More Related Content
What's hot
What's hot (20)
Similar to Sports Medicine and Doping.pptx
Similar to Sports Medicine and Doping.pptx (20)
More from VikramSharma288
More from VikramSharma288 (11)
Recently uploaded
Recently uploaded (20)
Sports Medicine and Doping.pptx
- 1. SPORTS MEDICINE AND DOPING DR. MAMTA YADAV Senior Resident, MAMC
- 2. Definitions • Sports Medicine : A field of medicine that relates to the prevention and treatment of injuries and other health problems that affect people who play sports. • Doping - Use of prohibited substances or methods that may enhance performance by athletes.
- 3. Role of Sports medicine • Formation & Evaluation of Physical Education Program. • Development of field and laboratory tests ( Fat %, lung capacity, heart volumes etc ). • Prevention of accidents & injuries. • Provides sound principles for sports training. • Treatment of sports injuries. • Provides guidelines on drugs and doping • Detection of drug use in sport • Provides guidelines and principles of exercise for different age groups. • Tells about the right food, nutrition and supplements. • Effect of environment.
- 4. Doping in Sports • It is the use of chemical substances or methods in order to alter the performance. • It is unethical and against the spirit of sports as well as poses a risk to the health of the sportsperson.
- 6. WADA • IOC established the WADA in 1999 to coordinate the fight against doping in sports. • WADA has support & participation of Govts, Intergovt Organizations, Public Authorities and other public & private bodies. • WADA code - any 2 of the following criteria- (i) Enhances performance (ii) Actual/ potential health risk to athlete (iii) Use must be against the “spirit of sport” • Newer techniques to expand the detection window of doping substances. • The legal actions vary among different nations. (France, Denmark, Austria)
- 7. Role of WADA • Its mission is : – To promote, co-ordinate and monitor the fight against doping in sports – To protect the athletes right to compete in a doping free sports – To promote the equality among the competitors worldwide • The main focus lies in promoting education, research and development of Anti-Doping policies for all countries.
- 8. • The WADA Code is adopted by more than 600 national and international sports organizations. • The main purpose of having the international standards is to harmonize different technical and operational processes of Anti-Doping programs among different countries. • It works on basis of 5 International standards – Testing procedures – Laboratories – List of prohibited substances and methods – Therapeutic use exemptions – Confidentiality of personal information
- 9. Doping Test • It is to be performed on routine basis for all the sportsperson. – Those who are participating in the competition – Those who won the medals in previous sports events • There is also provision for random drug testing of sportsperson by their governing body for in competition and out-of- competition testing. • The samples for testing includes blood and urine.
- 10. • Test methods:
- 11. • Stimulants • Narcotics • Cannabinoids • Glucocorticoids • Beta-blockers • Non approved substances • Anabolic Agents Anabolic Androgenic Steroids (AAS), Other Anabolic Agents • Peptide Hormones Growth Factors And Related Substances • Beta-2 Agonists • Hormone Antagonists & Modulators • Diuretics And Other Masking Agents Methods • Enhancement Of Oxygen Transfer • Chemical And Physical Manipulation • Gene Doping
- 14. Anabolic Steroids • Most commonly misused. Ana + Andro. RoA – Oral, i/m. – Testosterone undecanoate – Nandrolone decanoate – Oxymetholone – Stanozolol – Methandrostenolone – Clenbuterol • X 10-100 times Effects: • Increased muscle size, density and weight. • Faster muscle recovery. Inhibit muscle breakdown.
- 15. Adverse effects • Estrogenic - Gynecomastia, water retention • Androgenic- Acne, alopecia, body hair growth • Cardiovascular- Cardiomegaly, ↑ LDL, ↓ HDL • Hepatotoxicity • Suppression of natural testosterone production. • Mood disturbances • Tumors Associated sports • Contact sports • Weight lifting • Sprinting • Throwing shotput • Bodybuilding
- 17. Human Growth Hormone (hGH) • Anabolic hormone used with anabolic steroids. – Recombinant hGH Effects • IGF-1. • Increased protein synthesis. Procollagen type III amino-terminal propeptide (P-III-NP) • ↑ Bone density • Adipose tissue lipolysis.
