C30 Chromatographic Packing Separates High-Purity Coenzyme Q10 Test

 Coenzyme Q10 has the functions of improving human immunity, enhancing antioxidant, delaying aging and increasing human vitality, and is also widely used in health products or food additives. According to the requirements of the 2020 edition of the Chinese Pharmacopoeia [1] and the United States Pharmacopoeia USP28-NF23 [2], the total content of coenzyme Q10 should not be less than 98%; and the separation of coenzyme Q10 can meet the requirements after extraction, silica gel chromatography and crystallization, and the current annual production can reach the level of several hundred tons. However, the subsequent European Pharmacopoeia 8.8 and later editions, in addition to the main content of not less than 98.0%, increased the level of single heterogeneous content of not less than 0.1% [3].

 

In the present study, we tried to isolate the impurities by the traditional method, and found that the other impurities could meet the requirements after two silica gel chromatography and crystallization, but one of the impurities was difficult to be reduced to the level; the relative retention time of the impurity was about 1.45 by using the method of 2020 edition of the Chinese Pharmacopoeia, and the literature reported that this impurity was called coenzyme Q11 [4].

 

The aim of this paper is to establish a safe and energy-saving method for the purification of coenzyme Q10 by preparative liquid chromatography. Combined with the chromatographic analysis method of Pharmacopoeia, the most probable process for the separation and removal of coenzyme Q11 is reversed-phase chromatography (RPCC). The patent "A purification method of coenzyme Q10" (Patent No. 201810233454.1) reported that octadecyl- or octyl-bonded silica gel was used as the stationary phase, and acetone/methanol or acetone/ethanol was used as the mobile phase, and the whole system was separated in a heated state, with the volume of the sample above 10% and the recovery rate above 80%. The whole system must be separated at 45 ℃, which consumes too much energy for large-scale industrial production, increases the cost of equipment and the complexity of the system, and raises the risk of explosion or fire of organic solvents under pressure.

 

After analysis, the sample in acetone solubility is good, but in the C18 packing or C8 retention is poor, after adding a certain amount of methanol, the solubility of the sample is significantly reduced, so the use of heating to solve the problem of solubility. In this study, it was concluded that if the length of the carbon chain was increased, the retention of the sample in the stationary phase would be relatively increased, and the retention could be reduced by decreasing the proportion of methanol in the mobile phase, so that the separation of coenzyme Q11 could be realized at ambient temperature or a little higher than ambient temperature.

 

1 Materials and Methods

1.1 Reagents and Consumables    

Coenzyme Q10 crude (Batch No. 20210603C, provided by North China Pharmaceutical); Coenzyme Q10 control (Batch No. 20220112, homemade); Methanol (Batch No. 20200403, analytically pure, Komeo); Acetone (Batch No. 20200105, analytically pure, Concord); Chromatographic packing C30, 30 μm, 105Å (Batch No. NS30A12B210903, Bojun Bio); Biorain C18 (5 μm, 100Å, 4.6 × 150 mm, Bojun Bio); Chromatographic column Biorain C18 (5 μm, 4.6 × 150 mm, Bojun Bio). NS30A12B210903, Biorain biological); chromatographic column Biorain C18 (5 μm, 100 Å, 4.6 × 150 mm, Biorain biological).

 

1.2 Instruments   

An Agilent 1200-phase liquid chromatograph (with degasser, high-pressure dual-pump system, autosampler, column oven, DAD detector); TF-Prep liquid chromatography (100 mL-min-1 dual-pump system, 10 mL-min-1 high-pressure injection pump, 6-channel fraction collector); and a 100,000-part balance (EX125DZH, Ohaus Instruments Co., Ltd.) were used for the analysis of the analytes.

 

1.3 Experimental Methods

1.3.1 Sample Dissolution

Weighing coenzyme Q10 crude about 20 g, with about 160 mL of acetone ultrasonic dissolution, and then add about 40 mL of methanol, mixing, and then placed at room temperature away from light, the sample concentration of 100 g - L -1 .

