Read Reliability of the Acid Phosphatase Test for the Identification of Seminal Fluid text version

J Forensic Sci, Oct. 1978, Vol. 23, No. 4

A. F. SchfjYM.D. A . F. Schiff, 1 M . D .

Phosphatase Test Reliability of the Acid Phosphatase Test for the Identification of Seminal Fluid of Seminal

From the day in 1677 when a medical student named Johann Ham first discovered From day in 1677 when a medical student named Johann Ham first discovered

spermatozoa swimming about in a microscopic field to the present, it has been traditional spermatozoa swimming about in a microscopic field to the present, it has been traditional

to identify seminal fluid by finding spermatozoa in the specimen. It has been pointed identify seminal fluid by finding spermatozoa in the specimen. It has pointed out [1-4], conditions exist in which the ejaculate bears no spermatozoa. out [1-4J, however, that conditions exist in which the ejaculate bears no spermatozoa. Because of this situation, forensic scientists have been searching for a foolproof method Because of this situation, forensic scientists have been searching for a foolproof method

of identifying seminal fluid in the absence of spermatozoa. identifying seminal fluid in the absence spermatozoa. A successful method depends upon the presence of aa singular substance, different in A successful method depends upon the presence of singular substance, different in some way from any other material. Unfortunately, seminal fluid is a veritable repository some way from any other material. Unfortunately, seminal fluid is a veritable repository of chemicals, both organic and inorganic, running the gamut from amino acids, cholesterol, chemicals, both organic and inorganic, running the amino acids, cholesterol, fructose, lactic acid, and acid phosphatase to calcium, magnesium, potassium, and sodium. fructose, lactic acid, and acid phosphatase to calcium, magnesium, potassium, and sodium. 1896, Florence [5] first microchemical test for seminal stains. The reIn 1896, Florence [5] devised the first microchemical test for seminal stains. The reaction is based upon the presence of free choline, a degradation product of phosphorylbased the presence of free choline, degradation product of phosphorylcholine and glyceryiphosphoryicholine.These two products of the seminal vesicles liberate glycerylphosphorylcholine. These two products of the seminal vesicles liberate choline through catalytic conversion of phosphatase; after 66hh outside the body, high outside the body, high choline through catalytic conversion of phosphatase; after levels (2120 mg/100 ml) are produced. The test was enthusiastically received in medicolegal levels (2120 mg/100 ml) are The test was enthusiastically received in medicolegal circles and within a few decades was recognized as the classical test for the identification circles and within a few decades was recognized as the classical test for the identification of seminal fluid. Nevertheless, some individuals detected the test's fallibility. Kind [6] seminal fluid. Nevertheless, some individuals detected the test's fallibility. Kind [6] has shown that occasionally the test remained negative in the presence of definite seminal definite seminal shown occasionally the test remained negative in the presence stains; conversely, it has occasionally turned positive in the absence of semen. Pollack [ 1] stains; conversely, it has occasionally turned positive in the absence of semen. Pollack [11 believed results with the Florence test could be ambiguous, thereby making the test unbelieved results with the Florence test could be ambiguous, thereby making the test unreliable and hence, useless. Gradwohl [7] phrased it thus: "Weisman states that a positive useless. Gradwohl [7] phrased "Weisman a positive reaction is not significant since other body fluids give such results with this test. Our own reaction is not significant since other body fluids give such results with this test. Our own experience confirms this statement. A negative test, however, informs the investigator experience confirms this statement. A negative test, however, informs the investigator that the suspected substance is not semen and this is the only value of the Florence test." value of the Florence test." the suspected substance semen and Nevertheless,the Florence test continued to be used. However, because of the persistent Nevertheless, the Florence test continued to be used. However, because of the persistent increase in sex crimes [8] through the ensuing years, a search for a better and more increase sex crimes [8] the ensuing years, a search for a better and more affirmative means of identifying seminal fluid has pushed forward. affa'mative means identifying seminal fluid has pushed forward. As early as 1923, Robison [9] demonstrated the presence of an enzyme capable of As early as 1923, Robison [9] demonstrated the presence of enzyme capable hydrolyzingcalcium hexosemonophosphate inin bone.The name given to this enzyme was hydrolyzing calcium hexosemonophosphate bone. The name given to this enzyme was phosphate esterase or phosphatase. A study was initiated to learn where else in nature phosphate esterase or phosphatase. A study was initiated to learn where else in nature this newly discovered enzyme could be located. In short order, phosphate esterase was newly discovered enzyme could be located. In short order, phosphate esterase was found in kidney, intestine, liver, striated muscle, and lung in addition to the teeth of young kidney, intestine, liver, striated muscle, and addition animals, milk, floral plants. 1935, Kutscher Wolbergs animals, milk, and the floral parts of plants. In 1935, Kutscher and Wolbergs [10] made a major breakthrough in the identification of seminal fluid by discovering that normal major breakthrough in the identification of seminal fluid by discovering that normal human prostatic tissue exceptionally rich in phosphatase, having optimal activity at human prostatic tissue is exceptionally rich in phosphatase, having optimal activity at

Sept. 1977; revised manuscript received 14 Nov. 1977; accepted for Received for publication 21 Sept. 1977; revised manuscript received 14 Nov. 1977; accepted for for publication publication 17 April 1978. publication 17 April 1978. 1Deputy 1Deputy medical examiner, Dade County, Florida.

833 833

834 JOURNAL OF FORENSIC SCIENCES JOURNAL OF FORENSIC SCIENCES

a pH of 4.9. A year later, Gutman Ct al [11] cautiously associated increased serum phos4.9. year later, a pH et al cautiously associated increased phatase activity with metastasizing carcinoma of the prostate gland; in 1938, both Gutphatase activity with metastasizing carcinoma of the prostate gland; in 1938, both Gut[12] 11 patients metastasizing prostatic carcinoma "significantly mans [12] found in 11 of 15 patients with metastasizing prostatic carcinoma "significantly increased new and brilliant increased 'acid' phosphatase activity of the serum." Gomori [13] added a new and brilliant of the serum." Gomori [13] chapter the story in 1941 by introducing a histochemical method for demonstrating chapter to the story in 1941 by introducing a histochemical method for demonstrating acid phosphatase in tissue. In 1943, Pollack [1] declared unequivocallythat "the chemical phosphatase in tissue. 1943, Pollack [1] unequivocally that composition of human semen gives little hope that a suitable test will be detected." Nevercomposition of human semen gives little hope that a suitable test will be detected." Never-

