Function of Garcinia Cambogia Extract,HCA or Hydroxycitric Acid Calcium Salt?

  seminal trace...Garcinia Cambogia Extract,HCA Hydroxycitric Acid Calcium Salt.CAS-No.6205-14-7.Formula: CaHCA50.Chemical Name.1,2-dihydroxy-1,2,3-Propanetricarboxylic Acid...

  Garcinia Cambogia Extract.Hydroxycitric Acid Calcium Salt

 Botanical Source: GARCINIA CAMBOGIA;
 Botanical Synoms:(in india)r kokum, uppage huli;(in English)Brindel Berry/Uppagi, Malabar tamarind,Brindal Berry, Gorikapuli, HCA, Hydroxycitric acid, Malabar Tamarind.
 Plant Part Used: Fruit Pulp
 Content Synoms: ((-)-HCA).
 CAS-No.:6205-14-7
 Formula: CaHCA50
 Chemical Name:1,2-dihydroxy-1,2,3-Propanetricarboxylic Acid
 

 The herb Garcinia cambogia, a diminutive purple fruit native to India and southeast Asia, has garnered a lot of attention of late as a popular natural weight loss aid. The reason is that the rind of this pumpkinlike fruit is rich in a substance called hydroxycitric acid, or HCA, which is closely related to the citric acid found in grapefruits and oranges.

 HCA has been used in connection with the following conditions (refer to it for complete information):

  Weight loss:

 Reliable and relatively consistent scientific data showing a substantial health benefit.
 Contradictory, insufficient, or preliminary studies suggesting a health benefit or minimal health benefit.
 An herb is primarily supported by traditional use, or the herb or supplement has little scientific support and/or minimal health benefit.

 Who is likely to be deficient? Since it is not an essential nutrient, HCA is not associated with a deficiency state.
 How much is usually taken? Optimal amounts of HCA remain unknown. Although dieters sometimes take 500 mg of HCA three times per day (before each meal), this amount is far below the levels used in animal research (figured on a per-pound body weight basis). The effect of HCA is enhanced when used in conjunction with a low-fat diet, because HCA does nothing to reduce the caloric effects of dietary fat. Since HCA's mechanism of action seems to be at least partially a blockade of conversion of simple sugars into fats,13 it is likely to work best in conjunction with a high simple sugar diet. HCA may therefore be less useful if it only offsets the negative effects of an otherwise unhealthy diet. High-fiber diets may impair absorption of HCA as noted above. HCA supplements are available in many forms, including tablets, capsules, powders, snack bars, and chewing gum.
 Are there any side effects or interactions? HCA has not been linked to any adverse effects.
 At the time of writing, there were no well-known drug interactions with Hydroxycitric Acid.


 Garcinia Cambogia Extract.Hydroxycitric Acid Calcium Salt
 Garcinia and Its Physiology:
 Weight Loss Effects:
 Fat-Burning Capability of HCA:
 Other Functions and Mechanisms:
 Three collectors' case histories and India Culture:
  Indian Tree Gum Extract Studied for Weight Loss: Clinical Trial:
 Process for the production of potassium hydroxy citric acid, and compositions containing the potassium hydroxy citric acid
 How Search engine think about Garcinia:


  Garcinia and Its Physiology:

 The active ingredient, hydroxycitric acid(HCA), is extracted from the rind of the fruit as a substance to promote weight loss. Studies have shown that HCA decreases the activity of an essential enzyme in metabolism of fat. The conversion of carbohydrates into fat requires an enzyme called citrate lyase. HCA temporarily reduces the action of this enzyme, blocking the production of fatty acids making less fat available for cellular storage.

 

  Where is it found?

 hydroxycitric acid(HCA) is found in only a few plants, with one rich source being the rind of a little pumpkin-shaped fruit called Garcinia cambogia, which is native to Southeast Asia. This fruit (also called Malabar tamarind) is used as a condiment in dishes such as curry.

 Garcinia cambogia is the fruit of the garcinia indica tree also known as Brindle berry, grown in southern India. Hydroxycitric acid (HCA), the active ingredient of garcinia cambogia, is synthesized from the rind of the cambogia fruit.

 This product is often sold in weight loss supplements in various forms and under various brand names such as Brindleberry Extract, HCA, and the standardised form which is marketed as Citrimax
 HCA is a substance which has been studied extensively for more than two decades.
 
 Various studies have shown that Hydroxycitric acid helps prevent the liver from forming fats from dietary carbohydrates. HCA increases the production and storage of glycogen while reducing both appetite and weight gain. HCA also causes extra calories to be consumed as thermogenisis is enhanced.

 Some studies have shown that hydroxycitric acid(HCA) derived from garcinia cambogia promotes weight loss when combined with a high carbohydrate diet, while others have found it ineffective.

 Early research by Hoffman-La Roche found that HCA interferes with the break down of fatty acids in the Krebs Cycle resulting in increased energy expenditure, reduced appetite, decreased plasma cholesterol, and inhibited fat synthesis from excess carbohydrate calories.

 The most recent findings on HCA, published in the November 11 issue of the Journal of the American Medical Association (JAMA), concluded that HCA was no more effective in promoting weight loss than diet and exercise alone.

 It cannot be concluded from just one study that HCA is not effective in assisting weight loss.

 The high fibre diet prescribed in the study inhibited HCA absorption The low calorie diet prevented evaluation of HCA ability to reduce food intakeAt present it appears that further research is needed in order to collect more information as to the effectiveness of HCA in assisting weight loss.

 What does it do? (-)-hydroxycitric acid(HCA) is a compound found in Garcinia cambogia, a type of fruit. HCA has a chemical structure similar to that of citric acid (the primary acid in citrus fruits). Preliminary research in the laboratory and in animal research, suggests that HCA may be a useful weight loss aid. HCA has been demonstrated in the laboratory (but not yet in trials with people) to reduce the conversion of carbohydrates into stored fat by inhibiting certain enzyme processes. Animal research indicates that HCA suppresses appetite and induces weight loss.One case report found that eating 1 gram of the fruit containing HCA before each meal resulted in the loss of 1 pound per day.
 HCA (hydroxycitric acid) is a close relative of citric acid, the agent that gives citrus fruits their characteristic tart flavor. HCA is obtained as a 50% standardized extract of Garcinia cambogia, a small fruit from southern India, where it has been used for centuries as a food preservative, flavoring agent and digestive aid. Studies show that HCA can curb appetite, reduce food intake and inhibit the production of fats and cholesterol.
 HCA exerts its anti-obesity effects through its inhibition of the enzyme ATP citrate lyase, playing a critical role in energy storage, and affecting the appetite. Appetite comes from feedback signals between the stomach and brain, making you feel hungry. When you eat, your food is reduced to the simple sugar glucose, which is then converted into energy. When calorie intake exceeds the body's energy needs, the excess glucose is converted into glycogen, which is stored in the liver and muscles for future conversion into energy. Weight gain occurs after the body's capacity for glycogen storage is reached. At this point, glucose from excessive calorie intake is converted into acetyl coenzyme A via a metabolic pathway involving ATP-citrate lyase and then into fat molecules which are stored in fat cells. HCA inhibits this process by binding to ATP-citrate lyase to reduce the production of acetyl coenzyme A, reducing the body's production of fat and cholesterol.
 HCA also increases the ability of the liver and muscles to synthesize and store glycogen, thereby suppressing appetite.
 A double-blind trial that provided either 1,500 mg of HCA or a placebo per day to 135 overweight men and women, who also were on a calorie-restricted diet, found after 12 weeks that the HCA supplementation did not produce a significant change in weight loss.10 Uncontrolled and/or preliminary evidence from several other human trials suggests the possibility that weight loss might occur;however, none of these studies is as methodologically strong as the negative trial previously mentioned. These less-rigorous studies used a similar calorie-restricted diet and a similar amount of HCA as the negative trial. However, the double-blind study used a high-fiber diet not used in the prior studies. It has been suggested that such a diet might limit absorption of HCA.Future studies that measure blood levels of HCA (to check whether or not the supplement was absorbed) are necessary to resolve this issue. At the present time, the effectiveness of HCA for weight loss remains unclear and unproven.
 


