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Syntax Idea: p–ISSN: 2684-6853 e-ISSN: 2684-883X

Vol.4, No.5, May 2022

ANALYSIS OF DRUG COMPOUNDS IN BIOLOGICAL SAMPLES OF BLOOD PLASMA

Angel Novia Fransiska, Diba Masyrofah, Gita Kurniawati Putri, Lhidya Halizah Malik,
Putri Wulanbirru, Tintia Rafika Putri
Singaperbangsa University Karawang (UNSIKA) West Java, Indonesia
Email: [email protected], [email protected],
[email protected], [email protected],
[email protected], [email protected]

Abstract
The drug compounds used in this research were plasma with amoxicillin, glyburide,
vancomycin, amlodipine, and paracetamol. Chromatography-Mass Spectrometry
(GC-MS). The first compound used was amoxicillin, plasma concentrations were
measured at 250 g/mL and 500 g/mL, and each concentration was repeated 3 times.
Glyburide, plasma concentration was determined with 3 concentrations, namely 1 g/
mL, 2 g/mL and 4 g/mL, and each concentration was repeated 5 times. For
Vancomycin, 4 concentrations were used for determination of plasma concentration,
namely 3 g/mL, 15 g/mL, 31.5 g/mL and 48 g/mL concentrations that were suitable
for determination of amoxicillin, namely the level of 500 g/mL. All concentrations are
suitable for the determination of glyburide. The most accurate method is the liquid
chromatography/mass spectrometry (LC/MS) method, and acetaminophen using the
gas chromatography-mass spectrometry (GC/MS) method.-MS), paracetamol levels
in the blood samples obtained is 175.2 ppm.

Keywords: levels; blood plasma; drug compounds

Abstract
The drug compounds used in this study were plasma with amoxicillin, glyburide,
vancomycin, amlodipine, and paracetamol. Chromatography-Mass Spectrometry (GC-
MS). The first compound used was amoxicillin, plasma concentrations were measured
at 250 g/ mL and 500 g/ mL, and each concentration was repeated 3 times.
Gliburides, plasma concentrations were determined in 3 concentrations, namely 1 g/
mL, 2 g/ mL and 4 g/ mL, and each concentration was repeated 5 times. For Vancomycin,
4 concentrations were used for determination of plasma concentrations, namely 3 g/
mL, 15 g/ mL, 31.5 g/ mL and 48 g/ mL concentrations suitable for determination of
amoxicillin, ie levels of 500 g/ mL. All concentrations were suitable for the determination
of glyburide the most accurate methods were liquid chromatography/ mass spectrometry
(LC/ MS), and acetaminophen using gas chromatography-mass spectrometry (GC/ MS)
method.-MS), levels of paracetamol in blood samples obtained is 175.2 ppm.

Keywords: rate; blood plasma; medicinal compounds

How to cite: Fransiska, A, N., Masyrofah, D., Putri, G, K., Malik, L, H., Wulanbirru, P., Putri, T, R., (2022)
Analysis of Drug Compounds in Biological Samples of Blood Plasma, Syntax Idea, 4(5), https://doi.org/
10.36418/syntax-idea.v4i5.1834
E-ISSN: 2684-883X
Ridwan Institute
Published by:
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Angel Novia Fransiska, Diba Masyrofah, Gita Kurniawati Putri, Lhidya Halizah Malik,
Wulanbirru's daughter, Tintia Rafika Putri

Received: 2022-04-22; Accepted: 2022-05-05; Published: 2022-05-11

Introduction
Validation of the analytical method is a process that ensures the analysis is accurate,
specific, reproducible and resistant to the range of analytes to be analyzed and is appropriate for
the desired use (Fauziah et al., 2017). Several analytical parameters that must be considered in
validation of analytical methods are described and defined as well as how they are determined.
These parameters include accuracy, precision, selectivity, linearity and range, detection limit (LOD)
and quantity limit (LOQ), method robustness, method strength (Nyoman et al., 2015). An analytical
method can be used if it has been validated and adapted to laboratory conditions and available
equipment, even though the method to be used has been published in an official journal or textbook
(Uno et al., 2015).

