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Electrochemical activity of glassy carbon electrode modified with ZnO nanoparticles prepared Via Senna Alata L. leaf extract towards antiretroviral drug

Electrochemical activity of glassy carbon electrode modified with ZnO nanoparticles prepared Via... The phytosynthesis method was used to prepare ZnO nanoparticles (ZnO NPs) via Senna alata L. leaf extract (SALE) by involving alkaloids, which play an essential role as a source of weak bases during the formation reaction of NPs. ZnO NPs on glassy carbon electrodes (GCE/ZnO NP) have been introduced to investigate its electrochemical activity towards the antiretroviral drug, lamivudine (3TC). Several characterization techniques, such as Fourier Transform Infra- Red (FTIR), X-Ray Diffraction (XRD), Scanning Electron Microscopy (SEM), Energy Dispersive X-Ray Spectroscopy (EDS), and Dynamic Light Scattering (DLS) techniques were employed to analyze the properties of GCE/ZnO NPs. As a result, ZnO NPs in spherical shape showed a high purity crystalline hexagonal wurtzite structure with a particle diameter of 40–60 nm. A Cyclic Voltammetry (CV ) measurement confirmed that the electrochemical reduction of 3TC on GCE/ ZnO NPs exhibited an excellent linear range of 10–300 µM with a detection limit of 1.902 µM, quantitation limit of 6.330 µM, and sensitivity of 0.0278 µA/µM. Thus, this research suggests a facile method for the preparation of material- based ZnO NPs as a promising antiretroviral drug sensors due to their excellent electrochemical properties. Keywords: ZnO nanoparticles, Lamivudine, Electrochemical sensor, Phytosynthesis, Green synthesis Introduction rt180, and rt204 are rapidly selected by 3TC monother- Lamivudine (2′-deoxy-3′-thiacytidine, 3TC) is a nucleo- apy of HBV, occurring in 15% of treated people after side analog reverse transcriptase inhibitor (NARTI) drug one year and up to 80% after 3  years. In addition, selec- that is actively against retroviruses, including human tion of mutants not identified by hepatitis B surface anti - immunodeficiency virus type 1 (HIV-1), human immu - gen (HBsAg) testing may obscure the detection of HBV nodeficiency virus type 2 (HIV-2), and hepatitis B virus infection after 3TC treatment. Individuals with these (HBV) [1]. 3TC inhibits the virus growth through the mutations may spread HBV that is resistant to the immu- DNA chain-breaking by forming intracellular triphos- nological responses generated by the HBV vaccine [5]. phate to prevent the multiplication of viral DNA in the Based on their medicinal and pharmacological signifi - host [2–4]. However, the use of 3TC in long-term use cance, evaluating appropriate levels of 3TC in biological leads to the emergence of mutant HBV resistance. Drug- fluids and pharmaceutical preparations is tremendously resistant genotypes with mutations at codons rt80, rt173, essential. The analytical methods for assessing 3TC lev - els have been repeatedly reported, such as spectropho- tometry [6, 7], chromatography [8], electrochemistry [9–12], etc. The chromatography and spectrophotometry *Correspondence: harits@staff.gunadarma.ac.id methods have presented good selectivity and detection Department of Pharmacy, Universitas Gunadarma, Depok 16424, Indonesia limits. However, several flaws, such as time-consuming, Full list of author information is available at the end of the article © The Author(s) 2022. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http:// creat iveco mmons. org/ licen ses/ by/4. 0/. Ariyanta et al. Micro and Nano Systems Letters (2022) 10:5 Page 2 of 7 relatively expensive, requiring a complicated analysis step 250  mL of methanol for 7  days by continuously stirring. have been investigated. Therefore, well-qualified person - Afterward, the filtrate was filtered using Whatman no. 40 nel are required to develop other analytical methods such filters and fractionated in n-hexane with a volume ratio as electrochemistry due to its low cost, easy to handle, of 1:1. The result was evaporated using a vacuum rotary and better reproducibility [13, 14]. Additionally, in the evaporator and dissolved into 100  mL of distilled water. electrochemical method, modification of the electrode Finally, the obtained SALE was stored