Digestion 19: 48-51 (1979)
Pancreatic Enzymic Activities of Commercial Pancreatic Enzyme Preparations Incubated in Human Small Intestinal Juice I. Ihse and P. Lilja Department of Surgery, University of Lund, Lund
Key Words. Amylase • Lipase • Phospholipase • Pancreatic juice • Pancreatic insufficiency • Intestinal secretion Abstract. The activities of amylase, lipase, phospholipase and trypsin of twelve commercial
pancreatic enzyme preparations were measured under identical conditions. Human small intestinal juice was chosen as incubation medium. A wide variation of enzymic activities was found in preparations in tablet form contrary to preparations in granulated form. A prerequisite for a successful therapy is enzymically potent pancreatic extracts with high enzyme content per tablet, capsule or recommended dose of granulated preparation. Therefore, some preparations seem to be preferable to others for clinical purposes.
Material and Methods All preparations were obtained from the hospital pharmacy. One tablet or 10 ml o f the preparations in granulat ed form was crushed in a mortar and incubated for 40 min at a temperature of 37 °C in 50 ml human duodenal juice obtained from normal subjects. Six tablets or six 10-ml dosages from 2 different batches o f each preparation were analyzed in duplicate. The activities of amylase, lipase, phospholipase and trypsin o f the incubation medium (human duodenal juice) were estimated before addition o f the pancreatin preparation and after incubation with it. The enzyme activities of the different preparations were defined as the activities estimated after incubation minus those
Downloaded by: Univ. of California Santa Barbara 128.111.121.42 - 3/26/2018 1:49:21 AM
Pancreatic enzyme preparations are used in malabsorption caused by exocrine pancreatic insufficiency. Comparison of in vitro digestive capacity of such preparations, as reported by the manufacturers, is difficult due to the con siderable variation in the enzyme assays and units used. The purpose of the present study was to measure and compare enzyme activities of 12 commercial supplements under identical condi tions. In order to imitate physiological cir cumstances as close as possible, human small intestinal juice was chosen as the incubation medium.
In vitro Activities of Pancreatic Supplements
49
estimated before addition of the pancreatin prepara tions to the medium. Amylase was determined according to Dahlquist (1), and lipase by a scaled-down version o f the method described by Erlanson and Borgström (3). Phospho lipase was determined according to Ihse and Arnesjo (6) and trypsin modified after Hummel using TAME (p-toluene-sul phony l-I.-arginineme thy lester) as the sub strate (5). As human duodenal juice was used for the incubation, no further activators of trypsinogen or prophospholipase were needed.
Results
The activities of the lipolytic enzymes of the different preparations are shown in table I. As can be seen, Pancreon forte and Pancreon comp
forte had the highest activity of both lipase and phospholipase. Furthermore, it is obvious that the lipase as well as the phospholipase activity of the different tablets varied considerably. On the other hand, the activities of lipase and ¡hospholipase per milliliter of all three prepara tions in granulated form showed less variation. Also, in the amylase and trypsin activities of the tablets, a wide range was found (table II). Here too, Pancreon forte and Pancreon comp forte revealed the highest activities of amylase and trypsin. The variation of the trypsin acti vities of the different preparations in granu lated form was not as marked as for the tablets, while the amylase activity in Pancreon granulate was only half of that of Pancreatin
Name
Manufacturer
Lipase activity per tablet
Lipase activity of maximum recommended dosage per meal
Phospho lipase activity per tablet
Phospholipase activity of maximum recommended dosage per meal
Pancreon forte Pancreon comp forte Festal Combizym comp Pancreozym Helopanzym Supplezym Combizym Luizym
(T)
Kali-Chemie
11,861
23,729 (2 tablets)
1,816
3,632
(T, B) (T. B)
Kali-Chemie Hoechst
10,825 6,900
21,650 (2 tablets) 13,800 (2 tablets)
1,784 459
3,568 918
(T, B) (T) (T, B) (T) (T) (T)
Luitpold-Werke Tika Helopharm Tika Luitpold-Werke Luitpold-Werke
5.589 5,483 3,610 1,387 1.259 515
5,589(1 32,989 (6 7,220 (2 8,322 (6 2,518 (2 1,030 (2
598 685 223 788 838 114
598 4,110 446 4,728 1,676 228
Pancreatin Pancreon Panteric
(G) (G) (G)
Rosco AS Kali-Chemie Parke-Davies
1,781' 1,398' 1,966'
26,715 (15 ml) 13,980 (10 ml) 4,915 (2.5 ml)
550' 568' 728'
8,250 5,680 1,820
tablet) tablets) tablets) tablets) tablets) tablets)
Lipase activity expressed as micromoles fatty acids released per milliliter per minute; phospholipase activity expressed as micromoles fatty acids released per milliliter per minute. T = Tablets; G = granules; B = the preparation contains bile acids. ' Lipase activity and phospholipase activity, respectively, per milliliter of granules.