Biological matrices that are often used to analyze levels of drugs or other compounds are
serum, urine, blood and saliva. Biological matrix (mainly blood) is obtained invasively through a
vein (Venipuncture) (Supandi, 2015).
Chromatography is a mixture separation technique based on differences in the distribution of
components in the mobile phase and stationary phase (Rizalina et al., 2018).
Examination of drug levels in blood plasma is an appropriate method for ensuring drug
quality and optimizing drug use in pharmaceutical services, therefore a valid analytical method is
needed to determine the levels of drug compounds in blood plasma (Tania et al., 2016). The drug
compounds used are different, while the biological samples are the same, namely blood plasma
using the High Performance Liquid Chromatography method for determining Amoxicillin,
Glibenclamide, vancomycin, Gas Chromatography for determining Amlodipine, and Gas
Chromatography-Mass Spectrometry (GC-MS) for Paracetamol.

The first drug compound, Amoxicillin, is a semisynthetic penicillin antibiotic that has a B-
lactam ring, usually used in phenomia therapy (Sofyani et al., 2018). Amoxicillin is an antibiotic
whose concentration in the blood is very small so a sensitive, selective and valid analytical method
is needed for analysis
(Sofyani et al., 2018). According to the Food and Drug Administration (FDA), bioequivalence
testing is one of the drugs that must be tested. This bioequivalence test relates to the quality of the
drug in blood plasma, which is important to carry out to ensure that the drug being tested has
efficacy and safety for patients (Nyoman et al., 2015),
(Tania et al., 2016).
The second drug compound, Glibenclamide, is an antidiabetic drug from the sulfonylurea
class which is difficult to dissolve in water (Rohayati et al., 2015). One of the classes of oral
antidiabetics that is often used is the glibenclamide group or often called glyburide which is
included in the sulfonyl urea class of drugs (Tresnawati & Saputri, 2017). Glibenclamide works by
stimulating insulin release by inhibiting the attachment of sulfonyl urea receptors in Langhears islet
ÿ cells.

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Analysis of Drug Compounds in Biological Samples of Blood Plasma

and finally causes a voltage to open calcium channels which ultimately results in an increase in
intra-cell calcium. Analysis of glibenclamide in biological fluids can reportedly be carried out using
High Performance Liquid Chromatography (HPLC), ultraviolet-visible spectrophotometry, capillary
electrophoresis, and liquid chromatography mass spectrometry (Tresnawati) . & Saputri, 2017).

The third drug compound, Vancomycin is one of the first-line antibiotic therapies that is
widely indicated in invasive multi-resistant gram-positive bacterial infections, especially in
methicillin-resistant Staphylococcus aureus (MRSA) in many countries, vancomycin is used for
injection complications such as meningitis, pneumonia, osteomyelitis and endocarditis (Suardi et
al., 2016), (Wibowo, Maulidina, Fitri, & Ningrum, 2019). When compared to other antibiotics,
vancomycin has a narrow therapeutic index with a high degree of pharmacokinetic variability.
Several methods for determining vancomycin levels in human plasma use High Performance
Liquid Chromatography (HPLC) with a UV fluorescence detector, and mass spectrometry (Wibowo,
Maulidina, Fitri, & Ningrum, 2019).

The fourth drug compound, Amlodipine, is a frequently used anti-hypertension (lowering

high blood pressure) drug, which works by relaxing blood vessels (Zulfiah, 2019). Users of this
drug are usually dependent and cannot stop using the drug. The use of this drug is usually lifelong.
Amlodipine provides pharmacological effects as an antihypertensive agent with a Calcium Channel
Blocker (CCB) mechanism of action. Analysis of amlodipine in blood plasma is aimed at
Therapeutic Drug Monitoring (TDM) (Alawiyah & Mutakin, 2017).

