Int J App Pharm, Vol 14, Issue 1, 2022, 246-250Original Article



1Bhakti Pertiwi, School of Pharmacy Science, Palembang 30128 Indonesia, 2Departement of Pharmaceutics, Faculty of Pharmacy, Gadjah Mada University, Yogyakarta 55281 Indonesia, 3Departement of Pharmacy, Faculty of Mathematics and Natural Sciences, Sriwijaya University, Indralaya 30622 Indonesia
Email: [email protected]

Received: 24 Aug 2021, Revised and Accepted: 14 Oct 2021


Objective: This study aims to increase the solubility of simvastatin (SIM), a hydrophobic drug, by incorporating it into PCL-PEG-PCL triblock copolymer micelles and validating the assay method used, namely Uv-Vis spectrophotometric.

Methods: The shake flask method was used to determine the increase in solubility experienced by SIM after being incorporated into the micellar system. The values ​​of maximum wavelength (λmax), linearity, LOD, LOQ, accuracy, and precision were used as parameters measured to assess the validity of the assay method used.

Results: The results showed that PCL-PEG-PCL triblock copolymer micelles could increase SIM solubility by 9.7 times (89.49±5.75 µg/ml) compared to SIM without modification (9.19±0.24 µg/ml). The validation results show the λmax value of 239 nm, a linear calibration curve with an R-value of 0.9994, LOD and LOQ of 0.33 µg/ml and 1.00 µg/ml, accurate measurement with recovery at concentrations of 80%, 100%, and 120% were 102.93±1.32%, 100.78±0.40%, and 104.58±0.79% and also had good precision ​​with RSD<2%.

Conclusion: The PCL-PEG-PCL triblock copolymer micelles can increase SIM solubility and the Uv-Vis spectrophotometric method has been validated successfully for the quantitative analysis of SIM in PCL-PEG-PCL triblock copolymer micelles.

Keywords: Simvastatin, Triblock copolymer, PCL, PEG, Validation, Uv-Vis spectrophotometric


Solubility is one of the physicochemical properties of drugs that need to be considered because it can affect the formulation and effectiveness of therapy. Drugs with low solubility (hydrophobic drugs) will provide low bioavailability so that the desired therapeutic effect is not perfect [1, 2].

SIM (C25H38O5) is an anticholesterol drug of the statin class with the mechanism of action of inhibiting the enzyme 3-hydroxy-3-methyl glutaryc-coenzyme A reductase (HMG-CoA reductase). SIM belongs to the class II biopharmaceutical classification system (BCS) with a solubility of 0.01 g/l (practically insoluble) and a bioavailability of<5% [3-6].

Many attempts have been made to increase the solubility of SIM, including hydrogels [7], complexes with arginine [8], solid dispersions [9, 10], micellar polymers with derivatives of tocopherol [11], spherical crystal [12], and co-crystal formation [13, 14]. In this study, the increase in the solubility of SIM was carried out by being incorporated into PCL-PEG-PCL triblock copolymer micelles which would then form a micellar polymer. PCL-PEG-PCL triblock copolymer micelles are an ideal drug carrier candidate for SIM with an entrapment efficiency of 87.74% [15].

To determine the increase in solubility experienced by SIM, it is necessary to determine the concentration of SIM in the PCL-PEG-PCL triblock copolymer micelle. According to the pharmacopeia, SIM levels were determined by the High-Performance Liquid Chromatography (HPLC) method. However, a simpler method, UV-Vis spectrophotometry, has been reported to be used for the assay of SIM in several pharmaceutical preparations showing results that meet the required acceptance criteria [16-19].



SIM is provided free by Dexa Medica (Palembang-Indonesia). All other chemicals and reagents used in this study met the criteria for an analytical grade.

Preparation of PCL-PEG-PCL triblock copolymer and SIM loaded PCL-PEG-PCL triblock copolymer micelles

The preparation of PCL-PEG-PCL triblock copolymer and incorporated SIM into the micelles system was obtained from our previous study. Where PCL-PEG-PCL triblock copolymer is made by reacting 5 g of PEG and 10 g of ɛ-CL using Sn (Oct)2 0.5% w/w as a catalyst by the ring-opening polymerization method (ROP). While SIM was incorporated into the polymeric micelles by the solvent evaporation method (film formation), 1 ml of SIM stock solution in dichloromethane (100 mg/10 ml) was mixed with 50 mg of PCL-PEG-PCL triblock copolymer [15].