Downloaded by: Univ. of California Santa Barbara 128.111.121.42 - 3/26/2018 1:49:21 AM
Table I. Mean enzyme activities of 6 different units (tablets, 1 ml of granules) of 12 different pancreatic enzyme preparations
Ilise/Lilja
50
Table II. Mean enzyme activities of 6 different units (tablets, 1 ml of granules) o f 12 different pancreatic enzyme preparations Name
Amylase activity per tablet
Amylase activity of maximum recommended dose per meal
Pancreon forte Pancreon comp forte Festal Combizym comp Pancrcozym Helopanzym Supplezym Combizym Luizym
(T) (T, B) (T, B) (T, B) (T) (T, B) (T) (T) (T)
24,850 33,793 10,070 15,340 7,188 2,806 3,492 10.225 187
49,700 67,586 20,140 15,340 43,128 5,612 20,952 20.450 374
Pancreatin Pancreon Panteric
(G) (G) (G)
13,598' 6,400' 12.190'
203.970 64,000 30.475
Trypsin activity per tablet
Trypsin activity of maximum recommended dosage per meal
3,764 5,735 1,125 912 875 1,050 375 1.259 515
7,528 11,740 2,250 912 5,250 2,100 2,250 2,518 1,030
858' 756' 866’
12,870 7,560 2,160
Amylase activity expressed as micromoles maltose released per milliliter per minute; trypsin activity expressed as micromoles TAME hydrolyzed per milliliter per minute. T = Tablets; G = granules; B = the preparation contains bile acids. ' Amylase activity and trypsin activity, respectively, per milliliter of granules.
Discussion
The present paper reports the activities of amylase, lipase, phospholipase and trypsin of 12 commercial pancreatic enzyme preparations. From a clinical point of view tire most im portant criterion of efficiency of such a pre paration is its ability to augment pancreatic enzyme levels in intestinal contents. A pre requisite for this objective is enzymically potent pancreatic extracts with high enzyme content per tablet, capsule or recommended dose of preparation in granulated form. The results of the present study confirm previous reports of wide variations in enzymic activity of different commercial pancreatic extracts (2,4). Some preparations, therefore, are preferable to others for clinical purposes.
Downloaded by: Univ. of California Santa Barbara 128.111.121.42 - 3/26/2018 1:49:21 AM
granules and Panteric. However, the amylase activity of all three preparations in granulated form was high. When calculating the highest recommended dose per meal it was obvious that the variation of enzyme activities between the different pre parations was still more marked (table I, II). For instance two tablets of Pancreon forte contained more than 20 times as much lipase as two tablets of Luizym. Best overall results were obtained with Pancreon forte and Pancreon comp forte among the tablets, while the three preparations in granulated form (Pancreatin, Pancreon, Panteric) all contained adequate amounts of enzymes. Finally, it was found that there was only a slight variation in enzyme activity of tablets or granules from the same batch.
In vitro Activities of Pancreatic Supplements
In this study we used human duodenal juice as the incubation medium in order to imitate physiological conditions as close as possible. Thus, the pancreatic extracts were incubated in a milieu containing naturally occurring activa tors and inhibitors of pancreatic enzymes, and bile acids. Another reason why we used human duodenal juice was that it gives a more reliable comparison between in vitro activities and in vivo activities (human small intestinal juice aspirated after oral administration of pancreatic extracts). Studies on these problems are at the moment under way. The results of the present study are also of interest from an economical point of view. For instance 10,000 U of lipase cost from 1 dime to 1 dollar. Thus, cost-benefit analyses of this aspect might be of great value. Although there are a number of pitfalls which can influence the therapeutic outcome of pancreatic extracts such as inactivation of enzymes by acid and pepsin in the stomach or unsatisfactory disintegration within the small intestine, an adequate enzyme content of the preparations is a primary basis for a successful treatment of exocrine pancreatic insufficiency.