- 18. Adverse effects • Long term: acromegalic changes (jaw overgrowth, frontal bossing, dental malocclusion, etc) • Arthralgias, myalgias • Insulin resistance leading to diabetes, cardiovascular problems (hypertension, dilated cardiomyopathy) • Cancer risk (colorectal, prostate, breast, thyroid cancers). Associated sports • Sprinting • Bodybuilding
- 19. Detection Direct - Isoform Differential Immunoassays (blood) • Differentiate b/w Exogenous & endogenous GH based on the available isoforms. • Drawback - difficulty in its detection after 24 hr of last dose. Indirect - hGH Biomarkers Test (blood) • Levels of IGF-I (1 week ) and P-III-NP (4-6 weeks) in blood by immunoassay. • Drawback - less specific, associated variations of age and sex among different individuals. • Also the out-of-competition testing needs to be done.
- 20. Erythropoiesis Stimulating Agents • Recombinant Human EPO (rHuEPO) (8-16,000 U/ Week) • Darbepoietin (40-80 Mcg/Week) • Methoxy Polyethylene Glycol • RoA – s/c, i/v or i,/p Effects • Increases the amount of red-blood cells entering the body • Increase in Haemoglobin • Increase in 02 intake • Greater levels of endurance
- 21. Adverse effects • Thromboembolism • Stroke, MI • Kidney/Liver failure • Increased BP • Risk of bone marrow tumors • Increased risk of HIV and Hepatitis through the use of needles and transfusion. Associated sports • Cycling • Long-distance runners • Cross-country skiing • Triathlon • Bi-athletes
- 22. Detection Blood testing – Hematocrit level – Hemoglobin level – Reticulocyte count Urine testing • Less acidic when compared to endogenous EPO. (isoelectric focusing) • Difficult to detect when the drug usage has ceased for more than 7 days.
- 23. Human Chorionic Gonadotropin (hCG) & Luteinizing Hormone (LH) Effects • Mediate the release of testosterone from Leydig cells which can help in muscle building and performance. • Banned in male athletes alone. • Highly expensive, several injections in a week. Detection • Detection in Urine: • hCG Immunoassays (screening tests) MS (confirmation). • LH immunological assays are done.
- 24. CNS Stimulants • Dual way: Cognition enhancer or performance enhancement. • Cocaine, caffeine, ephedrine, pseudoephedrine and amphetamine. Effects • Releasing neurotransmitters - Dopamine, Serotonin and Noradrenaline that increase the metabolism thereby reducing fatigue. Speed up reactions, Overcome tiredness.
- 25. Associated sports • Swimming, sprinting, contact sports and weight lifting. Adverse Effects • Over-training, hyperthermia, dehydration, hypertension, strokes, coronary insufficiency, cutaneous vasoconstriction and even death. Extremely addictive. Detection • Urine samples - confirmed with GC or LC-MS/MS.
- 26. Narcotics and Analgesics Morphine, Codeine, Heroin, Methadone. Effect • Mask the pain associated with injuries. Associated sports • Violent sports Adverse effects • Addictive , Feeling less pain can cause long term injuries. Lead to constipation and low blood pressure. Detection • Urine : Morphine to Codeine ratio <1 (only codeine); >1 (morphine or heroin). ?CYP2D6
- 27. Beta- 2 Agonists • Clenbuterol, Salbutamol, Terbutaline Effect • Improvement in lung function. Associated sports • Cycling, running, skiing, swimming, cricket. • Asthmatic athletes, TUE
- 28. Beta blockers • Propranolol, atenolol, acebutolol Effect • Helps in reducing tremor and anxiety of the athletes during competition. Associated sports • Precision sports - archery, shooting, golf and billiards. Adverse effects • Dizziness, Fainting, Bradycardia, disturbed sleep. Detection • In urine using GC-MS.
- 29. Diuretics • Bumetanide • Amiloride • Canrenone • Adverse effects • Cramps, dehydration and dizziness. • Detection • In urine using LC-MS.
- 30. Blood doping • Banned method. Previously allogenic, nowadays autologous blood transfusion. • Blood withdrawn, stored (6 – 8 weeks) Body naturally replaces over 2-3 weeks blood transfused back increased Hb enhanced muscle oxygen delivery improved aerobic capacity and performance. • Difficult to identify. – RBCHb/ RetHb ratio. – CO rebreathing method. • Perfluorocarbon emulsions improve tissue oxygenation, enhancing performance. A/E flu like symp, febrile reactions, thrombocytopenia. Difficult to detect.