The fifth drug compound, paracetamol, is a class of non-opioid analgesic drugs that are
sold freely (Sudarma & Subhaktiyasa, 2021b). Paracetamol is used to relieve mild or moderate
pain and mild fever conditions
(Octaviana et al., 2019). This drug is a relatively safe analgesic choice if consumed correctly
according to the instructions for use. The purpose of reviewing this journal is to determine the
levels of several drug compounds in blood plasma.

Research methods
This journal review uses primary references in the form of journals and is sourced from
Google Scholar which has been published and is trusted. The method used in the literature review
is to collect several journals about determining the levels of drug compounds in blood plasma.

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Angel Novia Fransiska, Diba Masyrofah, Gita Kurniawati Putri, Lhidya Halizah Malik,
Wulanbirru's daughter, Tintia Rafika Putri

Results and Discussion

A. Results

Medicinal Compounds Method Results

Amoxcillin High Performance Liquid Chromatography 500 µg/mL with a

(KKT) retention time of 2.71
minutes

Glibenclamide Resolution 2.35 with a

retention time of 1.14
minutes and 2.5 with a
retention time of 1.15
minutes. (>1.5)

Vancomycin 3 µg/mL with a retention

time of 5,613 to 5,811
minutes

Amlodipine Liquid Chromatography/Mass 0.5 µg/mL

Spectrophotometry (LC/MS)

Paracetamol Gas Chromatography- Mass 175.2 ppm.

Spectometry (GC - MS)

B. Discussion

Determination of plasma levels used in this journal was carried out using 2 different
concentrations. For the first compound Amoxicillin, blood levels were determined using concentrations
of 250 µg/mL and 500 µg/mL, each concentration was repeated 3 times. For a concentration of 250
µg/mL, the average peak area was 441847.33, the average retention time was 2.76 and the average
measured concentration was 1.67. Meanwhile, for a concentration of 500 µg/mL the average peak
area was 2069980.33, the retention time was 2.71 and the measured concentration was 3.88.

Previously, validation of the determination of Amoxicillin levels in plasma in Vitro was carried out
using a validated method, and showed that Amoxicillin had very low levels, and therefore high
measurable levels were needed so that Amoxicillin in plasma could be measured. And it can be
proven that the concentration that has a high measurable level value is the second concentration,
namely 500 µg/mL, which means that the level suitable for determining amoxicillin is the level at a
concentration of 500 µg/mL (Nyoman et al., 2015).

Meanwhile, for the second compound, namely Glibenclamide, blood levels were determined
using 3 concentrations, namely, 1 ÿg/mL, 2 ÿg/mL and 4 ÿg/mL, each concentration was repeated 5
times. The precision values obtained from each concentration sequentially were 16.90%, 10.92%
and 4.91%, while for SPE-MIP MAA it was 1.95%; 4.54%; and 4.56%.

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Analysis of Drug Compounds in Biological Samples of Blood Plasma

The accuracy values for each concentration are 95.99%, 99.2%, and 105.17% respectively.
Meanwhile for SPE-MIP MAA is 92.28%; 106.02%; and 97.39%. To determine the selectivity
of the method used, it can be seen from the separability (resolution) of the two peaks. From
the discussion, it can be seen that the three concentrations are suitable or can be used in
determining levels of Glibenclamide combined with Metformin in the blood. It is said that
because the resolution value obtained meets the requirements, namely >1.5, which can be
seen from the resolution results, namely 2.35 with a retention time of 1.14 minutes and 2.5
with a retention time of 1.15 minutes (Rohayati et al. al., 2015).

