Creatinine related reference ranges for urinary homovanillic acid and vanillylmandelic acid at 6 months of age.

The relationship between homovanillic acid (HVA), vanillylmandelic acid (VMA), and creatinine in the urine of 6 month old babies has been studied and reference ranges in the form of centiles constructed for HVA and VMA against creatinine. Over 10,000 urine samples were collected from babies in four health districts in the north of England. HVA and VMA concentration, either independently or when divided by creatinine concentration, were dependent upon the absolute concentration of creatinine in the sample. After adjustment for creatinine significant differences in the mean concentration of HVA were found between sexes. No such differences were found for VMA. HVA and VMA were also found to be age dependent. Centiles were constructed using a procedure which makes no distributional assumptions about the data. The net effect of utilising these centiles was to increase the predictive value of a positive screening test from 20% to 40% without any increase in the false negative rate.     

False positive results in a neuroblastoma screening programme

Twenty thousand, eight hundred and twenty-nine babies were screened for neuroblastoma at 6 months of age by measuring homovanillic (HVA) and vanillylmandelic (VMA) acid in urine and ratioing these to creatinine. Using a ?cut off”? of the mean + 3 SD, 10 were found to be positive. Two were found on evaluation to have neuroblastoma and in the remaining 8 the raised levels of HVA and/or VMA returned to normal. Only one of the 8 false positive babies was absolutely normal, most having a chronic disorder or illness. Utilising new centiles which relate HVA and VMA to creatinine, only 3 of the 8 would have remained positive, a false positive rate of 0.01%. The false negative rate would have remained unchanged.     

Urinary phenolic acid and alcohol excretion in the newborn.

Mean urinary excretion values of some phenolic acids and alcohols have been measured by gas chromatography in 44 neonates (36 males, 6 females) during the first 2 days and days 3-7 of life, and the effect of prematurity and jaundice assessed. 4-Hydroxy-3-methoxymandelic acid (VMA) output rises immediately after birth in term but not in preterm infants. A similar increase in homovanillic acid (HVA) output was restricted to nonjaundiced term babies; in nonjaundiced preterm babies there was a steady rise during the first week. The ratio of HVA to VMA output was higher in these infants than in adults, suggesting a more rapid turnover of dopamine than adrenaline and noradrenaline. Unlike adult values, both HVA and VMA excretion values were directly related to urine volume, an observation perhaps related to renal immaturity. An unexplained reduction in HVA output in jaundiced as opposed to nonjaundiced infants was observed in the first 2 days of life. The ratio of 4-hydroxy-3-methoxyphenylglycol to VMA was about the same as in the adult. p-Hydroxyphenyl-lactic acid (p-HPLA), because of its superior stability, was measured in preference to p-hydroxyphenylpyruvic acid as an index of tyrosyluria. An output of 1 mg p-HPLA/24 h is proposed as the upper limit of normal. Prematurity was associated with a significant rise in p-HPLA output. A dramatic increase in excretion of this acid was noted in jaundiced, compared with nonjaundiced infants, presumably a manifestation of general enzyme immaturity.     

随机尿VMA、HVA与肌酐比值的测定

目的 探讨随机尿ＶＭＡ、ＨＶＡ与肌酐比值测定的方法及意义。方法 高效液相色谱仪测定随机尿ＶＭＡ、ＨＶＡ浓度,Ｂｅｃｋｍａｎｎ自动生化分析仪测定随机尿肌酐（Ｃｒ）浓度,计算得出比值ＶＭＡ／Ｃｒ,ＨＶＡ／Ｃｒ。检测其与２４ｈ ＶＭＡ、ＨＶＡ总量测定的相关性和对神经母细胞瘤的诊断符合率。结果 随机尿ＶＭＡ／Ｃｒ、ＨＶＡ／Ｃｒ与２４ｈ尿ＶＭＡ、ＨＶＡ总量的相关性分别为ｒ＝０．９２和ｒ＝０．９１,均呈显著相     