 

1.3.2 Column Packing

Take about 50 g of C30 bonded packing and fill it into the 21.2 × 250 mm column tube by squatting: add the packing into the column tube one by one with a medicine spoon, 1 ~ 1.5 g at a time, squatting, and then add packing until the packing surface no longer falls; hang off the excess packing with a scraper, clean off the excess packing on the outer edge of the column tube, fasten the sieve plate with a polytetrafluoroethylene washer, weigh the weight of the packed material, and tighten the stainless steel screw threads with a wrench. Stainless steel threads, chromatographic column loading is complete, labeled mobile phase inflow direction for the opposite direction of filling.

 

1.3.3Column Equilibrium

The column was connected to a preparative liquid chromatography system and equilibrated with acetone/methanol (4/1, v/v) at a flow rate of 15 mL-min-1 for about 20 min.

 

1.3.4 Coenzyme Q10 Isolation

The sample was injected with the injection pump at a flow rate of 13~20 mL - min - 1, and the injection volume was set so that the sample volume was in the range of 5%~7%. The injection volume was recorded and the sample ratio was calculated; then acetone/methanol (4/1, v/v) was used as the mobile phase, and the flow rate was set at 15 mL - min - 1, and the detection wavelength was set at 275 nm for the DAD detector; the fractions were collected in full collection mode, and one fraction was collected every 5 min; the analysis was performed according to the conditions of Coenzyme Q10 in the 2020 Chinese Pharmacopoeia [4], and the percentage of impurity Coenzyme Q11 was determined by area normalization.

 

Fractions were collected in full collection mode, one fraction every 5 min; the chromatographic conditions of coenzyme Q10 in the 2020 edition of the Chinese Pharmacopoeia [4] were used to determine the percentage of the impurity coenzyme Q11 by area normalization, and the fractions with the content of coenzyme Q11 less than 0.1% were combined. Under the same operation mode, the preparative chromatographic separation was carried out with different sample volumes, and the qualified fractions were collected.

 

1.3.5 Post-processing

The qualified fractions were combined, and then concentrated to about 25% by volume under reduced pressure spinning at 30~35 ℃. The solution was transferred to a beaker, cooled at room temperature and allowed to stand for 18~24 h. The solid was then filtered through a sand-core funnel and allowed to air dry at room temperature for 18~24 h. The recoveries were calculated by weighing the solid, and solvent consumption was calculated according to the separation times of different sample volumes.

 

2.Results

After the above process, the typical separation liquid chromatogram was shown in Figure 1; the analytical analysis of the purified coenzyme Q10 fractions was carried out, and the typical analytical analysis of the qualified samples was shown in Figure 2; the separation recoveries and solvent consumptions of coenzyme Q10 under different sample sizes were shown in Table 1.

 

Table 1 Recovery and solvent consumption of coenzyme Q10 at different sample volumes.

 

Packing volume Sample volume Sample ratio Recovery rate				Coenzyme Q11 ( % )	Runtime solvent consumption
( g)	( g)	Example ( % )	( % )
					( min)	( L)
46.6	2.71	5.82	87. 31	0.04	160	2.40
	3.05	6.55	72.44	0.07	168	2.52
	2.45	5.26	88. 65	0.05	157	2.36

 

3.Discussion

3.1 Sample Analysis

Fractions were analyzed with reference to the 2020 edition of the Chinese Pharmacopoeia for Coenzyme Q10 chromatographic conditions. The received fraction contained acetone, which had strong absorption at 275 nm, so it was necessary to take the fraction and dissolve it with anhydrous ethanol after blowing dry under nitrogen gas to a concentration of about 0.5 mg -mL-1, and injected into the sample for 20 μL; the mobile phase was methanol-anhydrous ethanol (1∶1); the column temperature was set at 35 ℃; the detection wavelength was 275 nm; the flow rate was 1.5 mL - min-1; the method could separate the main components from the key impurities, and the separation degree was more than 2.0; in addition, the impurities were all related to coenzyme Q10, and the separation degree was more than 2.0, and the impurities were all related to coenzyme Q10. This method can separate the main components and key impurities, and the separation degree is more than 2.0; in addition, the impurities are similar to the structure of coenzyme Q10, only the number of polymerized isoprene is different, so the corresponding intensity of UV is not much different, and it can be used as a method for the detection of the impurity content.