theless, many researchers in the field were spurred on by Gomori's work. theless, researchers Two years later, Lundquist but historical paper, reviewed some of the Two years later, Lundquist [2], in a brief but historical paper, reviewed some of the older methods including the classical Florence test and listed their shortcomings. Noting methods including the classical Florence test and listed their shortcomings. Noting "the colossal amounts of a phosphatase with aapH optimum in the acid region," he proposed colossal amounts of a phosphatase with pH optimum in the acid region," he proposed that these "extraordinary amounts" be used, as the basis for the identification of seminal these "extraordinary be used as the basis identification of seminal fluid "independent of the presence of sperm cells." Others [14--16] were quick to pick up of the presence of sperm cells." Others [14-16] to pick up suggestion and by the mid-century, answers to the problem had been worked out. the suggestion and by the mid-century, answers to the problem had been worked out. In 1945, Rasmussen [17] speculated that "in those cases where no spermatozoa can 1945, Rasmussen speculated "in those cases where no spermatozoa be demonstrated in the stain, a determination of sF [acid phosphatase] may be of value demonstrated in a determination of sF [acid phosphatase] of value for deciding the origin of the stain." In 1949, Kaye [3] recommended "a microchemical deciding the origin of the stain." In 1949, Kaye [3] microchemieal test for the identification of semen and seminal stains by an acid phosphatase reaction the identification semen and seminal stains acid phosphatase reaction that appears to be both specific and sensitive" which "may answer the need for a reliable appears to be both specific and sensitive" which "may answer the need for reliable test." He added that "none of the tests so far proposed can positively indicate or exclude He added that "none of the tests so far proposed can positively indicate or exclude the presence of human semen." He analyzed 37 substances including vaginal fluids, urine, human vaginal fluids, urine, presence He analyzed substances feces, blood, saliva, and beet juice; in these substances, he found less than King-Armfeces, blood, saliva, and beet juice; in these substances, he found less than 5 King-Armstrong (K-A) units of acid phosphatase per millilitre of concentrated pure substance. On (K-A) units of acid phosphatase per millilitre of concentrated pure substance. On fresh seminal the other hand, fresh seminal fluid contained from 2000 to 2800 K-A units/ml. Arbitrarily 2000 to 2800 K-A units/mi. Arbitrarily established by comparison and observation, an acid phosphatase activity of 25 K-A established by comparison and observation, an acid phosphatase activity of 25 K-A units/mi of extract from an area of approximately 1 cm2 should be considered positive units/ml of extract from an area of approximately 1 cm 2 should considered positive for seminal stains, according to Kaye. One of his conclusionswas that the reaction "is seminal stains, according to Kaye. One of his conclusions was that the reaction "is very sensitive and appears to be specific for seminal fluid of man or monkey only." sensitive and appears to be specific for seminal fluid of man or monkey only." In the same year, Seligman and Manheimer [18] produced a test based on the acid the same year, Seligman and Manheimer produced test based acid phosphatase activity of seminal fluid. They obtained highly colored product at the phosphatase activity of seminal fluid. They obtained aa highly colored product at the of 4.9 by employing stabilized couple freely with the optimum pH of 4.9 by employing stabilized diazonium salts which couple freely with the liberated radical of a suitable substrate. suitable substrate. radical year later, Walker acknowledged need a test to identify seminal stains A year later, Walker [19] acknowledged the need for a test to identify seminal stains independent of the presence or absence of spermatozoa and introduced the procedure independent of the presence absence spermatozoa and introduced the procedure based on the histochemical technic of Seligman and Manheimer. He wrote, "With the based histochemical technic of Seligman and Manheimer. He wrote, "With the discoveryof the intense acid phosphatase activity of prostatic secretion, new definitive discovery of the intense phosphatase activity prostatic secretion, new definitive test methods for semen have become available." This procedure, which came to be known semen have become available." This procedure, which came to be known methods as the Walker test, "is considered today to be the most conclusive test for the presence of Walker "is considered most conclusive test for the presence of semen; in the absence of spermatozoa, this test has sometimes been regarded as being as semen; in absence spermatozoa, this sometimes been regarded as being as confirmatory as if spermatozoa had actually been found" [20]. confirmatory as if spermatozoa had actually been found" [20]. Riisfeldt [151 demonstrated increased phosphatase activity in vaginal fluid 6 and 12 h [15] demonstrated increased phosphatase activity in vaginal fluid 6 and 12 h after coitus, but not after 24 h. Fischer [21 ] prophesized, "There is, however, little reason 24 h. Fischer [21] prophesized, coitus, but is, however, little reason to doubt that the test will be used frequently, and no difficulty is anticipated in having doubt the test will be used frequently, and no difficulty anticipated having such evidence accepted by the Court, provided it is introduced by a qualified expert." such evidenceaccepted by the Court, provided it is introduced by a qualified expert." my opinion, the phrase most important thought whole paragraph; I In my opinion, the last phrase is the most important thought of the whole paragraph; I will elaborate on this later. will elaborate on this later. In 1955, focusing on the presence of large quantities of acid phosphatase in seminal fluid, 1955, phosphatase in seminal fluid, Berg [22] procedure which he used a substrate and suitable diazonium Berg [22] described a procedure in which he used a substrate and a suitable diazonium salt to produce an intense violet color. In discussing the Walker test, he "admitted ititto salt to produce an intense violet color. In discussing the Walker test, he "admitted to be suitable for medicolegal use. However, it did not give such clear results as I have just suitable medicolegal use. However, it did not give such clear results as I have just described." described." various tests developed since 1950 have usually centered about the acid phosphatase The various tests developed since 1950 have usually centered about the acid phosphatase

SCHIFF ON ACID PHOSPHATASE TEST 835 SCHIFF ON ACID PHOSPHATASE TEST

contents of seminal fluid. In general, the techniques use a substrate, usually a phenolic contents seminal fluid. In general, techniques use substrate, usually phenolic

ester, which is hydrolyzed in the presence of acid phosphatase. The reaction is occult. ester, which hydrolyzed in the presence of acid phosphatase. The reaction is occult. Therefore, a diazonium salt which reacts quickly in a medium adjusted to the right degree which reacts quickly in a medium adjusted to the right degree Therefore, diazonium acidity is added. "The coupling reaction," Fieser [23] written, "is spectacular of acidity is added. "The coupling reaction," Fieser [23] has written, "is spectacular because of the rapid formation of brightly colored products from colorless components." because of the formation of brightly colored products from colorless components." Investigators began searching for an ideal substrate and perfect coupling agent. Investigatorsbegan searching for an ideal substrate and a perfect coupling agent. Babson et al [241 evaluated six different substrates and concluded that for prostatic acid Babson et [24] different substrates and concluded that for prostatic acid

