d3, trimethoprim-d3, ciprofloxacin-d8, and sulfamethoxazole-d4 were used as internal standards (ISs). For each analyte, two transitions were monitored using protonated molecules as precursor ions. The extraction of analytes from herbal products was performed using a simple methanol : water : formic acid (90 : 10 : 0.05, v/v) extraction solvent. Chromatographic separation was done in a gradient of 0.01% formic acid in methanol and 0.01% formic acid in MilliQ water. The calibration curves were linear (r2 $ 0.996) over the range of 0.005–2.5 mg mL 1 for all compounds except metronidazole, whose range was 0.005–1 mg mL 1. The limit of detection (LOD) ranged from 0.012 to 0.046 mg mL 1, while the limit of quantification (LOQ) ranged from 0.066 to 0.153 mg mL 1. The accuracy, expressed as the recovery of spiked herbal products, ranged from 45% to 114%. The precision, expressed as relative standard deviation (RSD) at two concentration levels, ranged from 1.6% to 15.9%. The matrix effect expressed as the matrix factor (MF) ranged from 0.79 to 0.92. The developed method was applied to powder herbal products purchased in Tanzania. Amoxicillin, ampicillin, trimethoprim, sulfamethoxazole, and ciprofloxacin were not detected in all samples. Metronidazole was detected in eight samples with the highest concentration of 1.38 mg g 1. The developed method is suitable for the detection of all the studied antibiotic adulterants in herbal products. Quantification can be performed for all the compounds except ciprofloxacin due to its lower recovery.
Analytical methods for screening and determination of conventional drugs adulterated in herbal products
(Sokoine University of Agriculture, 2023) Mwankuna Christopher Johnson
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.
Analytical methods for screening and determination of conventional drugs adulterated in herbal products
(Sokoine University of Agriculture, 2023-05) Mwankuna Christopher Johnson
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, 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. The developed thin layer chromatography and high performance liquid chromatography–tandem mass spectrometry methods are considered valuable tools for a better understanding of the adulteration of herbal products by addition of conventional drugs. The thin layer chromatography methods can be used for preliminary screening of herbal products prior to confirmation by other techniques such as high performance liquid chromatography–tandem mass spectrometry. On the other hand, confirmation and quantification of the selected antibiotic and antimalarial adulterants in herbal products can be achieved using the developed high performance liquid chromatography–tandem mass spectrometry methods.
Analytical methods for screening and determination of conventional drugs adulterated in herbal products
(Sokoine University of Agriculture (SUA), 2023-05) Mwankuna Christopher Johnson
Herbal products are popular worldwide. Their popularity is 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 chromatographic method for detection of conventional drug adulterants in herbal products
(MDPI, 2022-12-31) Mwankuna Christopher Johnson; Mariki Eliapenda Elisante; Malebo Hamisi Masanja; Styrishave Bjarne; Mdegela Robinson Hammerton
Commercially available conventional drugs have been used to adulterate herbal products. Considering the rapid growth of herbal products’ market, it is essential to screen herbal products for the presence of conventional drugs. Simple analytical methods are needed for the rapid screening of conventional drugs that are likely to be adulterated in herbal products. Thin layer chromatography (TLC) methods for screening twelve conventional drugs in herbal products have been developed and applied. The analytes were extracted from herbal products using acetonitrile:methanol:acetic acid:water (4:4:1:1, v/v). Solvent mixture of dichloromethane:ethyl acetate:methanol (75:15:10, v/v) separated well trimethoprim, sildenafil, paracetamol, and sulfamethoxazole while 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 found to be highly reproducible, and more than 10 samples can be analysed in one run. The method was applied in the screening of 229 herbal products. Consequently, 24.0% of the samples contained one adulterant, while 21.4% contained at least two adulterants. All conventional drugs detected in herbal products were not mentioned on the labels and therefore the consumers are kept unaware of their side effects and health problems. Further studies for confirming and quantitatively determining the adulterants in a wide range of products as well as a systematic toxicological analysis of the adulterants in herbal products are recommended.