at 4 ℃ for the surface using electroconductive materials should be preparation of ZnO NPs by the phytosynthesis method. taken into account to optimize electron transfer between the analyte and the sensing surface. Also, electroconduc- Phytosynthesis of ZnO NPs tive materials have enhanced the surface area of the elec- ZnO NPs were fabricated via phytosynthesis method trode to produce superior sensitivity [15, 16]. through a sol–gel process. SALE was added to Zn(NO ) ZnO NPs are materials with good electrical conduc- 3 2 0.015 M to obtain 100 mL of the mixture with a volume tivity, negligible toxicity, high electron transfer kinetics, ratio of 1:9 (v/v). The SALE sol was formed by heating at and excellent electrocatalytic activity against biomol- 60 ℃ for 2 h. The sol was further heated at 120  ℃ for 6 h ecules [17]. Currently, phytosynthesis has been consid- and centrifuged to form a gel. Finally, the gel was calcined ered as one of the prospective methods to prepare ZnO at 500 ℃ for 4 h to obtain white powders of ZnO NPs. NPs compared to the conventional one due to its harm- less route [18] since it uses no toxic chemicals [19] such as sodium dodecyl sulfate (SDS), tetramethylammonium Fabrication of the GCE/ZnO NPs bromide (CTAB) [20, 21], and polyvinylpyrrolidone The GCE surface was manually polished with alumina (PVP) [22]. Additionally, this method utilizes the sec- slurry for a minute and washed with ultrapure water. ondary metabolite content of plant extracts as a source Then, the GCE was sonicated in methanol for 10  min, of weak bases, chelating agents, stabilizing agents, and then rinsed with ultrapure water. ZnO NPs were dis- reducing agents [23, 24]. persed in ethanol (4 mg/ml). Afterward, it was deposited In this work, ZnO NPs were synthesized using leaf on the surface of GCE (2 mm diameter) by the drop-cast- extract of Senna Alata L. (SALE), an angiosperm belong- ing technique. Finally, GCE/ZnO NPs were dried at 60 ℃. ing to the family of Fabaceae [25], which contains sec- ondary metabolites in the form of alkaloids, flavonoids, and saponins that act as sources of weak bases, chelating Electrochemical measurement agents, and stabilizing agents during the reaction. Fur- The three-electrode system was used for the electro - thermore, ZnO NPs were used to modify the surface of chemical study of prepared electrodes towards 3TC. The the glassy carbon electrode (GCE). The electrochemical GCE/ZnO NPs, platinum wire, and Ag/AgCl served as activity of GCE/ZnO NPs was analyzed against 3TC. The the working, counter, and reference electrodes, respec- sensitivity and detection limit were also investigated by tively. The electrolyte was a 0.1  M phosphate buffer electrochemical measurements. solution (PBS) at pH ranging from 5 to 9 prepared by mixing quantitative amounts of KH PO and K HPO in 2 4 2 4 Experimental section ultrapure water. The electrochemical measurements were Materials performed using CV techniques at a range potential of Senna Alata L. leaves were obtained from Cirebon dis- − 1.2 to 0.5 V. trict, West Java, Indonesia, and further determined at the Research Center for Biology, LIPI, Indonesia. Zinc nitrate hexahydrate (Zn(NO ) .6H O), potassium hydrogen Characterization 3 2 2 phosphate (K HPO ), potassium dihydrogen phosphate An Autolab PGSTAT204 was used to perform a three- 2 4 (KH PO ), methanol, and n-hexane were purchased from electrode system for voltammetric measurements. The 2 4 Merck. Lamivudine was obtained from the Indonesian morphologies of the sample were characterized by Scan- Pharmacopoeia Comparative Standard (BPFI). The dou - ning Electron Microscopy-Energy Dispersive X-ray Spec- ble distilled water used in this research was purified with trometer (SEM–EDS, JEOL JSM 6510 LA). In contrast, Millipore Direct-Q 5 UV. All chemicals were used as the sample structure was analyzed using Powder X-Ray received without purification. Diffraction (XRD, Miniflex 600-Rigaku X-Ray Analytical Instrument) and Fourier Transform Infra-Red spectros- Preparation of SALE copy (FTIR, IR Prestige-21 Shimadzu). Malvern Zetasizer Senna Alata L. leaf was washed, dried, and mashed Nano ZSP, a particle size analyzer, determined the parti- to obtain powder. 