a-naphthyi phosphate was twice as specific as/3-glycerophosphate. phosphatase, a-naphthyl phosphate was twice as specific as 3-glycerophosphate. In addition, it was 40 to 100 times as specific as the other substrates investigated. Nine years later, specific as the other substrates investigated. Nine years later, was 40 100 times Babson [25] held the same opinion. quoted Bruhn and Keller [26] [26] saying, Babson [25] still held the same opinion. He He quoted Bruhn and Keller as saying, "Because of its greater specificityalpha naphthyl phosphate is the substrate of choice for "Because of its greater specificity alpha naphthyl phosphate is the substrate choice for the estimation of prostatic acid phosphatase." Babson and his group employed diazodiestimation prostatic acid phosphatase." Babson and his group employed diazodiorthanisidine reagent or Brentamine fast Blue B salt as the chromogen. orthanisidine reagent Brentamine Blue B salt chromogen. A whole new field of investigationcentered about the identification of seminal fluid A whole new field of investigation centered about the identification of seminal fluid through the use of a substrate, generally a-naphthyl phosphate, and aacolor developer, through the of a substrate, generally ct-naphthyl phosphate, and color developer, usually Brentamine fast Blue B. Shupe [27] attempted to popularize the Babson and usually Brentamine fast Blue B. Shupe [27] attempted to popularize the Babson and co-workers [24] assay method by using two tablets (Tablet 1 was the "buffer substrate" co-workers [24] assay method by using two tablets (Tablet 1 was the "buffer substrate" and Tablet 2 the "color developer") and a small, 25-mm (1-in.) long test tube called a and Tablet the "color developer") and small, 25-mm (1-in.) long test tube called a "Teswell." This method is time-consuming when compared to the test described below "Teswell." This method is time-consuming when compared to the test described below and takes at least 42 mm to complete. Reinstein et al [28] (disclosing that Fishman and least 42 min to complete. Reinstein et al [28] (disclosing Fishman and Lerner prostatic acid phosphatase is inhibited by tartrate but that blood acid Lerner [29] noted prostatic acid phosphatase is inhibited by tartrate but that blood acid phosphatase is not) devised a method that they believed was specific for acid phosphatase phosphatase not) devised a method they believed was specific for acid phosphatase concluded, "The method extremely sensitive. As little as 0.001 ml of prostatic origin. They concluded, "The method isis extremelysensitive. As little as 0.001 ml of seminal fluid was detected on aafabric on which itit had been dried."Kind [6] introduced seminal fluid was detected on fabric on which had been dried." Kind [6] introduced a stable form of acid phosphatase reagent containing in a spray bottle a-naphthyl phosstable acid phosphatase reagent containing in spray bottle ct-naphthyl phosphoric acid and diazodiorthanisidine as the major components. In his experience, the soluadd diazodiorthanisidine components. his experience, solution was stable for several months at 4~ Raju and Iyengar [30] tested 100 stains of variwas stable for several months at 4°C. Raju Iyengar [30] tested 100 of varianimal and origins ranging from asparagus, garlic, emblic myrobalan, date, and ous animal and plant origins ranging from asparagus, garlic, emblic myrobalan, date, and fig to secretions of crow, duck, and hen. They dissolved cr-naphthyl phosphoric acid and secretions of crow, duck, hen. They dissolved ct-naphthyl phosphoric acid and the diazonium salt in acetate buffer of pH 5. Like Kind, they pressed dampened filter diazonium salt in acetate buffer of pH 5. Like Kind, they pressed dampened filter paper firmly on the suspected stain, leaving it in position for 5 min. They then removed the on the suspected stain, leaving it in position for S miii. They then removed the filter paper and sprayed it with their prepared reagent; they looked for an intense purplish paper and sprayed it with their prepared reagent; they looked for an intense purplish color to appear on the filter paper. This process reminds one of lifting fingerprints from color to appear on filter paper. This process reminds one of lifting fingerprints from smooth surfaces. In a short communication, Babson and Phillips [31 ] wrote of using the substrate they short communication, Babson and Phillips [31] using the substrate they previously recommended [24], but coupling agent, substituting 5-nitro-o-anisidine had previously recommended [24 ], but as a coupling agent, substituting 5-nitro-o-anisidine (fast Red B salt). They concluded the sensitivity was almost twice as great as in their (fast Red salt). They concluded the sensitivity was almost twice as great as in their original method and the end color was more stable. was more stable. original method and Schiff test in the following manner: he impregnated vaginal swabs Schiff [32] modified the test in the following manner: he impregnated vaginal swabs with an acidified, saturated solution of the relatively stable sodium a-naphthyl phosphate acidified, solution relatively stable sodium a-naphthyl phosphate and, after allowing them to dry, placed them in sterile test tubes and stored them in and, after allowingthem to dry, placed them in sterile test tubes and stored them in aa refrigerator 4.4~ In use, the swab was rubbed lightly against suspected material, refrigerator at 4.4°C. In use, the swab was rubbed lightly against any suspected material, either in the vagina, rectum, vulva, or moistened stains, and then treated only with either in the vagina, rectum, vulva, or moistened stains, and then treated only with aa coupling agent. The purpose was to eliminate two steps from the usual method; the coupling agent. The purpose was to eliminate two steps from the usual method; the chemical reaction, however, was the same as with the other methods. After six years, chemical reaction, however, was the same as with the other methods. After years, the treated swabs are still serviceable. swabs are still serviceable. Sivaram the test in yet another fashion. Assuming prostatic acid phosSivaram [33] modified the test in yet another fashion. Assuming prostatic acid phosphatase is "almost completely inhibited by 0.04M L-(+)-tartrate," he claimed he was phatase is "almost completely inhibited by 0.04M L-(+)-tartrate," he claimed he was able to differentiate human semen from other acid phosphatase-bearing substances. Two differentiate human semen other acid phosphatase-bearing substances. Two years later Willott [34] produced evidence that L-tartrate-inhibitable acid phosphatase years later Willott [34] produced evidence that L-tartrate-inhibitable acid phosphatase is found in vaginal secretions as well as in seminal fluid. Gomez et al [35] agreed with found in vaginal secretions as well as in seminal fluid. Gomez et al [35] agreed Willott's findings when they concluded, "Inasmuch as endogenous vaginal ACP (acid Willott's findings when they concluded, "Inasmuch as endogenous vaginal ACP (acid

836 JOURNAL OF FORENSIC SCIENCES JOURNAL OF FORENSIC SCIENCES

inhibited by to the same extent as seminal ACP, this inhibitor is phosphatase] is inhibited by tartrate to the same extent as seminal ACP, this inhibitor is

useless in resolving the two types of activity." of activity."

Material Method Material and Method

The materials and method were the same as previously used [321: method were same as previously used [32]:

(1) a buffer solution, 0.07M, prepared by dissolving 14.7 g citric acid monohydrate in buffer solution, 0.07M, prepared by dissolving 14.7 g citric acid monohydrate in 700 ml of distilled water and adjusting the pH to 5.6 with 2N sodium hydroxide; 700 nil of distilled water and 5.6 with 2N (2) substrate, sodium a-naphthyl phosphate (Borden Chemical Co., Phila., Pa.), (2) a substrate, sodium a-naphthyl phosphate (Borden Chemical Co., Phila., Pa.),

20 mg in 2 cm 3 distilled water (the substrate should be freshly prepared); mg in 2 cm3 distilled water (the substrate should be freshly prepared); (3) a chromagen, Brentamine fast Blue B (Borden Chemical Co.), 20 mg in 22cm33distilled Brentamine fast Blue B (Borden Chemical Co.), 20 mg in cm distilled water (this should be freshly prepared); (this should be freshly prepared);

(4) disks made from Whatman No. 1 ffdter paper upon which seminal fluid has been (4) disks made from Whatman No. 1 filter paper upon which seminal fluid has been smeared; after drying, the disks are punched out with standard paper punch (the author smeared; after drying, the disks are punched out with aastandard paper punch (the author has found these disks to be serviceable at least six years after having been made); and has found these disks to serviceable at least six years after having been made); and (5) control disks made in the same fashion as above except that the paper had not been control disks made in the same fashion as above except that the paper had not been

smeared with seminal fluid.

Vaginal aspirates were collected from 100 alleged rape victims. An acid phosphatase Vaginal aspirates were collected from 100 alleged rape victims. An acid phosphatase (AP) test was first performed on each aspirate. Then a careful search was made of at least was first performed on each aspirate. Then a search was made of at least two slide preparations for spermatozoa. two method testing for AP is simple: three glass slides are placed side by side on The method of testing for AP is simple: three glass slides are placed side by side on aa sheet of white paper and labeled known (K), unknown (U), and control (C). A disk having white paper and labeled known (K), unknown (U), and control (C). A disk having seminal fluid on it is positioned on the K slide while untreated disks are positioned on seminal fluid on it positioned on the slide while untreated disks are positioned on the U and C slides. Onto the U disk is dropped a minute quantity of the vaginal aspirate. C slides. Onto disk minute vaginal aspirate.