51
References 1 Dahlqvist, A.: A method for the determination of amylase in intestinal content. Scand. J. clin. Lab. Invest. 14: 145-151 (1962). 2 Drummond, S.; Saunders, J.H.B.; Leach, R., and Wormsley, K.G.: F.nzymic activities o f commercial preparations o f pancreatic enzymes. Scot. med. J. 22: 221-224 (1977). 3 Erlanson, C. and Borgstrom, B.: Tributyrine as a substrate for determination o f lipase activity of pancreatic juice and small intestinal content. Scand. J. Gastroent. 5: 293-295 (1970). 4 Giulian, B.B.; Lokendra, M.S.; Mansfield, A.O.; Pairent, F.W., and Howard, J.M.: Treatment of pancreatic exocrine insufficiency. In vitro lipolyticactivities of pancreatic lipase and fifteen com mercial pancreatic supplements. Ann. Surg. 165: 5 6 4 -5 7 0 (1967). 5 Hummel, B.C.W.: A modified spectrophotometric determination o f chymotrypsin, trypsin and trombin. Can. J. Biochem. Physiol. 37: 1393— 1399 (1957). 6 Ihse, 1. and Arnesjo, B.: The phospholipase A: activity o f human small intestinal contents. Acta chem. scand. 27: 2 7 49-2756 (1973).
Acknowledgement Received: May 22, 1978 Accepted: September 27, 1978 Per Lilja, MD, Department of Surgery, University o f Lur.d, S 221 85 Lund (Sweden)
Downloaded by: Univ. of California Santa Barbara 128.111.121.42 - 3/26/2018 1:49:21 AM
This work was supported by the Swedish Medical Research Council (project No. 17X -03012), the Medical Faculty of Lund and the Albert Pahlsson Foundation.
Pancreatic Enzymic Activities of Commercial Pancreatic Enzyme Preparations Incubated in Human Small Intestinal Juice I. Ihse and P. Lilja Department of Surgery, University of Lund, Lund
Key Words. Amylase • Lipase • Phospholipase • Pancreatic juice • Pancreatic insufficiency • Intestinal secretion Abstract. The activities of amylase, lipase, phospholipase and trypsin of twelve commercial
pancreatic enzyme preparations were measured under identical conditions. Human small intestinal juice was chosen as incubation medium. A wide variation of enzymic activities was found in preparations in tablet form contrary to preparations in granulated form. A prerequisite for a successful therapy is enzymically potent pancreatic extracts with high enzyme content per tablet, capsule or recommended dose of granulated preparation. Therefore, some preparations seem to be preferable to others for clinical purposes.
Material and Methods All preparations were obtained from the hospital pharmacy. One tablet or 10 ml o f the preparations in granulat ed form was crushed in a mortar and incubated for 40 min at a temperature of 37 °C in 50 ml human duodenal juice obtained from normal subjects. Six tablets or six 10-ml dosages from 2 different batches o f each preparation were analyzed in duplicate. The activities of amylase, lipase, phospholipase and trypsin o f the incubation medium (human duodenal juice) were estimated before addition o f the pancreatin preparation and after incubation with it. The enzyme activities of the different preparations were defined as the activities estimated after incubation minus those
Downloaded by: Univ. of California Santa Barbara 128.111.121.42 - 3/26/2018 1:49:21 AM
Pancreatic enzyme preparations are used in malabsorption caused by exocrine pancreatic insufficiency. Comparison of in vitro digestive capacity of such preparations, as reported by the manufacturers, is difficult due to the con siderable variation in the enzyme assays and units used. The purpose of the present study was to measure and compare enzyme activities of 12 commercial supplements under identical condi tions. In order to imitate physiological cir cumstances as close as possible, human small intestinal juice was chosen as the incubation medium.