50  g of powders were macerated in cle size distribution. A riyanta et al. Micro and Nano Systems Letters (2022) 10:5 Page 3 of 7 The SEM images in Fig.  2a depict the ZnO NPs pre- Results and discussion pared from SALE in a spherical shape with a particle Characterization of SALE and ZnO NPs size range of 40–60  nm. The presence of zinc and oxy - Figure  1a shows that the FTIR spectrum of ZnO NPs gen by elemental mapping was studied using EDS with an exhibit absorption peaks at wavenumbers of 3442, 2911, −1 atomic composition ratio of 53.6: 46.4. No other elements 1625, 1054, and 489  cm , which correspond to the from the impurities were observed, which indicates the vibration of O–H stretching, C–H (sp ) stretching, N–H formation of ZnO NPs with high purity in a homogene- bending, C–N stretching, and Zn–O stretching, respec- ous spherical shape (Fig.  2a,b). In addition, the particle tively [26, 27]. The O–H stretching vibration is assigned distribution curve obtained by the dynamic light scatter- to the residual secondary metabolites of SALE in the ing technique shows that the hydrodynamic size of the form of flavonoids, saponins, and polyphenols and the ZnO NPs was determined to be 68.061 nm (Fig. 2c). adsorption of H O from the atmosphere [1, 23]. The N–H bending and C–N stretching vibrations indicate the presence of alkaloids that play an essential role as a The electrochemical activity of GCE/ZnO NPs towards 3TC source of weak bases in the phytosynthesis of ZnO NPs The electrochemical activity of GCE/ZnO NPs against [23, 28]. Also, Zn–O stretching vibrations indicate that 3TC in 0.1 M PBS (pH 7) was determined through cyclic ZnO NPs have been successfully formed [29–32] through voltammetry (CV). Figure  3 demonstrates that GCE and the phytosynthesis method using SALE. GCE/ZnO NPs both had a reduction peak current of 3TC Moreover, the XRD diffraction pattern in Fig.  1b clearly at a reduction peak potential of −  0.906 and −  1.095  V, shows that all the diffraction peaks at (100), (002), (101), respectively, under the same experimental conditions. (102), (110), (103), (200), (112), and (201) planes were The improved conductivity and electrochemical charac - indexed to the crystalline ZnO hexagonal wurtzite struc- teristics of ZnO NPs resulted in greater electron trans- ture (JCPDS Card No. 36–1451). The average crystal - port, leading to a higher current response of GCE/ZnO lite size of ZnO NPs was calculated from the following NPs when compared to GCE. Scherrer’s formula [33]: The CV response of 3TC in 0.1 M PBS at the surface of the prepared electrode with different pH from 5 to 9 was 0.9 D = performed in Fig.  4a. The highest current response was βcosθ obtained at an optimal pH level of 7 through a reduction where, D is the crystallite size average, λ is the wave- peak of −  1.095  V (Fig.  4b). In addition, Fig.  4c shows length of the X-rays (0.15406 nm), ß is the FWHM and θ that the more alkaline in the pH system, the peak of the is the diffraction angle. The average value of the crystal - cathodic current shifts to a more negative potential, indi- lite size of ZnO NPs was found to be 18.87 nm. cating the involvement of protons in the redox reaction Fig. 1 a FTIR spectra of ZnO NPs in comparison with SALE b XRD pattern of ZnO NPs Ariyanta et al. Micro and Nano Systems Letters (2022) 10:5 Page 4 of 7 Fig. 2 a SEM image and EDX mapping, b EDX spectra and c size distribution curve of ZnO NPs The kinetics of electron transfer at the electrode sur - face was determined using the Randles–Sevcik Eq. (1) in the scan rate range of 50–150 mV/s [36]: 0.5 nFνD I = 0.4463 nF Ac (1) RT where Ip is the peak current (A), n is the number of elec- trons transferred peroxidation or reduction of one ana- lyte molecule; A is the electrode surface area (cm ), D is the analyte diffusion coefficient (cm /s), v is the scan rate (V/ s), c is the concentration of analyte in bulk solution (mol/cm ), R is the ideal gas constant, and T is the tem- perature (K). The reduction peak current of 3TC was increased Fig. 3 CV curves of 3TC in 0.1 M PBS (pH 7) on GCE and GCE/ZnO linearly with different scan rates (Fig.  4d). The plot NPs at the scan rate of 100 mV/s between anodic and cathodic peak currents versus the root of the scan rate in the Randles–Sevcik equa- tion shows a linear