To each disk is added a drop of the buffer solution, then a drop of substrate solution, each disk added drop of the buffer solution, then drop of substrate solution, and, finally, a drop chromagen. The disk develops a rich purple color, the U and, finally, a drop of the chromagen. The K disk develops a rich purple color, the U disk may or may not depending upon whether AP present in the aspirate, and the C disk may or may not depending upon whether AP isispresent in the aspirate, and the C

disk remains a pale yellow. a pale yellow.

Results

Table 1 reflects that the series consists of 61 white and 39 black females. The victims Table reflects that series consists of 61 white and 39 females. The victims range in age from 8 to 85 years old, with a mean of 25. Fifty-five percent are in the range in age from 85 years old, with a mean of 25. Fifty-five percent are in

16- to 26-year-old age group. The time interval between the assault and the examination 16- to 26-year-old age group. The time interval between the assault and examination was anywherefrom 1 hh to 4 days. It was a tribute to the cooperative efforts of the police was anywhere from 1 to 4 days. It was a the cooperative efforts of the police and the rape treatment center (RTC) that 45% of the victims were examined within 3 h rape treatment c e ~ e r (RTC) 45% of the victims were examined within 3 h

of the assault, and an additional 25% within 6 h. Eight percent reported to the RTC the assault, and an additional 25% within 6 Eight percent reported the RTC more than 24 h after the assault. Both AP and spermatozoa have been recorded in each more than 24 h after the assault. Both AP and spermatozoa have been recorded in each

case. A definite correlation has been noted between the presence of AP and spermatozoa case. A definite correlation has been noted between the presence of AP and spermatozoa or the absence of both in the same cases. In almost half the cases (49%) no spermatozoa absence of both in the same cases. In almost half the cases (49%) no spermatozoa was discovered; in Table 1 explanations are given: one assailant used condom, or AP was discovered; in Table 1 explanations are given: one assailant used aa condom, one did not have time to complete the sexual act, and several victims denied coitus had one have time complete the sexual act, several victims denied coitus had

been performed. It is interesting to note regarding the assailant who used a condom been performed. It interesting to note regarding the assailant who used condom

that this situation was encountered only one other time in more than 1500 alleged sexual one other time this situation was encountered 1500 alleged sexual assaults. This is far less than the 4% reported by Amir [36] in 173 rapes. than 4% reported by Amir [36]

Given the time factor and the various explanations in Table 1, there appears to be Given the time factor and the various explanations in Table 1, there appears to be an absolute parallel between the presence and absence of AP and the presence and absolute parallel between the presence and absence of AP and presence and

absence of spermatozoa.

TABLE 1--Summary of cases. TABLE l--Summa,y of cases.

Number Number

Spermatozoa Spermatozoa Comments Comments

Case Case

AP AP

0 0

strong strong strong strong strong strong

Race Race

Age Age

Time After Time After Intercourse, h Intercourse, h

11

w w

0 0

2 2

2/2 2~A

33 2 2

51/2 51/2

2/z 2%

numerous nonmotile numerous nonmotile numerous nonmotile numerous nonmotile numerous motile numerous motile

0 0

forced fellatio; mouth swabs negative forced fellatio; mouth swabs negative

b b

0 0

33

4 4 5 5

moderate moderate

nonmotile nonmotile

0 0

21/4 2~A 51/2 51/2

325 325 327 327 333 333 334 334 335 335

w w w w

34 34 22 22 27 27

b b

0 0

14 14 15 15

20 20

2 2 3 3

11/z 1 t/2

6 6 7 7

338 338 339 339

w w w w

19 19

8 8 9 9

strong strong strong strong strong strong

341 341 342 342

10 10

b b b b b b

motile motile motile motile nonmotile nonmotile

0 0

motile motile

11 11

31/2 31/2

4 4

0 0 3 3

31/2 31/2

12 12

345 345 347 347 348 348

weak weak strong strong strong strong strong strong

13 13

349 349 3 3 33 0 0 0 0 0 0 0 0 0 0 0 0 33

w w w w w w

14 14

b b

r (0 C) 0

15 1S

358 358 360 360

nonmotile nonmotile nonmotile nonmotile nonmotile nonmotile

I ii

victim assaulted by males victim assaulted by 33males not certain whether ejaculation occurred not certain whether ejaculation occurred

"1-

16 16

361 361

b b w w

43 43 25 25 65 65 35 35 22 22 45 45 25 25 18 18 55 55 24 24

11

"11 -n 0 Z

0 z

20 20

33

... ... ... questionable as to whether assault questionable as to whether assault actually occurred actually occurred all 3 attackers had been smoking " p o t all 3 attackers had been smoking "pot"" victim and assailants had been smoking victim and assailants had been smoking "pot" "pot" ... victim was a drug addict victim was a drug addict victim was intoxicated victim was intoxicated victim was "bombed out" victim was " b o m b e d out" assailant may have used condom assailant may have used condom penetration uncertain penetration uncertain ... ... although there was penetration, assailalthough there was penetration, assailant did not have time to ejaculate ant did not have time to ejaculate no penetration occurred no penetration occurred

17 17 18 18 19 19

b b b b

14 14

motile motile nonmotile nonmotile nonmotile nonmotile

0 0

19 19

> ()

C)

C

20 20

w w w w

"13

21 21

b b 7 7 2¾0 29

10 10

3 3

22 22

w w

37 37 26 26 13 13 36 36

23 23

368 368 369 369 374 374 376 376 378 378 379 379 380 380 nonmotile nonmotile

0 0

nonmotile nonmotile

b b 4 4

48 48

21 21

381 381

26 26

I 0 I -I

0 0 0 0

15 15

26 26

24 24 25 25 26 26 27 27 28 28

386 386 387 387 390 390 392 392

21 21

6 6

5 5

0 09 U, "13 -D "1" -4 > CO t/) m m -I ---I i11 ni Cl) O) --I -4

29 29

393 393

w w w w w w b b w w w w

22 22 20 20

over 24 over 24

nonmotile nonmotile nonmotile nonmotile

0 0

30 30

394 394

b

12 12

strong strong strong strong moderate moderate 0 0 moderate moderate 0 0 strong strong 0 0 0 0 moderate moderate strong strong 0 0

... ... ... ... ... sexual assault not completed sexual assault not completed victim had bathed and douched victim had bathed and douched victim was assaulted by two men victim was assaulted by two men ... ...