Fig. 1 Preparative liquid chromatogram of coenzyme Q10 separated by C30 at 275 nm (the red box is the qualified fraction collection window).

Fig. 2 Chromatogram of qualified fraction samples

(14.606 min and 20.186 min for coenzyme Q10 and coenzyme Q11, respectively, with area percent contents of 99.82% and 0.043%, respectively)

 

3.2 Selection of Preparation Conditions   

Based on the patent "a method of purification of coenzyme Q10" (invention patent No. 201810233454.1), the separation conditions of reversed-phase chromatography (RPCC) are C18-bonded silica gel as the chromatographic packing material, methanol-acetone (4:1) as the mobile phase and dissolved samples as the solvent, and the separation is carried out under the condition of about 45 ℃, and the separation effect of the process has been measured, and the effect of the process can be achieved, and coenzyme Q11 can be separated from the sample.

 

In this study, the separation effect of this process was also tested, and it was found that the separation and removal of coenzyme Q11 could be achieved. However, the heating method requires additional electric power and heat source, which is a safety hazard in general laboratory operation, and the same problem exists in industrial production.

 

Considering that coenzyme Q10 is soluble in acetone but poorly soluble in methanol, and that the increase of acetone content increased the solubility but the retention of coenzyme Q10 in C18 was relatively weak, we used to increase the carbon chain of the bonded silica gel from C18 to C30, which is because C30 is more nonpolar and the retention of coenzyme Q10 is relatively better, and the increase of the acetone content could achieve the isolation of coenzyme Q10 in the ambient temperature. This is because C30 is more non-polar and retains coenzyme Q10 better. After adjustment, it was found that the retention time of coenzyme Q10 was 45~120 min when the ratio of acetone to methanol was 4∶1; this retention time was very favorable to increase the sample volume and improve the separation effect.

 

3.3 Selection of Sample Volume   

With reference to the patent requirements, we compared the sample volume of 5.2%, 5.8% and 6.6% of the filler volume, and found that the recovery of qualified samples decreased with the increase of the sample volume, this is due to the overloading of the sample volume sample retention will be significantly reduced, the enrichment of coenzyme Q11 in the immobilization of coenzyme Q10 in a competitive relationship between coenzyme Q11 and coenzyme Q10 has become more obvious, that is, the peaks of Q11 broadened, resulting in a reduction in the qualified sample fraction.

 

Considering that the increase of sample volume from 5.8% to 6.6% resulted in a significant decrease of recovery from 87% to 72%, while the increase of sample volume from 5.2% to 5.8% only resulted in a decrease of recovery from 89% to 87%, we considered that the optimal volume of sample was 5.8%; although the volume of sample was only 50% compared with 10% of the volume of sample of the method reported in the patent, we still considered that this method of separation was relatively easy to operate at room temperature. Although the sample size is only 50% compared with the 10% sample size of the patent method, we still think that this method is relatively safe and energy-saving at room temperature, and it is also an alternative separation process.

 

References:

[1] National Pharmacopoeia Commission, ed. Chinese Pharmacopoeia (2020 Edition, Part II) [S]. Beijing: China Pharmaceutical Science and Technology Press ,2020 :1459.

[2] European Pharmacopoeia, ed. Ubidecarenone [S]:USP38 NF33 :6243.

[3] European Pharmacopoeia Editorial Committee Ubidecarenone [S]: EUROPEAN PHARMCO- POEIA 8.8 :6065.