Preparation of SIM stock solution

SIM was weighed as much as 10 mg, put into a 25 ml volumetric flask, and methanol was added to the mark and then homogenized to obtain a concentration of 400 ppm [16].

Determination of the λmax of SIM

The 0.05 ml of the stock solution is pipetted, put into a 5 ml volumetric flask, distilled water is added to the limit mark and homogenized to obtain a solution with a concentration of 4 ppm, then the solution is measured using a UV-vis spectrophotometer over a 200-300 nm wavelength range. The λmax of SIM is indicated by the wavelength that gives the highest absorbance [16].

Preparation of SIM calibration curve

The stock solution was pipetted as much as each 0.050, 0.075, 0.100, 0.125, 0.150, and 0.175 ml were put into a 5 ml volumetric flask, and then distilled water was added to the mark and homogenized to obtain a serial solution with a concentration of 4, 6, 8, 10, 12 and 14 ppm. The series solution was measured with a UV-vis spectrophotometer at the λmax of SIM [16].

Solubility enhancement test of SIM in the PCL-PEG-PCL triblock copolymer micelles

SIM excess (10 mg) and SIM loaded into PCL-PEG-PCL triblock copolymer was dissolved in 10 ml of distilled water and shaken for 24 h at 25±1 °C. After 24 h, the solution was filtered with a 0.45 µm membrane filter and measured using a UV-vis spectrophotometer at the λmax of SIM and the dissolved content was calculated using a calibration curve that had been prepared [20]. The instrument used was a UV-vis spectrophotometer (Thermo Scientific, Genesys 10S UV).

Method validation

Linearity test

The linearity test was carried out by analyzing the measurement results of the serial solution that had been made (4, 6, 8, 10, 12, and 14 ppm) then made a relationship between the absorbance and the concentration of the serial solution, a linear regression equation (y = ax+b) and correlation coefficient (R) was obtained [16].

Determination of limit of detection and limit of quantification (LOD and LOQ)

The LOD and LOQ was determined by measuring the absorbance of the serial solution on the calibration curve for 3 replications and then the standard deviation (SD) is determined [20].

LOD was calculated using the following equation:


LOQ was calculated using the following equation:


Accuracy and precision test

The stock solution was pipetted as much as each 0.100, 0.125, and 0.150 ml, were put into a 5 ml volumetric flask containing 0.125 ml of PCL-PEG-PCL triblock copolymer solution in water, then distilled water was added to the mark, and then homogenized. The test solution was measured with a UV-vis spectrophotometer at the λmax of SIM. The accuracy test is assessed based on the % recovery, while the precision test is determined based on the relative standard deviation (RSD) value [16, 21].


Increased solubility of SIM in PCL-PEG-PCL triblock copolymers micelles

Theoretically, SIM has a water solubility of 10 g/ml. The solubility of the modified SIM into the PCL-PEG-PCL triblock copolymer micelles was 89.49±5.75µg/ml, while the solubility of SIM without modification was 9.19±0.24µg/ml. These results indicate that there has been an increase in the solubility of SIM after being made into micelles [22]. The complete test results for increasing the solubility of SIM can be seen in table 1.

Table 1: The results for increasing solubility of SIM in triblock copolymer

Sample Concentration of SIM (µg/ml)* Increased solubility
SIM without modification 9.19±0.24 -
SIM in micelles 89.49±5.75 9.7 times

*All values are expressed as mean of n=3±standard deviation (SD)

SIM is a drug that has low solubility in water. Drugs with low solubility in water will be difficult to absorb into the gastrointestinal tract where the main component is water so that it will cause low bioavailability. Therefore, one way that can be used to increase the bioavailability of a drug is to increase its solubility in water [1, 3]. A solubility enhancement test was carried out to examine the ability of PCL-PEG-PCL triblock copolymer to increase the solubility of SIM. A similar study was conducted by Alami-milani et al. using a hydrophobic drug model of dexamethasone, in that study, it was reported that the use of PCL-PEG-PCL triblock copolymers could increase the solubility of dexamethasone by 11.7 times (1.17 mg/ml) [23].