  Weight Loss Effects:

 Studies indicate that hydroxycitric acid(HCA) helps people, including problematic "plateau" dieters, lose weight. Plateau dieters find weight loss especially frustrating because they do lose some weight, but reach a point at which they cannot lose more weight. In a 1995 Israeli study, HCA was gien to eight subjects who had been on a standard calorie-restricted diet for two months and had stopped losing weight. In the following two months they were given 250 mg. of HCA three times a day and continued to follow a moderate calorie-restricted diet. In the second two months, they lost between 3.3 and 17.6 more pounds (1.5 and 8 kg).
 HCA also promotes stable weight loss - what you lose stays off. In a 1994 Danish study, 28 subjects took 750 mg. of HCA and 125 of chromium for six weeks. Over this time period, the subjects lost an average of 8.21 pounds (3.73 kg). The last two weeks of the study were conducted without the HCA. During this period, there was further weight loss of 1.7 pounds (0.8 kg). No weight gain was seen.
 In a 1997 study, subjects took 2.6 grams (2600 mg.) of HCA per day for two months as part of a 1200 calorie-per-day, low-fat diet and exercise program. After the two months, the subjects continued taking the HCA for another year, but with no food restrictions. At the end of the year and two months, the subjects had lost 15% of their original weight, with an average weight loss of 30.4 pounds (13.8 kg).
 


  Fat-Burning Capability of HCA:

 To understand how HCA both inhibits us from producing fat and helps us burn fat, we should consider a simplified explanation of what happens when we eat too many carbohydrates. The following numbers and letters correspond to those on the illustration.

  1. When we over consume carbohydrates, we have an over abundance of a substance known as acetyl coenzyme A (acetyl CoA).
  2. Acetyl CoA cannot pass out of the mitochondrion, whiich is the cell's "energy plant." Because of this, the body transforms acetyl CoA into something called citrate. Citrate does pass out of the mitochondrion and into the cell cytosol, which is the fluid section of the cell.
  3. Here, the enzyme ATP citrate lyase separates the citrate into two components, acetyl CoA and oxaloacetate.
  4.+ 5. The acetyl CoA is then converted into a substance known as malonyl coenzyme A (malonyl CoA). Malonyl CoA is the "base" from which fatty acids (and thus fat) and cholesterol are formed from carbohydrates. You can see that if malonyl CoA is formed, the body is able to produce fat (and cholesterol).
  6. Malonyl CoA also blocks the activity of the enzyme carnitine acyltransferase. This enzyme transports existing fats back into the mitrochondrion where they can be burned. When malonyl CoA blocks its activity, it is harder to burn fat (and lose weight). We could say that when the body creates fat (due to malonyl CoA), it does not burn fat (because malonyl CoA does not let fat be transported to the mitochondrion).
 A.Research indicates that HCA may work by blocking ATP citrate lyase from separating Acetyl CoA from citrate. If there is no acetyl CoA, it cannot be converted into malonyl CoA. If there is no malonyl CoA, fats and cholesterol cannot be easily created.
 B.Just as importantly, the absence of malonyl CoA means that carnitine acyltranferase can transport existing fat into the mitochondrion where fats can be more easily burned. You might say that if the body does not create fat (there is no malonyl CoA to form fatty acids), it can burn fat (there is no malonyl CoA to prevent fat from being transported to the mitochondrion.
 

  Safety and mechanism of appetite suppression by a novel hydroxycitric acid extract (HCA-SX).: Mol Cell Biochem. 2002 Sep;238(1-2):89-103.Ohia SE, Opere CA, LeDay AM, Bagchi M, Bagchi D, Stohs SJ.Department of Pharmacy Sciences, Creighton University School of Pharmacy and Allied Health Professions, Omaha, NE 68178, USA. seohia@creighton.edu
 A growing body of evidence demonstrates the efficacy of Garcinia cambogia-derived natural (-)-hydroxycitric acid (HCA) in weight management by curbing appetite and inhibiting body fat biosynthesis. However, the exact mechanism of action of this novel phytopharmaceutical has yet to be fully understood. In a previous study, we showed that in the rat brain cortex a novel HCA extract (HCA-SX, Super CitriMax) increases the release/availability of radiolabeled 5-hydroxytryptamine or serotonin ([3H]-5-HT), a neurotransmitter implicated in the regulation of eating behavior and appetite control. The aim of the present study was 2-fold: (a) to determine the effect of HCA-SX on 5-HT uptake in rat brain cortex in vitro; and (b) to evaluate the safety of HCA-SX in vivo. Isolated rat brain cortex slices were incubated in oxygenated Krebs solution for 20 min and transferred to buffer solutions containing [3H]-5-HT for different time intervals. In some experiments, tissues were exposed to HCA-SX (10 microM - 1 mM) and the serotonin receptor reuptake inhibitors (SRRI) fluoxetine (100 microM) plus clomipramine (10 microM). Uptake of [3H]-5-HT was expressed as d.p.m./mg wet weight. A time-dependent uptake of [3H]-5-HT occurred in cortical slices reaching a maximum at 60 min. HCA-SX, and fluoxetine plus clomipramine inhibited the time-dependent uptake of [3H]-5-HT. At 90 min, HCA-SX (300 microM) caused a 20% decrease, whereas fluoxetine plus clomipramine inhibited [3H]-5-HT uptake by 30%. In safety studies, acute oral toxicity, acute dermal toxicity, primary dermal irritation and primary eye irritation, were conducted in animals using various doses of HCA-SX. Results indicate that the LD50 of HCA-SX is greater than 5,000 mg/kg when administered once orally via gastric intubation to fasted male and female Albino rats. No gross toxicological findings were observed under the experimental conditions. Taken together, these in vivo toxicological studies demonstrate that HCA-SX is a safe, natural supplement under the conditions it was tested. Furthermore, HCA-SX can inhibit [3H]-5-HT uptake (and also increase 5-HT availability) in isolated rat brain cortical slices in a manner similar to that of SRRIs, and thus may prove beneficial in controlling appetite, as well as treatment of depression, insomnia, migraine headaches and other serotonin-deficient conditions.


  Other Functions and Mechanisms:

  Appetite Suppression:   HCA also suppresses appetite. It does this not through an "aversion" mechanism - it does not make food taste bad - but by making your body think it has ben fed small meals throughout the day. And if your body thinks this, it is not going to feel hungry. The result? Less cravings to eat!

  Increased Energy:   Many HCA users also notice a greater energy level. This may prove true for many, because HCA helps your body burn fat it had not been able to burn. If your body is suddenly burning fat that it had not been able to burn before, it is logical that this would add to your energy.

  How It Works:   HCA research has been ongoing since the 1960s. In the three decades since then, much has been both theorized and determined. A summary of the most telling research brings out the following.