- 31. Gene doping • “Non-therapeutic use of cells, genes, genetic elements, or modulation of gene expression, having the capacity to improve athletic performance” (WADA) • “Sports genes” • Mutn discovered in MSTN & EPOR affect performance. • German coach Repoxygen (EPO gene - AAV vector) to improve the performance of his trainees. EPOR increased Hb (>200 g/L) and Hct levels (to 68%) ↑ delivery of O2 to muscles; implicating advantage. • Other targets - VEGF, IGF-I and FST. • Other vectors – rAAV, lentivirus systems.
- 32. Designer Drugs In Sports • Manufactured illegally - clandestine laboratories - novel modifications - low cost. Evade doping tests. • Major concern - safety profile unknown, unpredictable. • BALCO scandal. Drug Class Designer Drug Anabolic Steroids Tetrahydrogestrinone (THG) “clear”, Norbolethone, Testosterone cream (“The cream”) desoxymethyltestosterone Selective Androgen-Receptor Modulators (SARM) Ostarine (MK-2866), Andarine (serious side- effects, black market) Synthetic Cannabinoid Receptor Agonists (SCRA) Dronabinol, Nabilone, Nabiximols. Others Hypoxia-Inducible Factor stabilizer (FG-2216)
- 33. Therapeutic Use Exemption • Allows an athlete to use banned substance for therapeutic purposes only. • The athlete can obtain a TUE for the use of prohibited substance or method for the treatment of a legitimate medical condition. • In such cases, athletes should first check their physician to look if any alternatives are available. • If there is no alternative, then athletes should apply for a TUE.
- 34. Doping Control in India • The National Anti-Doping Agency (NADA) is the National Independent Organization become operational in 2009 in India to promote, coordinate & monitor the fight aganist doping in Sports in India. • India is one of the foundation members of World Anti- Doping Agency. • It has accepted the World Anti-Doping Code in march 2008. • The Anti Doping rules of NADA have been modified in compliance to WADA code 2009.
- 35. • The primary functions of NADA are : – Adopting and implementing Anti-doping rules and policies according to the World Anti Doping Code. – To work in coordination with other sports organizations and Anti Doping Organizations – Promoting anti doping research & education – Sample collection and testing of sportsperson – Imposing sanction or ban on athletes who violates the Anti- Doping Rules
- 36. • The samples collected from the athletes are tested in National Dope Testing Laboratory (NDTL), New Delhi. • NDTL is accredited by WADA for testing of blood and urine samples. • NADA also conducts various educational and awareness program about the Anti Doping rules and regulation.
- 37. Athlete Biological Passport (ABP) • An electronic record of an individual athlete’s biological attributes, developed over time from multiple sample collections. • To monitor the selected biological variable (doping biomarkers) rather than the doping substance or method itself. • Thus over a period of time it indirectly reveals the effects of Doping rather than the substance or the method used. • The first version of ABP contains the hematological or blood module.
- 38. • Passport data is interpreted on a regular basis identify the specific targets refinement of available analytical methods. • The data storage, reporting and sharing is done by WADA’s ADAMS (Anti-Doping Administration and Management System) that gives a review till date of an athlete’s passport. • ABP contains two modules (analysed in WADA accredited lab) – the Haematological module and the Steroid module.
Editor's Notes
- GC- gas chromatography, NPD/MSD- Nitrogen phosphorus detector and Mass Spectrometric detector LC- Liquid chromatography, MS- Mass spectrometry CERA-
- Most commonly misused class of drugs for performance enhancement. which have combined anabolic and androgenic action; The anabolic action promotes muscle building while the androgenic action is responsible for the development of masculine characters like facial hair and deepening of voice, aggressiveness. The anabolic agents include the Synthetic variants of male sex hormone (Testosterone) -Testosterone undecanoate is the most common form abused by athletes. Nandrolone decanoate, Testosterone Oxymetholone, Bodlenone, Trenbolone, Methandrostenolone (dianabol), Clenbuterol. Clenbuterol has anabolic and β2 agonistic action. Most commonly misused for performance enhancement. Upto 10-100 times the medicinal dose. Effects: Increased muscle size, density and weight. Faster muscle recovery. Inhibit muscle breakdown. (Allow athletes to train harder).