[4] Huang Yuqing. Research on the extraction process of coenzyme Q10 [D]. Master's thesis of Zhejiang University ,2017.

 

 

Mechanism of Algal Blue Protein in Reducing Myocardial Ischemia-Reperfusion Injury in Rats

Abstract: Objective: To observe the effects of phycocyanin on myocardial ischemia-reperfusion injury in rats and analyze the possible mechanisms. Methods Thirty-six rats were selected and divided into blank control group, model group and treatment group by random number table method. The model and treatment groups were modeled by repeating silk ligation 3 times and reperfusion of the anterior descending branch of the left coronary artery for 45 min. After modeling, the treatment group was treated with 10% alginate suspension at 200 mg/(kg - d) by gavage for 7 days, and the blank control group and the model group were treated with an equal volume of saline by gavage. Rat coronary artery cell cycle protein-dependent kinase inhibitor p27 and p21 were detected by image analysis; inducible nitric oxide synthase (iNOS) was detected by immunohistochemical staining; and intracardiac artery serum superoxide dismutase (SOD), creatine kinase isoform (CK-MB), malondialdehyde (MDA), and nitric oxide (NO) were detected by enzyme-linked immunosorbent assay. ).

 

Results: In the treatment group, the rat coronary artery cell cycle protein-dependent kinase inhibitor p27 and p21 were highly expressed, iNOS-positive cells and the levels of SOD and NO were elevated, and the levels of CK-MB and MDA were significantly reduced, and the differences were statistically significant when compared with that of the model group (P＜0.05). Conclusion Alginin has the effect of inhibiting myocardial ischemia-reperfusion injury.

 

The main cause of myocardial ischemia-reperfusion injury is not ischemia per se, but rather the massive production of oxygen free radicals after re-flow, and these lipid peroxidation products often cause irreversible damage to cardiomyocytes, which is also the main cause of cardiac mortality [1-2]. It has been shown that the coronary cell cycle protein-dependent kinase inhibitors p27 and p21 play a key role in the regulation of ischemic myocardial tissue remodeling [3-4].

 

In this process, nitric oxide (NO) is also involved in the migration, proliferation and neovascularization of vascular endothelial cells[5] . As the NO rate-limiting enzyme, changes in the activity of inducible nitric oxide synthase (iNOS) will affect NO production and indirectly regulate vascular endothelial function[6] . Thus, any factors affecting cell cycle protein-dependent kinase inhibitors p27, p21, and iNOS can improve or worsen myocardial ischemia-reperfusion injury. In view of this, our group selected Spirulina extract phycocyanin, which has the ability to inhibit lipid peroxidation and antioxidant activity, as a research target and observed its effects on myocardial ischemia-reperfusion injury in rats.

 

1 Materials and Methods 1 .1 Materials

1 .1 .1 Animals and groupings  

Thirty-six male rats, SPF grade, 5 weeks old, weighing 150.0 ± 10.5 kg were selected. The rats were purchased from Wuhan Jintangxin Breeding Technology Co., Ltd [Certificate of Conformity: SCXK (E) 2014-0015] and kept in the SPF laboratory of Hubei Medical College [Certificate of Conformity: SYXK (E) 2018-0031]. The rats were divided into blank control group, model group and treatment group, each with 12 rats after being numbered by random number table before modeling. The experiments were conducted in accordance with the 3R principle to provide animals with humane care and fully satisfy their welfare: during the experiments, the animals were allowed to eat and drink freely, and the temperature of the incubation room was controlled at 22-26 ℃, the relative humidity was controlled at 50%-75%, and the animals were exposed to light and darkness alternately every 12 hours.

 

1 .1 .2 Major equipment and reagents  

BW-BM1103 small animal respirator and integrated informatization signal acquisition and processing system were purchased from Chengdu Taimeng Software Co. Mouse anti-p27 and p21 polyclonal antibodies were purchased from Wuhan Three Eagles Biotechnology Company Limited (Lot No. LS-C211591 and LS-C211597, respectively). iNOS, NO, superoxide dismutase (SOD), creatine kinase isoform (CK-MB) and malondialdehyde (MDA) test kits were purchased from Nanjing Jianjian Institute of Biological Engineering (Lot No. LZ087), (batch numbers: LZ087, H1098, H1012, H1054, H1104).