Three years of experience with random urinary homovanillic and vanillylmandelic acid levels in the diagnosis of neuroblastoma

During the last 3 years, random urine samples from 408 patients were tested for elevated homovanillic acid (HVA) and vanillylmandelic acid (VMA) levels to rule out the diagnosis of neuroblastoma. Thirty-seven of these patients had elevated HVA and/or VMA levels, and neuroblastoma was subsequently diagnosed. In three additional patients with negative test results (normal HVA and VMA levels), tumors were subsequently diagnosed (false-negative rate of 7.5%). Ten percent of the patients with neuroblastoma had normal HVA and 27.5% had normal VMA levels at the time of diagnosis. Only one patient (2.5%) with neuroblastoma had elevated VMA levels in the presence of normal HVA levels. More than 60% of the patients with neuroblastoma had urinary HVA and/or VMA levels higher than twice the upper limit of normal. No false-positive results were encountered. Age and stage distributions of the patients are shown, and the significance of the results is discussed.     

Reduced sympathetic metabolites in urine of obese patients with craniopharyngioma

Severe obesity is a major problem in patients suffering from craniopharyngioma (CP), a benign tumor located in pituitary and hypothalamic regions. In this study, the hypothesis that hypothalamic damage leads to a reduction in overall sympathetic tone was tested. Catecholamines, as well as their metabolites homovanillic acid (HVA) and vanillylmandelic acid (VMA), markers of catecholamine turnover, were measured in morning voided urine of 109 patients participating in a German pediatric CP study, and their physical activity was analyzed using a questionnaire. HVA and VMA results were compared with age-matched HVA and VMA in urine of patients proven to not have a catecholamine-secreting tumor. Patients with the most severe obesity displayed the lowest urine HVA and VMA values. Patients with hypothalamic CP had 3.2-fold higher BMI values (p<0.0001), lower HVA (0.72-fold, p<0.001), and VMA (0.84-fold, p<0.01) values, and significantly lower activity scores than those without hypothalamic involvement, but their epinephrine- and norepinephrine/creatinine ratios were not significantly different, possibly due to low levels. The low HVA and VMA values suggest decreased sympathetic outflow contributing to reduced physical activity and severe obesity, especially in patients with a hypothalamic tumor. In further studies investigating treatment options for hypothalamic obesity, disturbed sympathetic tone should be considered.     

库仑阵列电化学测定尿儿茶酚胺代谢物方法的建立及其在神经母细胞瘤诊断中的应用

目的 建立以反相液相色谱-库仑阵列电化学联用紫外检测法同时测定随机尿液中儿茶酚胺代谢产物方法,并应用于0～10岁儿童正常值范围的确立及神经母细胞瘤的诊断.方法 反相液相色谱梯度洗脱法分离样品,库仑阵列电化学定量检测VMA和HVA的浓度,紫外检测器检测Cr的浓度.以VMA/Cr和HVA/Cr表示尿儿茶酚胺代谢物的含量.对该法进行系统性评价后应用于120例健康儿童和15例神经母细胞瘤患儿随机尿样的测定中.结果 VMA、HVA和Cr的日内变异系数和日问变异系数均小于5%,线性相关系数均大于0.9990.回收率均大于95%.健康儿童VMA/Cr(2.99±1.77)μg/mg和HVA/Cr(3.40±1.50)μg/mg与年龄有显著相关性(P＜0.001),0～2岁组[VMA/Cr(4.04±1.24)μg/mg,HVA/Cr(4.87±1.81)μg/mg]与其他年龄组[2～4岁:VMA/Cr(3.02±0.81)μg/mg,HVA/Cr(3.46±1.06)μg/mg;4～6岁:VMA/Cr(2.68±0.95)μg/mg,HVA/Cr(2.75±1.04)μg/mg;6～10岁:VMA/Cr(2.32±0.95)μg/mg,HVA/Cr(2.61±0.84)μg/mg]差异均有统计学意义(P＜0.001).以低于(-x)+3s作为正常值,超过此范围诊断为神经母细胞瘤,对患儿诊断的符合率达93%.结论 本研究建立了同时快速准确检测随机尿样VMA、HVA和Cr的方法,并分年龄组建立了0～10岁儿童的正常值范围,为神经母细胞瘤的诊断和筛查提供了有效的参考指标和简单易行、准确灵敏的生化检验手段,也可用于与儿茶酚胺异常分泌相关的其他研究领域.     