  HCA is safe to use:   HCA inhibits carbohydrate lipogenesis. Simply put, this means that the carbohydrates we eat are not as easily converted to fat and potential weight gain. HCA not only inhibits the manufacturing of fat from carbohydrates, it also helps the body burn off fat more efficiently.

  Suggestion for Capsule:   500 mg of Garcinia cambogia fruit powder extract standardized (50%) to supply 250 mg of hydroxycitric acid (HCA).

 Although HCA can remain active for several hours after ingestion, trials have shown that it is up to 8 times more effective when taken 2-3 times daily. HCA is even more effective when taken with chromium.
 


  Three collectors' case histories and India Culture:

 This case study describes the collection and simple processing of uppage, the fruit of an evergreen tree in the south Indian state of Karnataka. The collection of uppage is the traditional occupation of a group of women of a particular Brahmin caste who collect the fruit for its seed and use it to prepare a substitute for ghee (clarified butter). Dried uppage fruit rinds are now experiencing a commercial boom with a growing market in the neighbouring state of Kerala where they are used as a condiment.
 This study examines the nature of the transition that takes place when a traditional, locally consumed product develops a new market and new end use. In addition to a substantial increase in the number of people involved in collection and processing, transition leads to a new set of production relations with a chain of intermediaries. Access to produce becomes restricted and workers face a new set of circumstances. The short-lived efforts of a marketing cooperative are also examined.
 Uppage (pronounced 'oopahjey') is the most commonly used name of Garcinia cambogia, a medium sized (10 m) evergreen tree, which grows in tropical moist evergreen, semi-evergreen, and wet temperate forest types in Southern India. In Karnataka, uppage is found in the forests of Sirsi, Siddapur, Honnavar, Yellapur, and Sagar within Uttara Kannada and Shimoga Districts. Occasionally uppage trees grow along road sides, in residual forest patches and on wastelands around villages.
 In order to gain a better understanding of the role of uppage collection in the individual lives of Karnataka's women, three brief case histories of women living in an uppage producing village in Siddapur are provided.

  Case history 1:

 Kamala Kamala is a young, vivacious 25 year old mother of three. She lives in the village of Birlamakhe. Her husband works as a labourer with the Public Works Department earning Rs 300 per month. Kamala works as a labourer in an Areca plantation, earning Rs 8 per day including lunch and tea. During the slack season she works for the Forest Department planting saplings, digging pits and trenching. She also undertakes this kind of heavy labour for local landlords. If the uppage yield is good Kamala collects uppage every day for at least a month. She finds the work arduous but worthwhile. She can collect up to 2 kg of uppage in an 8 hour day walking five or six km in the process. In 1986 she collected 40 kg which she sold for Rs 4/kg. Her only regret is that her need for money forces her to sell the uppage as soon as it is dried, rather than stocking it as richer people do, to be able to sell at a higher price.

  Case history 2:

 Ganapi Ganapi is a 50 year old Nayak women who lives in a small thatched house in the all-Nayak village, Surgal. Ganapi has no children of her own but she shares her home with 5 other members of her joint family including her husband who is an alcoholic. Her family has no arable land. They earn their livelihood as agricultural labourers. Ganapi works in a neighbouring areca plantation in the summer months and on paddy fields during the monsoon. The rest of the year she is only marginally employed, spending much of her time making mats out of echalu grass which she sells for Rs 20 per mat.

 Ganapi has collected uppage from the forests surrounding her village for the last five years. If the uppage yield is good she forgoes agricultural work in the peak paddy transplanting season. Instead she sets out each morning at about 8:30 a.m. alone or with a few women. She throws a blanket-like cape over her shoulders and carries a wicker basket in her hand. Although she passes through betta lands she is careful not to collect uppage, as she has no traditional rights to the land.

 Once in the hilly evergreen forest she may cover two or three kilometers searching for soft fallen uppage fruits and seeds. In 1986, a good yield year, Ganapi collected 1 quintal (100 kg) of uppage which she sold to the local landlord/agent for Rs 6/kg. She used this money to buy herself clothes and utensils. Ganapi had no objection to large uppage dryers and standardized trade.

  Case history 3:

 Omi Omi is a 65 year old Marathi woman from the village of Marathikoppa, which is entirely inhabited by Marathis. She is a spokeswomen for Marathis, a minority in the state. Omi lives in a clean little thatched house with her son, his wife and their children. The family owns a small plot of land which they cultivate. They also work for the local landlord as agricultural labourers. Omi spends most of the year making mats and bamboo baskets which she sells to Seva Sagar Sanga, a local society which markets the products. Omi rarely ventures out of the village. The Marathi men collect uppage; Omi and the rest of the women help process the rinds and produce other NTFPs. The Marathi women also began to extract and use uppage ghee five years ago when collection became popular. Omi learned the art of ghee extraction from women in neighbouring villages and the local landlord's family. In Marathikoppa all the uppage and other collected NTFPs are sold to the local landlord/agent, to whom almost every household owes money. Uppage sometimes helps repay loans or buy provisions, for the landlord is also the local shopkeeper.
 


   Indian Tree Gum Extract Studied for Weight Loss: Clinical Trial:

 October,1999,A double-blind, randomized, placebo-controlled trial of a potential weight/fat loss formula consisting of phosphate and extracts of 2 Indian medicinal plants was recently reported to cause significant results in decreasing both fat mass and body weight. The purpose of the study was investigate the effects of the formula on mood and body composition in overweight individuals with a body-mass index of > 25 kg/m. The extracts were made from the fruit rinds of the Malabar tamarind or Garcinia cambogia Desrouss., and the gum resin of guggul or Commiphora wightii (Arn.) Bhandari (formerly C. mukul). Traditionally, Garcinia fruits are used in curries and the treatment of indigestion, chronic diarrhea, and constipation, and the gum resin of the guggal or guggul tree is prescribed in treating rheumatism and for reducing serum cholesterol.

 Researchers Jose Antonio at the University of Nebraska, High Performance Laboratory, in collaboration with Carlon M. Colker at Beth Israel Medical Center in New York and coworkers at the Peak Wellness in Greenwich, CT and Brink Training Systems of Newtown, PA, referred to the formula as "guggulsterone phosphate compound." It consists of the following ingredients per capsule, each having a theoretical basis for inclusion as "weight-loss agents": 125 mg each of L-tyrosine, calcium phosphate (23%), guggal extract (10%), and Garcinia cambogia fruits (50% hydroxycitric acid), along with 75 mg potassium phosphate, 37.5 mg each of sodium phosphate monobasic and sodium phosphate dibasic, and 12.5 mg phosphatidylcholine. The daily dosage in 20 hospital- and physician-referred outpatients was 6 capsules/day (2 capsules per main meal) for 6 wks. Patients were randomly assigned to a placebo group (maltodextrin), a no-formula "control" group, and a formula treatment group. In addition, patients exercised 3 days/wk. for 45 mins. (weight training and step aerobics), and adhered to the 1,800-kcal American Heart Association Step One diet, which was regularly checked for compliance by a registered dietician.

