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29PHARMACEUTICAL SCIENCES
DEVELOPMENT OF METHODS FOR QUANTITATIVE DETERMINATION OF SILVER AND GOLD IN SEMI-SOLID PREPARATIONS WITH METAL NANOPARTICLES
Bilous S.B.
Danylo Halytsky Lviv National Medical University,
Lviv, Ukraine Sheina T. V.
State Scientific Institution "Institute for Single Crystals " NAS of Ukraine,
Kharkiv, Ukraine Belikov K.M.
State Scientific Institution "Institute for Single Crystals " NAS of Ukraine,
Kharkiv, Ukraine Kalynyuk T.H.
Danylo Halytsky Lviv National Medical University,
Lviv, Ukraine
ABSTRACT
Recently, with the active progress of nanotechnology, studies on the development of medicines based on metal nanoparticles is being carried out. For their further introduction into manufacture, it is necessary to develop quality control methods, in particular quantitative determination, which makes it possible to establish storage conditions and expiration date of the medicines and characterizes compliance with the conditions of the technological process in their manufacture. For the quantitative determination of silver and gold nanoparticles in semi-solid preparations atomic absorption spectrometry with method of standard additions was proposed.
Keywords: silver nanoparticles, gold nanoparticles, medicines, quantitative determination, atomic absorption spectrometry.
Actuality. Pharmaceutical development of medicinal products inextricably linked with methods of quality control. Without the development of quality control methods, in particular such indicator as quantitative determination method, it is not possible to investigate the stability of medicinal products during storage and, accordingly, establish their shelf-life. The quantity of active pharmaceutical ingredients is a key indicator of the medicines stability. Therefore, one of the priority tasks is to develop methods for quality control of medicines with metal nanoparticles, which will enable the intensification of studies on their creation.
The aim. To develop a method for quantitative determination of silver and gold nanoparticles in semisolid preparations.
Materials and methods. Five dosage forms were used for investigation: 3 - with silver nanoparticles -ointment, cream and gel, which could be promising for use in dermatology and 2 based on silver and gold na-noparticles for use in dentistry and surgery for the treatment of infectious, purulent- inflammatory diseases of the skin and mucous membranes of oral cavity.
For quantitative determination of silver and gold nanoparticles atomic absorption spectrometry with method of standard additions was used.
Results and discussion. Metal nanoparticles have been actively investigated in recent years due to their high biological activity. Of particular interest are silver and gold nanoparticles that exhibit antimicrobial [1, 2] and anti-inflammatory [3] effects, and can stimulate tissue regeneration [4, 5, 6].
At the Department of Drug Technology and Bio-pharmaceutics of Danylo Halytsky Lviv National Med-
ical University are conducted studies considering pharmaceutical development of antimicrobial preparations in which metal nanoparticles are used as active pharmaceutical ingredients [7, 8, 9].
The investigated metal nanoparticles obtained by two methods: the method of colloidal-chemical synthesis developed at F.D. Ovcharenko Institute of Biocol-loidal Chemistry and the method of electron-beam technologies developed at E.O.Paton Electric Welding Institute of the National Academy of Sciences of Ukraine [10, 11]. According to the physicochemical properties nanomaterials are nanostructured powders and colloidal solutions.
For further introduction of investigated medicinal products into manufacture it is necessary to develop quality control methods, in particular quantitative determination, which makes it possible to establish storage conditions and expiration date of the medicines and characterizes compliance with the conditions of the technological process in their manufacture.
There is currently no single approach to the analytical determination of metal nanoparticles. For determining metals spectral analysis methods are most often used: atomic absorption, atomic fluorescence, atomic emission with inductively coupled plasma and mass spectrometry with inductively coupled plasma, and others.
The most advanced and highly sensitive method for determining metals is the atomic absorption method due to such advantages as: high selectivity, accuracy and productivity, the ability to automate the analysis process, and availability for a wide range of laboratories [12, 13]. Modern spectrophotometers give an opportunity to define these elements both in the field of
low and high concentrations. Flame atomic absorption method makes it possible to measure the amount of gold and silver in solution within the linearity range from 0.1 mg/l to 20 mg/l, silver - from 0.01 mg/l to 10 mg/l.
In this paper, iCE 3500 spectrometer Thermo Fisher Scientific Inc. equipped with a single-slotted nozzle (length of absorbing layer 50 mm) for reception and registration of atomic absorption of gold and silver was used.