Japanese experience of screening

In Sapporo City, the mass-screening for neuroblastoma concerning the urine of infants at about 6 months of age, started in 1981. After 1984, the quantitative determination of VMA, HVA in the urine was carried out using high performance liquid chromatography from the first step. About 104,000 babies have received the screening during the last 7 years, and 19 patients were found (incidence of detection 1:5,500). Except for one, who died of the duodenal perforation shortly after the surgery, all other patients are surviving free from disease. Among 10,000 neonates, we determined urinary concentration of VMA, HVA, and no positive cases were found. However, from this group, 2 patients we detected by the mass-screening at 6 months of age (1:5,000). After the start of mass-screening, the number of the patients with this tumor in much earlier stages and younger ages increased, clarifying that the main cause for improvement of the survival rates of neuroblastoma in Sapporo.     

Value of random urinary homovanillic acid and vanillylmandelic acid levels in the diagnosis and management of patients with neuroblastoma: comparison with 24-hour urine collections

Urinary homovanillic acid (HVA) and vanillylmandelic acid (VMA) levels were determined in random samples and in 24-hour collections from 13 patients with neuroblastoma and 22 patients without neuroblastoma. Random sample levels were compared with levels in 24-hour collections and showed a positive correlation of 95% for HVA (N = 59) and 93% for VMA (N = 52). No false positives or false negatives occurred using random samples for diagnosis. Nonneuroblastoma (normal) HVA (N = 126) and VMA (N = 119) levels are reported for different age groups. Sequential random HVA and VMA determinations in patients with neuroblastoma during and after therapy are shown. Random urinary HVA and VMA levels are shown to be adequate for utilization in the diagnosis of neuroblastoma and sequential determinations of random HVA and VMA are shown to be helpful in the follow-up of those patients.     

高效液相色谱法检测尿中儿茶酚胺代谢产物含量在神经母细胞瘤早期诊断中的价值

目的　尿中香草扁桃酸 (VMA)及高香草酸 (HVA)的检测是目前早期诊断神经母细胞瘤 (NB)的重要指标 ,以前的检测方法干扰因素多 ,结果不够准确。该文探讨高效液相色谱法 (HPLC)检测随机尿中VMA、HVA的含量在NB诊断和早期诊断中的价值。方法　用HPLC方法 (流动相 pH4 .3,甲醇含量 2 % ,正辛烷磺酸钠溶液 3.0mmol/L)检测 5 0例正常儿童 (正常对照组 )、2 7例NB患儿 (NB组 )及 1 5例急性淋巴细胞性白血病患儿(ALL组 )随机尿中VMA、HVA浓度。用比色法测定同一样品中肌酐浓度。以VMA、HVA与肌酐的比值表示VMA、HVA含量。结果　NB组患儿随机尿VMA/Cr及HVA/Cr的含量 (5 1 .6 0± 4 .5 3μmol/ μmolCr,5 8.0 0±3.75 μmol/ μmolCr)明显高于正常对照组 (8.4 2± 3.6 1 μmol/ μmolCr,1 0 .1 2± 3.88μmol/ μmolCr)及ALL组患儿(8.78± 3.5 0 μmol/ μmolCr,1 1 .5 0± 2 .6 8μmol/ μmolCr) ,其差异均有显著性 (P <0 .0 5 )。以正常对照组儿童随机尿中VMA/Cr及HVA/Cr均值加 2 .5个标准差为异常判定值 ,分别为 1 7.5、1 9.8μmol/ μmolCr,超过此值为异常 ,则尿VMA/Cr或HVA/Cr异常对NB的诊断效率为 96 .3%。结论　HPLC同时测定随机尿中VMA、HVA含量 ,能为NB的早期诊断提供可靠的实验依据 ,并且可用于NB的诊断     

