1. THE LEVEL OF FREE AND BIOAVAILABLE TESTOSTERONE IN MEN AGED ABOVE 40 YEARS OLD
Oyun-Erdene R. ; Nansalmaa N. ; Munkhtsetseg J.
Mongolian Pharmacy and Pharmacology 2013;2(1):11-
Introduction: With the average longevity in men and women, sexual health concerns have become more and more important and demands for help are far more common than in the past. The percentage of aging population is increasing also. A metabolic and hormonal change occurs in male during aging.The level of total, free and bioavailable testosterone decline with aging and it leads to decrease in sexual activities, metabolism and also the life quality.The aim of this initial study was the determination of free testosterone and bioavailable testosterone and it was the novelty of our study. Data obtained from our research can be used as basic information for hormone replacement therapy in late onset hypogonadism.Research goal: To study the free testosterone and bioavailable testosterone level in aging malesMaterials and Methods: This study is a part of study: “Androgen status of aging males” which was supported by Asian Research Center, Korean Foundation for Advanced Studies. The study was approved by IRB of MoH and written consent was obtained from all participants.Fasting blood samples were collected in the morning between 8.00-10.00 AM. We used commercial ELISA kits from Magiwel CoLtd (USA) for determining the total testosterone, sex hormone binding globulin levels. Bromcresol green method was used in determination of serum albumin level. Bioavailable and free testosterone were calculated by Alex Vermeulen, Lieve Verdonk and M. Kaufman’s formula, which was recommended by International Society for the Study of Aging Male.We studied 114 healthy males aged above 40 years old, all undergone the General and Urological examination.Result and discussion: The average age was 57.48±10.48 years in our study participants. In group of 40-49 years were 29% (n=33), in 50-59 age group 23% (n=26), in 60-69 age group 27% (n=38) and in age group over 70-s were 15% (n=17).Mean total testosterone was 6.04±1.83 ng/ml, in 40-49 age group it was 6.14±1.65 ng/ml, in 50-59 age group 6.04±2.36 ng/ml, in 60-69 age group 6.05±1.80 ng/ml, and over 70’s it was 5.85±1.43 ng/ml.Mean sex hormone binding globulin was 50.22±29.97 nmol/l, in 40-49 age group 37.60±23.03 nmol/l, in 50-59 age group 47.08±29.61 nmol/l, in 60-69 age group 57.24±33.91 nmol/l, and over 70’s it was 59.22±25.38 nmol/l.Mean albumin was 40.86±6.89 g/l, in 40-49 age group 44.55±5.93 g/l, in 50-59 age group 41.85±6.93 g/l, in 60-69 age group 38.92±6.85 g/l, and over 70’s was 36.55±4.77 g/l.Mean free testosterone was 0.112±0.064 ng/ml, in 40- 49 age group 0.124±0.058 ng/ml, in 50-59 age group0.114±0.077 ng/ml, in 60-69 age group 0.107±0.072 ng/ml, and over 70’s it was 0.097±0.044 ng/ml.Mean bioavailable testosterone was 2.53±1.48 ng/ ml, in 40-49 age group 2.76±1.37 ng/ml, in 50-59 age group 2.60±1.70 ng/ml, in 60-69 age group 2.51±1.56 ng/ml, and over 70’s it was 2.04±1.05 ng/ml.Conclusion:1. In our participants aged above 40 years old, the average mean of free testosterone was 0.112±0.066 ng/ml, free testosterone index was 16.95±11.82. Free testosterone had inverse correlation with aging (r=-0.168, p=0.03) and had peer decline among aging groups.2. The average mean of bioavailable testosterone was 2.53±1.48 ng/ml, and had age related inverse correlation (r=-0.169, p=0.037), which decline was deeper in men aged over 70 years.Key words:Aging, total, free, bioavailable testosterone,free testosterone index
2.Bioavailable testosterone and age correlation in aging males
Oyun-Erdene R ; Nansalmaa N ; Munkhtsetseg J
Mongolian Medical Sciences 2011;172(2):17-19
Introduction: With the increasing longevity in men and women, sexual health concerns have become more and more important and demands for help are far more common than in the past. The percentage of aging population is increasing also. The of this study in aging men. Late onset hypogonadism will need the testosterone replacement therapy and we hope that our study result will help to get basic information of testosterone among over 40 yearsold Mongolian men.Goal: To determine the bioavailable testosterone (BT) of aging males and correlate with aging process.Materials and Methods: This study is a part of ongoing study: “Androgen status of aging males” which was supported by Asian Research Center, Korean Foundation for Advanced Studies. We studied 114 healthy males aged above 40 years old, all undergone the General and Urological examination. Bioavailable testosterone was determined by formula suggested by ISSAM.Result: The average bioavailable testosterone level was 2.53±1.48 ng/ml, in 40-49 age group 2.76±1.37 ng/ml, in 50-59 age group 2.60±1.70 ng/ml, in 60-69 age group 2.51±1.56 ng/ml, and over 70’s it was 2.04±1.05 ng/ml. If consider the bioavailable testosterone 100%, in 40-49 age than it is decreasing 94.2% in 50-59, 90.9% in 60-69 ages and 73.9% decreased in over 70s. Respectively, it decreases approximately by 0.9% every year after 40’s.Conclusion: The bioavailable testosterone level was 2.53±1.48ng/ml in aging males and has reverse correlation with aging (r=-0.169, p=0.037).