The PCL-PEG-PCL triblock copolymer was composed of PCL as a hydrophobic block and PEG as a hydrophilic block. In aqueous media, PCL-PEG-PCL triblock copolymer will spontaneously form micelles with the hydrophilic part on the outside and the hydrophobic part on the inside as the core [24, 25]. The ability to increase drug solubility by triblock micellar copolymers is influenced by the length of the hydrophobic block and the ratio of the constituent polymers. The longer the hydrophobic block that makes up the triblock copolymer, the greater the solubility of the drug because more hydrophobic drugs can be loaded into the micellar system [26].

Analysis method validation

Method validation was carried out on the parameters of the λmax, correlation coefficient (R) of the obtained calibration curve, LOD, LOQ, accuracy, and precision. The complete validation test results for various parameters can be seen in table 2.

Table 2: The summary of validation

Parameter Result
λmax 239 nm


y = ax+b

a: slope

b: intercept

Y = 0.0499x–0.0249
Coeficient correlation (R) 0.9994
LOD (µg/ml) 0.33
LOQ (µg/ml) 1.00
Accuracy (% recovery)* 80% 102.93±1.32
100% 100.78±0.40
120% 104.58±0.79
Precision (% RSD) Intra-day 1.371 (8 µg/ml)
0.418 (10 µg/ml)
0.786 (12 µg/ml)
Inter-day (10 µg/ml) 0.418 (1st day)
0.525 (2nd day)

*All values are expressed as mean of n=3±standard deviation (SD)

λmax of SIM measurement results

The λmax is the wavelength that gives the maximum absorption of SIM. The determination of the λmax of SIM aims to provide maximum sensitivity of samples containing SIM, a calibration curve that is linear and produces fairly constant data if repeated measurements are made. Determination of the λmax was carried out at a concentration of 4 ppm using a Uv-Vis spectrophotometer, the fig. 1 showed the λmax of SIM was 239 nm, the results were not much different from the λmax of SIM in the literature, which was 238 nm. The λmax shift of the measurement results compared to the literature can be caused by several factors, such as differences in the source of materials and the tools used. However, the wavelength shift is not more than 3% of the λmax in the literature so it can be said that the results of the measurements carried out meet the requirements for use for analysis [16].

Fig. 1: The λmax of SIM

The results of the calibration curve and linearity test

The calibration curve was used to determine the linear regression equation that would be used to calculate SIM levels in the PCL-PEG-PCL triblock copolymer. The linear regression equation was obtained from the relationship between the concentration of the prepared SIM series solution and its absorbance measured at a wavelength of 239 nm which is the λmax of SIM used in this study, which was presented in table 3. SIM calibration curve graph can be seen in fig. 2, with intercept value =-0.0249 and slope value = 0.0499, so that the linear regression equation y = 0.0499x-0.0249 with R = 0.9994 is obtained. The linearity test can be determined based on the correlation coefficient (R) of the obtained linear regression equation, where the acceptance criteria of the linearity test are R 0.9994. When viewed from the R-value obtained in the test, it shows that the method used has good linearity [16, 17, 27].

Accuracy and precision test results

One of the fundamental requirements in the analysis is accuracy and precision. Accuracy indicates the closeness of the measurement results to the actual value which is expressed as % recovery, while precision indicates the degree of suitability of the test results as measured by the distribution of the results from the average when repeated measurements are made which will produce an average value that is very close to the true value. The measuring parameter to determine precision is the percent relative standard deviation (% RSD) [16, 17, 28].

Table 4 presents that the average % recovery obtained in determining the accuracy is 102.93±1.32%, 100.78±0.40%, and 104.58±0.79%. % recovery is acceptable because it is in the range of 80-110% [17, 21, 28].

Table 3: The result of absorbance measured of the series solution

Concentration (ppm) Absorbance*
4 0.181±0,002
6 0.264±0,008
8 0.379±0,003
10 0.468±0,007
12 0.580±0,006
14 0.672±0,004

*All values are expressed as mean of n=3±standard deviation (SD)

Fig. 2: The graph of SIM calibration curve

Table 4: The results of accuracy test of SIM in PCL-PEG-PCL triblock copolymer micelles

Concentration (%) Theoretical level (µg/ml) Calculated level (µg/ml) Recovery (%)* % RSD
80 8 8.234 102.93±1.32 1.269
100 10 10.078 100.78±0.40 0.398
120 12 12.549 104.58±0.79 0.755

*All values are expressed as mean of n=3±standard deviation (SD)

The precision test results are shown in table 5 and 6 show that the % RSD of the average SIM absorbance obtained was 1.371, 0.418, and 0.786 for the intra-day measurement, while 0.418 for the first day and 0.525 for the second day on the inter-day measurement with a concentration of 10 g/ml. The value of % RSD<2 indicates that the method shows good precision [13, 14].