00 a, C,)

,.4

TABLE 1--(Continued) TABLE 1--(Continued)

03

C-

Number Number

Spermatozoa Spermatozoa Comments Comments assailant (about 15 or 16 years old) assailant (about 15 or 16 years old) had smell of transmission fluid on had smell of transmission fluid on his breath; may not have ejaculated his breath; may not have ejaculated

21/2 / 2 21

21/2 21/2

Case Case

0 0

Race Race

0 0

Age Age

Time After Time After Intercourse, h Intercourse, h

AP AP

0 C C "11

31 31

395 395

b b

nonmotile nonmotile

0 0 nonmotile nonmotile

20 20

z >

Z

F1

0 0 ·0

32 32 33 33 34 34 2 2 5 S

moderate moderate 0 0 strong strong

396 396 397 397 398 398

w w twice t~ce

no penetration no penetration

b b b b

27 27

17 17 20 20

0

assailant had intercourse with victim assailant had intercourse with victim

m 0 :0 m m

z z

C')

b b

C~ C)

ci) O'J C)

r mn

399 399 400 400

13 13 3 3 3 3 11 8 8 31 3/2 / 2 3 3

strong strong motile motile 0 0 0 0

w w

z C) o

z

35 35 36 36 37 37 38 38 40 40 41 41 42 42 0 0 strong strong strong strong 0 0 0 0 moderate moderate moderate moderate 0 0 nonmotile nonmotile nonmotile nonmotile 0 0 0 0 nonmotile nonmotile 0 0

401 401 402 402 403 403 404 404 405 405

b b b b b b w w b b

14 14 28 28 25 25 24 24 58 58 55 55 18 18

assailant may not have ejaculated assailant may not have ejaculated no ejaculation no ejaculation

rrl m

C))

three men allegedly had intercourse with three men allegedly had intercourse with

victim who had had sexual intervictim who had had sexual inter0 0

w w

strong strong strong strong strong strong

43 43 44 44 45 45

406 406 407 407 409 409

w w

nonmotile nonmotile nonmotile nonmotile

w w

16 16 21 21 18 18

4 4 4 4 4 4

course with boyfriend 12 hhpreviously course with boyfriend 12 previously two men assaulted victim two men assaulted victim manual manipulation only manual manipulation only

b b w w w w

3 3 6 6 14 14 3 3 3 3 3 3 0 0 0 0 strong strong moderate moderate

diluted diluted three men involved three men involved

two men involved; vaginal aspirate two men involved; vaginal aspirate

w w

46 46 47 47 48 48 49 49 50 50 51 51

413 413 414 414 415 415 418 418 419 419 422 422

w w

w w

23 23 16 16 24 24 77 77 18 18 15 1S

w w

b b w w

victim was not certain whether assailant victim was not certain whether assailant had ejaculated had ejaculated false allegation; no intercourse occurred false allegation; no intercourse occurred condom was used condom was used

victim had heavy menses victim had heavy menses

w w

52 52 53 53 54 54 55 SS 56 56 57 57 5 S 2 2 4 4 5 S 3 3 18 18 0 0 0 0 strong strong 0 0 strong strong 0 0

423 423 424 424 425 425 427 427 428 428 429 429

w w

w w

13 13 12 12 22 22 25 25 23 23 11 11

0 0 0 0 nonmotile nonmotile both motile and both motile and nonmotile nonmotile 0 0 0 0 motile motile 0 0 nonmotile nonmotile 0 0

18 18

0 0

0 0

victim unaware if assailant ejaculated victim unaware if assailant ejaculated

...

20 20

moderate moderate

false allegation false allegation

58 58 59 59 60 60 61 61 62 62

430 430 431 431 432 432 433 433 444 444

w w w w b b b b w w

nonmotile nonmotile

0 0 0 0 0 0

11 11 16 16 16 16

10 10 13 13 19 19 96 96 29 29 0 0 0 0 0 0

assailant supposed to have had vaassailant supposed to have had vasectomy sectomy Victims 63 and 64 were sisters Victims 63 and 64 were sisters

...

b b b b

16 16 16 16

moderate moderate

8 8 8 8 4 4 4 4 4 4 5 5 0 0

strong strong doubt as to validity of story doubt as to validity of story 3 men had sexual intercourse 3 men had sexual intercourse doubt as to validity of story doubt as to validity of story

63 63 64 64 65 65 66 66 67 67 68 68

11

445 445 446 446 448 448 450 450 451 451 452 452

w w w w w w w w

16 16 21 21 19 19 30 30

0 0 0 0 0 0

453 453

0 0

strong strong strong strong strong strong

no penetration; no ejaculation no penetration; no ejaculation

..~

4 4 2 2

21/2 21/2

victim unaware if assailant ejaculated victim unaware if assailant ejaculated

,,~

69 69 70 70 71 71 72 72 73 73

455 455 457 457 459 459 460 460

w w w w w w w w w w

30 30 18 18 27 27 23 23 85 85

8 8 8 8

111/2 1/2

0 0

nonmotile nonmotile nonmotile nonmotile

0 0 0 0 0 0 nonmotile nonmotile 0 0 some motile, some motile, some nonmotile some nonmotile 0 0 motile motile nonmotile nonmotile nonmotile nonmotile 0 0

not penetrate not penetrate

,..

assailant could not get erection; did assailant could not get erection; did

b b

strong strong strong strong

74 74 75 75

461 461 463 463

w w

31 31 18 18

victim claimed to have been assaulted victim claimed to have been assaulted

C) 0

O~ -i"11 '1

by two men, but did not believe by two men, but did not believe ejaculation occurred; she may have ejaculation occurred; she may have had intercourse with aaman who was had intercourse with man who was not an offender not an offender no penetration no penetration

~

0 o Z

victim showered before reporting to victim showered before reporting to

police police

Z

76 76

b b

28 28

2189 21/2

464 464 466 466

12 12

0 0 0 0

strong strong

0 0

0 0 0 0 0 0

half motile, half motile, half nonmotile half nonmotile nonmotile nonmotile

o_

467 467

77 77 78 78 79 79

468 468

w w b b w w

16 16 44 44 17 17 29 29

27 27

0 0

-u -T-

0

0 0

80 80

474 474 strong strong strong strong moderate moderate

b b

24 24

2 2 3 3 10 10

O r "U "1"

478 478

81 81 82 82

480 480

b b w w

18 18 17 17

victim did not know whether sexual victim did not know whether sexual

assault had occurred because she had assault had occurred because she had been knocked out been knocked out

motile and nonmotile motile and nonmotile

0 0 nonmotile nonmotile

,., .~

m -'4 m O9 -4 {20 r.~ r

w W w W

2 2 6 6 11/2 1V2

83 83 84 84 85 85

481 481 482 482 485 485

w W

75 75 35 35 38 38

strong strong strong strong moderate moderate

victim assaulted sexually by 33males victim assaulted sexually by males

TABLE 11--(Continued) TABLE --(Continued)

C.-

Number Number

Spermatozoa Spermatozoa

Case Case

Race Race

Age Age

Time After T i m e After Intercourse, h Intercourse, h

AP AP

Commentss Comment

0 0 0 0 0 0 nonmotile nonmotile 0 0 moderate moderate

o 0

0 C c z z >

rI-

86 86

487 487

w w

3 3 15 1S

58 58

26 26

0

87 87 88 88

488 488 489 489

b b w w

15 15 15 15

0 o T1 -n -Il --n o m m z

z

C,,

0 c3

0, 0~

C) _o

89 89 90 90

490 490

24 24 32 32

victim h a d been intoxicated; allegation victim had been intoxicated; allegation open to suspicion open to suspicion allegation open to suspicion allegation open to suspicion allegation open to suspicion; voluntary allegation open to suspicion; voluntary sexual intercourse m a y have occurred sexual intercourse may have occurred later later victim had bathed andd douched victim h a d bathed a n douched