3.Free testosterone and age correlation in mongolian aging males
Oyun-Erdene R ; Nansalmaa N ; Munkhtsetseg J
Mongolian Medical Sciences 2011;155(1):20-22
Materials and Methods: We studied 114 healthy males aged above 40 years old, who undergone urologists examination and General practitioner. All men answered the Aging Males’ Symptom Scale questionnaire. This is part of the ongoing study Mongolian males Andrological Status sponsored by Asian Research center, Korean Foundation for Advenced Studies. Formal concent permission was obtained from all participants, which approved by Ethical Committee of MoH. We took 4 ml blood from vien between 8-11am and determined testosterone, SHBG by ELISA and albumin by liquicolor reagent. Bioavailable testosterone was calculated, using previously described mathematical modeling, suggested by ISSAM.Result: The average free testosterone level was 0.112±0.06 ng/ml, in 40-49 age group 0.124±0.05 ng/ml, in 50-59 age group 0.114±0.07 ng/ml, in 60-69 age group 0.107±0.07 ng/ml, and over 70’s it was 0.097±0.04 ng/ml. If consider the free testosterone 100%, in 40-49 age than it is decreasing 91.6% in 50-59, 83.3% in 60-69 ages and 75% decreased in over 70s. Respectively, it decreases approximately by 0.8% every year after 40’s.Conclusion: The free testosterone level was 0.11±0.06 ng/ml in aging males and has reverse correlation with aging (r=-0.168, p= 0.03).
4.The study of androgen receptor gene cag and ggn polymorphism in relation with androgen status
Munkhtogtokh J ; Buyankhuu T ; Tuul B ; Oyun-Erdene R ; Munkhtsetseg J
Mongolian Medical Sciences 2013;166(4):6-8
Background: Discrepancies in the sensitivity to biological effects of the androgens, exerted through the binding of the hormone to the androgen receptor (AR), may also be involved in the inter-individual variation of T as well as in age related decline. The human androgen receptor (AR), located on chromosome Xq11-12, is a transcription factor regulating the development of male reproductive organs in the fetus and secondary sex characteristics at puberty in response to testosterone (T) and 5a-dihydrotestosterone (DHT). The AR contains two polymorphic regions, the (CAG)nCAA repeat encoding polyglutamine, and the (GGT)3GGG(GGT)2(GGC)n repeat encoding polyglycine, commonly referred to as the CAG and GGN repeats respectively. The aim of this study is to investigate the effect of the human androgen receptor genes CAG and GGN repeat polymorphisms in relation with androgen level.Materials and Methods: Sample collection: 180 male, the medical data of these volunteers were obtained and determined some androgen hormones at first phase of study in 2010-2011 (total testosterone (TT), free testosterone (FT) and bioavailable testosterone (BAT)). To determine CAG/GGN repeats length in exon of androgen receptor gene, using frozen serum as a source of deoxyribonucleic acid (DNA). DNA was extracted from blood samples (200 ml) using High PurePCR Template Preparation Kits.Results: The 180 men whose age is at least 40 were involved in our research and their average age was 55.1±10.3. The 46.7% (84) of the participants presents CAG gene, the 6.1% (11) of the participants presents GGN gene while the 25.5% (46) of the participants presents both CAG and GGN genes. However, the 21.7% of 39 men not presents CAG and GGN genes.Conclusion: The free testosterone level was significantly decreasing with aging. However, the appearance of CAG gene polymorphism was significantly higher in more aged people. Decline of free testosterone level in participants with CAG and [CAG+GGN] combined form was stronger than in people with GGN gene polymorphism and CAG, GGN both undetected people. But the level of bioavailable testosterone was decreasing with aging and the appearance of CAG gene polymorphism (r=-0.425, p=0.01) and [CAG+GGN] combined form (r=-0.491, p=0.028) was also increasing.
