Eisei Shikenjo hokoku. Bulletin of National Institute of Hygienic Sciences最新文献

Raw thiamine hydrochloride material was tested for preparation of the "Thiamine Hydrochloride Reference Standard (Control 931)". Analytical data obtained were as follows: melting point, 242.7 degrees C (decomposition); infrared spectrum, the same as that of the JP Thiamine Hydrochloride Reference Standard; thin-layer chromatography, one impurity was detected; high-performance liquid chromatography (HPLC), a trace amount of one impurity was detected; assay results, 100.4% by UV spectrophotometry and 100.0% by HPLC, respectively. Based on the above findings, the raw material was authorized as the JP Thiamine Hydrochloride Reference Standard (Control 931).

Raw testosterone propionate material was tested for preparation of the "Testosterone Propionate Reference Standard (Control 881)". Analytical data obtained were as follows: loss on drying, 0.1%; melting point, 120.6 degrees C; optical rotation [alpha]20D = + 85.4 degrees; ultraviolet spectrum, lambdamax = 241 nm and specific absorbance E 1cm1% (241 nm) = 483; infrared spectrum, the same as that of the NIHS Testosterone Propionate Reference Standard; thin-layer chromatography, no impurities were detected; high-performance liquid chromatography (HPLC), one impurity was detected; assay result, 100.6% by UV spectrophotometry. Based on the above findings, the raw material was authorized as the Testosterone Propionate Reference Standard (Control 881) of the National Institute of Health Sciences.

The Pharmacopoeia of Japan (JP) has played an important leading role in establishing standards of quality and official test methods for drugs and drug preparations in Japan. The JP XII's second supplement and the JP XIII are scheduled to come out by the end of 1994 and on April of 1996, respectively. To provide a more open revision process for JP and to announce the revision and future conception of the JP committees, the outlines of the current progress of each JP sub-committee were introduced.

The Limulus test, which has been adopted as the Test for Bacterial Endotoxins in the JP XII, can detect or quantitate endotoxins of Gram-negative bacterial origin using blood corpuscle extracts (Limulus amebocyte lysate, LAL) of horseshoe crabs (Limulus polyphemus, Tachypleus tridentatus, etc.). It may be conducted by the gel-clot or spectrophotometric (turbidimetric and colorimetric) techniques, the former being based on gel formation due to the activation of LAL by endotoxins. The turbidimetric technique is based on the LAL turbidity change during the gel formation and the colorimetric technique on activation of peptide hydrolytic enzymes in LAL. The Limulus test has been unofficially utilized as a simple and highly sensitive method for the determination of endotoxins in parenteral drugs in lieu of the in vivo Pyrogen Test using rabbits. For the Bacterial Endotoxins Test of the JP XII, the gel-clot technique alone was adopted, the technique being only allowed for Injection. Although most parenteral drugs show inhibition or enhancement in practice, this test can be most easily conducted by eliminating interfering effects through dilution of specimens by a factor not exceeding the maximum valid dilution (MVD) with water. Since MVD is dependent on the sensitivity of applied methodology, the turbidimetric and colorimetric techniques, which are more sensitive than the gel-clot technique, have a distinct advantage. The JP, as the leading Pharmacopoeia for the international harmonization of Bacterial Endotoxins Testing, has presented a "Draft towards International Harmonization of Bacterial Endotoxins Test", whose main purpose is the introduction of supplementary turbidimetric and colorimetric techniques. Under these circumstances the following subjects are discussed: (1) the proposal that, with a view towards international harmonization of the technical requirements of Pharmacopoeias, both the turbidimetric and colorimetic techniques should be included together with the gel-clot technique, (2) the differences in the testing principles and/or conditions prescribed in the current Bacterial Endotoxins Test of JP, USP and EP, and (3) the worldwide situation for Endotoxin Reference Standards.