492 492 2 2 15 1S 2 2 3 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

b b b b 5 5

o 0

0 0 0 0

19 19

intact hymen; victim did not believe intact laymen; victim did not believe

penetration or ejaculation occurred penetration or ejaculation occurred victim believes assailant did not penevictim believes assailant did not penetrate, b u ejaculated outside body trate, but tejaculated outside body

m m

z z t') C)

91 91

493 493

w w

22 22

ni i-n co 0)

92 92

494 494

b b

15 1S

mother gave victim bath anddvinegar m o t h e r gave victim b a t h a n vinegar

douche douche

victim did not believe ejaculation ocvictim did not believe ejaculation occurred curred victim did not believe ejaculation ocvictim did not believe ejaculation occurred; Victims 93 and 94 were part curred; Victims 93 a n d 94 were part

93 93

496 496

w w

17 17

94 94

497 497

w w

19 19

of the same assault with assailant of the same assault with assailant

alternating between the two alternating between the two

41/2 41/2

95 95

498 498

w w

15 1S

0 0

0 0

there was penetration and injury, but there was penetration a n d injury, b u t victim did not believe known ofvictim did not believe known ofrare s p e r m rare sperm fender h a d ejaculated nor even penefender had ejaculated nor even penetrated very far since she was virgin trated very far since she was aavirgin victim did not know whether there was victim did not know whether there was penetration or ejaculation penetration or ejaculation

96 96 4 4

499 499

w w

16 16

weak weak

0 0

strong strong strong strong

500 500

2 2 51/2 51/2 12 12 0 0

w w

97 97 98 98 99 99 100 100

b b b b

25 25 20 20 18 18

0 0

nonmotile nonmotile nonmotile nonmotile

0 0

501 501 502 502 504 504

w w

14 14 64 64

victim assaulted by two men victim assaulted by two m e n victim assaulted by two m e n victim assaulted by two men victim slightly intoxicated victim slightly intoxicated

SCHIFF ON ACID PHOSPHATASE TEST 841 SCHIFF ON ACID PHOSPHATASE TEST 841

Discussion

When the AP test first became known and used, it was greeted with unreserved When the AP test first became known and used, it was greeted with unreserved

acclaim by many medieolegal experts. Lundquist [37] declared, "We hesitate to acclaim by many medicolegal experts. Lundquist [37] declared, "We do not hesitate to consider the positive acid phosphatase test as proof of the presence of semen as valid consider the positive acid phosphatase test as proof of the presence semen valid as the demonstration of spermatozoa." Jones [38] stated, "Many workers are now condemonstration of spermatozoa." Jones [38] stated, workers now con-

vinced that the fairly rapid appearance of an intense colour on applying the test is vinced that the fairly rapid appearance of an intense colour on applying the test is

almost certainly diagnostic of the presence of human semen or, more accurately, human almost certainly diagnostic of the presence of human semen or, more accurately, human prostatic secretion. The word 'almost' is used in order to include the possibility, however secretion. The word is used in order to include the possibility, however remote, of monkey semen." Levonen [39] is generally considered to have remote, of monkey semen." Levonen [39] affirmed, "It is generally considered to have almost the same specificity as the microscopic recognition of spermatozoa." Schumann almost same specificity as microscopic recognition of spermatozoa." Schumann et al [40] concluded that vaginal fluid AP "is a reliable and sensitive method for the al [40] that vaginal fluid AP a reliable and sensitive method for the identification of semen." identification of semen." On the other hand, some workers disagreed. Gradwohl [7] stated, "With these points other hand, some workers disagreed. Gradwohl [7] stated, "With these points in mind, it will be found that this test is very useful as an indicative or screening test mind, it will found that this test is very useful as an indicative or screening test

and cannot, under any circumstances, be used unconfirmed as a conclusive test for cannot, under any circumstances, used unconfirmed as a conclusive test semen." PoIson [41] agreed, "It cannot, however, displace the microscopical demonsemen." Poison [41] agreed, cannot, however, displace the microscopical demonstration of sperms as proof of seminal fluid." Sivaram [33] wrote that although the stration sperms proof of seminal fluid." Sivaram [33] that although the

presence phosphatase activity was strongly suggestive of the presence of semen, presence of acid phosphatase activity was strongly suggestive of the presence of semen, was nevertheless not specific because of widespread occurrence of enzyme it was nevertheless not specific because of the widespread occurrence of the enzyme in nature. [42] pointed "few medical examiners willing to testify on the basis of Evrard [42] pointed out that "few medical examiners are willing to testify on the basis of

elevated vaginal acid phosphatase without the presence of sperm." vaginal acid phosphatase Kind [6] stated bluntly that "in the present state of knowledge acid phosphatase Kind [6] stated bluntly "in the present state of knowledge acid phosphatase activity per se is not proof of semen." His laboratory, however, has used the method as activity per se is semen." His laboratory, however, has used the method as a screening method. screening method. Henningsen2 has wrirten as recently as 1976 that the AP test "is very sensitive and is Henningsen 2 has written as recently as 1976 that the AP test very sensitive and is conclusive evidence of semen, but negative generally accepted disproof not conclusive evidence of semen, but if negative it is generally accepted as disproof or anyway as sufficient evidence against semen to omit further examination." Yet, conanyway as sufficient evidence against semen to omit further examination." Yet, he coneluded, "Summarizing, we generally use the acid phosphatase test as screen test but in cluded, "Summarizing, we generally use the acid phosphatase test as aa screen test but in certain cases and subject to special criteria, it is used as a reliable indication of the presence cases and subject to special criteria, is used as a reliable indication of the presence of seminal fluid." seminal Pinto [43] stated quite definitely, "We not, in any circumstances, advocate the Pinto [43] stated quite definitely, "We do not, in any circumstances, advocate the substitution of this test for the microscopic identification of spermatozoa." Then, he substitution of this test for the microscopic identification of spermatozoa." Then, he became slightly ambiguous by commenting on what is done in a legal case where the became slightly ambiguous by commenting on what is done in a legal case where the test is positive and yet "an exhaustive examination" reveals no spermatozoa: "The test is positive and yet "an exhaustive examination" reveals no spermatozoa: "The testimony in Court should be that a positive chemical reaction for semen was obtained testimony in Court should be a positive chemical reaction for semen was obtained which was not confirmed by the microscopic identification of spermatozoa. It would which was not confirmed the microscopic identification of spermatozoa. It would then be left to the Court to assess the value of this evidence in the light of the experibe left to the Court to assess the value of this evidence the light of the experience of the expert with the phosphatase test." This is hardly a fair proposition to place ence of the expert with the phosphatase test." This is hardly a fair proposition to place upon the shoulders of a judge or a jury, laymen, the onus of making a decision which, upon the shoulders of a judge jury, laymen, the onus of making a decision which, here, is the function and responsibility of the expert. The court has the right to expect here, is the function and responsibility of the expert. The court has the right expect relativelydirect and authoritative answers regarding opinion as well as fact from one relatively direct and authoritative answers regarding opinion as well as fact from one who is qualified as an expert. qualified as an expert. After 14 year's experience with the AP test, the author has found it to have great merit as 14 year's experience with the AP test, the author has found it to have great merit as a test for the identification of seminal fluid in the absence of spermatozoa. Relying on test for the identification of seminal in the absence of spermatozoa. Relying on the test, the author has had no hesitancy in testifying in court regarding the presence or the author has had no hesitancy in testifying court regarding presence or absence of seminal fluid. Because he has used the test qualitativelyrather than quantiabsence of seminal fluid. Because he has used the test qualitatively rather than quantitatively, he has established no arbitrary, numerical cutoff as to when the test is to be detatively, he has established no arbitrary, numerical cutoff as to when the test is to be dedared clared positive and when negative.

personal communication, 15 Sept. 1976. 2K. Henningsen, personal communication, 15 Sept. 1976.