- Most sports require strength and these anabolic agents are therefore abused almost in every sport. Contact sports are sports that require physical contact between players. Rugby union, American football, kabaddi, boxing, wrestling, martial arts like: sumo, Muay Thai, judo, jiu-jitsu etc. Weight lifting. Sprinting/ throwing. Even young people going to the gym - Sources for obtaining Anabolic Steroids: 25% from friend or relative; 22% from Website/mail order; 17% from coach or trainer; 15% from teammate or other athlete.
- Detection is the most important slide from the point of view of a pharmacologist. Testosterone undecanoate: After its oral or intramuscular administration, the glucuronidated metabolites like androsterone, 5-dihydrotestosterone and etiocholanolone are elevated in plasma. The ratios of Testosterone Glucuronide(TG)/unconjugated Testosterone(T), TG/17- Hydroxyprogesterone(17OHP) and TG/Luteinizing Hormone(LH) in plasma are considered to be the most sensitive markers to detect exogenous testosterone administration. Also, the ratio of urine testosterone to LH more than 30 is considered to be a positive finding in the doping test of athletes as exogenous testosterone suppresses LH. Another way is to calculate the ratio of TG to Epitestosterone Glucuronide (EG). A cutoff value of 6 is considered to be positive as the usage of anabolic steroids suppresses the endogenous testosterone secretion. Epitestosterone is also normally synthesized in the body and it has been shown that exogenous administration of testosterone does not affect levels of epitestosterone in the body. Epitestosterone has not been shown to enhance athletic performance, although administration of epistestosterone can be used to mask a high level of testosterone if the standard T/E ratio test is used. As such, epitestosterone is banned by many sporting authorities as a masking agent for testosterone. As testosterone is an endogenous compound, illicit use of such compounds is rather a tough job to prove. So, the isotope ratio mass spectrometry techniques (12C: 13C) were developed to distinguish between the exogenous and endogenous steroids. And also, as they are used routinely during the entire training period, an out-of-competition testing is quite beneficial rather than its testing only during the event.
- It is an anabolic hormone used commonly along with anabolic steroids. use recombinant HGH by athletes for muscle building and to improve their physical performance. Anabolic effects of GH mediated by IGF-1. Insulin-like growth factor 1 (IGF-1, or sometimes with a Roman numeral as IGF-I) is mainly secreted by the liver as a result of stimulation by growth hormone (GH). Increased protein synthesis - promoting protein synthesis leading to muscle growth. causes a release of procollagen type III amino-terminal propeptide (P-III-NP) that further enhances the muscle growth and strength. IGF-1 also has stimulatory effects on osteoblast and chondrocyte activity to promote bone growth. ↑ Bone density It also acts on the adipose tissue causing lipolysis; and diverting the energy that is liberated for the synthesis of proteins and carbohydrates.
- Detection can be done in 2 ways – Direct and Indirect Direct - Isoform Differential Immunoassays (blood) This was framed to differentiate between the exogenous and endogenous GH in blood based on the available isoforms (recombinant HGH exist in a single isoform whereas the endogenous form exists in 3 different isoforms that vary in their molecular weight). The drawback associated with this method is the difficulty in its detection after 24 hr of last dose due to short half-life. Indirect - hGH Biomarkers Test (blood) This has led to the development of specific biomarker tests to measure the levels of IGF-I and P-III-NP in blood by immunoassay. The levels of IGF-I can be detected till one week and those of P-III-NP for about 4-6 weeks. So, there is a wide opportunity for their detection in the doping tests but are comparatively less specific with limitations in the available assays and also due to the associated variations with respect to age and sex among different individuals. Also the out-of-competition testing needs to be done.
- EPO is a glycoprotein produced by kidney (juxtaglomerular cells) and macrophages. Erythropoietin Receptor Agonists Used Are - Recombinant Human EPO (rHuEPO) (8000-16,000 units/week), Darbepoietin (40-80 mcg/week) It can be given subcutaneously, intravenously or intraperitoneally and is indicated to treat anaemia in conditions of chronic renal failure, in cancer chemotherapy and in AIDS patients. Effects EPO causes the stimulation of erythroid precursors to generate red blood cells that increase the delivery of oxygen to tissues. Increase in the oxygen carrying capacity of the blood. Increases the amount of red-blood cells entering the body Increase in Haemoglobin Increase in 02 intake Higher V02 Max All this leads to Greater levels of endurance as Increase in the oxygen carrying capacity of the blood.