relationship, indicating the kinetics of electron transfer controlled by the diffusion process [34]. According to the reaction mechanism (Scheme  1), [37]. Figure 4e presents a linear plot between the reduc- the reduction peak results from the reduction of 3TC tion peak current of 3TC and the root of the scan rate in GCE/ZnO NPs, involving two electron transfers. The with a correlation coefficient of 0.9903. This result indi - C = N bond from 3TC is reduced, followed by the deami- cates that the electron transfer process at the electrode nation process. [35]. A riyanta et al. Micro and Nano Systems Letters (2022) 10:5 Page 5 of 7 Fig. 4 a CV curves of GCE/ZnO NPs at different pH values (5–9), b plot of pH versus the reduction peak current, c plot of pH versus the reduction 0.5 peak potential, d CV curves of GCE/ZnO NPs at various scan rates (50–150 mV/s), e plots of v versus the reduction current (B), plots of log v versus log I Scheme 1 Electrochemical reduction mechanism of 3TC is diffusion controlled. The plot between the logarithm as seen in Fig.  5a. The calibration curve equation was of the scan rate versus the logarithm of the peak cur- obtained to be y = −  0.0278×–81 (R = 0.9987), indi- rent in the Randles–Sevcik equation was also investi- cating excellent linearity of GCE/ZnO NPs towards gated to understand the possibility of an adsorption 3TC in the low concentration range (10–75  µM) to process on the electrode surface. If the gradient value high (300  µM) with a remarkable detection limit of in the resulting linear equation is close to 0.5, then the 1.902  µM, a quantitation limit of 6.330  µM, and a redox reaction is completely diffusion controlled. How - sensitivity of 0.0278 µA/µM. The detection limit was ever, if the gradient value is more than 0.5, it indicates obtained from three times the standard deviation devi- an adsorption process [38]. Figure  4f shows the linear ded by the slope of the calibration curve, whereas the equation between log v and log I at a gradient value of limit quantitation was obtained using the ten times. 0.2162. This result suggests that the 3TC reduction pro - Also, the sensitivity is determined from the slope on cess on the surface of GCE/ZnO NPs is not controlled the calibration curve [39]. by adsorption. The repeatability of GCE/ZnO NPs was investigated For the calibration curve, CV measurement was car- at 40  µM 3TC using the same fabricated electrode ried out in the 3TC concentration range of 10–300 µM, (Fig.  5b). The calculated relative standard deviation Ariyanta et al. Micro and Nano Systems Letters (2022) 10:5 Page 6 of 7 Fig. 5 a Calibration curve of the 3TC concentration versus the reduction peak current and b repeatability studies of GCE/ZnO NPs for 3TC in 0.1 M PBS (pH 7) Declarations (RSD) was found to be 1.26% (n = 6). This result shows the good repeatability with GCE/ZnO NPs for 3TC in Competing interests 0.1 M PBS (pH 7). The authors declare that they have no competing interests. Author details Department of Pharmacy, Universitas Gunadarma, Depok 16424, Indonesia. Conclusions Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Indonesia, Depok 16424, Indonesia. In this work, ZnO NPs have been successfully syn- thesized by phytosyntesis method using SALE by Received: 2 April 2022 Accepted: 3 May 2022 involving alkaloids which play an important role as a source of weak bases. GCE modified with ZnO NPs showed excellent electrochemical activity toward 3TC at a good linear range of 10–300  µM with a detec- References 1. Ariyanta HA, Chodijah S, Roji F, Kurnia A, Apriandanu DOB (2022) The tion limit of 1.902  µM, quantitation limit of 6.330  µM, role of Andrographis paniculata L. modified nanochitosan for lamivudine and sensitivity of 0.0278 µA/µM. The excellent elec - encapsulation efficiency enhancement and in vitro drug release study. trochemical activity of the prepared electrode might J Drug Deliv Sci Technol 67:103016. https:// doi. org/ 10. 1016/j. jddst. 2021. be attributed to the good electrical conductivity and 2. Ngilirabanga JB, Aucamp M, Samsodien H (2021) Mechanochemical high electron transfer kinetics of ZnO NPs. 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Electrochemical activity of glassy carbon electrode modified with ZnO nanoparticles prepared Via Senna Alata L. leaf extract towards antiretroviral drug