842 JOURNAL OF FORENSIC SCIENCES JOURNAL OF FORENSIC SCIENCES

Attorneys, judges, and other legal authorities recognize well that the polygraph is only and other legal authorities recognize that the polygraph is only as unquestioned as the skill of a particular operator. This same belief holds true for the unquestioned as the skill of a particular operator. This same belief holds true for the AP test. It is only as dependable as the physician, chemist, or pathologist who performs AP test. is only as dependable as the physician, chemist, or pathologist who performs it. Certain guidelines must be set down and adhered to. First, the reagents must be freshly Certain guidelines must be set down and adhered to. First, the reagents must be freshly prepared. Table 2 shows that solutions left standing at room temperature for 44 h will prepared. Table shows that solutions left standing at room temperature for 44 will still be usable. However, a rare lawyer has questioned the author as to when the solutions be usable. However, a rare lawyer has questioned the author as to when the solutions were prepared, hinting at the fact, with an eye on the jury, that if 2 or 3 h had passed, were prepared, hinting at the fact, with an eye on the jury, that if 2 or 3 h had passed, the test would be void. The diazo-coupling agent that originally was clear and lightly the test would void. The diazo-coupling agent that originally was clear and lightly yellowtinted begins to precipitate after 12 h. This m a y leave doubt in an inexperienced yellow tinted begins to precipitate after 12 h. This may leave doubt in an inexperienced

examiner's mind as to its acceptability. Second, the examiner must follow the same protocol acceptability. Second, the examiner must follow the same protocol in every case. If he places three drops of buffer solution on each disk, then he must do every case. If he places three drops of buffer solution on each disk, then he must do the same with each case. If two drops of Brentamine fast Blue B are used in one case, the same with each case. If two drops of Brentamine fast Blue B are used in one case, then two drops must be used in every case. Third, and most important, the examiner then two drops must be used every case. Third, most important, the examiner must not deviate from his method of reading the test. An intense purple color must appear deviate from his method of reading the test. An intense purple color must immediatelyor up to S s after the application of the chromagen.Then the test is positive. immediately or up to 5 s after application chromagen.Then test is positive. The RTC with which the author was formerly associated listed four classifications under RTC which the was formerly associated listed four classifications under AP: negative, weak, medium, and strong. Under these circumstances, it is obvious that negative, weak, medium, and strong. these circumstances, obvious that test results were meaningful only if all six examiners on the panel agreed on strict criteria were meaningful only if all six examiners on the panel agreed on strict criteria of uniformity in their reports and only if each stage was defined. This was not done; what uniformity in their reports and only if each stage was defined. This was not done; what one physician would interpret as medium, another might read as strong. physician would interpret as medium, another might read strong. It must be emphasized that the author is not comparing a qualitative with a quantitative with a quantitative must emphasized author is not comparing method, but, rather, is underscoring his belief that the AP test employed in many centers method, rather, is underscoring his belief the AP test employed in many centers reliable presence seminal fluid one follows the three guidelines is a reliable indicator of the presence of seminal fluid if one followsthe three guidelines detailed herein. The test described is very simple and can be performed in several minutes. By comparison, very simple and can be the least involved test making the most unsophisticated attempt at quantification consumes involved test making the most unsophisticated attempt at quantification consumes a minimum of 15 min and involvesincubation of the unknown material. Hence, one must minimum 15 min and involves incubation of the unknown material. Hence, one must have access to an incubator. After incubation, one makes comparisons with three different have access to an incubator. comparisons with three colors: normal (about 0.6 Bodansky units), questionable (about 1.0 Bodansky unit), and colors: normal (about 0.6 Bodansky units), questionable (about 1.0 Bodansky unit), and markedly elevated (over 2.5 Bodansky units). A much more sophisticated and accurate method of determining AP quantitatively A much more sophisticated and accurate method of determining AP quantitatively employs aa spectrophotometer, which is not customarily found in an RTC. Only aa trained employs spectrophotometer, which is not customarily found in an RTC. Only trained and knowledgeableperson can use this laboratory apparatus. In addition, under ideal knowledgeable person can use this laboratory apparatus. In addition, under ideal conditions, the test consumes approximately 40 rain. 40 mm. Some believe the quantitative method is more accurate than the qualitative one and Some believe the quantitative method is more accurate than the qualitative one and assists in pinpointing the time of the assault, but the vagaries of the enzyme in vivo must assists in pinpointing the time of the assault, the vagaries enzyme in vivo must be reckoned with. For example, one must question why, in one woman, with either method, reckoned with. For example, one must question why, in one woman, with either method, the enzyme has a high value 10 h after coitus, yet in another woman 10 h after coitus the enzyme has a high value 10 h after coitus, yet in another woman 10 h after coitus

TABLE 2--Stability of reagents. TABLE 2--Stability of reagents.

Time, h

0 1 6

12 12 16 24 30 36 44

Blank Disk

0 0

Seminal Fluid Disk

0 0 0

0

50

+ +

0 0 0

++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++

SCHIFF ON ACID PHOSPHATASE TEST 843 SCHIFF ON ACID PI4OSPHATASE TEST 843

no enzyme is found. If a victim had to hike 10 km (six miles) to the nearest police station, km (six miles) to the nearest police station, no enzyme is found. victim had very tittle AP would be found in the vaginal barrel because of her vigorous exercise and, very little AP would be found in the vaginal because vigorous exercise and, possibly, the loss of much of the seminal fluid. It is also possible that the vaginal environpossibly, the loss of much of the seminal fluid. possible that environment might be so hostile that--as sometimes in the case of spermatozoa--the enzyme ment might so hostile t h a t - - a s sometimes the case of spermatozoa--the enzyme might be destroyed within aa few hours. There would be little difference as to whether aa might be destroyed within few hours. There would be little difference as to whether qualitative or a quantitative method were employed. IfIfaatrue quantitative test, with aa true quantitative test, with qualitative or a quantitative method were employed. spectrophotometer and established standards, were to be performed in the RTC and if, spectrophotometerand established standards, were to be performed in the RTC and if, for example, the test should yield a value of 2000 K-A units, then one would certainly example, test should yield a value of 2000 then one would certainly have to agree that sexual intercourse had occurred recently. Then, in this context, the have to agree sexual intercourse had occurred recently. Then, in this context, word "recently" would have to be defined. To one examiner "recently" might be within word "recently" would have to be defined. To one examiner "recently" might be within 5 h; to another, the word might extend to twice that time. With the entire concept being another, the word extend to twice that time. With the entire concept being so nebulous, one must concede that a qualitative AP test resulting in an intense purple nebulous, one must concede that a qualitative AP test resulting in an intense purple color would serve the same purpose. To fix the approximate time of the assault, the author would serve the same purpose. To fix the approximate time of the assault, the author draws upon three sources: (1) official police reports, (2) the victim's statement, and draws upon three sources: (1) official police reports, (2) the victim's statement, and (3) the qualitative AP test. AP

Sunumu~ Summary from 100 cases alleged victims demonstrate why the author believes the Data from 100 cases of alleged rape victims demonstrate why the author believes the AP test to be a reliable tool, when employed by an expert for the identification of seminal be a reliable tool, when employed by an identification seminal fluid.