- Effects hCG and LH act by binding to Choriogonadotropin (CG) and LH receptor respectively and mediate the release of testosterone from Leydig cells which can help in muscle building and performance. Their usage is banned in male athletes alone as testosterone production in females was found to be negligible. Also, the usage of other substances like anti-estrogens and aromatase inhibitors that can release endogenous LH are also prohibited. They are highly expensive and need the administration of several injections in a week. Detection Their illicit use can be detected from the urine samples. To detect hCG in urine, immunoassays are carried out initially as screening tests followed by their confirmation with mass spectrometry. If hCGα/β heterodimer levels are >5 IU/L by immunoassays or >2 IU/L by mass spectroscopic assays, it is said to be positive. For LH, immunological assays are done and if it is >60 IU/L by Immulite assay or >4 IU/L by Delfia assay, it is considered to be a positive finding.
- This class of drugs can be used in dual ways either as a cognition enhancer (nootropics or smart drugs) or for athletic performance enhancement in athletes. They are found to improve performance in non-endurance sports like swimming, sprinting and weight lifting. Contact sports like boxing because it speeds up reactions The most common drugs abused include cocaine, caffeine, ephedrine, pseudoephedrine and amphetamine. They mainly act by releasing neurotransmitters like dopamine, serotonin and noradrenaline that increase the metabolism thereby reducing fatigue and causing aggressiveness and competitiveness. Speed up your reactions, Make you feel LESS pain, Overcome tiredness. Urine samples of the athletes are confirmed with doping by techniques like gas chromatography or Liquid Chromatography (LC) Mass Spectrometry (LC-MS/MS). The common adverse effects include hyperthermia, dehydration, hypertension, coronary insufficiency, cutaneous vasoconstriction and even death. Over-training. High blood pressure, heart and liver problems and strokes. Extremely addictive. Stimulate Adr. Receptors directly:- Adrenaline, NA , Isoprenaline , Phenylephrine Release NA from presynaptic terminals:- Amphetamines Inhibit Reuptake:- Cocaine
- Effect- So the pain threshold is set to a higher level by narcotics and cannabinoids acting on the CNS. This increases the pain tolerance level and the exercise tolerance level. Urine samples: (codeine is metabolized to morphine). So, the morphine to codeine ratio is taken as the most common marker to assess its misuse in athletes. If the ratio is <1, it is due to the intake of only codeine; and if it is >1, it is due to the intake of either morphine or heroin. However, a cut-off value of 1 is not always absolute as some of the athletes with ultra-rapid CYP2D6 metabolism already have levels of >1 even with the intake of codeine alone. So, the pharmacogenomics testing for CYP2D6 metabolism should be done.
- Clenbuterol was originally developed to treat asthma in horses but is now used to increase muscle mass and reduce body fat. Classified by WADA (also banned by it), there are growing concerns more of the drug is making its way into meat in places like China and Mexico. There is an improvement in lung function characterized by an increase in FEV1 with mild bronchodilator effect. Inhaled β2 agonists were tried as an ergogenic aid in athletes like cyclists, runners, skiers and swimmers. However, they are still included in the list of prohibited substances with the exception of salbutamol, terbutaline, salmeterol and formeterol usage in asthmatic athletes. In special situations (asthmatic athletes), TUE (Therapeutic Use Exemption) signed by the athlete and the physician needs to be submitted to the concerned anti-doping authorities along with the lung function reports. If the medication an athlete is required to take to treat an illness or condition happens to fall under the Prohibited List, a Therapeutic Use Exemption (TUE) may give that athlete the authorization to take the needed medicine. Checklists For Therapeutic Use Exemption (Tue) Applications
- Helps in reducing tremor and anxiety of the athletes during competition help improve their performance. When hands don’t shake, the sportsman can target more easily and iprove performance.