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Abstract

The phytosynthesis method was used to prepare ZnO nanoparticles (ZnO NPs) via Senna alata L. leaf extract (SALE) by involving alkaloids, which play an essential role as a source of weak bases during the formation reaction of NPs. ZnO NPs on glassy carbon electrodes (GCE/ZnO NP) have been introduced to investigate its electrochemical activity towards the antiretroviral drug, lamivudine (3TC). Several characterization techniques, such as Fourier Transform Infra- Red (FTIR), X-Ray Diffraction (XRD), Scanning Electron Microscopy (SEM), Energy Dispersive X-Ray Spectroscopy (EDS), and Dynamic Light Scattering (DLS) techniques were employed to analyze the properties of GCE/ZnO NPs. As a result, ZnO NPs in spherical shape showed a high purity crystalline hexagonal wurtzite structure with a particle diameter of 40–60 nm. A Cyclic Voltammetry (CV ) measurement confirmed that the electrochemical reduction of 3TC on GCE/ ZnO NPs exhibited an excellent linear range of 10–300 µM with a detection limit of 1.902 µM, quantitation limit of 6.330 µM, and sensitivity of 0.0278 µA/µM. Thus, this research suggests a facile method for the preparation of material- based ZnO NPs as a promising antiretroviral drug sensors due to their excellent electrochemical properties. Keywords: ZnO nanoparticles, Lamivudine, Electrochemical sensor, Phytosynthesis, Green synthesis Introduction rt180, and rt204 are rapidly selected by 3TC monother- Lamivudine (2′-deoxy-3′-thiacytidine, 3TC) is a nucleo- apy of HBV, occurring in 15% of treated people after side analog reverse transcriptase inhibitor (NARTI) drug one year and up to 80% after 3  years. In addition, selec- that is actively against retroviruses, including human tion of mutants not identified by hepatitis B surface anti - immunodeficiency virus type 1 (HIV-1), human immu - gen (HBsAg) testing may obscure the detection of HBV nodeficiency virus type 2 (HIV-2), and hepatitis B virus infection after 3TC treatment. Individuals with these (HBV) [1]. 3TC inhibits the virus growth through the mutations may spread HBV that is resistant to the immu- DNA chain-breaking by forming intracellular triphos- nological responses generated by the HBV vaccine [5]. phate to prevent the multiplication of viral DNA in the Based on their medicinal and pharmacological signifi - host [2–4]. However, the use of 3TC in long-term use cance, evaluating appropriate levels of 3TC in biological leads to the emergence of mutant HBV resistance. Drug- fluids and pharmaceutical preparations is tremendously resistant genotypes with mutations at codons rt80, rt173, essential. The analytical methods for assessing 3TC lev - els have been repeatedly reported, such as spectropho- tometry [6, 7], chromatography [8], electrochemistry [9–12], etc. The chromatography and spectrophotometry *Correspondence: harits@staff.gunadarma.ac.id methods have presented good selectivity and detection Department of Pharmacy, Universitas Gunadarma, Depok 16424, Indonesia limits. However, several flaws, such as time-consuming, Full list of author information is available at the end of the article © The Author(s) 2022. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http:// creat iveco mmons. org/ licen ses/ by/4. 0/. Ariyanta et al. Micro and Nano Systems Letters (2022) 10:5 Page 2 of 7 relatively expensive, requiring a complicated analysis step 250  mL of methanol for 7  days by continuously stirring. have been investigated. Therefore, well-qualified person - Afterward, the filtrate was filtered using Whatman no. 40 nel are required to develop other analytical methods such filters and fractionated in n-hexane with a volume ratio as electrochemistry due to its low cost, easy to handle, of 1:1. The result was evaporated using a vacuum rotary and better reproducibility [13, 14]. Additionally, in the evaporator and dissolved into 100  mL of distilled water. electrochemical method, modification of the electrode Finally, the obtained SALE was stored at 4 ℃ for the surface using electroconductive materials should be preparation of ZnO NPs by the phytosynthesis method. taken into account to optimize electron