Table 5: The results of intra-day precision test of SIM in PCL-PEG-PCL triblock copolymer micelles

Concentration (µg/ml) Absorbance* % RSD
8 0.386±0.005 1.371
10 0.478±0.002 0.418
12 0.601±0.005 0.786

*All values are expressed as mean of n=3±standard deviation (SD)

Table 6: The results of inter-day precision test of SIM in PCL-PEG-PCL triblock copolymer micelles

Concentration (µg/ml) Days- Absorbance* % RSD
10 1st 0.478±0.002 0.418
2nd 0.479±0.003 0.525

*All values are expressed as mean of n=3±standard deviation (SD)


The solubility of SIM after being incorporated into PCL-PEG-PCL triblock copolymer micelles was successfully increased by 9.7 times compared to SIM without modification. The Uv-Vis spectrophotometer used to measure dissolved SIM levels has been successfully validated. The validation results show the λmax value of 239 nm, a linear calibration curve with an R-value of 0.9994, LOD and LOQ of 0.33 µg/ml and 1.00 µg/ml, accurate measurement with % recovery at concentrations of 80%, 100%, and 120% were 102.93±1,32%, 100.78±0.40% and 104.58±0.79% and also has a good precision value with RSD<2%.


The authors would like to thank the research grand of Yayasan Notari Bhakti Pertiwi Palembang for funding this research.


All of the authors listed in this manuscript have contributed equally.


The author declares that there is no conflict of interest related to this report.


  1. He G, Ma LL, Pan J, Venkatraman S. ABA and BAB type triblock copolymers of PEG and PLA: A comparative study of drug release properties and ”stealth” particle characteristics. Int J Pharm. 2007;334(1-2):48-55. doi: 10.1016/j.ijpharm.2006.10.020, PMID 17116377.

  2. Murtaza G. Solubility enhancement of simvastatin: a review. Acta Pol Pharm. 2012;69(4):581-90. PMID 22876598.

  3. Jiang T, Han N, Zhao B, Xie Y, Wang S. Enhanced dissolution rate and oral bioavailability of simvastatin nanocrystal prepared by sonoprecipitation. Drug Dev Ind Pharm. 2012;38(10):1230-9. doi: 10.3109/03639045.2011.645830, PMID 22229827.

  4. Rohilla A, Khan MU, Khanam R. Cardioprotective potential of simvastatin in the hyperhomocysteinemic rat heart. J Adv Pharm Technol Res. 2012;3(3):193-8. doi: 10.4103/2231-4040.101018, PMID 23057007.

  5. Yulianita R, Sopyan I, Muchtaridi M. Forced degradation study of statins: a review. Int J Appl Pharm. 2018;10(6):38-42. doi: 10.22159/ijap.2018v10i6.29086.

  6. Verma N. Introduction to hyperlipidemia and its treatment: a review. Int J Curr Pharm Sci. 2017;9(1):6-14. doi: 10.22159/ijcpr.2017v9i1.16616.

  7. Rosyida NF, Pudyani PS, Nugroho AK, Ana ID, Ariyanto T. Solubility enhancement of simvastatin through surfactant addition for development of hydrophobic drug-loaded gelatin hydrogel. Indones J Chem. 2019;19(4):920-7. doi: 10.22146/ijc.38153.

  8. Meor Mohd Affandi MM, Tripathy M, Shah SA, Majeed AB. Solubility enhancement of simvastatin by arginine: thermodynamics, solute–solvent interactions, and spectral analysis. Drug Des Devel Ther. 2016;10:959-69. doi: 10.2147/DDDT.S94701, PMID 27041998.

  9. Essa EA, Dwaikat M. Enhancement of simvastatin dissolution by surface solid dispersion: effect of carriers and wetting agents. J Appl Pharm Sci. 2015;5:46-53.

  10. Borawake PD, Arumugam K, Shinde JV. Formulation of solid dispersions for enhancement of solubility and dissolution rate of simvastatin. Int J Pharm Pharm Sci. 2021;13:94-100. doi: 10.22159/ijpps.2021v13i7.41205.