Refegences References

[ 1 ] Pollack, O. J., "Semen and Seminal Stains," Archives of Pathology, Vol. 35, No. 1, Jan. 1943, [11 Pollack, 0. J., "Semen and Seminal Stains," Archives of Pathology, Vol. 35, No. 1, Jan. 1943, 140-196. pp. 140-196. [2] Lundquist, F., "Om Forensisk Paavsisk af Sperma og Spermaplatter," Nordisk Medicin, (2] Lundquist, F., "Om Forensisk Paavsisk af Sperma og Spermaplatter," Nordisk Medicin, 1945, pp. 2131-2132. Vol. 28, No. 42, Oct. 1945, pp. 2131-2132. Kaye, S., "Acid Phosphatase Test for Identification of Seminal Stains," Journal of Laboratory [3] Kaye, S., "Acid Phosphatase Test for Identification of Seminal Stains," Journal of Laboratory and Clinical Vol. No. May 1949, pp. 728-732. and Clinical Medicine, Vol. 34, No. 5, May 1949, pp. 728-732. [4] Griffiths, P. D. and Lehmann, H., "Estimation ofof Creatine Phosphokinase as an Additional Griffiths, P. D. and Lehmann, H., "Estimation Creatine Phosphokinase as an Additional Method for Jan. Method for Identification of Seminal Stains," Medicine, Science and the Law, Vol. 4, Jan. of Seminal Stains," Medicine, Science and the Law, 1964, pp. 32-34. [5] Florence, A., "Du Sperme et des Taches de Sperme en Medicine Legale," Archives d'AnthroFlorence, A., "Du Sperme ct des Taehes de Sperme en Medicine Legale," Archives d'AnthroVol. 11, 1896, p. 37. pologie Criminelle, Vol. 11, 1896, p. 37[ 6 Kind, S. S., "The Acid Phosphatase Test," Methods of Forensic Science, Vol. 3, A. S. Curry, [6]] Kind, S. S., "The Acid Phosphatase Test," inin Methodsof Forensic Science, Vol. 3, A. S. Curry, Interscience, London, 1965, 267-288. Ed., Interscience, London, 1965, pp. 267-288. [ 7] Gradwohl, R. B.,Legal Medicine, C. V. Mosby Co., St. Louis, Mo., 1954. [7] Gradwohl, R. B., Legal Medicine, C. V. Mosby Co., St. Louis, Mo., 1954. Kelley, M., "Crime in the United States," Uniform Crime Reports, U.S. Government [8] Kelley, C. M., "Crime in the United States," Unjform Crime Reports, U.S. Government Printing Office, Washington, D.C., 1975. Printing Office, Washington, D.C., 1975. [9] Robison, R., "The Possible Significance of Hexosephosphode Esters in Ossification," Bio[9] Robison, R., "The Possible Significance of Hexosephosphoric Esters in Ossification," Biochemicallournal, 1923, chemical Journal, Vol. 17, No. 2, Feb. 1923, pp. 286-292. [10] Kutscher, W. and Wolbergs, H., Zeitschrifl ffLr Physiologische Chemie, Vol. 236, 1935, [10] Kutseher, W. and Wolbergs, H., Zeitschnft filr Physiologische Chemie, Vol. 236, 1935, 237-240. pp. 237-240. [11] Gutman, E. B., Sproul, E. E., and Gutman, A. B., "Significance of Increased Phosphatase [ii] Gutman, E. B., Sproul, E. E., and Gutman, A. B., "Significance of Increased Phosphatase Activity of Bone at the Site of Osteopathic Metastases Secondary to Carcinoma of the Prostate of Bone at the Metastases Secondary Carcinoma Prostate Gland," American Journal of Cancer, Vol. 28, No. Nov. 1936, pp. 485--495. Gland," American Journal of Cancer, Vol. 28, No. 3,3, Nov. 1936, pp. 485-495. [ 12] Gutman, B. and Gutman, E. B., "'Acid' Phosphatase Occurring in Serum of Patients (12] Gutman, A. B. and Gutman, E. B., "'Acid' Phosphatase Occurring in Serum of Patients with Metastasizing Carcinoma of Prostate Gland," Journal Clinical Investigation, with Metastasizing Carcinoma of the Prostate Gland," Journal ofof Clinical Investigation, Vol. 17, No. 4, July 1938, pp. 473-378. [13] Gomori, G., "Distribution of Acid Phosphatase in the Tissues Under Normal and Pathologic [13] Gomori, G., "Distribution of Acid Phosphatase in the Tissues Under Normal and Pathologic Conditions," Archives Pathology, 1941, 189-199. Conditions," Archives of Pathology, Vol. 32, No. 8, Aug. 1941, pp. 189-199. 32, I-Iansen, P. F., "Determination of the Prostatic Acid Phosphatase as New Method for the [14] Hansen, P. F., "Determination of the Prostatic Acid Phosphatase as aaNew Method for the Medico-Legal Demonstration of Sperm Spots," Acta Pathologica etMicrobiologica Scandinavica, Demonstration Spots," Pathologica et Microbiologica Scandinavica, 23, 1946, pp. 187-211. Vol. 23, 1946, pp. 187--211. Riisfeldt, O., "Acid Phosphatase Employed as New Method for Demonstrating Seminal [15] Rilsfeldt, 0., "Acid Phosphatase Employed as a aNew Method for Demonstrating Seminal Spots in Forensic Medicine," Acta Pathologica atet MicrobiologicaScandinavica Supplement, Forensic Medicine," Aeta Pathologica Microbiologica Seandinavica Supplement, Vol. 58, 1946, pp. 1-78.

844 JOURNAL OF FORENSIC SCIENCES 844 JOURNAL OF FORENSIC SCIENCES

Benotti, J., Rosenberg, Dewey, B., Gutman Method the [16] Benotti, J., Rosenberg, L., and Dewey, B., "Modification of the Gutman and Gutman Method Estimating 'Acid' Phosphatase Activity," Journal of Laboratory and Clinical Medicine, of Estimating 'Acid' Phosphatase Activity," Journal of Laboratory and Clinical Medicine, Vol. No. 3, March 1946, pp. 357-360. Vol. 31, No. 3, March 1946, pp. 357--360. Rasmussen, P. S., "Om Bestemmelse Fosfatase Spermapletter Toj," Nordisk [17] Rasmussen, P. S., "Om Bestemmelse af 'sur' Fosfatase ii Spermapletter paa Toj," Nordisk Medicin, Vol. 49, No. 28, 7 Dec. 1945, pp. 2523-2525. Medicin, Vol. 49, No. 28, 7 Dec. 1945, pp. 2523-2525. [18] Seligman, A. M. and Manheimer, L. H., "New Method for Histochemical Demonstration of Seligman, M. and Manheimer, L. 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Reliability of the Acid Phosphatase Test for the Identification of Seminal Fluid

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