The value of catecholamine metabolite determination on untimed urine collections in the diagnosis of neural creast tumours in children

Abstract There is still controversy regarding the relative merits of catecholamine metabolite estimations on 24 h versus untimed urine collections. The former has the advantage of taking into account diurnal variation in the rate of metabolite excretion but has the disadvantages of delaying results and of being affected by errors in collection. In this study percentile values were established for a reference population of 181 children for urinary 4-hydroxy-3-methoxyphenylacetic acid (HVA) creatinine and 163 children for 4-hydroxy-3-methoxymandelic acid (VMA)/creatinine, using untimed urine collections. Results of similar determinations performed as part of the diagnostic work up of 23 consecutive children subsequently proven to have neural crest tumours showed that all patients had the value of at least one metabolite concentration at or above the highest reference value. In neuroblastoma all patients' VMA/creatinine exceeded the highest reference value and in neural crest tumors overall, this ratio was greater than the highest reference value in 96% of patients. These results are as good as, or better than, previously published results and demonstrate the practical value of using catecholamine metabolite determinations expressed as 'creatinine equivalents' on untimed urine specimens in the diagnosis of neuroblastoma and related tumours in children.     

The value of catecholamine metabolite determination on untimed urine collections in the diagnosis of neural crest tumours in children

There is still controversy regarding the relative merits of catecholamine metabolite estimations on 24 h versus untimed urine collections. The former has the advantage of taking into account diurnal variation in the rate of metabolite excretion but has the disadvantages of delaying results and of being affected by errors in collection. In this study percentile values were established for a reference population of 181 children for urinary 4-hydroxy-3-methoxyphenylacetic acid (HVA)/creatinine and 163 children for 4-hydroxy-3-methoxymandelic acid (VMA)/creatinine, using untimed urine collections. Results of similar determinations performed as part of the diagnostic work up of 23 consecutive children subsequently proven to have neural crest tumours showed that all patients had the value of at least one metabolite concentration at or above the highest reference value. In neuroblastoma all patients' VMA/creatinine exceeded the highest reference value and in neural crest tumours overall, this ratio was greater than the highest reference value in 96% of patients. These results are as good as, or better than, previously published results and demonstrate the practical value of using catecholamine metabolite determinations expressed as 'creatinine equivalents' on untimed urine specimens in the diagnosis of neuroblastoma and related tumours in children.     

Fluctuation in the concentrations of vanillylmandelic acid and homovanillic acid in mass screening for neuroblastoma

Fluctuations in the amount of vanillylmandelic acid (VMA) and homovanillic acid (HVA) were studied in random urine samples from 13 infants with neuroblastoma. In patients with a small tumour, many samples contained amounts below the cut off values, suggesting that detection of a patient with neuroblastoma depended on mathematical probability. Using high performance liquid chromatography a patient with a tumour of about 10 g may well be overlooked, whereas a patient whose tumour weighs over 30 g would probably be detected. With qualitative screening, although more likely to miss a patient with low VMA excretion, a patient with a tumour weighing over 50 g would be detected. In a thorough hospital examination care should be exercised in interpreting borderline values of VMA and HVA; excretion information suggesting the existence of a tumour may aid localization by radiological imaging procedures.     

Optimising urinary catecholamine metabolite diagnostics for neuroblastoma

Introduction

The analysis of urinary catecholamine metabolites is a cornerstone of neuroblastoma diagnostics. Currently, there is no consensus regarding the sampling method, and variable combinations of catecholamine metabolites are being used. We investigated if spot urine samples can be reliably used for analysis of a panel of catecholamine metabolites for the diagnosis of neuroblastoma.