 Compared to initial values before the trial, fat mass in the formula group showed a significant decrease of 4.3 kg, accompanied by a significant decrease in mean body weight of 2.6 kg. The placebo group showed a decrease in fat mass of 1.4 kg, an amount that came close to being significant, and the control group showed a significant decrease in fat mass of 1.5 kg. Percentage-wise, all 3 groups showed a significant decrease in body fat compared to initial values, although the decrease was still greater in the formula group (-20.6%) than either the control (-8.6%) or placebo group (-9.5%). Lean body mass, however, showed no significant change in any group, and when the groups were compared to each other, percentage fat loss was not more significantly decreased in any one group, indicating a significant effect from the diet and exercise component alone. As for changes in mood, fatigue scores according to the Profile of Mood States (POMS) questionnaire showed a significant decrease of fatigue (-63.7%) in the formula group, and no significant change in the control or placebo group; the latter showing an unexplainable increase in fatigue scores of 58.8%. The scores for "vigor" showed a significant increase in the formula group (32.3%, p< 0.01) compared to initial values, whereas no significant improvement was evident in either of the other groups. No side effects were noted by any of the participants in the formula group. Antonio and colleagues suspect that the weight loss found in the formula group was partly the result of an increase in levels of thyroid hormone and an elevated metabolic rate; however, too small a sample was tested to deduce such an effect with certainty.

 When it comes to weight management, Garcinia cambogia is often combined with other herbs and compounds such as St. John's wort and chromiusm.

 Studies concerning hydroxycitric acid(HCA) found in Garcinia cambogia suggests that HCA may be a useful weight loss aid. As mentioned above, the HCA reduces the action of citrate lyase blocking the conversion of carbohydrates into fat. Cheema-Dhadli S, Harlperin ML, Leznoff CC. Inhibition of enzymes which interact with citrate by (-)hydroxycitrate and 1,2,3,-tricarboxybenzene. Eur J Biochem 1973;38:98¨C10. Animal research indicates that HCA suppresses appetite and induces weight loss. Greenwood MRC, Cleary MP, Gruen R, et al. Effect of (-)-hydroxycitrate on development of obesity in the Zucker obese rat. Am J Physiol 1981;240:E72¨C8. Sullivan AC, Triscari J. Metabolic regulation as a control for lipid disorders. Am J Clin Nutr 1977;30:767¨C76.

 Normal Dosages:The usual dosage is one pill that provides 500 mg of HCA, the active ingredient in Garcinia cambogia, 3 times a day. Take it 30 to 60 minutes before each meal along with plenty of water. Garcinia cambogia is also added to certain snack bars and chewing gums. Normally suggestive dosage of HCA now fashionable internationally varies 250mgs three times daily, for the HCA, or 500mgs three times daily for 50% concentration garcinia cambogia extracts, this is just for single formula, but for mixed formula, normally no need such more. For example, if mixed with Hoodia,50mgs three times daily proved good function from our slim marketing and depend on different cases.

 General Interaction:There are no known drug or nutrient interactions associated with Garcinia cambogia. More studies are needed to determine whether a high-fiber diet or other nutritional factors may alter the effectiveness of the supplement, however.
 


  Cautions:

 The pumpkin-shaped Garcinia cambogia fruit is a traditional food of southeast Asia and has long been used as a seasoning for curry. Supplements containing the active ingredient, HCA, are not known to cause any adverse side effects.
 As with many supplements, Garcinia cambogia has not been well tested in pregnant women. If you are pregnant or plan to become pregnant or are breast-feeding, check with your doctor before using this supplement.
  More about HCA from Garcinia Cambogia and the possible substitution:

 hydroxycitric acid(HCA) is obtained as a 50% standardized extract of Garcinia cambogia, a small fruit from southern India, where it has been used for centuries as a food preservative, flavoring agent and digestive aid. HCA is a close relative of citric acid, the agent that gives citrus fruits their characteristic tart flavor. Studies show that HCA can curb appetite, reduce food intake and inhibit the production of fats and cholesterol.
 hydroxycitric acid(HCA) exerts its anti-obesity effects through its inhibition of the enzyme ATP citrate lyase, playing a critical role in energy storage, and affecting the appetite. Appetite comes from feedback signals between the stomach and brain, making you feel hungry. When you eat, your food is reduced to the simple sugar glucose, which is then converted into energy. When calorie intake exceeds the body's energy needs, the excess glucose is converted into glycogen, which is stored in the liver and muscles for future conversion into energy. Weight gain occurs after the body's capacity for glycogen storage is reached. At this point, glucose from excessive calorie intake is converted into acetyl coenzyme A via a metabolic pathway involving ATP-citrate lyase and then into fat molecules which are stored in fat cells. HCA inhibits this process by binding to ATP-citrate lyase to reduce the production of acetyl coenzyme A, reducing the body's production of fat and cholesterol. HCA also increases the ability of the liver and muscles to synthesize and store glycogen, thereby suppressing appetite.
 


  Precautions and/or adverse effects:

 Individuals with preexisting medical conditions should consult with their physician particularly if they are taking prescription or over-the-counter-medications or supplements. Women who are pregnant or breast feeding and children should also consult with there physician prior to taking any new supplements. There has been no reported significant side effects associated with the use of Garcinia cambogia at the time of this writing.
   In recent years, Garcinia cambogia has been promoted for weight loss due to various possible effects it may have on the body. First, it is believed to interfere with an enzyme needed to store fat, possibly causing more fat from foods to be eliminated from the body. In addition, Garcinia cambogia may cause the body to use existing fat stores for energy during prolonged exercise. Ordinarily, carbohydrates are used first during exercise. In animal studies, hydroxycitric acid (HCA), a major component of Garcinia cambogia, also seemed to reduce appetite by raising the amount of serotonin in the body. Serotonin is a neurotransmitter that is thought to affect appetite control. In clinical studies of humans, however, individuals who took Garcinia cambogia while following a weight-reduction diet lost no more weight, on average, than members of a control group who followed the same diet without taking a supplement. Other human studies have had mixed results on appetite reduction; some showed little or no effect, while others reported a 15% to 30% decrease in food intake. More studies are needed to prove or disprove the effectiveness of Garcinia cambogia in weight control.
 


  Safety and Acute toxicity:(-) HCA

 Acute toxicity(LD50):
 LD50-Lethal dose,50 percent kill. Intraperitonial.Rodent-mouse. > 2000 mg/kg.
 LD50-Lethal dose,50 percent kill. Oral.Rodent-mouse. > 4000 mg/kg.
  It does not affect respiration, phosphorylation, or the citrate synthesis of the reconstituted mitochondria. It neither penetrates the mitochondria nor affects normal energy production. Therefore (-) HCA decreases food intake, diminishes appetite, and without adverse side effects.


  Process for the production of potassium hydroxy citric acid, and compositions containing the potassium hydroxy citric acid:

   

Claims:

We claim: 1. A process for the production of potassium hydroxy citric acid, which potassium hydroxy citric acid is not in the form of a lactone, comprising the steps of: a) providing Garcinia fruit; b) extracting the Garcinia fruit with an alkyl alcohol to obtain an extract; c) repeating step b) to obtain another extract; d) combining the extracts of steps b) and c) to obtain a combined extract; e) treating the combined extract with potassium hydroxide to obtain a treated extract; f) refluxing the treated extract to obtain potassium hydroxy citrate precipitate; g) isolating the precipitate; h) washing the precipitate with an alkyl alcohol to obtain a washed precipitate; and thereafter i) drying the washed precipitate to obtain dried potassium hydroxy citric acid. 2. The process of claim 1 comprising: a) providing Garcinia fruit; b) extracting the Garcinia fruit with methanol at reflux temperature and collecting the extract; c) repeating step b) an additional two times; d) combining the three extracts of steps b) and c); e) treating the combined extracts with methanolic potassium hydroxy at about pH 10 and reflux for about three hours to precipitate potassium hydroxy citrate; f) filter the precipitate; g) wash with methanol and dry under vacuum; and h) mill, sift, blend, and pack the dried product under nitrogen. 3. A new technological process for commercial manufacturing of potassium hydroxy citric acid from natural source, which potassium hydroxy citric acid is not in the form of a lactone, said process comprising the steps of: a) providing Garcinia fruit; b) extracting the Garcinia fruit with an alkyl alcohol to obtain an extract; c) repeating step b) an additional two times to obtain another extract; d) combining the extracts of steps b) and c) to obtain a combined extract; e) treating the combined extract with potassium hydroxide to obtain a treated extract; f) refluxing the treated extract to obtain potassium hydroxy citrate precipitate; g) isolating the precipitate; h) washing the precipitate with an alkyl alcohol to obtain a washed precipitate; and thereafter i) drying the washed precipitate to obtain dried potassium hydroxy citric acid. 4. The new technological process according to claim 3, further comprising milling, sifting, blending and packing the dried potassium hydroxy citric acid under nitrogen. 5. The process of claim 3, wherein the Garcinia fruit is Garcinia cambogia or Garcinia indica fruit. 6. The process of claim 5, wherein the Garcinia fruit is Garcinia cambogia.

Description:

BACKGROUND OF THE INVENTION 1. Field of the Invention The invention is directed to a new process for making hydroxy citric acid in a form that is stable and biologically active. Compositions containing the potassium hydroxy citric acid are useful as natural appetite suppressants. 2. Description of Related Art During the 1970s, scientists at Brandeis University and at Hoffman LaRoche demonstrated that synthetic hydroxycitric acid, when blended with the diet, had a marked suppressive effect on weight gain in rats. The researchers noted that the HCA-treated rats tended to eat less; food consumption was suppressed by 10% or more on optimal HCA intakes, that is, when HCA constituted 1% or more of the diet. The mechanism by which the HCA affected weight gain was not known. Since the brain uptake of HCA appeared to be negligible, scientists speculated that the appetite-suppressive effect of HCA was exerted not through central nervous system (CNS) action, but rather by directly affecting the metabolic processes of the organism. One of the most metabolically active organs in the body is the liver. One important function of the liver is to insure that the blood maintains adequate concentrations of glucose to fuel the body's energy requirements. The liver can store dietary glucose in the form of the polysaccharide glycogen, and release glucose when blood glucose levels are low. The liver can also synthesize glucose in a complex process known as gluconeogenesis, from either amino acids or lactic acid as starting material. This newly synthesized glucose can either be released into the blood stream to supply energy requirements of body tissues, or can be stored as glycogen for future use. The direct parasympathetic connection between the liver and the CNS monitors the level of glucose and glycogen in the liver. A high level of glycogen, as a result of high glucose supply, is translated by the CNS as the state of satiety, which results in decreased craving for food. Since increased glycogen in the liver aids satiety, the effect of HCA on gluconeogenesis in rat liver has been studied. It has been found that the rate of gluconeogenesis, from lactate or the amino acid alanine, was approximately doubled in HCA-treated rats. This result provides support for the idea that HCA causes the observed appetite suppression via altering the rate of gluconeogenesis. However, it appears unlikely that reduction of food intake can entirely account for the substantial reductions in weight gain seen in HCA-treated rats. For example, in one study, the net reduction in food consumption during the 80 day study period amounted to only 4%--and yet the rats had gained 78% less weight than the controls over this period. Other studies, providing less dramatic results, suggested that the reduction of weight gain was disproportionately large compared to the reduction in food consumption. Because of this discrepancy between considerable weight loss versus a meager appetite suppression, it has been postulated that HCA exerts a mechanism which increases fat burning, which in turn could decrease body weight, in addition to affecting gluconeogenesis. Fat burning, or oxidation, plays a prominent role in liver metabolism. Liver metabolic activity accounts for over a quarter of the total body oxygen consumption in a subject at rest. The substantial energy needs of the liver are met largely by oxidation of fat. The dietary fat is absorbed by liver cells, which oxidize or burn it for energy in the mitochondria. The fats are transported from the cell cytoplasm into the liver mitochondria, by linking them to the special transporting molecule L-carnitine. This reaction is facilitated by the enzyme carnitine acyltransferase. Carnitine acyltransferase is inhibited by malonyl CoA, which can be obtained from conversion of acetyl CoA. Malonyl CoA, can not only inhibit the fat burning process, but also increase the body fat synthesis, since it is the direct precursor for the synthesis of fat and cholesterol. The acetyl CoA is synthesized in mitochondria, but it has to be transported to the cell cytoplasm to exert its biochemical action. However, it cannot be transported to the cytoplasm from mitochondria before it is converted to citric acid. Thus, citric acid is a transportable form of acetyl CoA. Citric acid, once in the cytoplasm, is converted to acetyl CoA with the help of the enzyme--citrate lyase. HCA was found to be an extremely potent competitive inhibitor of citrate lyase (K.sub.i =0.15 .mu.M). The affinity of the enzyme for HCA was over a hundred times greater than the affinity of the enzyme for citric acid. This action was afforded only by a HCA in a pure acid form, but not in the lactone form. The significance of citrate lyase inhibition by HCA is that without active citrate lyase, little acetyl CoA could reach the cytoplasm. This in turn would limit the availability of malonyl CoA and slow the synthesis of fats and cholesterol, while disinhibiting the metabolic breakdown of fat, or oxidation of fat. In light of the considerations noted above, it is likely that the ability of HCA to promote fat loss in humans results primarily from the stimulation of fat oxidation. Activation of fat oxidation in the liver also tends to stimulate gluconeogenesis, primarily due to increased activity of the key enzyme pyruvate carboxylase. This in turn may replenish the stores of liver glycogen, and send a message of satiety to the brain center. The drawbacks of HCA use as a weight loss compound stem from the following problems: 1. The poor technology of HA extraction from the fruit of Garcinia cambogia often provides HCA in lactone form, which is inactive, or less active, in inhibiting the citrate lyase; 2. The HCA, if not stabilized chemically, has natural propensity to be converted to the lactone form in aqueous solutions and in the gastrointestinal tract, i.e., without absorption of HCA in pure acid form, the HCA can hot inhibit the citrate lyase; and 3. High concentrations of HCA, that is, 1% or more (by weight) of the daily dietary intake, are required to exert the metabolic activity, because of poor cellular uptake. Without absorption of HCA, and the presence of HCA in the cytoplasm in pure acid form, HCA can not inhibit citrate lyase and exert its inhibitory activity on acetyl CoA formation. In the past, it has been difficult to isolate hydroxy citrate in a form which is both stable and biologically active. Hydroxy citric acid exists in two forms, the free acid form and the lactone form. The free acid form is biologically active and the lactone form is inactive. However, the free acid form is not stable and gets converted to its lactone form, which is stable but inactive. One prior art isolation procedure, that of Y. S. Lewis et al., in Phytochem 1965, Vol. 4, pp. 610-625, results in the isolation of hydroxy citric acid lactone. I. WATER EXTRACT OF (-) HYDROXYCITRIC ACID FROM FRUIT OF GARCINIA CAMBOGIA (Lewis, Y. S. and Neelakantan, S., phytochemistry (1965) Vol. 4; pp. 619-625) The prior art procedure to obtain (-)HCA from Garcinia cambogia on a large scale included the following procedure: 1. The dried rind was cooked with about three volumes of water in an autoclave (10 lb/in.sup.2) for 15 minutes; 2. The resulting extract was filtered through a cloth and then through a paper filter; 3. The obtained filtrate was concentrated to a small volume, and the alcohol precipitation method removed pectin contamination; 4. The clear filtrate was then treated with potassium hydroxide (alkali) to form viscous, dark, heavy liquid; this treatment resulted in formation of a hygroscopic material consisting of potassium salt of hydroxycitric acid; 5. The clear supernatant was decanted off and the oily liquid washed with 60% alcohol several times; 6. By repeated treatment with absolute alcohol, the material could be dried to a pale yellow hygroscopic powder, which formed pure alkali salt; 7. Aqueous solutions of the alkali salt were passed through a cation-exchange resin (Zeocarb 215) for recovery of the acid; 8. The obtained (-)HCA was chemically unstable, and upon evaporation formed lactone. Another process for isolation of lactone was reported by Y. S. Lewis. II. ACETONE EXTRACT OF (-) HYDROXYCITRIC ACID FROM THE FRUIT OF GARCINIA CAMBOGIA (Lewis, Y. S. (1981) Methods in Enzymology, Vol. 77; Published by Academic Press; pp. 613-619). 1. One kg of fruit of Garcinia cambogia is kept in 1500 ml of acetone in an overnight; 2. The fruit is re-extracted in a similar manner; 3. Acetone is removed from the combined extracts by distillation; 4. The viscous residue is stirred with 1 liter of water at 45-50.degree. C.; 5. The mixture is filtered through cheesecloth; 6. The precipitated insoluble material is removed by filtration; 7. The reddish brown filtrate is treated with activated charcoal at 80-90.degree. C. and concentrated to a thick syrup; 8. The syrup is "seeded" with a few crystals of the lactone and left overnight; 9. The yield is vigorously extracted with 3 liters of ether; 10. The combined extracts are dried over anhydrous sodium sulfate; 11. Ether is then removed, and the remaining material is white solid, consisting mainly of lactone. The yield is approximately 150 gm. SUMMARY OF THE INVENTION The principle of the present invention is to provide technology for extraction of HCA in pure acid form, and technology for chemical modification of HCA to afford chemically stable product, which will not convert into lactone form, which will not be hygroscopic, and which is soluble in aqueous solutions and easily absorbable by the gastrointestinal tract. The invention provides HCA by combining it with potassium into potassium hydroxycitrate--a water soluble salt. Potassium is an ion primarily found in the cell cytoplasm, and it can easily cross from outside the cell to inside the cell. The cell membrane permeability for potassium is 100 times higher than for sodium and 25 times higher than for chloride. Potassium salt of HCA acts as a transporter of HCA inside the cell, where the biochemical action of HCA is exerted. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is an infrared spectrum of potassium hydroxy citrate. FIG. 2 is a thermogram of potassium hydroxy citrate. FIG. 3 is a NMR spectrum of potassium hydroxy citrate. FIG. 4 is an HPLC chromatogram of Potassium Hydroxy Citrate DESCRIPTION OF THE PREFERRED EMBODIMENTS Process Outline The process of the present invention is used to isolate hydroxy citric acid as potassium hydroxy citrate from a natural source of Garcinia species. Preferred sources include Garcinia cambogia and Garcinia indica. Briefly, fruit of the Garcinia species is extracted with an alkyl alcohol. Preferred alcohols include methyl alcohol, ethyl alcohol, propyl alcohol, and isopropyl alcohol. Especially preferred is methanol. The extract is treated with a suitable alkali to precipitate the potassium hydroxy citrate. Preferred alkalis include potassium hydroxide, potassium carbonate, etc. Most preferred alkali is potassium hydroxide. The general process includes the following steps. Garcinia fruit is extracted with an alkyl alcohol at above ambient temperature. This is done at or above atmospheric pressure. The extract is collected. The extraction step is repeated at least three times. The extracts are combined and treated with an alcoholic solution containing alkali. The resultant mass is heated to above ambient temperature and pH is adjusted to make the solution alkaline. The pH of the solution is normally between 8 to 11.5. The product is filtered and washed with alcohol. The product is dried at or above 25.degree. C. under vacuum or at atmospheric pressure or under inert atmosphere, like nitrogen. The dried product is milled, sifted, blended and packed under nitrogen blanket to obtain product. The yield from 500 kgs of garcinia fruit ranges from 60 to 150 kgs of potassium hydroxy citrate based on the hydroxy citric acid content present in the fruit. Unique Aspects of our Process Hydroxy citric acid exists in two forms, i.e., free acid form and lactone form. The free acid form is biologically active and the lactone form is inactive. However, the free acid form is not stable and this gets converted to its lactone form, which is stable but inactive. In our process, the free acid form is isolated and stabilized as potassium salt to retain the activity. This is one of the unique aspects of our process. Another unique aspect of our process is that our potassium hydroxy citrate is water soluble and therefore, it is readily available in the biological system for its bioefficacy. EXAMPLE The detailed procedure used to obtain the product trademarked at CITRIN.RTM.-K is as follows: 1. The 500 kg of Garcinia fruit is extracted with 1500 l of methanol at about reflux temperature for 3 hours; 2. This is filtered through the cloth filter to collect the first extract; 3. Additional 1500 l of methanol is added to the Garcinia fruit and refluxed for about 3 hours; 4. This is filtered to collect the second extract; 5. The 1500 l of methanol is added again to the Garcinia fruit and refluxed for 3 hours; 6. This is filtered, and the third extract is collected; 7. All the three extracts are combined; 8. The combined extracts are treated with methanolic potassium hydroxide at pH 10; 9. This is again refluxed for about 3 hours to attain constant pH 10 to precipitate potassium hydroxycitrate; 10. The precipitate is filtered and washed with 500 l of methanol; 11. The precipitate is dried under vacuum at about 70.degree. C.; 12. The dried product is milled, sifted, blended and packed under nitrogen blanket to obtain product trademarked at CITRIN.RTM.-K; 13. The methanolic mother solution is distilled to recover methanol; 14. The yield from 500 kg of Garcinia fruit is about 150 kg of potassium hydroxycitrate. The specifications of the product CITRIN.RTM.-K is given below: Specifications Molecular Structure Molecular Formula C.sub.6 H.sub.5 K.sub.3 O.sub.8 H.sub.2 O Molecular weight 340.41 Description Beige to pale brown colored powder Solubility Soluble in water, acids and aqueous alcohols. Insoluble in solvents like methanol, alcohol, chloroform, benzene, etc. Loss on Drying Not less than 3% and not more than 6.0% pH of 5% solution 7.0 to 9.0 in water Specific Rotation -18.degree. to -25.degree. on anhydrous basis Potassium content not less than 30% by weight on anhydrous basis Hydroxy citric Not less than 50% on anhydrous basis acid content Lactone content less than 2% by weight (HPLC) Identification a) By IR Spectrum The infrared absorption spectrum of a potassium bromide dispersion of potassium hydroxy citrate, previously dried, exhibits maxima only at the same wavelength as that of similar preparation of working Standard. IR Spectrum of Potassium Hydroxy Citrate Working Standard is shown in FIG. 1. b) For Potassium Produces yellow or orange-yellow precipitate with sodium cobaltinitrite solution. Dissolve 50 mg of 1 ml of water, add 1 ml of dilute acetic acid and 1 ml of freshly prepared 10% w/v solution of sodium cobaltinitrite. A yellow or orange-yellow precipitate forms immediately. c) For Citrate Dissolve 0.5 g in a mixture of 10 mL of water and 2.5 mL of 2 N nitric acid. Add 1 mL of mercuric sulphate solution heat to boiling, and add 1 mL of potassium permanganate solution: a white precipitate is formed. d) By Paper Chromatography Mobile Phase Butanol (4): Acetic Acid (1): Water (5) Prepare 100 mL of mobile phase in separator and mix well. Allow it to separate and use the upper layer as mobile phase. Stationary Phase: Whatman filter Paper No. 1 Sample Preparation Dissolve 100 mg of the sample in 1 mL of water and dilute to 10 mL with methanol in a volumetric flask. Standard Preparation Dissolve 100 mg of the Working Standard in 1 mL of water and dilute to 10 mL with methanol in a volumetric flask. Procedure Apply separately equal volume (10 .mu.l) of sample and standard preparation and develop the chromatogram in the chamber previously saturated with mobile phase. After developing the chromatogram to 3/4, the paper is removed and dried in a current of air. Detection The paper is sprayed with sodium metavanadate solution (5% w/v) and observed for the orange spot. The Rf value of the spot obtained from the sample solution is same as that of the Standard solution. Loss on Drying Limit: Not less than 3.0% and not more than 6.0% The material shows weight loss of about 5% when dried at 150.degree. C. under vacuum for four hours. This weight loss is due to the release of water of hydration from the molecule. Thermal Analysis Potassium hydroxy citrate is analyzed by Thermogravimetry. This technique is used to estimate the presence of water of hydration in the product. The details of the methods are given below: In this method, the sample is heated under nitrogen/argon atmosphere and the weight loss is recorded continuously. Limit: The weight loss is not more than 6.0% Analysis is carried out using about 3 mg of the sample accurately weighed. The temperature setting is from 30.degree. C. to 400.degree. C. with the rate of heating as 10.degree. C. per minute. The heating of the sample is done under nitrogen/argon atmosphere flowing at a flow rate of 40 mL/min. From the TGA thermogram, it is observed that there is weight loss between 180.degree. C. and 250.degree. C. to a level of about 5% which indicates the presence of water of hydration. The percentage loss corresponds to one molecule of water. A typical thermogram is given in FIG. 2. pH of Solution pH of 5.0% w/v solution is between 7.0 and 9.0. Dissolve 2.5 g in 50 ml of water and determine the pH using suitable calibrated pH meter. Specific Rotation Between -18.0.degree. and -25.0.degree. calculated on anhydrous basis. Weigh accurately about 1 g of the sample and transfer into a 100 ml volumetric flask, dissolve in water, dilute to volume and mix. Measure the rotation using suitable polarimeter at about 25.degree. C. Assay Assay of the product is estimated by estimating the content of HYDROXY CITRIC ACID and POTASSIUM. For determination of HYDROXY CITRIC ACID, the following methods are employed: i) TITRATION METHOD ii) HPLC METHOD The details of the methods are given below: Limit: Content of HCA is not less than 50.0% calculated on anhydrous basis Titration Method Weigh accurately about 200 mg of the sample and transfer into a beaker. Add 100 ml of water and dissolve. Pass the solution through cation ion exchange resin column and collect the affluent into a 1 L flask. Rinse the beaker with water and pass the washings through the column. Wash the column with distilled water until the elute shows a pH of 4.0 to 4.5. Adjust the volume to about 500 ml and titrate with 0.1 N sodium hydroxide solution using phenolphthalein solution as indicator. Perform a blank titration after eluting 500 ml of water through the column. Calculation: (Titre value-blank value).times.0.1 N of NaOH.times.0.006933.times.100.times.100 0.1.times.Weight of the sample.times.(100--LOD) Note 1 Column Preparation and Regeneration: About 75 g of cation exchange is packed in a column of 2 cm diameter. Soak the column for 30 minutes in 2 N Hcl. Wash thoroughly with distilled water to get a pH of 4.0 to 4.5 After the analysis, the cation exchange resin is soaked with 2 N Hcl for 3 hours. It is then washed well with distilled water until the pH of the washings shows 4.0 to 4.5 Note 2 The above method is based on the published research paper titled "Chemical Constituents of Kokum Fruit Rind" by CFTRI, Mysore. Note 3 Specification of the cation exchange resin is given in FIG. 3 Note 4 The factor of 0.006933 is arrived at by the following calculation (C.sub.6 H.sub.8 O.sub.8)+6NaOH --Na.sub.6 (C.sub.6 H.sub.5 O.sub.8)+6H.sub.2 O [2 moles of HCA]. 