In vitro Activities of Pancreatic Supplements
49
estimated before addition of the pancreatin prepara tions to the medium. Amylase was determined according to Dahlquist (1), and lipase by a scaled-down version o f the method described by Erlanson and Borgström (3). Phospho lipase was determined according to Ihse and Arnesjo (6) and trypsin modified after Hummel using TAME (p-toluene-sul phony l-I.-arginineme thy lester) as the sub strate (5). As human duodenal juice was used for the incubation, no further activators of trypsinogen or prophospholipase were needed.
Results
The activities of the lipolytic enzymes of the different preparations are shown in table I. As can be seen, Pancreon forte and Pancreon comp
forte had the highest activity of both lipase and phospholipase. Furthermore, it is obvious that the lipase as well as the phospholipase activity of the different tablets varied considerably. On the other hand, the activities of lipase and ¡hospholipase per milliliter of all three prepara tions in granulated form showed less variation. Also, in the amylase and trypsin activities of the tablets, a wide range was found (table II). Here too, Pancreon forte and Pancreon comp forte revealed the highest activities of amylase and trypsin. The variation of the trypsin acti vities of the different preparations in granu lated form was not as marked as for the tablets, while the amylase activity in Pancreon granulate was only half of that of Pancreatin
Name
Manufacturer
Lipase activity per tablet
Lipase activity of maximum recommended dosage per meal
Phospho lipase activity per tablet
Phospholipase activity of maximum recommended dosage per meal
Pancreon forte Pancreon comp forte Festal Combizym comp Pancreozym Helopanzym Supplezym Combizym Luizym
(T)
Kali-Chemie
11,861
23,729 (2 tablets)
1,816
3,632
(T, B) (T. B)
Kali-Chemie Hoechst
10,825 6,900
21,650 (2 tablets) 13,800 (2 tablets)
1,784 459
3,568 918
(T, B) (T) (T, B) (T) (T) (T)
Luitpold-Werke Tika Helopharm Tika Luitpold-Werke Luitpold-Werke
5.589 5,483 3,610 1,387 1.259 515
5,589(1 32,989 (6 7,220 (2 8,322 (6 2,518 (2 1,030 (2
598 685 223 788 838 114
598 4,110 446 4,728 1,676 228
Pancreatin Pancreon Panteric
(G) (G) (G)
Rosco AS Kali-Chemie Parke-Davies
1,781' 1,398' 1,966'
26,715 (15 ml) 13,980 (10 ml) 4,915 (2.5 ml)
550' 568' 728'
8,250 5,680 1,820
tablet) tablets) tablets) tablets) tablets) tablets)
Lipase activity expressed as micromoles fatty acids released per milliliter per minute; phospholipase activity expressed as micromoles fatty acids released per milliliter per minute. T = Tablets; G = granules; B = the preparation contains bile acids. ' Lipase activity and phospholipase activity, respectively, per milliliter of granules.
Downloaded by: Univ. of California Santa Barbara 128.111.121.42 - 3/26/2018 1:49:21 AM
Table I. Mean enzyme activities of 6 different units (tablets, 1 ml of granules) of 12 different pancreatic enzyme preparations
Ilise/Lilja
50
Table II. Mean enzyme activities of 6 different units (tablets, 1 ml of granules) o f 12 different pancreatic enzyme preparations Name
Amylase activity per tablet
Amylase activity of maximum recommended dose per meal
Pancreon forte Pancreon comp forte Festal Combizym comp Pancrcozym Helopanzym Supplezym Combizym Luizym
(T) (T, B) (T, B) (T, B) (T) (T, B) (T) (T) (T)
24,850 33,793 10,070 15,340 7,188 2,806 3,492 10.225 187
49,700 67,586 20,140 15,340 43,128 5,612 20,952 20.450 374
Pancreatin Pancreon Panteric
(G) (G) (G)
13,598' 6,400' 12.190'
203.970 64,000 30.475
Trypsin activity per tablet
Trypsin activity of maximum recommended dosage per meal
3,764 5,735 1,125 912 875 1,050 375 1.259 515
7,528 11,740 2,250 912 5,250 2,100 2,250 2,518 1,030
858' 756' 866’
12,870 7,560 2,160
Amylase activity expressed as micromoles maltose released per milliliter per minute; trypsin activity expressed as micromoles TAME hydrolyzed per milliliter per minute. T = Tablets; G = granules; B = the preparation contains bile acids. ' Amylase activity and trypsin activity, respectively, per milliliter of granules.