Methods

Twenty-four-hour urine or spot urine samples were collected from patients with and without neuroblastoma at diagnosis. Homovanillic acid (HVA), vanillylmandelic acid (VMA), dopamine, 3-methoxytyramine, norepinephrine, normetanephrine, epinephrine and metanephrine were measured by high-performance liquid chromatography coupled with fluorescence detection (HPLC-FD) and/or ultra-performance liquid chromatography coupled with electrospray tandem mass spectrometry (UPLC-MS/MS).

Results

Catecholamine metabolite levels were measured in urine samples of 400 neuroblastoma patients (24-hour urine, n = 234; spot urine, n = 166) and 571 controls (all spot urine). Excretion levels of catecholamine metabolites and the diagnostic sensitivity for each metabolite were similar in 24-hour urine and spot urine samples (p > .08 and >.27 for all metabolites). The area under the receiver-operating-characteristic curve (AUC) of the panel containing all eight catecholamine metabolites was significantly higher compared to that of only HVA and VMA (AUC = 0.952 vs. 0.920, p = .02). No differences were observed in metabolite levels between the two analysis methods.

Conclusion

Catecholamine metabolites in spot urine and 24-hour urine resulted in similar diagnostic sensitivities. The Catecholamine Working Group recommends the implementation of spot urine as standard of care. The panel of eight catecholamine metabolites has superior diagnostic accuracy over VMA and HVA.     

Olive ingestion causing a false suspicion of relapsed neuroblastoma: A case of “oliveblastoma?”

Measurement of the urine catecholamine metabolites homovanillic acid (HVA) and vanillylmandelic acid (VMA) are the standard method for detecting disease recurrence in neuroblastoma. We present a case of abnormal concentrations of catecholamine metabolites that prompted investigations for relapsed neuroblastoma. However, further study revealed that the abnormal biochemistry was likely due to ingestion of olives. Olive ingestion should be considered when interpreting urine HVA and VMA results, and excluded if concentrations are unexpectedly abnormal.     

Catecholamine metabolites in ganglioneuroma

With rare exception, ganglioneuroma (GN) is a benign lesion which presents as a localized mass without metastatic potential and which is chemotherapy resistant. Thus, its distinction from neuroblastoma (NB) may be important. The diagnosis of GN implies the absence of neuroblastic elements. Incomplete resection prevents complete microscopic examination and raises the possibility that focal NB was not sampled. In an attempt to determine what features other than histology distinguish these two entities, we reviewed the charts of 25 patients with GN with regard to patient age and sex, tumor location and size, and urine catecholamine metabolite levels. One patient with GN (5%) and gross total resection had elevated quantitative vanillylmandelic acid (VMA) and homovanillic acid (HVA) levels (2.4 × upper limit of normal for age), and two others had positive spot analyses for VMA. An additional patient with a large mass, multiple biopsies of which documented GN, also had greatly elevated (approximately 5 × normal) VMA and HVA levels. However, a subsequent attempt at resection disclosed several gross foci of NB. Even excluding this patient, there was a trend for elevated values in GN patients to correlate with tumor size (P = .07 and .14 for VMA and HVA, respectively). The incidence of elevated values appears to increase as a function of tumor size, and small tumors are not likely to result in positive urinary measurements. We conclude that while elevations of VMA and HVA are consistent with a well-documented diagnosis of GN, extreme elevations (>3 × nl) should prompt careful serial evaluation for occult NB.© 1994 Wiley-Liss, Inc.     