  11. Pescina S, Sonvico F, Clementino A, Padula C, Santi P, Nicoli S. Preliminary investigation on simvastatin-loaded polymeric micelles in view of the treatment of the back of the eye. Pharmaceutics. 2021;13(6):1-15. doi: 10.3390/pharmaceutics13060855, PMID 34207544.

  12. Shital SP, Rakesh M, Shirolkar SV. Spherical agglomeration a novel approach for solubility and dissolution enhancement of simvastatin. Asian J Pharm Clin Res. 2016;9:65-72.

  13. Sopyan I, Syah ISK, Nurhayti D, Budiman A. Improvement of simvastatin dissolution rate using derivative non-covalent approach by solvent drop grinding method. Int J Appl Pharm. 2020;12:21-4. doi: 10.22159/ijap.2020v12i1.35865.

  14. Sopyan I, Fudhol A, Puspitasari MM I. A simple effort to enhance solubility and dissolution rate of simvastatin using co-crystallization. Int J Pharm Pharm Sci. 2016;8:342-6.

  15. Sulaiman TNS, Patmayuni D, Zulkarnain AK. Optimization of PCL-PEG-PCL triblock copolymer micelles as hydrophobic drug carrier with a 22 full factorial design. Int J Appl Pharm. 2019;11:42-7.

  16. Birari AE. Development and validation of UV spectrophotometric method for estimation of simvastatin in bulk and solid dosage form. Int J Pharm Sci Res. 2015;6:85-9.

  17. Dey S, Pradhan PK, Upadhayay UM, Patel C, Lad B. Method development and validation of simvastatin by UV spectrophotometric method. J Pharm Res. 2012;5:5380-2.

  18. Sharma M, Kaur R, Singh S, Kharb V, Jain UK. Development and validation of Uv spectroscopic method for the estimation of simvastatin. World J Pharm Pharm Sci. 2013;3:763-71.

  19. Panamasha AJ, Tejaswi K, Parimala SS. Spectrophotometric estimation of simvastatin in bulk and tablet dosage form. Int J Innov Pharm Res. 2013;4:284-7.

  20. Sandeep K, Suresh P, Gupta GD. Effect of non-ionic surfactant on the solubility and dissolution of simvastatin. Int Res J Pharm. 2011;2:100-2.

  21. AOAC. Peer verified program, manual on policies and procedures. Washington DC. Arlington; 1993.

  22. Moffat C, Osselton M, Widdop B. Clarke’s analysis of drugs and poisons. 3rd ed. London: Pharmaceutical Press; 2005.

  23. Alami-milani M, Zakeri-milani P, Valizadeh H, Salehi R, Jelvehgari M. Preparation and evaluation of PCL-PEG-PCL micelles as potential nanocarriers for ocular delivery of dexamethasone. Iran J Basic Med Sci. 2018;21(2):153-64. doi: 10.22038/IJBMS.2017.26590.6513, PMID 29456812.

  24. Cho HK, Cheong IW, Lee JM, Kim JH. Polymeric nnanoparticles, mmicelles, and ppolymersomes ffrom aamphiphilic bblock ccopolymer. Kor J Chem Eng. 2010;27:731-40.

  25. Zamani S, Khoee S. Preparation of core-shell chitosan/PCL-PEG triblock copolymer nanoparticles with ABA and BAB morphologies: effect of intraparticle interactions on physicochemical properties. Polymer. 2012;53(25):5723-36. doi: 10.1016/j.polymer.2012.09.051.

  26. Rajeshwar BR, Gatla A, Rajesh G, Arjun N, Swapna M. Polymeric micelles: A nanoscience technology. Indo American J Pharm Res. 2011;1:351-63.

  27. Doolaanea AA, Mawazi SM, Hadi HAB, Al-mahmood SMAMawazi SM, Hadi HAB, Al-mahmood SMA, Doolaanea AA. Development and validation of UV–vis spectroscopic method of assay of carbamazepine in microparticles. Int J Appl Pharm. 2019;11(1):34-7. doi: 10.22159/ijap.2019v11i1.26256.

  28. Prasad AR, Thireesha B. Uv-spectrophotometric method development and validation for the determination of lornoxicam in microsponges. Int J Appl Pharm. 2018;10(1):74-8. doi: 10.22159/ijap.2018v10i1.22357.