- • Side-Effects of Blood Doping include Increased blood viscosity, Heart attack, Stroke, risk of Infections, hypertension, weight gain and chest pain. • Blood doping is most commonly used by endurance athletes, such as distance runners and cyclists. It comes under banned methods Previously, allogenic blood transfusion was widely followed by many athletes. However, with the development of flow cytometry techniques, it was very easy to identify the athletes who have received even one unit of allogenic blood. Because of these concerns, athletes started using their own blood (autologous blood transfusion) in which about 1-2 litres of blood was withdrawn and stored at suitable conditions 6 – 8 weeks before competition, to be infused before the competition. Body naturally replaces this over two to three weeks . Prior to competition, the blood is transfused back into the body. This is found to increase the level of haemoglobin and in turn enhancing the oxygen delivery to the muscles, thereby improving their aerobic capacity and performance. It is rather difficult to identify this illicit practice when compared to allogenic blood doping. But presently, the ratio of mass of Haemoglobin (Hb) in the mature erythrocyte to that in the reticulocytes (RBCHb/RetHb ratio) is considered to be one of the best indicators for detecting autologous blood doping. This ratio gets markedly increased. Increased in EPO abuse also. Another method for detection is by Carbon Monoxide (CO) rebreathing method in which the athlete is asked to inhale oxygen-carbon monoxide mixture to calculate the Hb mass. However, it is not acceptable as the inhalation of CO before the competition affects their performance. Artificial oxygen carriers like perfluorocarbon emulsions (oxygen, oxyfluor) were used as a PED. On intravenous administration, the droplets are taken by the reticuloendothelial system and improved the tissue oxygenation, thereby enhancing the performance. The common side effects include flu like symptoms and febrile reactions; but with higher doses, transient thrombocytopenia is evident in a few cases. It is difficult to detect this substance in doping tests as it is mainly expelled out from the body by breathing; and there are no changes observed in the blood or urine tests. Exploited by cyclists, skiers, marathon runners.
- Defn- “Non-therapeutic use of cells, genes, genetic elements, or modulation of gene expression, having the capacity to improve athletic performance” (WADA) Some genes have effects on body weight, size and aerobic capacity; and are referred to as sports genes. This was supported with the discovery of Mutations in Myostatin Gene (MSTN) and Erythropoietin (EPO) receptor gene (EPOR) that can affect the performance. EPOR increased the haemoglobin mass (>200 g/L) and haematocrit levels (to 68%) enhancing the delivery of oxygen to muscles; implicating its advantage in elite sports. The other targets for gene doping include Vascular Endothelial Growth Factor (VEGF), insulin-like growth-factor-I (IGF-I) and follistatin (FST). The first thought of its misuse in sports arose when a German coach had tried to acquire a drug called Repoxygen containing EPO gene linked with the Adeno-Associated Virus (AAV) vector to improve the performance of his trainee. He was found guilty of giving athletes performance enhancing drugs without their knowledge. Repoxygen was under preclinical development by Oxford Biomedica as a possible treatment for anemia but was abandoned. The authorities had then put a ban on the delivery of genes, genetic elements and nucleic acids for doping. The other vector systems commonly used are the Recombinant AAV (rAAV) and lentivirus systems; and do have a good efficacy and safety profile in trials.
- These drugs are manufactured illegally in the clandestine laboratories with some novel modifications in the already existing drugs at a low cost. This has led to the advent of designer drugs that are specifically engineered to evade the doping tests. Another major concern with the designer drugs is their safety profile which is not known and at times is unpredictable as there are lack of studies confirming their efficacy and safety. The most common drug classes targeted are the anabolic steroids, CNS stimulants, cannabinoids in human sports; and the sildenafil analogs in animal sports. The anti-doping authorities were first aware of the usage of designer steroids by athletes with the Balco scandal. The Bay Area Laboratory Co-operative (BALCO) was a San Francisco Bay Area business which supplied anabolic steroids to professional athletes. During this scandal, a new drug Tetrahydrogestrinone (THG), also commonly called as clear; came into limelight. The main intention of designing this is to escape their detection in the doping tests with the currently available methods. It was later found out that THG is not the only compound abused, but also others like the T/ET cream (the cream), erythropoietin, human GH, insulin, modafinil and liothryonine were also given to enhance the performance of the athletes. The program was so planned that they were not detected even in the out-of-competition testing. SARM they were rejected for further pharmaceutical development. The usage of designer drugs has widely increased in the recent years due to their availability even on the internet. They are illegally sold by labelling them as “not for human consumption” or as “plant products” so as to avoid the legal issues. And many of these drugs do not actually contain the active ingredient that is claimed in the products. So, for their detection, a variety of newer techniques were being developed in MS, immunoassays, specific biomarkers.