416 g of HCA.ident.6000 ml of 1 N NaOH or 6000 ml of 1 N NaOH.ident.416 g of HCA ##EQU1## HPLC Method In this method, normally, (-) Threo Hydroxy Citric Acid Ethylene Diamine Salt (Fluka Standard) is used as a Standard to estimate Hydroxy Citric Acid content in Potassium Hydroxy Citrate. This Standard is not readily available, and therefore an alternate standard, Potassium Hydroxy Citrate is preferred. A pure sample of Potassium Hydroxy Citrate has been synthesized and validated against the Fluka Standard (FIG. 4). In the method given below, Potassium Hydroxy Citrate is used as a Working Standard (WS). Mobile Phase Prepare 0.01 N sulfuric acid, filter and degas. Sample Preparation 50 mg of the sample is accurately weighed, dissolved in water and diluted to 25 ml with water. Standard Preparation 50 mg of Potassium Salt of Hydroxy Citric Acid (WS) is dissolved in 10 ml of water, and diluted to 25 ml with water. Chromatographic System The liquid chromatograph is equipped with 210 nm detector and a 4.6.times.250 mm organic acid column (Vydac make). The flow rate is about 1 ml per minute. Chromatograph the standard preparation and calculate the Relative Standard Deviation (RSD) for replicate injections. The RSD is not more than 2.0%. Procedure Separately inject equal volume (20 .mu.l) of sample and standard preparation and record the responses obtained for the major peaks. Calculation: Area of the sample.times.standard weight.times.(100-water content of STD.times.HCA Content of Standard/Area of the standard.times.sample weight (100-LOD of Sample)=Hydroxy Citric Acid content in the sample Estimation of Potassium This estimation is done by two methods: i) FLAME PHOTOMETRY ii) ATOMIC ABSORPTION The details of the methods are given below: Limit of Potassium: Not less than 30.0% calculated on anhydrous basis Flame Photometry Standard Stock Solution Weigh accurately about 1.84 g of Potassium Chloride, previously dried at 105.degree. C. for 2 hours and transfer into a 250 ml volumetric flask, add water to volume and mix. Lithium Diluent Solution Transfer 1.04 g of Lithium Nitrate to a 1000 ml volumetric flask, add a suitable nonionic surfactant, add water to volume and mix Standard Preparation Pipette 5 ml of stock solution into a 50 ml of volumetric flask, dilute to volume with water and mix. Transfer 5 ml of this solution to a 100 ml volumetric flask and dilute with lithium diluent solution to volume and mix. Assay Preparation Weigh accurately about 3 g of the sample and transfer into a 250 ml volumetric flask, add water to dissolve and dilute to volume and mix. Pipette 5 ml of this solution into a 50 ml volumetric flask, add water to volume and mix. Transfer 5 ml of this to a 100 ml volumetric flask, dilute with lithium diluent solution to volume and mix. Procedure Using a suitable flame photometer adjust to read zero with lithium diluent solution concomitantly determine the emission readings for Standard and Sample preparations at about 766 nm Calculate the content of Potassium as follows: Emission Reading of Standard.times.Weight of Standard.times.39.1/Emission Reading of Sample.times.Weight of Sample.times.74.55 Atomic Absorption Potassium Stock Solution Dissolve 190.7 mg of potassium chloride, previously dried at 105.degree. C. for 2 hours, in water. Transfer to a 500 ml volumetric flask, dilute with water to volume and mix, transfer 5 ml of this solution to a 100 ml volumetric flask, dilute to volume with water and mix. Standard Preparation To separate 100 ml volumetric flask, transfer 10, 15 and 20 ml respectively of Potassium stock solution. To each flask, add 2 ml of sodium chloride solution (1 in 5) and 1 ml of hydrochloric acid, dilute with water to volume and mix. Assay Preparation Weigh accurately about 1 g of the sample and transfer into a 500 ml volumetric flask dissolve in water, dilute to volume and mix. Transfer 5 ml of this to a 100 ml volumetric flask, dilute to volume with water and mix. Transfer again 5 ml of this solution to a 100 ml volumetric flask, add 2 ml of sodium chloride solution (1 in 4) and 1 ml of hydrochloric acid, Dilute with water to volume and mix. Procedure Concomitantly determine the absorbencies of the Standard preparations and assay preparation at the potassium emission line of 766.5 nm, with a suitable atomic absorption spectrophotometer equipped with a potassium hollow cathode lamp and an air acetylene flame, using water as the blank. Plot the absorbance of standard preparation versus concentration in .mu.g per ml of potassium and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration, in .mu.g per ml of potassium in the assay preparation. Calculate the content of potassium in mg as follows: 200.times.C where `C` is concentration in .mu.g per ml Calculate the percentage of potassium as follows: 200.times.`C`.times.100/Wt. taken in mg Microbial Assay Total Plate Count, E. coli, Salmonella, yeasts and molds are estimated as per procedures described in "OFFICIAL METHODS OF ANALYSIS--ASSOCIATION OF OFFICIAL ANALYTICAL CHEMISTS" (14th Edition, 1990) The limits are given below: Total Plate Count 10000 cfu/g E. coli Absent Salmonella Absent Yeasts/Molds 1000 cfu/g Aflatoxins are estimated by the following procedure, which is based on the methods described in "OFFICIAL METHODS OF ANALYSIS--ASSOCIATION OF OFFICIAL ANALYTICAL CHEMISTS" (15th Edition 1990) Limit: Aflatoxins--Not more than 20 parts per billion Procedure for Estimated Aflatoxins Apparatus High speed stirrer (1400-1600 rpm with stainless steel shaft and propeller blade) Ultra Violet Light Long wave UV with intensity of 430.mu. watt/cm.sup.2 at 15 cm at 365 nm Minicolumn Borosilicate std wall tubing, Ca 6(id).times.200 nm tapered at one end to ca 2 mm Densitometer With fluorometry attachment Reagents Distilled water or deionized water may be used. a) Solvents CHCl.sub.3 and acetone AR grade must be used b) Potassium hydroxide wash solution 0.02 N KOH with 1% Kcl. Dissolve 1.12 g KOH pellets and 10 g Kcl in 1 L H.sub.2 O c) Sodium hydroxide solution 0.02 N 8.00 g of NaOH/LH.sub.2 O d) Sulphuric acid solution--0.03% Dilute 0.3 ml H.sub.2 SO.sub.4 to 1 L e) Precipitating reagents i) Copper carbonate, basic ii) Ferric chloride slurry: Mix 20 g anhydrous FeCl.sub.3 with 300 ml H.sub.2 SO.sub.4 f) Diatomaceous earth Hyflo Super-Cel or equivalent g) Column Packing Silica Gel G 60-100mesh; Florisil 100-200 mesh; Alumina neutral 80-200 mesh; CaSO.sub.4 anhydrous 20-40 mesh h) Aflatoxin Standard solutions Standards from Sigma Chemicals, USA, are used. Preparation of Mini Column Tamp small plug of glass wool into tapered end of column. To column, add to height indicated in following order: 30 mm silica gel; 10 mm neutral alumina and 10 mm CaSO.sub.4. Tamp small plug of glass wool on top of column. Tamp column after each addition to settles packing and maintain interfaces as level as possible. After packing, apply pressure to top glass wool plug with 5 mm diam. rod. Activate packed columns at 110.degree. C. for 1-2 hours and store in vapor tight container. Extraction of Aflatoxins Weigh 50 g sample into stirrer, add 250 ml CHCl.sub.3 --H.sub.2 O(85+15) and stir it for 30 minutes. Filter through Whatman No. 4 filter paper. Collect 150 ml filtrate and transfer to 500 ml beaker. Purification To 50 ml beaker, add 170 ml 0.2 N NaOH and 30 ml FeCl.sub.3 slurry and mix well. Add 3 g basic CuCO.sub.3 to sample extract in 500 ml beaker and mix well, add both 1 and 2 mixtures and mix well. Filter the mixture through Whatman No. 4 filter paper in a Buchner funnel using Hyflo supercel bed. Transfer 150 ml filtrate to 500 ml separator, add 150 ml 0.03% H.sub.3 SO.sub.4 and 10 ml CHCl.sub.3. Shake vigorously for 5 minutes and allow to stand for 30 minutes. Transfer lower CHCl.sub.3 layer (13-14 ml) to 125 ml separator. Add 100 ml KOH wash solution, swirl gently for 30 seconds and allow to stand. (If emulsion occurs, drain emulsion into 10 ml test tube add 1 g anhydrous Na.sub.2 SO.sub.4, stopper, shake 30 seconds and allow to stand (CHCl.sub.3 phase need not be completely clear). If emulsion is not broken, transfer emulsion to 125 separator and wash with 50 ml 0.03% H.sub.2 SO.sub.4. Collect 3 ml CHCl.sub.3 layer in 10 ml test tube. Column Chromatography Transfer 2 ml ChCl.sub.3 solution (extract) to minicolumn, using 5 ml syringe. Hold the column vertically and apply slight air pressure (with the help of a rubber bulb) to force solvent through column at rate .ltoreq.10 cm/min until solvent appears at tip. Remove rubber bulb and add about 5 ml of elution solvent containing CHCl.sub.3 -acetone (9:1). Collect the fractions. Examine column under UV lamp for blue fluorescent band at top of Florisil layer (Ca 2.5 cm from bottom of column) indicative of aflatoxin. Collect the fractions corresponding the blue band separately and concentrate to a residue. TLC Dissolve the residue in minimum quantity of CHCl.sub.3 and carry out the TLC testing along with authentic sample of Aflatoxins. Solvent system--Benzene:Methanol:Acetic Acid (95:5:5). Quantify the aflatoxin by using TLC densitometer. STABILITY The stability of the product was evaluated in solid state and in aqueous solution in temperature and humidity conditions as specified below. The following parameters of the product were considered: physical appearance, specific rotation, HCA content by HPLC, lactone content by HPLC. 1. In solid state: a) Room temperature, b) 37.degree..+-.2.degree. C. and 75%.+-.2 relative humidity c) 45.degree..+-.2.degree. C. and 75%.+-.2 relative humidity 2. In solution form (5% in water) a) Room temperature b) 37.degree..+-.2.degree. C. c) 45.degree..+-.2.degree. C. Conclusion The product is found to be stable under stress conditions (higher temperature and higher humidity) for a minimum of 90 days. These results indicate that the product will be stable for about 5 years under normal storage conditions.