transfer between the analyte and the sensing surface. Also, electroconduc- Phytosynthesis of ZnO NPs tive materials have enhanced the surface area of the elec- ZnO NPs were fabricated via phytosynthesis method trode to produce superior sensitivity [15, 16]. through a sol–gel process. SALE was added to Zn(NO ) ZnO NPs are materials with good electrical conduc- 3 2 0.015 M to obtain 100 mL of the mixture with a volume tivity, negligible toxicity, high electron transfer kinetics, ratio of 1:9 (v/v). The SALE sol was formed by heating at and excellent electrocatalytic activity against biomol- 60 ℃ for 2 h. The sol was further heated at 120  ℃ for 6 h ecules [17]. Currently, phytosynthesis has been consid- and centrifuged to form a gel. Finally, the gel was calcined ered as one of the prospective methods to prepare ZnO at 500 ℃ for 4 h to obtain white powders of ZnO NPs. NPs compared to the conventional one due to its harm- less route [18] since it uses no toxic chemicals [19] such as sodium dodecyl sulfate (SDS), tetramethylammonium Fabrication of the GCE/ZnO NPs bromide (CTAB) [20, 21], and polyvinylpyrrolidone The GCE surface was manually polished with alumina (PVP) [22]. Additionally, this method utilizes the sec- slurry for a minute and washed with ultrapure water. ondary metabolite content of plant extracts as a source Then, the GCE was sonicated in methanol for 10  min, of weak bases, chelating agents, stabilizing agents, and then rinsed with ultrapure water. ZnO NPs were dis- reducing agents [23, 24]. persed in ethanol (4 mg/ml). Afterward, it was deposited In this work, ZnO NPs were synthesized using leaf on the surface of GCE (2 mm diameter) by the drop-cast- extract of Senna Alata L. (SALE), an angiosperm belong- ing technique. Finally, GCE/ZnO NPs were dried at 60 ℃. ing to the family of Fabaceae [25], which contains sec- ondary metabolites in the form of alkaloids, flavonoids, and saponins that act as sources of weak bases, chelating Electrochemical measurement agents, and stabilizing agents during the reaction. Fur- The three-electrode system was used for the electro - thermore, ZnO NPs were used to modify the surface of chemical study of prepared electrodes towards 3TC. The the glassy carbon electrode (GCE). The electrochemical GCE/ZnO NPs, platinum wire, and Ag/AgCl served as activity of GCE/ZnO NPs was analyzed against 3TC. The the working, counter, and reference electrodes, respec- sensitivity and detection limit were also investigated by tively. The electrolyte was a 0.1  M phosphate buffer electrochemical measurements. solution (PBS) at pH ranging from 5 to 9 prepared by mixing quantitative amounts of KH PO and K HPO in 2 4 2 4 Experimental section ultrapure water. The electrochemical measurements were Materials performed using CV techniques at a range potential of Senna Alata L. leaves were obtained from Cirebon dis- − 1.2 to 0.5 V. trict, West Java, Indonesia, and further determined at the Research Center for Biology, LIPI, Indonesia. Zinc nitrate hexahydrate (Zn(NO ) .6H O), potassium hydrogen Characterization 3 2 2 phosphate (K HPO ), potassium dihydrogen phosphate An Autolab PGSTAT204 was used to perform a three- 2 4 (KH PO ), methanol, and n-hexane were purchased from electrode system for voltammetric measurements. The 2 4 Merck. Lamivudine was obtained from the Indonesian morphologies of the sample were characterized by Scan- Pharmacopoeia Comparative Standard (BPFI). The dou - ning Electron Microscopy-Energy Dispersive X-ray Spec- ble distilled water used in this research was purified with trometer (SEM–EDS, JEOL JSM 6510 LA). In contrast, Millipore Direct-Q 5 UV. All chemicals were used as the sample structure was analyzed using Powder X-Ray received without purification. Diffraction (XRD, Miniflex 600-Rigaku X-Ray Analytical Instrument) and Fourier Transform Infra-Red spectros- Preparation of SALE copy (FTIR, IR Prestige-21 Shimadzu). Malvern Zetasizer Senna Alata L. leaf was washed, dried, and mashed Nano ZSP, a particle size analyzer, determined the parti- to obtain powder. 