 

1 .2 Experimental Methods

1 .2 .1 Modeling and Treatment  

The model and treatment groups were anesthetized with 3% sodium pentobarbital 30 mg/kg intraperitoneally. After the rats were anesthetized, they were fixed on the operating table and connected to the BW-BM1103 small animal ventilator and the BL-420S Biofunctional Experiment Recording System (to record the Ⅱ-lead electrocardiograms), and then underwent open-heart surgery. After the open-heart surgery, the anterior descending branches of the left coronary artery were located, and the left ventricle was blocked with a silk thread to cause acute ischemia, 45 min later, the silk thread was released and reperfused for 45 min, which was repeated three times. After 45 min, the wire was loosened and reperfused for 45 min, and this procedure was repeated for three times, and the successful replication of the model was based on the appearance of presystole, T-wave elevation, and ST-segment elevation or upward shift in the Ⅱ-lead electrocardiogram after reperfusion [7]. After the model was completed, the chest cavity was closed, sutured in layers, sterilized, and the animals were kept separately. The blank control group was not anesthetized and did not undergo the above procedures. Immediately after modeling, the treatment group was treated with 10% alginate suspension at 200 mg/(kg - d) by gavage for 7 days, while the blank control group and the model group were treated with equal volume of saline by gavage.

 

1 .2 .2 Detection indicators and methods  

Coronary artery cell cycle protein-dependent kinase inhibitor p27 and p21 were detected by image analysis; iNOS expression and the number of positive cells were detected by immunohistochemical staining; and the levels of SOD, CK-MB, MDA and NO were detected by enzyme-linked immunosorbent assay.

1.3 Statistical processing The data were analyzed using SPSS21.0 statistical analysis software. Measurement data were expressed as x ± s. Comparison of group means was performed by one-way analysis of variance (ANOVA), and comparison of group means was performed by two-sample paired t-test, with P < 0.05 indicating that the differences were statistically significant.

 

2 Results

2 .1 Cell cycle protein-dependent kinase inhibitory factors in rats of all groups

Comparison of the expression levels of p27 and p21 The rats in the model group expressed low levels of cell cycle protein-dependent kinase inhibitory factors p27 and p21, and the differences were statistically significant compared with those in the blank control group (P<0.05). The rats in the treatment group expressed high levels of cell cycle protein-dependent kinase inhibitory factors p27 and p21, and the differences were statistically significant (P<0.05) when compared with the model group. The differences were statistically significant (P < 0.05) compared with the model group.

 

2.2 Comparison of iNOS and NO levels in rats of each group  

The levels of iNOS and NO in the model group were lower than those in the blank control group, and the differences were statistically significant (P<0.05). The levels of iNOS and NO in the treatment group were significantly higher than those in the model group, and the differences were statistically significant (P < 0.05). See Table 2.

 

2.3 Comparison of myocardial enzyme indexes of rats in each group SOD level of rats in the model group was lower than that of the blank control group, while CK-MB and MDA levels were significantly higher than that of the blank control group, and the differences were statistically significant (P< 0.05); SOD level of rats in the treatment group was higher than that of the model group, while CK-MB and MDA levels were significantly lower than that of the model group, and the differences were statistically significant.

(P < 0 .05). See Table 3 .