Cochineal (C), a scarlet material extracted from the powdered pregnant insect, Dactylopius Coceus Costa, is used as a color food additive in the form of aluminum lakes. A 13 week subchronic toxicity study was conducted to investigate the effects of simultaneous administration of C and aluminum potassium sulfate (A). Male and female Wistar rats (5-weeks-old, 15 rats/group) were given diets containing 0.75%A and 0.75%C (1.5%AC), 1.5%A and 1.5%C (3%AC), 3%C alone or 3%A alone. The following results were obtained. 1) No toxic symptoms or death occurred in any treated group. Body weight gain in male rats of the 3%A group decreased significantly. 2) Serum levels of phospholipids, triglycerides (TG) and total cholesterol in male rats and TG in female rats fed 3%C, 3%A or 3%AC were significantly decreased at the 13th week. The serum level of glutamate dehydrogenase (GIDH) in male rats treated with 1.5% or 3%AC was increased at the 4th week but no difference from control was observed at the 13th week. 3) No histopathological changes attributable to A and/or C administration were observed. In this 13-week oral toxicity study, no dose-dependent synergistic effects of simultaneous administration of C and A were found except for an increase in serum GIDH.

Raw dexamethasone material was tested for preparation of the "Dexamethasone Reference Standard (Control 931)". Analytical data obtained were as follows: melting point, 245.1 degrees C (decomposition); infrared spectrum, the same as that of the JP Dexamethasone Reference Standard; optical rotation, -alpha-20D = + 76.75 degrees; thin-layer chromatography, two impurities were detected; high-performance liquid chromatography (HPLC), one impurity was detected; loss on drying, 0.14%; assay result, 99.4% by HPLC. Based on the above findings, the raw material was authorized as the JP Dexamethasone Reference Standard (Control 931).

Liquid secondary ion mass spectrometry (LSIMS) was proven to have advantage for analysis of chemically synthesized peptides. The molecular weights of large peptides were determined accurately, without difficulty in spite of only small sample quantities (50 pmol-10 nmol). This approach was also useful in comparing actual isotopic cluster-distribution of protonated molecular-ion with that derived theoretically for ascertaining peptide integrity. Sometimes LSIMS spectra showed abundant fragment ions from which partial peptide sequences were determinable and unexpected synthetic product could be easily identified from informative fragment ions in LSIMS spectra. The analytical procedure combining LSIMS and high performance liquid chromatography (HPLC) was very beneficial for detecting impurities having slight structural differences. Moreover, LSIMS proved excellent for use in analysis of tyrosine-O-sulfate [Tyr(SO3H)] containing peptides. Constant neutral loss (80 amu) spectra, measured in view of the specific fragmentation (-SO3), showed the complete sequence of a Tyr(SO3H) containing peptide. These results document the usefulness of LSIMS for analysis of peptide samples where difficulty is encountered in evaluation by traditional analytical methods.

Fumonisin has no particular cromophore groups in its chemical structure applicable for trace analysis, and therefore requires derivatization prior to measurement. Currently, the analysis of fumonisin B1 (FB1), B2 (FB2) is generally carried out after derivatizing FB1 mainly with o-phthalaldehyde to make a fluorescent compound (OPA-F). The OPA-F is, however, not sufficiently stable for precise analysis, and we have thus examined other fluorescent derivatives including fluorescamine, FMOC, NBD-F and DBD-F. The analytical method using FMOC as fluorescent agent required column chromatography after the derivatizing reaction, while reaction of FB1 with fluorescamine gave two chromatographic peaks. FMOC and fluorescamine are accordingly not desirable as fluorescent agents for analysis of small quantities of FB1 and FB2 in foods. In conclusion, for precise analysis of such trace amounts of FB1 and FB2, NBD-F or DBD-F should be used as fluorescent agents.

Studies on the stability of ferrous ion in the feeds added iron lactate were conducted for a chronic toxicity test. No significant changes in contents of ferrous ion were observed in all samples when feeds containing 0.2 and 5% of iron lactate were stored in the dark at room temperature (25 +/- 2 degrees C) for 72 hours, or feeds containing 1 and 2% iron lactate stored at 4 degrees C for 90 days.

The number of official inspections of coal-tar dyes and their aluminum lakes from April 1993 till March 1994 were 734 in total, with the quantity which passed inspection amounting to 213 tons in Japan. Data for production by color for each month are summarised in Table 1, and by each producing company in Table 2. The food coal-tar dye produced in the largest quantity was Food Yellow No. 4, occupying a 42.9% proportion of the total during this period.