Discussion
The present paper reports the activities of amylase, lipase, phospholipase and trypsin of 12 commercial pancreatic enzyme preparations. From a clinical point of view tire most im portant criterion of efficiency of such a pre paration is its ability to augment pancreatic enzyme levels in intestinal contents. A pre requisite for this objective is enzymically potent pancreatic extracts with high enzyme content per tablet, capsule or recommended dose of preparation in granulated form. The results of the present study confirm previous reports of wide variations in enzymic activity of different commercial pancreatic extracts (2,4). Some preparations, therefore, are preferable to others for clinical purposes.
Downloaded by: Univ. of California Santa Barbara 128.111.121.42 - 3/26/2018 1:49:21 AM
granules and Panteric. However, the amylase activity of all three preparations in granulated form was high. When calculating the highest recommended dose per meal it was obvious that the variation of enzyme activities between the different pre parations was still more marked (table I, II). For instance two tablets of Pancreon forte contained more than 20 times as much lipase as two tablets of Luizym. Best overall results were obtained with Pancreon forte and Pancreon comp forte among the tablets, while the three preparations in granulated form (Pancreatin, Pancreon, Panteric) all contained adequate amounts of enzymes. Finally, it was found that there was only a slight variation in enzyme activity of tablets or granules from the same batch.
In vitro Activities of Pancreatic Supplements
In this study we used human duodenal juice as the incubation medium in order to imitate physiological conditions as close as possible. Thus, the pancreatic extracts were incubated in a milieu containing naturally occurring activa tors and inhibitors of pancreatic enzymes, and bile acids. Another reason why we used human duodenal juice was that it gives a more reliable comparison between in vitro activities and in vivo activities (human small intestinal juice aspirated after oral administration of pancreatic extracts). Studies on these problems are at the moment under way. The results of the present study are also of interest from an economical point of view. For instance 10,000 U of lipase cost from 1 dime to 1 dollar. Thus, cost-benefit analyses of this aspect might be of great value. Although there are a number of pitfalls which can influence the therapeutic outcome of pancreatic extracts such as inactivation of enzymes by acid and pepsin in the stomach or unsatisfactory disintegration within the small intestine, an adequate enzyme content of the preparations is a primary basis for a successful treatment of exocrine pancreatic insufficiency.
51
References 1 Dahlqvist, A.: A method for the determination of amylase in intestinal content. Scand. J. clin. Lab. Invest. 14: 145-151 (1962). 2 Drummond, S.; Saunders, J.H.B.; Leach, R., and Wormsley, K.G.: F.nzymic activities o f commercial preparations o f pancreatic enzymes. Scot. med. J. 22: 221-224 (1977). 3 Erlanson, C. and Borgstrom, B.: Tributyrine as a substrate for determination o f lipase activity of pancreatic juice and small intestinal content. Scand. J. Gastroent. 5: 293-295 (1970). 4 Giulian, B.B.; Lokendra, M.S.; Mansfield, A.O.; Pairent, F.W., and Howard, J.M.: Treatment of pancreatic exocrine insufficiency. In vitro lipolyticactivities of pancreatic lipase and fifteen com mercial pancreatic supplements. Ann. Surg. 165: 5 6 4 -5 7 0 (1967). 5 Hummel, B.C.W.: A modified spectrophotometric determination o f chymotrypsin, trypsin and trombin. Can. J. Biochem. Physiol. 37: 1393— 1399 (1957). 6 Ihse, 1. and Arnesjo, B.: The phospholipase A: activity o f human small intestinal contents. Acta chem. scand. 27: 2 7 49-2756 (1973).
Acknowledgement Received: May 22, 1978 Accepted: September 27, 1978 Per Lilja, MD, Department of Surgery, University o f Lur.d, S 221 85 Lund (Sweden)
Downloaded by: Univ. of California Santa Barbara 128.111.121.42 - 3/26/2018 1:49:21 AM
This work was supported by the Swedish Medical Research Council (project No. 17X -03012), the Medical Faculty of Lund and the Albert Pahlsson Foundation.