 

3 Discussion

In the research of myocardial ischemia-reperfusion injury, in addition to targeted basic research on pathophysiology and physiology, medical practitioners should also devote themselves to searching for or screening highly effective and low-toxicity drugs for the treatment or prevention of ischemia-reperfusion injury. Phycocyanin, a photosynthetic pigment protein extracted from Spirulina, was first found to have many pharmacological effects in animal experiments, such as antioxidant, reducing oxidative stress, maintaining homeostasis, detoxification, anti-inflammatory, anti-allergic, anti-tumor and so on[8-10] . In the present study, based on its ability to scavenge hydroxyl radicals and hydroperoxyl radicals, we analyzed its mechanism of action to reduce the effects of myocardial reperfusion injury by observing the factors that can reflect the remodeling of myocardial tissue in myocardial ischemia-reperfusion injury.

 

The results of this study showed that coronary artery p27 and p21 were highly expressed in the treatment group, and the levels of SOD, iNOS, and NO were significantly higher than those in the model group, and the levels of CK-MB and MDA were significantly lower than those in the model group. It has been shown that cell cycle protein-dependent kinase inhibitor p21 and p27 can regulate the proliferation cycle of myocardial and vascular smooth muscle cells, and the high expression of cell cycle protein-dependent kinase inhibitor p27 and p21 can promote the proliferation of myocardial and vascular smooth muscle, thus affecting the reconstruction of damaged myocardial tissues after myocardial ischemia/reperfusion.11-12 NO is involved in this process, and NO is also involved in the reconstruction of damaged myocardial tissues after myocardial ischemia/reperfusion. In this process, NO is involved in the migration and proliferation of vascular endothelial cells, and is an important initiator of myocardial tissue remodeling after ischemia-reperfusion[13] .

 

Under normal physiological conditions, NO and iNOS remain in homeostasis, but when myocardium suffers ischemia-reperfusion injury, iNOS activation is increased, and its high expression induces NO synthesis, which plays a role in regulating vascular endothelial function to promote the remodeling of damaged myocardial tissues[14-15] . From the results of the present study, phycocyanin acts on this link and promotes the expression of cyclin-dependent kinase inhibitors p27 and p21 and iNOS in ischemia-reperfusion myocardial cells, and this effect directly induces NO synthesis, which regulates vascular endothelial function and plays a key role in ischemia-reperfusion myocardial tissue remodeling. In addition, this study also observed that alginate has the effect of scavenging free radicals and inhibiting lipid peroxidation, which is reflected in the increase of SOD activity in the treatment group and the decrease of the levels of CK-MB, a product of cell death, and MDA, a product of lipid peroxidation, in the ischemia-reperfusion group compared with that in the model group.

 

In conclusion, the mechanism by which phycocyanin reduces myocardial ischemia-reperfusion injury in rats may be related to its ability to promote the expression of cell cycle protein-dependent kinase inhibitors p27 and p21 and to enhance the synthesis of iNOS-induced NO, which can regulate the function of the vascular endothelium, promote the remodeling of damaged myocardial tissues, and alleviate the destruction of myocardial cells by lipid peroxidation products to mitigate the effects of myocardial ischemia-reperfusion injury.

 

References:

[1 ] Feng Shuo, Zhang Ruiyan . Factors influencing the occurrence of left ventricular remodeling after acute myocardial infarction[J ] . International Journal of Cardiovascular Disease, 2019 , 46(5 ): 267-269 .

[2 ] WU Xiaoyan, MIAO Lin, ZHENG Rui, et al. Research progress of myocardial ischemia-reperfusion injury [J ] . Chinese Journal of Clinical Pharmacology, 2016 , 12(11):1043-1045 .

[3 ] Cao Fai, Xu Liqin, Liu Qingzhong, et al. Protective effect of phycocyanin on acute myocardial ischemia-reperfusion injury in rats[J ] . China Medicine Herald, 2017 , 14(14):25-28 .

[4 ] LIU CZ , ZHENG HZ , XIE L , et al . Decreased miR-208 induced is chemia myocardial and reperfusion injury by targeting p21 [J ] . Pharm azie , 2016 , 71(12):719-723 .