Meanwhile, the third compound, namely Vancomycin, was determined in blood levels
using 4 concentrations, namely, 3 ÿg/mL, 15 ÿg/mL, 31.5 ÿg/mL and 48 ÿg/mL, each
concentration was tested 5 times. The results of this HPLC method obtained a CV value of
1.21% at a concentration of 3 ÿg/mL with a retention time of 5,613 to 5,811 minutes. Then,
accuracy and precision testing of vancomycin levels was also carried out, the results of
accuracy and precision testing of the levels intra-day vancomycin was found to be close to
known levels and tests from each replication were close to each other. In addition to intra-
day accuracy and precision testing, Vancomycin levels were also tested for inter-day accuracy
and precision. In the inter-day test (between run) it was obtained and stated that the % diff
and CV values met the requirements, namely less than 15%. At a concentration of 48 ÿg/mL,
a % diff was obtained with a range of -13.65% - 13.20% and a CV of 5.97%. Which means
that in the Vanskomin test in spiked plasma between days, the replication of the level value
is close to other levels, namely at a concentration of 48 ÿg/mL (Wibowo, Maulidina, Fitri, &
Dwi, 2019).

Next, namely the fourth compound, Amlodipine binds 98% to plasma proteins,
therefore causing the levels in it to be small and a very accurate analysis method is needed.
The methods used are Ultra Performance Liquid Chromatography-Electrospray Ionization
Mass Spectrometry and Liquid Chromatography/ Mass Spectrophotometry instrumentation.

(LC/MS). Of these two methods, the most accurate is the Liquid Chromatography/ Mass
Spectrophotometry (LC/MS) method because it is able to detect smaller concentrations so it
is used in the analysis of amlodipine in blood plasma fluid (Alawiyah & Mutakin, 2017).

The last compound is Paracetamol. Qualitative analysis of paracetamol levels in

blood plasma can be seen from the chromatogram in the form of RT (Retention time) which
is the same or close to the paracetamol standard which shows that the paracetamol
compound was detected. The retention time value for blood plasma samples is 15,056. In
analyzing paracetamol levels in blood plasma using the Gas Chromatography-Mass
Spectrometry (GC-MS) method, the level of paracetamol in blood samples was 175.2 ppm.

(Sudarma & Subhaktiyasa, 2021a).

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Angel Novia Fransiska, Diba Masyrofah, Gita Kurniawati Putri, Lhidya Halizah Malik,
Wulanbirru's daughter, Tintia Rafika Putri

Conclusion
Blood levels of amoxicilin were determined using concentrations of 250 µg/mL and 500 µg/mL,
the concentration suitable for determining amoxicilin was 500 µg/mL. Determination of glibenclamide
levels in blood uses concentrations of 1 ÿg/mL, 2 ÿg/mL and 4 ÿg/mL, all concentrations are suitable
for determining glibenclamide levels. Determination of vanskomine levels in blood uses concentrations
of 3 ÿg/mL, 15 ÿg/mL, 31.5 ÿg/mL and 48 ÿg/mL, levels which are close to other levels, namely at a
concentration of 48 ÿg/mL. Amlodipine uses Ultra Performance Liquid Chromatography-Electrospray
Ionization Mass Spectrometry and Liquid Chromatography/ Mass Spectrophotometry (LC/MS)
instrumentation methods . The most accurate method is the Liquid Chromatography/ Mass
Spectrophotometry (LC/MS) method, because it is able to detect smaller concentrations so it is used in
the analysis of amlodipine in blood plasma fluid. Paracetamol using the Gas Chromatography-Mass
Spectrometry (GC-MS) method, obtained paracetamol levels in blood samples, namely 175.2

ppm.
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Syntax Idea, Vol.4, No.5, May 2022 911

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Angel Novia Fransiska, Diba Masyrofah, Gita Kurniawati Putri, Lhidya Halizah Malik,
Wulanbirru's daughter, Tintia Rafika Putri

Copyright holder:
Angel Novia Fransiska, Diba Masyrofah, Gita Kurniawati Putri, Lhidya Halizah
Malik, Putri Wulanbirru, Tintia Rafika Putri (2022)

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Idea Syntax

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912 Syntax Idea, Vol.4, No.5, May 2022