Cases of neuroblastoma missed by the mass screening programs

Twenty-two reported cases of neuroblastoma missed by the mass screening programs were reviewed (six cases in Sapporo City where a quantitative measurement for both vanillylmandelic acid (VMA) and homovanillic acid (HVA) is used and 16 cases in the districts of qualitative judgment only for VMA), and some potential factors for missing a patient were discussed. Most of the 22 patients were diagnosed over 12 mo and had advanced clinical stages at diagnosis (III or IV). Of the 17 cases whose outcome in prognosis was available, nine cases were dead and only three cases were alive without any trace of tumor. Fluctuation in the amounts of urinary VMA and HVA seemed to be one of the causes for missing a patient, especially with a small tumor. The comparison about tumor wt, VMA and HVA amounts, and clinical stage between the 22 true positive cases in Sapporo City and 46 ones in the districts of qualitative methods, and the difference in detection rate between the two groups suggested that the qualitative methods missed more patients with a small tumor. The patients who had abnormally high amounts of HVA with low amounts of VMA accounted for a fairly large part of the missed cases in Sapporo City as well as in the districts of qualitative methods. To the contrary, no patient of this type was found even among the 22 true positive cases in Sapporo City.     

Diagnostic and prognostic impact of urinary catecholamines in neuroblastoma patients

BACKGROUND: Urinary catecholamine metabolites are well-known to be elevated in patients with neuroblastoma. Some investigators have described different patterns in favorable and unfavorable cases. However, extended studies have not been published. PROCEDURE: We investigated urinary catecholamine patterns and their correlation to stage, biological features, and outcome in 114 consecutively clinically diagnosed neuroblastoma patients. RESULTS: Sensitivity of vanillylmandelic acid (VMA), homovanillic acid (HVA), and dopamine (DA) was 80.7, 71.9, and 61.3%, respectively. In 91.2% of patients at least one parameter was above normal. High VMA levels were associated with favorable biological features, high DA levels were predominantly found in biologically unfavorable disease. Whereas patients with normal HVA levels had a significant better outcome, the other parameters showed no significant association with prognosis. For disseminated neuroblastoma of infancy, DA/VMA ratio proved to be helpful for the discrimination of stage 4 versus stage 4s. CONCLUSION: Urinary catecholamines appear to be useful to give a first but important hint about the biological behavior and thus the prognosis of the underlying disease. Particularly DA/VMA ratio may serve as a tool for "biological grading"-especially in disseminated disease of infancy. In addition, it may be speculated that HVA negativity and low DA/VMA ratio may be helpful for the decision of a "wait and see" strategy in selected neuroblastoma patients with localized disease.     

Increased Urinary Hva Levels in Neuroblastoma Screens Related to Diet,Not Tumor

At present, urinary vanillylmandelic acid (VMA) and/or homovanillic acid (HVA), metabolites of catecholamines, are the most sensitive diagnostic markers of neuroblastoma. They can be measured by the high-performance liquid chromatographic method, which has been used to screen for neuroblastoma in infants in Japan. From 6486 urine samples of 18-month-old children, 103 samples showed high HVA levels over the cutoff point for neuroblastoma, and most of them also showed markedly high levels of 5-hydroxyindoleacetic acid (5-HIAA), a metabolite of serotonin. One of the causes of high HVA excretion might be due to the ingestion of banana, common weaning food in early childhood. Our study showed that it is better to restrict banana ingestion within 24 hours before taking a urine sample for diagnosis of neuroblastoma and that 5-HIAA is a good marker of a pseudopositive result due to banana ingestion.     

Screening procedures for neuroblastoma: false-negative results.

Since elevation of 3-methoxy-4-hydroxymandelic acid (VMA) excretion frequently occurs in patients with neuroblastoma, simpler methods have been developed to differentiate between high and low levels of VMA in the urine, namely the LaBrosse spot test and VMA test strips. Sensitivity of these two procedures depends upon the concentration of VMA in the urine rather than total amount of VMA excreted. VMA excretion data from patients with neuroblastoma show that even though the total amount of VMA excreted in a 24-hour period may definitely be elevated, the concentration of VMA in the 24-hour urine samples may vary greatly from one day to the next and from one patient to another. Therefore, these rapid screening methods for urinary VMA would appear to be too insensitive to use as the sole means of detection or follow-up of patients with neuroblastoma since these techniques require a high concentration of VMA.