[5 ] NAGASAKA Y , FERNANDEZ BO , GARCIA-SAURA MF , et al .Brief periods of nitric oxide inhalation protect against myocar - dial ischemia-reperfusion injury [ J ] .Anesthesiology , 2008 , 109 (4): 675-682 .

[6 ] LU Shenlu, WANG Ruiying, NIU Yaqiong, et al. Study on the trend of serum nitric oxide level in intermittent hypoxia mice[J ] . China Cardiovascular Disease Research, 2019 , 17(11):1028-1031 .

[7 ] WANG Wenjie, CHEN Hongying, PENG Jixia, et al. Effect of Rhodiola rosea glycosides on the expression of vascular endothelial growth factor after preconditioning for myocardial ischemia in rats [J ] . China Medical Journal, 2015 , 12(9 ): 29-33 .

[8 ] WU Xianjun, YANG Hong, SHENG Yi, et al . Cloning, expression and antioxidant activity of recombinant proteins of three cyanobacterial phycocyanin β-subunit deacylated protein genes [J ] . Jiangsu Journal of Agriculture, 2018 , 34(5):998-1004 .

[9 ] Liu Huihui, Jiang Liangqian, Wang Yujuan, et al. Current status of the application of algal blue protein against tumor[J ] . Tumor Control Research, 2018 , 45(6):420-424 .

[10] LIU Qi, LI Wenjun, TANG Zhihong, et al. Research progress on the role of algal blue protein in the prevention and treatment of oxidative stress-related diseases[J ] . Marine Science, 2017 , 41(10):132-138 .

[11 ] LI H , ZOU T , MENG S , et al . p21 protects cardio myocytes a- gainst ischemia-reperfusion injury by inhibiting oxidative stress [J ] .Mol Med Rep , 2018 . 17(3):4665-4671 .

[12] CHU Xian-Ming, LI Bing, DU Ri-Ying, et al. Effect of P27 on luminal stenosis after balloon injury in rat thoracic aorta[J ] . Chinese Journal of Arteriosclerosis, 2006(5):417-421 .

[13] YANG Shasha, ZHOU Li . Effect of alginate on myocardial ischemia-reperfusion injury in rabbits[J ] . Journal of Changchun University of Traditional Chinese Medicine, 2017 , 33(6):868-870 .

[14] ZHANG Wei, ZHAO Lijun, LI De'an, et al. Pigment epithelium-derived factor is involved in arsenic-induced vascular endothelial cell dysfunction[J ] . Chinese Journal of Endemic Diseases, 2019(2):107-110 .

[15] LIU Zhaoya, XU Xi, TANG Yixin, et al. Research progress on the regulation of endothelial-type nitric oxide synthase activity and cardiovascular diseases[J ] . Journal of Central South University (Medical Edition), 2016 , 41(61 ): 632

 

What is BCAA Amino Acids Powder?

 Branched-chain amino acids (BcAAs) are composed of leucine (Leu), isoleucine (I1e), and valine (Val), and are so named because of the prominent branched chains in their carbon skeletons. All three amino acids are essential amino acids that cannot be synthesized in the body and need to be provided by food protein. Studies have shown that branched-chain amino acids have the effect of resisting central fatigue and muscle fatigue. The enhancement of anti-fatigue ability can not only effectively ensure the quality of exercise, but also help to improve the ability to exercise, and at the same time can promote the recovery of fatigue, to avoid sports injuries.

 

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Product Name	BCAA Amino Acids Powder
Specification	‘2:1:1
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Appearance	White to Off-White Fine Powder
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 Alpha Lipoic Acid is an enzyme present in mitochondria, similar to vitamins, which can act as a coenzyme involved in acyl transfers in the metabolism of substances in the body, and can eliminate free radicals that lead to accelerated aging and pathogenicity. Lipoic acid is absorbed through the intestinal tract and enters the cells. It is both fat-soluble and water-soluble, so it can pass through the whole body without any obstruction and reach any cellular part, providing the human body with a full range of performance, it is universal antioxidant with both fat-soluble and water-soluble properties.

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