5.Some results of studies of “Монгол хунчир” injection
Tsendbadam B ; Lkhaasuren R ; Battulga B ; Baljinnyam I ; Tsetsegmaa S ; Khurelbaatar L ; Munkhtsetseg R
Mongolian Pharmacy and Pharmacology 2021;19(2):93-99
Introduction:
Astragalus is the largest member of the Fabaceae family of about 3,000 species on all continents except Australia, and the Astragalus mongholicus Bunge and the Astragalus membranaceus (Fisch.) Bge are studied and widely used. Astragalus contains polysaccharides, saponins, flavonoids, amino acids and trace elements, so it has a variety of pharmacological effects and is active in supporting the immune system and protecting the liver, heart and kidneys.
Objectives:
A joint research team of the “Tsombo Pharm” Co., LTD and the Drug research Institute is conducting an experiment to produce an injectable solution from Astragalus mongholicus Bunge. The aim is to expand these previous studies to determine the main parameters of the “Монгол хунчир” injection drug technology.
Methods:
The quality of the injection was assessed by the following parameters. These include: appearance, color of the injection solution, mechanical mixture sensing method, solution filling volume method, solution environment potentiometry method, solution clarity comparison method, insoluble particle size microscopy method, heavy metal mixture atomic absorption spectroscopy method and calicosine-7-o-β-d-glycoside content was determined by HPLC.
Results:
According to the results of the study, the injectable drug was weak yellow, clear, free of mechanical impurities and heavy metal content, filling 2 ± 0.001 ml, pH 6.5, insoluble particle size greater than 10 μm, 3 particles per 1 ml, small particles larger than 25 μm were present in 1 ml. Calicosin-7-o-β-d-glycosides were identified in the “Монгол хунчир” injection as having the same standard and sample peak times, with the standard substance being detected at 9.003 minutes and the sample solution at 9.016 minutes (Picture 1, 2). In addition, the injection sample contained 0.0477 ± 0.0021 mg / g of calicosin-7-o-β-d-glycoside, and 0.0451-0.0551 mg / g was considered appropriate for further standardization.
Conclusions
The “Монгол хунчир” injection meets the general requirements for injection in accordance with the Mongolian National Pharmacopeia 2011. This shows that the technological parameters developed by our research team are appropriate.
6.Method validation of hydroxysafflor yellow A in “Carthamus tinctorius” injection by High-performance liquid chromatography
Baljinnyam I ; Tsendbadam B ; Battulga B ; Munkhtsetseg R ; Badamtsetseg S ; Bayanmunkh A ; Tsetsegmaa S ; Lkhaasuren R
Mongolian Medical Sciences 2021;195(1):56-63
Introduction:
Carthamus tinctorius L. widely accepted as Safflower or false saffron, belongs to the Compositae
or Asteraceae family. Hydroxysafflor yellow A is the main active chemical compound present in
florets of Carthamus tinctorius L. A joint research team of the “Tsombo Pharm” Co., LTD and the
Drug research Institute is conducting an experiment to produce a solution of “Carthamus tinctorius”
injection prepared by Carthamus tinctorius L.
Goal :
The aim of this study was to develop the validation method of hydroxysafflor yellow A in “Carthamus
tinctorius” injection.