50  g of powders were macerated in cle size distribution. A riyanta et al. Micro and Nano Systems Letters (2022) 10:5 Page 3 of 7 The SEM images in Fig.  2a depict the ZnO NPs pre- Results and discussion pared from SALE in a spherical shape with a particle Characterization of SALE and ZnO NPs size range of 40–60  nm. The presence of zinc and oxy - Figure  1a shows that the FTIR spectrum of ZnO NPs gen by elemental mapping was studied using EDS with an exhibit absorption peaks at wavenumbers of 3442, 2911, −1 atomic composition ratio of 53.6: 46.4. No other elements 1625, 1054, and 489  cm , which correspond to the from the impurities were observed, which indicates the vibration of O–H stretching, C–H (sp ) stretching, N–H formation of ZnO NPs with high purity in a homogene- bending, C–N stretching, and Zn–O stretching, respec- ous spherical shape (Fig.  2a,b). In addition, the particle tively [26, 27]. The O–H stretching vibration is assigned distribution curve obtained by the dynamic light scatter- to the residual secondary metabolites of SALE in the ing technique shows that the hydrodynamic size of the form of flavonoids, saponins, and polyphenols and the ZnO NPs was determined to be 68.061 nm (Fig. 2c). adsorption of H O from the atmosphere [1, 23]. The N–H bending and C–N stretching vibrations indicate the presence of alkaloids that play an essential role as a The electrochemical activity of GCE/ZnO NPs towards 3TC source of weak bases in the phytosynthesis of ZnO NPs The electrochemical activity of GCE/ZnO NPs against [23, 28]. Also, Zn–O stretching vibrations indicate that 3TC in 0.1 M PBS (pH 7) was determined through cyclic ZnO NPs have been successfully formed [29–32] through voltammetry (CV). Figure  3 demonstrates that GCE and the phytosynthesis method using SALE. GCE/ZnO NPs both had a reduction peak current of 3TC Moreover, the XRD diffraction pattern in Fig.  1b clearly at a reduction peak potential of −  0.906 and −  1.095  V, shows that all the diffraction peaks at (100), (002), (101), respectively, under the same experimental conditions. (102), (110), (103), (200), (112), and (201) planes were The improved conductivity and electrochemical charac - indexed to the crystalline ZnO hexagonal wurtzite struc- teristics of ZnO NPs resulted in greater electron trans- ture (JCPDS Card No. 36–1451). The average crystal - port, leading to a higher current response of GCE/ZnO lite size of ZnO NPs was calculated from the following NPs when compared to GCE. Scherrer’s formula [33]: The CV response of 3TC in 0.1 M PBS at the surface of the prepared electrode with different pH from 5 to 9 was 0.9 D = performed in Fig.  4a. The highest current response was βcosθ obtained at an optimal pH level of 7 through a reduction where, D is the crystallite size average, λ is the wave- peak of −  1.095  V (Fig.  4b). In addition, Fig.  4c shows length of the X-rays (0.15406 nm), ß is the FWHM and θ that the more alkaline in the pH system, the peak of the is the diffraction angle. The average value of the crystal - cathodic current shifts to a more negative potential, indi- lite size of ZnO NPs was found to be 18.87 nm. cating the involvement of protons in the redox reaction Fig. 1 a FTIR spectra of ZnO NPs in comparison with SALE b XRD pattern of ZnO NPs Ariyanta et al. Micro and Nano Systems Letters (2022) 10:5 Page 4 of 7 Fig. 2 a SEM image and EDX mapping, b EDX spectra and c size distribution curve of ZnO NPs The kinetics of electron transfer at the electrode sur - face was determined using the Randles–Sevcik Eq. (1) in the scan rate range of 50–150 mV/s [36]: 0.5 nFνD I = 0.4463 nF Ac (1) RT where Ip is the peak current (A), n is the number of elec- trons transferred peroxidation or reduction of one ana- lyte molecule; A is the electrode surface area (cm ), D is the analyte diffusion coefficient (cm /s), v is the scan rate (V/ s), c is the concentration of analyte in bulk solution (mol/cm ), R is the ideal gas constant, and T is the tem- perature (K). The reduction peak current of 3TC was increased Fig. 3 CV curves of 3TC in 0.1 M PBS (pH 7) on GCE and GCE/ZnO linearly with different scan rates (Fig.  4d). The plot NPs at the scan rate of 100 mV/s between anodic and cathodic peak currents versus the root of the scan rate in the Randles–Sevcik equa- tion shows a linear relationship, indicating the kinetics of electron transfer controlled by the diffusion process [34]. According to the reaction mechanism (Scheme  1), [37]. Figure 