The method of metals determination has peculiarities of application depending on the type of dosage forms. In the case of semi-solid preparations, preliminary dissolution of samples of ointments, creams or gels is required. Sample dissolution was carried out in closed fluoroplastic vessels in a laboratory microwave
The resulting solutions after cooling to room temperature were transferred from a fluoroplastic vessel to a 25 ml volumetric flask and added ultra-pure water to the mark. In the case of gold determination, 0.25 ml of lanthanum (III) chloride solution (100 g/l) was added.
The method of standard additions was used to determine the concentration of silver and gold. Concentration of Ag in calibration solutions (mg/l): 0.4, 0.8, 1.6, Au - 2.0, 4.0, 8.0. In the work, Interstate standard samples of the aqueous solution were used: for Ag 1 mg/ml and Au 0.1 mg/ml. Graduated and analyzed solutions were sequentially sprayed into a flame. The registration of signals, the construction of calibration graphs and the calculation of the concentration of elements in the analyzed solutions were carried out by the software of the spectrometer.
oven MDS-2000 (CEM corporation, USA). The microwave oven equipped with a set of vessels operating at pressure 120 psi and a maximum temperature 220 °C; the power of ultrahigh-frequency radiation varies in the range 0-630 W with increments 50 W; the built-in computer allows to program a five-step process of processing samples, each stage of which does not exceed 60 minutes of operation at maximum values of pressure and temperature.
Samples weighing 0.5 g (exact weighting) were placed in a fluoroplastic vessel, 5 ml of concentrated nitric acid R was added for the determination of silver or 5 ml of a mixture (3:1) of concentrated hydrochloric acid and nitric acidR for the determination of gold. The dissolution was carried out in a microwave oven by the next 4-stage program (Table 1).
Table 1
Measurements were performed on an iCE 3500 spectrophotometer in absorption mode. As a source of spectral excitation, a flame of gas mixture acetylene-air was used. The measurement conditions are given in Table 2.
Table 2
The content of silver (gold) in samples of ointments, creams and gels was calculated according to the formula:
X, (^g / g) = C ■ 100/4 ■ m
C - concentration of silver (mg / l) in the investigated solution;
m - sample weight, g
The results of the study and the metrological characteristics of the method are presented in Table 3. The verification of correctness was performed by the method of varying the weight.
Parameters of microwave disintegration of samples
Stage No. 1 2 3 4
Oven power, % from the maximum 50 50 50 50
Pressure in the vessel, % from the maximum 20 40 60 70
Time, min 15 15 15 15
Hold-time at a given pressure, min 10 10 10 10
Optimal conditions for the determination of silver and gold
Element Analytical Height of The height of the Slit Bulb cur- Correction of
line, nm internal photometric area width, rent,0/« background ab-
cone of over the burner nm sorption
flame, mm base, mm
Ag 328,1 2 5 0,5 90 Deuterium lamp
Au 242,8 2 5 0,5 80 Deuterium lamp
Table З
Results of the study and metrological characteristics of the silver and gold determination in semi-solid __preparations (P=0,95; n=3)___
Dosage form Sample weight, g Silver found, mg/g Relative standard deviation, sr Gold found, VgJg Relative standard deviation, sr
Ointment with silver nanoparticles Q,26 Q,51 1,Q3 2,3±Q,1 2,4±Q,1 2,2±Q,1 Q,Q6 Q,Q5 Q,Q3
Cream with silver nanoparticles Q,25 Q,5Q 1,2 2,Q±Q,1 1,9±Q,1 2,1±Q,1 Q,Q7 Q,Q5 Q,Q4
Gel with silver nanoparticles Q,26 Q,52 1,1 2,6±Q,1 2,8±Q,Q9 2,7±Q,Q9 Q,Q8 Q,Q5 Q,Q5
Ointment with silver and gold nanoparticles Q,25 Q,55 1,2 Q,83±Q,7 Q,85±Q,4 Q,83±Q,4 Q,Q2 Q,Q1 Q,Q1 3Q,Q±Q,6 29,3±Q,3 29,7±Q,3 Q,Q2 Q,Q1 Q,Q1
Gel with silver and gold nanoparticles Q,25 Q,45 1,1 Q,88±Q,2 Q,8Q±Q,4 Q,82±Q,1 Q,Q1 Q,Q1 Q,QQ5 3Q,2±Q,6 29,3±Q,3 3Q,3±Q,3 Q,Q2 Q,Q1 Q,Q1
As can be seen from the results presented in Table 3, the technique of performing the analysis provides the required correctness of the definitions.
Conclusions: 1. For the quantitative determination of silver and gold nanoparticles in semi-solid preparations was proposed to use atomic absorption spec-trometry using the method of standard additions.
2. The correctness of the methods is verified by the method of variation of weight.
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