Material and Methods :
As a test sample “Carthamus tinctorius” injection was produced by “Tsombo pharma” Co., LTD. The
standard Hydroxysafflor yellow A was supplied from Sigma-Aldrich Co., Ltd. The reagent were high-performance liquid chromatography grade acetonitrile, phosphoric acid, methanol and purified water.
Shimadzu HPLC (CMB-20 A, UV detector Shimadzu SPD-20A was used as the analytical instrument
and the analysis conditions were as follows Table 1.
Results:
A Shimpack С18 column was used with methanol:acetonitrile:0.7% phosphoric acid as the mobile
phase under the condition of gradient elution. The hydroxysafflor yellow A were analyzed by using
a timed wavelength measure according to their maximum absorption wavelength. Accuracy and
precision were assessed by analyzing five sets of samples, independently prepared at low (50%)
middle (100%) and high (150%) concentrations. The intraday and interday precisions of the
investigated compound were less than 1.59 % and the average recoveries ranged from 81.9% to
101.5%.
There were good linear correlations between the concentrations of the hydroxysafflor yellow A and
its chromatographic peak areas (R2 = 0.998), the proposed method was successfully applied to
determine the hydroxysafflor yellow A in “Carthamus tinctorius” injection.
Conclusions
The results indicated that the proposed method is simple, stable, and accurate and could be readily
utilized as a quality control method for manufacturing process of “Carthamus tinctorius” injection.
7.Method validation of calycosin-7-O-β-D-glucoside in “Astragalus mongholicus” injection by High-performance liquid chromatography
Tsendbadam B ; Battulga B ; Lkhaasuren L ; Baljinnyam I ; Tsetsegmaa S ; Khurelbaatar L ; Munkhtsetseg R
Mongolian Medical Sciences 2021;195(1):43-50
Introduction:
Calycosin-7-O-β-D-glucoside is a glycosyloxyisoflavone that is calycosin substituted by a beta-D-glucopyranosyl residue at position at 7 via a glycosidic linkage. calycosin-7-O-β-D-glucoside, a
calycosin derivative compound derived from Astragali Radix, has protective effect against ischemia/
reperfusion injury as well as bacterial endotoxin-induced vascular cell injury. A joint research team of
the “Tsombo Pharm” Co., LTD and the Drug research Institute is conducting an experiment to produce
a solution of “Astragalus mongholicus” injection prepared by Astragalus mongholicus bunge.
Goal :
The aim of this study was to develop the validation method of Calycosin-7-O-β-D-glucoside in
“Astragalus mongholicus” injection.
Material and Methods:
As a test sample “Astragalus mongholicus” injection was produced by “Tsombo pharma” Co., LTD.
The starndard Calycosin-7-O-β-D-glucoside was supplied from Xilong Scientific Co., Ltd. The reagent
were high-performance liquid chromatography (HPLC) grade acetonitrile, formic acid, methanol and
purified water. Shimadzu HPLC (CMB-20 A, UV detector Shimadzu SPD-20A was used as the
analytical instrument and the analysis conditions were as follows Table 1.
Results:
The calibration curves for Calycosin-7-O-β-D-glucoside were made by plotting the peak area versus
the concentration for each analyte using regression analysis. Each calibration curve was obtained
using six levels of concentrations in the range 12.5-100µg/ml. The linear correlation coefficient (R2)
for all calibration curves was higher than 0.9981 for all analytes. The limit of detection and limit of
quantitation for Calycosin-7-O-β-D-glucoside were in 10.37 µg/ml and 31.45 µg/ml. Accuracy and
precision were assessed by analyzing five sets of samples, independently prepared at low (50%)
middle (100%) and high (150%) concentrations. The RSD values of both repeatability and intermediate precision were below 0.68% and 0.618% the accuracy remaining between 95.55 to 101.71%. The
resulting accuracy data were satisfactory for the quantitative analysis of Calycosin-7-O-β-D-glucoside
in “Astragalus mongholicus” injection.
Conclusions
Finally, this method can be employed conveniently, reliably and successfully for the estimation
of Calycosin-7-O-β-D-glucoside for routine quality contral and stability studies in “Astragalus
mongholicus” injection.