4e presents a linear plot between the reduc- the reduction peak results from the reduction of 3TC tion peak current of 3TC and the root of the scan rate in GCE/ZnO NPs, involving two electron transfers. The with a correlation coefficient of 0.9903. This result indi - C = N bond from 3TC is reduced, followed by the deami- cates that the electron transfer process at the electrode nation process. [35]. A riyanta et al. Micro and Nano Systems Letters (2022) 10:5 Page 5 of 7 Fig. 4 a CV curves of GCE/ZnO NPs at different pH values (5–9), b plot of pH versus the reduction peak current, c plot of pH versus the reduction 0.5 peak potential, d CV curves of GCE/ZnO NPs at various scan rates (50–150 mV/s), e plots of v versus the reduction current (B), plots of log v versus log I Scheme 1 Electrochemical reduction mechanism of 3TC is diffusion controlled. The plot between the logarithm as seen in Fig.  5a. The calibration curve equation was of the scan rate versus the logarithm of the peak cur- obtained to be y = −  0.0278×–81 (R = 0.9987), indi- rent in the Randles–Sevcik equation was also investi- cating excellent linearity of GCE/ZnO NPs towards gated to understand the possibility of an adsorption 3TC in the low concentration range (10–75  µM) to process on the electrode surface. If the gradient value high (300  µM) with a remarkable detection limit of in the resulting linear equation is close to 0.5, then the 1.902  µM, a quantitation limit of 6.330  µM, and a redox reaction is completely diffusion controlled. How - sensitivity of 0.0278 µA/µM. The detection limit was ever, if the gradient value is more than 0.5, it indicates obtained from three times the standard deviation devi- an adsorption process [38]. Figure  4f shows the linear ded by the slope of the calibration curve, whereas the equation between log v and log I at a gradient value of limit quantitation was obtained using the ten times. 0.2162. This result suggests that the 3TC reduction pro - Also, the sensitivity is determined from the slope on cess on the surface of GCE/ZnO NPs is not controlled the calibration curve [39]. by adsorption. The repeatability of GCE/ZnO NPs was investigated For the calibration curve, CV measurement was car- at 40  µM 3TC using the same fabricated electrode ried out in the 3TC concentration range of 10–300 µM, (Fig.  5b). The calculated relative standard deviation Ariyanta et al. Micro and Nano Systems Letters (2022) 10:5 Page 6 of 7 Fig. 5 a Calibration curve of the 3TC concentration versus the reduction peak current and b repeatability studies of GCE/ZnO NPs for 3TC in 0.1 M PBS (pH 7) Declarations (RSD) was found to be 1.26% (n = 6). This result shows the good repeatability with GCE/ZnO NPs for 3TC in Competing interests 0.1 M PBS (pH 7). The authors declare that they have no competing interests. Author details Department of Pharmacy, Universitas Gunadarma, Depok 16424, Indonesia. Conclusions Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Indonesia, Depok 16424, Indonesia. In this work, ZnO NPs have been successfully syn- thesized by phytosyntesis method using SALE by Received: 2 April 2022 Accepted: 3 May 2022 involving alkaloids which play an important role as a source of weak bases. GCE modified with ZnO NPs showed excellent electrochemical activity toward 3TC at a good linear range of 10–300  µM with a detec- References 1. Ariyanta HA, Chodijah S, Roji F, Kurnia A, Apriandanu DOB (2022) The tion limit of 1.902  µM, quantitation limit of 6.330  µM, role of Andrographis paniculata L. modified nanochitosan for lamivudine and sensitivity of 0.0278 µA/µM. The excellent elec - encapsulation efficiency enhancement and in vitro drug release study. trochemical activity of the prepared electrode might J Drug Deliv Sci Technol 67:103016. https:// doi. org/ 10. 1016/j. jddst. 2021. be attributed to the good electrical conductivity and 2. Ngilirabanga JB, Aucamp M, Samsodien H (2021) Mechanochemical high electron transfer kinetics of ZnO NPs. 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Journal

Micro and Nano Systems Letters – Springer Journals

Published: May 17, 2022

Keywords: ZnO nanoparticles; Lamivudine; Electrochemical sensor; Phytosynthesis; Green synthesis

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